U.S. patent application number 09/795783 was filed with the patent office on 2001-08-09 for treatment of autoimmune diseases, including aids.
This patent application is currently assigned to Advanced Biotherapy Concepts, Inc.. Invention is credited to Skurkovich, Boris, Skurkovich, Simon V..
Application Number | 20010012514 09/795783 |
Document ID | / |
Family ID | 25093085 |
Filed Date | 2001-08-09 |
United States Patent
Application |
20010012514 |
Kind Code |
A1 |
Skurkovich, Boris ; et
al. |
August 9, 2001 |
Treatment of autoimmune diseases, including AIDS
Abstract
The present disclosure concerns the treatment of a patient with
autoimmune disease, including AIDS, by neutralizing, removing or
inhibiting different types of interferons, tumor necrosis factor,
HLA class II antigens, IgE, and other pathological factors and/or
their receptors, as well as neutralizing, removing or inhibiting
autoantibodies, including antibodies to target cells, CD4 cells and
DNA. Treatment comprises administration of an autoimmune inhibitor,
or extracorporeal exposure of the patient's fluid to an
immunosorbent comprising an autoimmune inhibitor, followed by
return of the treated fluid to the patient; or it comprises a
combined therapy involving extracorporeal immunosorption in
conjunction with the administration of an autoimmune inhibitor.
Inventors: |
Skurkovich, Boris;
(Pawtucket, RI) ; Skurkovich, Simon V.;
(Rockville, MD) |
Correspondence
Address: |
MORGAN, LEWIS & BOCKIUS
1800 M STREET NW
WASHINGTON
DC
20036-5869
US
|
Assignee: |
Advanced Biotherapy Concepts,
Inc.
|
Family ID: |
25093085 |
Appl. No.: |
09/795783 |
Filed: |
February 28, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09795783 |
Feb 28, 2001 |
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08995730 |
Dec 22, 1997 |
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08995730 |
Dec 22, 1997 |
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08771831 |
Dec 23, 1996 |
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5888511 |
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08771831 |
Dec 23, 1996 |
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08025408 |
Feb 26, 1993 |
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5626843 |
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Current U.S.
Class: |
424/143.1 ;
424/145.1; 424/152.1; 424/153.1 |
Current CPC
Class: |
A61K 38/00 20130101;
A61K 39/395 20130101; A61K 39/395 20130101; C07K 16/2833 20130101;
C07K 16/249 20130101; C07K 16/241 20130101; A61K 2039/505 20130101;
A61K 2300/00 20130101 |
Class at
Publication: |
424/143.1 ;
424/145.1; 424/152.1; 424/153.1 |
International
Class: |
A61K 039/395 |
Claims
What is claimed is:
1. A method of treating autoimmune disease in a patient comprising:
administering to the patient an effective amount of a plurality of
at least two antibodies selected from the group consisting of:
antibodies to alpha interferon, antibodies to alpha interferon
receptor, antibodies to gamma interferon, antibodies to
gamma-interferon receptor, antibodies to tumor necrosis factor,
antibodies to tumor necrosis factor receptor, antibodies to HLA
class II antigen, and antibodies to HLA class II antigen
receptor.
2. The method of treating autoimmune disease according to claim 1,
further comprising removing antigens from the patient by the
additional steps comprising: drawing fluid from said patient;
passing said fluid through immunosorbent comprising a combination
of antibodies, comprising at least two antibodies selected from the
group consisting of: antibodies to alpha interferon, antibodies to
alpha interferon receptor, antibodies to gamma interferon,
antibodies to gamma interferon receptor, antibodies to tumor
necrosis factor and antibodies to tumor necrosis factor receptor,
antibodies to HLA class II antigen, and antibodies to HLA class II
antigen receptor; and returning said fluid to said patient.
3. A method of treating autoimmune disease according to claim 1,
wherein the plurality of antibodies comprises (a) at least one
antibody selected from the group consisting of antibodies to alpha
interferon and antibodies to alpha interferon receptor, and (b) at
least one antibody selected from the group consisting of antibodies
to gamma interferon and antibodies to gamma interferon
receptor.
4. The method of treating autoimmune disease according to claim 2,
wherein the immunosorbent comprises (a) at least one antibody
selected from the group consisting of antibodies to alpha
interferon and antibodies to alpha interferon receptor, and (b) at
least one antibody selected from the group consisting of antibodies
to gamma interferon and antibodies to gamma interferon
receptor.
5. The method of treating autoimmune disease according to claim 3,
wherein the plurality of antibodies further comprises at least one
antibody to tumor necrosis factor or to tumor necrosis factor
receptor.
6. The method of treating autoimmune disease according to claim 4,
wherein the immunosorbent further comprises at least one antibody
to tumor necrosis factor or to tumor necrosis factor receptor.
7. The method of treating autoimmune disease according to claim 5,
wherein the plurality of antibodies further comprises at least one
antibody to HLA class II antigen or to its receptor.
8. The method of treating autoimmune disease according to claim 6,
wherein the immunosorbent further comprises at least one antibody
to an HLA class II antigen or to its receptor.
9. The method of treating autoimmune disease according to claim 1,
wherein said antibody or antibodies are selected from the group
consisting of monoclonal antibodies, polyclonal antibodies, and
combinations thereof, including biologically active fragments,
functional equivalents, derivatives, or allelic or species variants
thereof.
10. The method of treating autoimmune disease according to claim 2,
wherein said antibody or antibodies in the additional steps are
selected from the group consisting of monoclonal antibodies,
polyclonal antibodies, and combinations thereof, including
biologically active fragments, functional equivalents, derivatives,
or allelic or species variants thereof.
11. The method of treating autoimmune disease according to claim 1,
wherein an effective amount of beta interferon is also
administered.
12. The method of treating autoimmune disease in a patient
according to claim 11, wherein said autoimmune disease is multiple
sclerosis.
13. The method of treating autoimmune disease according to claim 2,
wherein an effective amount of beta interferon is also
administered.
14. The method of treating autoimmune disease in a patient
according to claim 13, wherein said autoimmune disease is multiple
sclerosis.
15. The method of treating autoimmune disease according to claim 1,
wherein the plurality of antibodies further comprises antibody to
interleukin 6 or to its receptor.
16. The method of treating autoimmune disease according to claim 2,
wherein the immunosorbent further comprises antibody to interleukin
6 or to its receptor.
17. The method of treating autoimmune disease in a patient
according to claim 15, wherein said autoimmune disease is systemic
lupus erythematosus or insulin-dependent diabetes mellitus.
18. The method of treating autoimmune disease in a patient
according to claim 16, wherein said autoimmune disease is systemic
lupus erythematosus or insulin-dependent diabetes mellitus.
19. The method of treating autoimmune disease according to claim 2,
wherein the immunosorbent further comprises antibody to IgE or to
its receptor.
20. The method of treating autoimmune disease in a patient
comprising removing autoantibodies from the patient by the steps
comprising: drawing fluid from said patient; passing said fluid
through immunosorbent comprising at least one member of the group
consisting of: target cells, CD4 cells and DNA, to remove,
neutralize or inhibit autoantibodies in the patient's fluid; and
returning said fluid to said patient.
21. The method of treating autoimmune disease according to claim
20, wherein the immunosorbent further comprises at least one member
of the group consisting of: antibodies to alpha interferon,
antibodies to alpha interferon receptor, antibodies to gamma
interferon, antibodies to gamma interferon receptor, antibodies to
tumor necrosis factor, antibodies to tumor necrosis factor
receptor, antibodies to HLA class II antigen, and antibodies to HLA
class II antigen receptor.
22. The method of treating autoimmune disease according to claim 2,
wherein the immunosorbent further comprises a target cell or cells
to remove, neutralize or inhibit autoantibodies to target cells in
the patient's fluid.
23. The method of treating autoimmune disease according to claim 2,
wherein the immunosorbent further comprises DNA to remove,
neutralize or inhibit autoantibodies to DNA in the patient's
fluid.
24. The method of treating autoimmune disease according to claim 2,
wherein the immunosorbent further comprises CD4 cells to remove,
neutralize or inhibit autoantibodies to CD4 cells in the patient's
fluid.
25. The method of treating autoimmune disease in a patient
according to claim 24, wherein said autoimmune disease is AIDS.
26. A method of treating autoimmune disease in a patient comprising
administering to the patient antibodies to gamma interferon or to
gamma interferon receptor in an amount effective to neutralize or
reduce fluid activity levels of gamma interferon.
27. A method of treating autoimmune disease in a patient comprising
removing antigens from the patient by the steps comprising: drawing
fluid from said patient; passing said fluid through immunosorbent
comprising antibodies to gamma interferon or to gamma interferon
receptor; and returning said fluid to said patient.
28. A method of treating autoimmune disease in a patient comprising
administering to the patient antibodies to HLA class II antigen or
to HLA class II antigen receptor in an amount effective to
neutralize or reduce fluid activity levels of HLA class II
antigen.
29. A method of treating autoimmune disease in a patient comprising
removing antigens from the patient by the steps comprising: drawing
fluid from said patient; passing said fluid through immunosorbent
comprising antibodies to HLA class II antigen or to HLA class II
antigen receptor; and returning said fluid to said patient.
30. A pharmaceutical composition comprising in combination an
effective amount to treat a patient with autoimmune disease of a
plurality of two or more components selected from the group
consisting of: antibodies to alpha interferon, antibodies to alpha
interferon receptor, antibodies to gamma interferon, antibodies to
gamma interferon receptor, antibodies to tumor necrosis factor,
antibodies to tumor necrosis factor receptor, antibodies to HLA
class II antigen, and antibodies to HLA class II antigen receptor,
and a pharmaceutically acceptable carrier therefor.
31. A kit comprising an immunosorbent comprising an effective
amount to extracorporeally remove, reduce or neutralize one or more
autoimmunogens from the fluid of a patient with autoimmune disease
of at least one member of the group consisting of: antibodies to
alpha interferon, antibodies to alpha interferon receptor,
antibodies to gamma interferon, antibodies to gamma interferon
receptor, antibodies to tumor necrosis factor, antibodies to tumor
necrosis factor receptor, antibodies to HLA class II antigen,
antibodies to HLA class II antigen receptor, and antibodies to IgE
and to its receptor.
32. A kit comprising an immunosorbent comprising an effective
amount to extracorporeally remove, reduce or neutralize one or more
autoantibodies from the fluid of a patient with autoimmune disease
of at least one member of the group consisting of: target cells,
CD4 cells, and DNA.
Description
[0001] This application is a continuation-in-part of U.S.
application Ser. No. 08/025,408, filed Feb. 26, 1993.
BACKGROUND OF THE INVENTION
[0002] 1. Field of Invention
[0003] The present invention is a method to treat autoimmune
diseases and conditions in a patient, which are caused by the
disturbance of the synthesis of interferons (IFNs) and certain
other substances (e.g., tumor necrosis factors (TNFs)) and the
production of autoantibodies to target cells, including CD4 cells,
which damage the patient's immune system and have a direct
pathological action on the patient's cells.
[0004] 2. Description of Related Art
[0005] The ability of the immune system to discriminate between
"self" and "non-self" antigens is vital to the functioning of the
immune system as a specific defense against invading
microorganisms. "Non-self" antigens are those antigens on
substances entering or in the body which are detectably different
or foreign from the animal's own constituents, whereas "self"
antigens are those which, in the healthy animal, are not detectably
different or foreign from its own constituents. However, under
certain conditions, including in certain disease states, an
individual's immune system will identify its own constituents as
"non-self," and initiate an immune response against "self"
material, at times causing more damage or discomfort as from an
invading microbe or foreign material, and often producing serious
illness in an individual. Autoimmune disease results when an
individual's immune system attacks his own organs or tissues,
producing a clinical condition associated with the destruction of
that tissue, as exemplified by diseases such as rheumatoid
arthritis, insulin-dependent diabetes mellitus, acquired
immunodeficiency syndrome ("AIDS"), hemolytic anemias, rheumatic
fever, Crohn's disease, Guillain-Barr syndrome, psoriasis,
thyroiditis, Graves' disease, myasthenia gravis,
glomerulonephritis, autoimmune hepatitis, multiple sclerosis,
systemic lupus erythematosus, etc. Blocking, neutralizing or
inhibiting the immune response or removing its cause in these cases
is, therefore, desirable.
[0006] AD can come mostly from a genetic predisposition, alone or
from the influence of certain exogenous agents such as, viruses,
bacteria, or chemical agents, or from the action of both. Some
forms of autoimmunity come about as the result of trauma to an area
usually not exposed to lymphocytes, such as neural tissue or the
lens of the eye. When the tissues in these areas become exposed to
lymphocytes, their surface proteins can act as antigens and trigger
the production of antibodies and cellular immune responses which
then begin to destroy those tissues. Other autoimmune diseases
develop after exposure of the individual to antigens which are
antigenically similar to, that is cross-reactive with, the
individual's own tissue. For example, in rheumatic fever an antigen
of the streptococcal bacterium, which causes rheumatic fever, is
cross-reactive with parts of the human heart The antibodies cannot
differentiate between the bacterial antigens and the heart muscle
antigens, consequently cells with either of those antigens can be
destroyed.
[0007] Other autoimmune diseases, for example, insulin-dependent
diabetes mellitus (involving the destruction of the insulin
producing beta-cells of the islets of Langerhans), multiple
sclerosis (involving the destruction of the conducting fibers of
the nervous system) and rheumatoid arthritis (involving the
destruction of the joint lining tissue), are characterized as being
the result of a mostly cell-mediated autoimmune response and appear
to be due primarily to the action of T-cells (See, Sinha et al.,
Science 248: 1380 (1990)). Yet others, such as myesthenia gravis
and systemic lupus erythematosus, are characterized as being the
result of primarily a humoral autoimmune response (Id.).
Nevertheless, the autoimmune diseases share a common underlying
pathogenesis, resulting in the need for safe and effective therapy.
Yet none of the presently available drugs are completely effective
for the treatment of autoimmune disease, and most are limited by
severe toxicity.
[0008] In recent years, a new point of view on the pathogenesis of
autoimmune diseases, including AIDS, has developed, in which it has
been suggested that autoimmune disease is connected with a
disturbance in the synthesis of interferons and other cytokines
induced by interferons (Skurkovich et al., Nature 217:551-2 (1974);
Skurkovich et al., Annals of Allergy 35:356 (1975); Skurkovich et
al., J. IFN Res. 12, Suppl. 1:S110 (1992); Skurkovich et al., Med.
Hypoth. 41:177-185 (1993); Skurkovich et al., Med. Hypoth. 42:27-35
(1994); Gringeri et al., Cell. Mol. Biol. 41(3): 381-387 (1995);
Gringeri et al., J. Acquir. Immun. Defic. Syndr. 13:55-67 (1996)).
IFN has been found in the circulation of patients with autoimmune
diseases, and it has been neutralized in vivo with antibody to
leukocyte (alpha) IFN ("IFN.alpha."). Healthy people do not have
interferon in their blood (Skurkovich et al., 1975). In addition,
it has been shown that hyperproduced IFN.alpha. is found not only
in the circulation of patients with classic autoimmune diseases,
but also in patients with HIV infection (DeStefano et al, J. Infec.
Disease 146:451 (1982)), where its presence is a predictive marker
of AIDS progression (Vadhan-Raj et al., Cancer Res. 46:417 (1986)).
The IFN induced by HIV has low anti-(HIV) viral activity (Gendelman
et al., J. Immunol. 148:422 (1992)). It was shown that the
circulating IFN.alpha. possesses antigenic specificity like natural
IFN.alpha., which is pH stable, but this interferon is pH labile
like IFN.gamma. (Preble et al., Science 216:429 (1982)); thus, it
is known as aberrant IFN.alpha..
[0009] Investigators have also shown that tumor necrosis factors
(TNF.alpha. and TNF.beta.) also play a significant role in the
pathology of autoimmune diseases. For example, the presence of
TNF.alpha. has been correlated with rheumatoid arthritis (Brennan
et al., Brit. J. Rheum. 31(5):293-8 (1992)), and TNF.alpha. has
been found related to an increase in the severity of collagen
induced arthritis in animal models (Brahn et al., Lymphokine and
Cylokine Res. 11 (5):253 (1992)), while it has also been shown that
anti-TNF alpha antibody administration ameliorates collagen induced
arthritis (Williams et al., Clin. & Exp. Immunol. 87(2):183
(1992)). TNF-.alpha. is increased in the serum of RA patients (Holt
et al., Brit. J Rheum. 21(11):725 (1992); Altomonte et al., Clin.
Rheum. 11(2):202 (1992), and both the cytokine (Chu et al., Brit.
J. Rheum. 31(10):653-661 (1992)) and its receptors have been
identified in rheumatoid synovium, as well as at the
cartilage-pannus junction (Deleuran et al., Arthritis Rheum.
35(10):1180 (1992)).
[0010] In addition, increased circulating levels of TNF.alpha. have
been found to be associated with disease progression in patients
with multiple sclerosis (Shariff et al., N. Engl. J. Med.
325(7):467-472 (1992)); while increased serum levels of soluble TNF
receptor and interferon .gamma. ("INF.gamma.") have been
independently correlated with disease activity in individuals,
e.g., those with systemic lupus erythematosus (Aderka et al.,
Arthritis Rheum. 36(8):1111-1120 (1993); Machold et al., J.
Rheumat. 17(6):831-832 (1990)). The spontaneous release of
interferon and TNF in HIV-positive subjects (Vilcek et al., In
AIDS: The Epidemic of Karposi's Syndrome and Opportunistic
Infections, A. E. Friedman-Kien & L. J. Laubenstein, eds.
Masson Publishing, New York, N.Y., 1986; Hess et al., Infection 19,
Suppl. 2:S93-97 (1991); Biglino et al., Infection 19 (1):11/7-11/17
(1991)), and the decline seen in the serum levels of TNF-.alpha. in
RA patients following long term administration of the disease
modifying drug sulfasalazine (Danis et al., Ann. Rheum. Diseas.
51(8):946 (1992)), further suggest that the concentrations of
cytokines and/or their receptors is reflected in the clinical
course of autoimmune disease.
[0011] IFN is known to induce tumor necrosis factor (TNF) and its
receptors (Lau et al., AIDS Research and Human Retroviruses 7:545
(1991)), which enhances virus replication (Matsuyama et al., Proc.
Natl. Acad. Sci. USA 86:2365 (1989)). In addition to its presence
in the circulation, IFNs have also been found in the cerebrospinal
fluid in some patients with psychiatric and neurologic diseases
(Lebikova et al., Acta. Biol. Med. Germ. 38:879 (1979); Preble et
al., Am. J. Psychiatry 142:10 (1985)), as well as in patients with
rheumatoid arthritis. Therefore, since healthy people do not have
interferons in their spinal or synovial fluids, the inventors have
suggested that one or more alpha IFNs may be involved in the
development of the initial autoimmune disease response.
Consequently, the removal and/or neutralization of IFN.alpha. has
been proposed as a method of treatment of patients with autoimmune
disease, including AIDS. The appearance of cytokines and
autoimmunogens induced by IFN.alpha. and their prolonged
circulation in the body is an inseparable part of the development
of autoimmune disease, triggering immune dysregulation in
autoimmune disease, including AIDS. See, U.S. Pat. Nos. 4,824,432;
4,605,394; and 4,362,155, herein incorporated by reference.
However, it now appears that gamma IFN ("IFN.gamma.") can also play
a pathogenetic role since each participates in immune
regulation.
[0012] In addition to classic AD and AIDS, autoantibodies play a
pathogenic role in many other pathological conditions. For example,
after cell (or organ) transplantation or after heart attack or
stroke, certain antigens from the transplanted cells (organs) or
necrotic cells from the heart or the brain can stimulate the
production of autoantibodies or immune lymphocytes (Johnson et al.,
Sem. Nuc. Med. 19:238 (1989); Leinonen et al., Microbiol. Path.
9:67 (1990); Montalban et al., Stroke 22:750 (1991)), which later
participate in rejection (in the case of a transplant) or attack
cardiac or brain target cells, aggravating the condition. Moreover,
in human autoimmune disease certain cells express abnormally
elevated levels of HLA class II antigens, which is stimulated by
the disturbed production of cytokines, e.g., IFN.gamma. alone, or
IFN.gamma. in combination with TNF (Feldman et al., "Interferons
and Autoimmunity," In IFN 9. Academic Press, p.75 (1987).
[0013] Recognition of the important role of cytokines in autoimmune
disease has fostered the. development of a new generation of
therapeutic agents to modulate cytokine activity. Preliminary
results of trials in which anti-interferon monoclonal antibodies
were administered to a small group of rheumatoid patients suggest
improvement in both the clinical and the laboratory manifestations
of the disease (Skurkovich et al., Annals of Allergy 39:344-350
(1977)). Moreover, proteins, such as polyclonal antibodies and
soluble receptors targeted against interferons and TNF-.alpha. are
currently being evaluated in clinical trials for the treatment of
RA and other autoimmune diseases. The administration of monoclonal
antibodies to TNF-.alpha. has provided encouraging early results in
the treatment of patients with severe RA (Elliott et. al., J. Cell.
Biochem., Suppl 17B: 145 (1993); Elliott et al., Lancet 344:
1105-1110 (1994)). Also positive preliminary results were achieved
in AIDS patients given antibodies or other agents to reduce the
level of circulating IFN.alpha. in the body (Skurkovich et al.,
1994; Gringeri et al., 1996). However, because autoimmune diseases
are complex, often characterized by multiple cytokine
abnormalities, effective treatment appears to require the
simultaneous administration or utilization of several agents, each
targeting a specific cytokine pathway or its by-product. To meet
this need, the methods of treatment of the present invention
include not only the use of specific antibodies, but also provide
pleiotrophic autoimmune inhibitors, including antibodies to
cytokines and HLA class II antigens, and antigens for the removal
of autoantibodies to target cells or DNA. The use of these
antibodies and antigens as disclosed in the present invention
results in the removal, neutralization or inhibition of the
pathogenic cytokine(s), HLA class II antigens, and/or
autoantibody(ies) to target cells or DNA from the autoimmune
patient, thereby significantly improving the quality of life of the
individual.
SUMMARY OF THE INVENTION
[0014] The present invention concerns the treatment of autoimmune
diseases, including AIDS, by blocking, inhibiting, neutralizing, or
removing harmful interferons, tumor necrosis factors, and other
pathological immunogens or factors, or their receptors, and
antibodies to target cells, including CD4 cells, in a patient in
need of such treatment.
[0015] It is an object of the present invention to provide a method
of treating autoimmune disease in a patient comprising
administering to the patient an effective amount of anti-IFN.gamma.
antibodies and/or antibodies to IFN.gamma. receptor.
[0016] In addition, it is an object of the invention to provide a
method of treating autoimmune disease in a patient comprising
administering to the patient an effective amount of a plurality of
at least two antibodies selected from the group consisting of
anti-IFN.alpha. antibodies and antibodies to IFN.alpha. receptor,
anti-IFN.gamma. antibodies and antibodies to IFN.gamma. receptor,
anti-TNF antibodies and antibodies to TNF receptor, and antibodies
to an HLA class II antigen or its receptor. In particular, it is an
object of the invention to provide a method of treating autoimmune
disease, wherein a plurality of antibodies is administered
comprising (a) at least one anti-IFN.alpha. antibody and/or
antibody to IFN.alpha. receptor, and (b) at least one
anti-IFN.gamma. antibody and/or antibody to IFN.gamma. receptor. It
is also a particular object of the invention to provide a method of
treating autoimmune disease, wherein at least one anti-TNF antibody
and/or antibody to TNF receptor is administered, alone or in
conjunction with one or more of the following: anti-IFN.alpha.
antibody, antibody to IFN.alpha. receptor, anti-IFN.gamma. antibody
and/or antibody to IFN.gamma. receptor. It is further a particular
object of the invention to provide a method of treating autoimmune
disease, wherein at least one antibody to an HLA class II antigen
and/or its receptor is administered, alone or in conjunction with
one or more of the following: anti-IFN.alpha. antibody, antibody to
IFN.alpha. receptor, anti-IFN.gamma. antibody, antibody to
IFN.gamma. receptor, anti-TNF antibody and/or antibody to TNF
receptor.
[0017] It is also an object of the invention to provide a method of
treating autoimmune disease in a patient comprising, in conjunction
with administering to the patient an effective amount of a
plurality of at least two antibodies selected from the group
consisting of anti-IFN.alpha. antibodies and antibodies to
IFN.alpha. receptor, anti-IFN.gamma. antibodies and antibodies to
IFN.gamma. receptor, anti-TNF antibodies and antibodies to TNF
receptor, and antibodies to an HLA class II antigen or its
receptor, an extracorporeal treatment comprising removing antigens
from the patient by drawing fluid from the patient; passing said
fluid through an immunosorbent comprising a combination of at least
two antibodies selected from the same group; followed by returning
the treated fluid to the patient. In particular, it is an object of
the invention to provide a method of treating autoimmune disease,
wherein the extracorporeal treatment comprises passing the
patient's fluid through an immunosorbent comprising a combination
of at least two antibodies selected from (a) at least one
anti-IFN.alpha. antibody and/or antibody to IFN.alpha. receptor,
and (b) at least one anti-IFN.gamma. antibody and/or antibody to
IFN.gamma. receptor. It is also a particular object of the
invention to provide an extracorporeal method of treatment, wherein
the immunosorbent comprises at least one anti-TNF antibody and/or
antibody to TNF receptor, alone or in conjunction with one or more
of the following: anti-IFN.alpha. antibody, antibody to IFN.alpha.
receptor, anti-IFN.gamma. antibody and/or antibody to IFN.gamma.
receptor. It is further a particular object of the invention to
provide an extracorporeal treatment, wherein the immunosorbent
comprises at least one antibody to an HLA class II antigen and/or
its receptor, alone or in conjunction with one or more of the
following: anti-IFN.alpha. antibody, antibody to IFN.alpha.
receptor, anti-IFN.gamma. antibody, antibody to IFN.gamma.
receptor, anti-TNF antibody and/or antibody to TNF receptor.
[0018] Yet another object of the invention is to treat specific
autoimmune diseases by the administration of autoimmune inhibitor
to the patient. For example, a method of treatment is provided
comprising administering to the patient an effective amount of beta
interferon in addition to one or more antibodies selected from the
group consisting of anti-IFN.alpha. antibody and antibodies to
IFN.alpha. receptor, anti-IFN.gamma. antibodies and antibodies to
IFN.gamma. receptor, anti-TNF antibodies and antibodies to TNF
receptor, and antibodies to an HLA class II antigen or its
receptor. This method is particularly effective for the treatment
of multiple sclerosis. An additional method of treatment is
provided comprising administering to the patient an effective
amount of antibodies to interleukin ("IL"), preferably to IL-6, in
addition to one or more antibodies selected from the group
consisting of anti-IFN.alpha. antibodies and antibodies to
IFN.alpha. receptor, anti-IFN.gamma. antibodies and antibodies to
IFN.gamma. receptor, anti-TNF antibodies and antibodies to TNF
receptor, and antibodies to an HLA class II antigen or its
receptor. This method is particularly effective for the treatment
of systemic lupus erythematosus and insulin-dependent diabetes
mellitus.
[0019] It is also an object of the invention to provide a method of
treating specific autoimmune diseases by the extracorporeal
exposure of the patient's fluid to an immunosorbent comprising
autoimmune inhibitor, followed by the return of the treated fluid
to the patient. This method may be practiced alone, or in
conjunction with the administration of autoimmune inhibitor to the
patient. For example, a method of treatment is provided comprising
exposing the patient's fluid to an immunosorbent comprising an
effective amount of antibodies to interleukin, preferably anti-IL-6
antibody, in addition to one or more antibodies selected from the
group consisting of anti-IFN.alpha. antibody and antibodies to
IFN.alpha. receptor, anti-IFN.gamma. antibodies and antibodies to
IFN.gamma. receptor, anti-TNF antibodies and antibodies to TNF
receptor, and antibodies to an HLA class II antigen or its
receptor. This method is particularly effective for the treatment
of systemic lupus erythematosus and insulin-dependent diabetes
mellitus.
[0020] It is a further particular object of the invention to
provide a method of treating specific autoimmune diseases, wherein
the specific extracorporeal treatment comprises passing the
patient's fluid through an immunosorbent comprising antibody(ies)
to immuno globulin E ("IgE"), followed by the return of the treated
fluid to the patient. This method may be practiced alone, or in
conjunction with the administration of autoimmune inhibitor. For
example, for treating certain diseases related to hypersensitivity
of the immediate type, e.g., bronchial asthma, antibody to IgE is
used as an immunosorbent, alone or in conjunction with other
autoimmune inhibitors, such as antibodies to IFNs and/or to TNF or
to their receptors.
[0021] It is a further object of the invention to provide a method
of treating autoimmune disease in a patient comprising, alone or in
conjunction with administering to the patient an effective amount
of one or more antibodies (e.g., anti-IFN.alpha. antibodies,
antibodies to IFN.alpha. receptor, anti-IFN.gamma. antibodies,
antibodies to IFN.gamma. receptor, anti-TNF antibodies, antibodies
to TNF receptor, and antibodies to an HLA class II antigen or its
receptor), extracorporeal treatment comprising removing
autoantibodies from the patient by drawing fluid from the patient;
passing said fluid anti-TNF antibodies comprising an effective
amount of different target cells, CD4 cells and/or DNA, to remove,
neutralize or inhibit auto-antibodies in the patient's fluid;
followed by returning the treated fluid to the patient. The
immunosorbent for extracorporeal treatment may further comprise one
or more antibodies (e.g., anti-IFN.alpha. antibodies, antibodies to
IFN.alpha. receptor, anti-IFN.gamma. antibodies, antibodies to
IFN.gamma. receptor, anti-TNF antibodies, antibodies to TNF
receptor, and antibodies to an HLA class II antigen or its
receptor.
[0022] In particular, it is an object of the invention to provide a
method of treating specific autoimmune diseases, wherein the
specific extracorporeal treatment comprises passing the patient's
fluid through an immunosorbent comprising target cells. For
example, for the treatment of rheumatoid arthritis, target cell
antigens from joints, skin, collagen, and possibly other target
antigens are used as immunosorbents, alone or in conjunction with
other autoimmune inhibitors, such as antibodies to IFNs and/or
antibodies to TNF or their receptors and/or antibodies to HLA class
II antigens or their receptors. In addition, for the treatment of
rheumatic fever, the invention provides an immunosorbent comprising
antibodies to IFNs and/or to TNF and/or their receptors and/or to
HLA class II antigens and/or their receptors and/or other
substances, in conjunction with a second cardiac tissue sorbent for
removing autoantibodies against cardiac tissue. The second sorbent
can also include selected serotypes of Streptococcus (group "A"),
because certain antigens from cardiac tissue and some serotypes of
Streptococcus are antigenically similar. For the treatment of
autoimmune diseases of the central nervous system, target cell
antigens from the brain cells are used to absorb autoantibodies
formed against brain cells.
[0023] It is also a particular object of the invention to provide a
method of treating specific autoimmune diseases, wherein the
specific extracorporeal treatment comprises passing the patient's
fluid through an immunosorbent comprising DNA. For example, for the
treatment of systemic lupus erythematosus the immunosorbent
comprises DNA to remove, reduce or neutralize the patient's
anti-DNA autoantibodies.
[0024] Yet another particular object of the invention to provide a
method of treating specific autoimmune diseases, wherein the
specific extracorporeal treatment comprises passing the patient's
fluid anti-TNF antibodies comprising CD4 cells. For example, for
the treatment of AIDS, the immunosorbent comprises CD4 cells, alone
or in conjunction with other autoimmune inhibitors, such as
antibodies to IFNs and/or TNF and/or HLA class II antigen, or their
receptors.
[0025] It is also an object of the invention to provide
pharmaceutical compositions comprising in combination an effective
amount to treat a patient with autoimmune disease of a plurality of
two or more components selected from the group consisting of:
anti-IFN.alpha. antibodies, antibodies to IFN.alpha. receptor,
anti-IFN.gamma. antibodies, antibodies to IFN.gamma. receptor,
anti-TNF antibodies, antibodies to TNF receptor, and antibodies to
an HLA class II antigen or its receptor, and a pharmaceutically
acceptable carrier therefor.
[0026] It is a further object of the invention to provide kits or
compositions comprising an immunosorbent comprising an effective
amount to extracorporeally remove, reduce or neutralize one or more
autoimmunogens from the fluid of a patient with autoimmune disease
of at least one of the following antibodies: anti-IFN.alpha.
antibodies, antibodies to IFN.alpha. receptor, anti-IFN.gamma.
antibodies, antibodies to IFN.gamma. receptor, anti-TNF antibodies,
antibodies to TNF receptor, antibodies to an HLA class II antigen
or to its receptor, antibodies to IgE. While it is yet another
object to provide kits or compositions comprising an immunosorbent
comprising an effective amount to extracorporeally remove, reduce
or neutralize one or more auto-antibodies from the fluid of a
patient with autoimmune disease of at least one of the following:
target cells, CD4 cells, and DNA.
[0027] Additional objects, advantages and novel features of the
invention will be set forth in part in the description which
follows, and in part will become apparent to those skilled in the
art on examination of the following, or may be learned by practice
of the invention.
DESCRIPTION OF PREFERRED EMBODIMENTS
[0028] The present invention provides methods for treating various
autoimmune diseases (AD), including AIDS by blocking, neutralizing
or inhibiting different kinds of interferons, tumor necrosis
factor, HLA class II antigens, and other pathological factors,
which are common in most AD, and which participate in damage of the
immune system and the development of AD. In addition, it provides
methods for removing, reducing or neutralizing antibodies which may
destroy the DNA or target cells of AD patients and/or the CD4 cells
in patients with AIDS.
[0029] Interferon is now known to be not only an antiviral and
anti-proliferative cytokine, but it is also a factor which plays an
important role in normal and pathological immunity. For the normal
functioning of the immune system, it is necessary for an individual
to have a normally functioning cytokine system. The interferon
system in humans is a very stable system. Since healthy people do
not have interferon in their blood, prolonged hyperproduction of
interferon--primarily alpha but sometimes gamma
interferons--typically indicates the presence of immune
disease.
[0030] Upon observation of the diverse clinical pictures manifested
in patients with various AD, which includes hypersensitivity of the
immediate type (e.g., bronchial asthma, which is also an autoimmune
condition), and AIDS (a viral disease with autoimmune components),
it becomes apparent that these diseases have in common a large
number of similar laboratory characteristics. This suggests that a
similar disease mechanism is occurring in each autoimmune disease,
but in different target cells. Thus, it is the unique target (e.g.,
skin, joints, liver, and the like) of each autoimmune disease that
leads to its characterization in terms of clinical manifestations.
For example, an autoimmune attack destroying the insulin producing
beta-cells of the islets of Langerhans of an individual would be
diagnosed as diabetes (Type I), whereas autoimmune destruction of
the conducting fibers of the nervous system is characteristic of
multiple sclerosis, or autoimmune destruction of the joint lining
tissue is characteristic of rheumatoid arthritis. Likewise in the
case of skin transplantation, the skin area can be damaged. Yet in
each case the mechanism underlying the autoimmune response is
similar; a high level of IFNs, a detectable level of TNF, an
elevated level of HLA class II antigens in the blood or on the
surface of the cells, and antibodies to target cells. In addition,
cells taken from autoimmune patients show a decreased production of
IFNs in vitro, even after stimulation with an interferonogen.
Consequently, the method of treatment of the various ADs is similar
in principle, despite the apparent clinical differences among the
diseases.
[0031] The present invention is based upon the inventors'
conclusions that the optimal treatment of each different AD or
autoimmune condition involves the removal, neutralization or
inhibition of complex pathological agents (including hyperproduced
cytokines) from the patient, and/or the administration to the
patient of an effective amount of selected molecules or antibodies,
or their receptors, to bind to, neutralize or inhibit the
circulating pathological agents and/or those on the surface of the
cells targeted in the specific autoimmune response ("target
cells")>The primary indicator of each AD is the hyperproduction
of IFN.alpha. or, to be more exact, the disturbance of the
synthesis of one or more alpha IFNs (IFN.alpha. comprises at least
15 distinct subtypes). In most patients with AD, some level of
IFN.gamma. is also found. Patients with systemic lupus
erythematosus ("SLE") and AIDS appear to have the highest levels of
IFN.alpha., as compared with patients with other autoimmune
diseases (See, Skurkovich et al., Annals of Allergy 35:356 (1975);
DeStefano et al., 1982).
[0032] IFN.alpha. is secreted by somatic cells and leukocytes,
accumulating on the membranes of cells and entering the
bloodstream. In autopsies, IFN.alpha. has been found, for example,
on the surface of cells in the pancreas of patients with insulin
dependent diabetes (Foulis et al., Lancet 2:1423 (1987)), in skin
lesions of patients with psoriasis (Livden et al., Arch. Dermatol.
Res. 281:392 (1989)), on the surface of brain cells of patients
with the psychiatric complications of systemic lupus erythematosus
("SLE")((Shiozawa et al., Arthr. Rheum. 35:417 (1992)), and in the
circulating body fluids of animal and human patients with AD
((Skurkovich et al., 1975; DeStefano et al., 1982). For instance,
IFN.alpha. has been found circulating in the blood of autoimmune
NZB/W and mrl/lpr mice (Skurkovich et al, Ann. Internat'l. Congress
for Interferon Research (1981), and in the circulation of patients
with RA, SLE, Sjogren's syndrome, scieroderma, insulin-dependent
diabetes, bronchial asthma, AIDS, and other ADs (Skurkovich et al,
1975; Hooks et al., N. Engl. J. Med. 301:5 (1979); DeStefano et al,
1982). Of particular interest, is a recent discovery that
interferon is also found in the blood and spinal fluid of patients
with neurological diseases, including, e.g., schizophrenia
(Lebikova et al., Med. Microbiol. Immun. 166:355 (1978); Preble et
al., 1985), depression, and multiple sclerosis (Link et al., Ann.
Neurol. 36:379 (1994)).
[0033] The uninterrupted production of IFN.alpha. is apparently
connected with the weakening or absence of the IFN.alpha.
repressor. In general, hyperproduction of IFN.alpha. is an
indicator of immunological disintegration, and many scientists
consider IFN.alpha. to be a recognized marker of the presence of an
autoimmune condition ((Skurkovich et al., 1975; Hooks et al.,
1979). Moreover, the disturbance of IFN.alpha. production in an
individual changes the biological activity of the cells, bringing
about the production of autoantigens (Skurkovich et al., 1994;
Shattner et al., Am. J. Med. Sci. 295:532 (1988)). The
hyperproduction of IFN.alpha. also stimulates the production of
tumor necrosis factor and its receptors, particularly TNF.alpha.
(Lau et al., 1991). Increased production of autoantigens leads to
the activation of the T-cells, and to the production of IFN.gamma..
It is possible every autoantigen stimulates the induction of a
unique, specific IFN.gamma..
[0034] In addition, in human AD some cells express abnormally
elevated levels of HLA class II antigens, or in some cases HLA
class I or III antigens, which is stimulated by the disturbed
production of IFN.gamma., alone or in combination with TNF (Feldman
et al., 1987). This synthesis of HLA class II antigens (or HLA
class I or III antigens) plays an important role in the
pathogenesis of AD and AIDS. The disturbance of the production of
HLA class II antigen in an individual leads to a pathological
disturbance of the presentation of antigens to the T-cells, to
disrupted T/B cooperation, and to the dysregulation of the
interactions among T-cells.
[0035] Every antigen is an interferonogen; "self" cannot induce
IFN. Thus, the production of IFN signals the invasion by a foreign
antigen, or in this case the presence of an autoantigen. The
production of IFN and its prolonged circulation in the body is an
inseparable part of the development of AD, and triggers
immunological chaos. For example, antibodies to CD4 in patients
with HIV infection (Dorsett et al., Am. J. Med. 78:621 (1985)) can
cross-react with HLA class II antigen, which in turn are induced by
IFN.gamma., or by IFN.gamma. in combination with TNF, and possibly
by IFN.alpha., which induces TNF.
[0036] IFN.alpha. and IFN.gamma. are biologically dangerous
elements in certain people. If injected into a human or animal with
a genetic predisposition to develop an AD, the interferons can
trigger or exacerbate the AD in the recipient. For example,
administration of IFN.alpha., IFN.gamma., or an inducer of
IFN.alpha. to autoimmune NZB/W and MRL/lpr/lpr mice have resulted
in an aggravation of the autoimmune response in the animal,
augmented morbidity, and increased mortality (Carpenter et al., Lab
Invest. 23:628 (1970); Engleman et al., Arthr. Rheum. 24:1396
(1981); Heremans et al., Infect. Immun. 21:925(1978)). Injection of
one unit of r-IFN.gamma. into the thyroid gland of CBA mice caused
autoimmune thyroiditis (Remy et al., Immunol Today 8:73 (1987)).
Administration of IFN.alpha. to human patients with psoriasis (a
disease with an autoimmune component) was found to exacerbate,
rather than alleviate the clinical manifestations of the disease
(Quesada et al., Lancet 2:1466 (1986)). Injection of natural or
recombinant IFN.alpha. ("r-IFN.alpha."), and sometimes IFN.gamma.,
to cancer patients has reportedly triggered or exacerbated
autoimmune parotitis, epididymitis, and thyroiditis, SLE, RA,
Graves' disease, and other autoimmune conditions (See, e.g.,
Quesada et al., Clin. Oncol. 2:4234 (1986); Bevan et al., Lancet
2:561 (1985); Ronnblom, et al. J. Intern. Med. 227:207 (1990);
Conlon et al., Cancer 65:2237 (1990); Machold et al., J. Rheum.
17:831 (1990); Schilling et al., Cancer 68:1536 (1991); Ronnblom et
al., Ann. Intern. Med. 115:178 (1991)). IFN.alpha. injections in
patients with different types of viral hepatitis have induced
autoimmune hepatitis (See, e.g., Ohta et al., J. Gastroenterol.
88:209 (1991); Fattovich et al., Brit. J. Med. Virol. 34:132
(1991)). In addition, it has been reported that a patient with
multiple sclerosis ("MS") given r-IFN.alpha. subcutaneously (Larrey
et al., JAMA 261:2065 (1989)), and another given r-IFN.gamma.
(Paniteh et al., Lancet 1:893 (1987)) intrathecally, manifested
clinical relapses at rates significantly higher than expected.
[0037] On the other hand, the neutralization of individual
cytokines, such as INF.alpha. or TNF.alpha., from the blood has
been associated with a significant therapeutic effect, in patients
with RA and in patients with AIDS (Skurkovich et al., 1975;
Gringeri et al., 1996). Thus, it is a purpose of the present
invention to provide methods of treating autoimmune disease by the
use of pleiotrophic autoimmune inhibitors, acting on each of the
known aberrant cytokine pathways in the patient and/or removing
pathogenic cytokines, HLA antigens, or autoantibodies from the
autoimmune patient.
[0038] The terms "patient" and "individual" are interchangeably
used to mean a warm-blooded animal, such as a mammal, suffering
from a disease, such as an autoimmune disease or "graft versus
host" disease, or is in danger of rejection of a transplanted
allogeneic tissue or organ. It is understood that humans and
animals are included within the scope of the term "patient" or
"individual."
[0039] "Cytokines" are intercellular mediators secreted by the
lymphocytes and/or macrophages. For example, cytokines play a role
in the generation of an immune response, such as in an immune
response to an infection or infectious organism. Cytokines
including, for example, interferons (IFN.alpha. and IFN.gamma.) and
TNFs induce other cytokines which participate in the development of
different autoimmune conditions and diseases. In the development of
anti-cytokine therapy in accordance with the present invention,
considerable emphasis has been placed on these three cytokines,
because it appears that by neutralizing these key cytokines
(IFN.alpha., IFN.gamma. and TNF), it is possible to decrease, halt
or prevent the synthesis of the cytokines induced by them. However,
is certain autoimmune conditions or diseases, including IDDM and
SLE, the induction of another cytokine (interleukins, specifically
IL-6) is so great and exerts such a strong pathological influence,
that it is desirable to remove IL-6 together with the other
cytokines.
[0040] IL-6 is made by several cells, including T-cells, B-cells,
and others (Hirano et al., Clin. Immunol. 62:S60 (1992)), and
induces insulinitis in IDDM. In response to IFN.gamma. and TNF,
B-cells of the pancreas produce large quantities of IL-6. It is
also an important pathological factor in the pathogenesis of SLE,
where is has been found to be present at a high level. IL-6
stimulates differentiation in B-cells and hyperactivity of T-cells
(Snick et al., Ann. Rev. Immunol. 8:253 (1990)). The increase in
IL-6 parallels the increase of TNF.alpha. (Majer et al., Lupus
2:359-365 (1993)).
[0041] The term "autoimmune inhibitor" is used to refer to a
"compound" or "compounds," including one or more molecules,
antigens, and/or antibodies (alone or in combination), which when
administered in an effective amount to a patient, binds to,
neutralizes or inhibits circulating pathological agents and/or
those on the surface of target cells, and which when placed in
extracorporeal contact with the patient's body fluids effects the
removal, neutralization or inhibition of complex pathological
agents (including hyperproduced cytokines and autoantibodies). The
autoimmune inhibitor may also comprise antibodies to a receptor of
the autoantigen. A "receptor" is a protein found on the surface of
a target cell or in its cytoplasm, that has a binding site with
high affinity to a particular signaling substance (e.g., a
cytokine, hormone, neurotransmitter, etc.). By competitively
inhibiting the availability of the receptor with an analog or
antibody to the receptor, the immune response to the autoimmunogen
is modified or neutralized.
[0042] In accordance with the present invention, treatments
involving administration of an autoimmune inhibitor to a patient,
and treatments involving the extracorporeal exposure of the
patient's fluid to an autoimmune inhibitor, may be performed alone
or in combination.
[0043] Administered autoimmune inhibitor of the invention binds to,
neutralizes and/or inhibits the molecule(s) associated with or
causing the auto immune response in the patient. More specifically,
administration of the autoimmune inhibitor to a patient results in
suppression of pathological humoral and adaptive immunity in the
patient. In other words, in accordance with the method of the
present invention, treatment of a patient with the autoimmune
inhibitor causes the humoral and adaptive immune response of the
patient to be inhibited or neutralized over that which was, or
would have been, present in the absence of treatment.
[0044] A patient is in need of treatment with an autoimmune
inhibitor, when the patient is suffering from an autoimmune
disease, or "graft-versus-host" disease, or when treatment is
needed to prevent rejection of transplanted allogeneic tissues or
organs, or when the patient has produced autoantibodies.
[0045] The term "autoimmune disease" refers to those disease states
and conditions wherein the immune response of the patient is
directed against the patient's own constituents, resulting in an
undesirable and often terribly debilitating condition. As used
herein, "autoimmune disease" is intended to further include
autoimmune conditions, syndromes and the like. An "autoantigen" is
a patient's self-produced constituent, which is perceived to be
foreign or undesirable, thus triggering an autoimmune response in
the patient, which may in turn lead to a chain of events, including
the synthesis of other autoantigens or autoantibodies. An
"autoantibody" is an antibody produced by an autoimmune patient to
one or more of his own constituents which are perceived to be
antigenic. For example, in AIDS disease the patient eventually
produces autoantibodies to CD4 cells, in SLE autoantibodies are
produced to DNA, while in many other types of AD autoantibodies are
produced to target cells (see, Table I for examples of specific
target cells of AD).
[0046] Patients suffering from autoimmune diseases including, e.g.,
rheumatoid arthritis, insulin-dependent diabetes mellitus,
hemolytic anemias, rheumatic fever, thyroiditis, Crohn's disease,
myasthenia gravis, glomerulonephritis, autoimmune hepatitis,
multiple sclerosis, systemic lupus erythematosus and others, are in
need of treatment in accordance with the present invention.
Treatment of patients suffering from these diseases by
administration of autoimmune inhibitor and/or removal of
compound(s) by extracorporeal immunosorption in accordance with the
present invention will alleviate the clinical manifestations of the
disease and/or minimize or prevent further deterioration or
worsening of the patient's condition. Treatment of a patient at an
early stage of an autoimmune disease including, e.g., rheumatoid
arthritis, insulin-dependent diabetes mellitus, multiple sclerosis,
myasthenia gravis, systemic lupus erythematosus, or others, will
minimize or eliminate deterioration of the disease state into a
more serious condition.
[0047] For example, insulin-dependent diabetes mellitus (IDDM) is
an autoimmune disease which is believed to result from the
autoimmune response directed against the beta cells of the islets
of Langerhans which secrete insulin. Treatment of a patient
suffering from an early stage of IDDM prior to the complete
destruction of the beta cells of the islets of Langerhans would be
particularly useful in preventing further progression of the
disease, since it would prevent or inhibit further destruction of
the remaining insulin-secreting beta cells. It is understood that
treatment of a patient suffering from an early stage of other
autoimmune diseases will also be particularly useful to prevent or
inhibit the natural progression of the disease state to more
serious stages.
[0048] The method of the present invention is applicable to
autoimmune diseases, such as those given in the following Table 1
(which is intended to be exemplary rather than inclusive), and
autoimmune conditions, such as those listed following the
Table.
1TABLE 1 Autoimmune Diseases Disease Tissue Affected Addison's
disease adrenal Autoimmune diseases of the ear ear Autoimmune
diseases of the eye eye Autoimmune hepatitis liver Autoimmune
parotitis parotid glands Crohn's disease intestine Diabetes (Type
I) pancreas Epididymitis epididymis Glomerulonephritis kidneys
Graves' disease thyroid Guillain-Barr syndrome nerve cells
Hashimoto's disease thyroid Hemolytic anemia red blood cells
Systemic lupus erythematosus multiple tissues Male infertility
sperm Multiple sclerosis nerve cells Myasthenia Gravis
neuromuscular junction Pemphigus primarily skin Psoriasis skin
Rheumatic fever heart andjoints Rheumatoid arthritis joint lining
Sarcoidosis multiple tissues and organs Scleroderma skin and
connective tissues Sjogren's syndrome exocrine glands, and other
tissues Spondyloarthropathies axial skeleton, and other tissues
Thyroiditis thyroid Vasculitis blood vessels
[0049] Autoimmune conditions for which the method of the present
invention is applicable include, for example, AIDS, atopic allergy,
bronchial asthma, eczema, leprosy, schizophrenia, inherited
depression, transplantation of tissues and organs, chronic fatigue
syndrome, Alzheimer's disease, Parkinson's disease, myocardial
infarction, stroke, autism, epilepsy, Arthus's phenomenon,
anaphylaxis, and alcohol and drug addiction. In the
above-identified autoimmune conditions, the tissue affected is the
primary target, in other cases it is the secondary target. These
conditions are partly or mostly autoimmune syndromes. Therefore, in
treating them, it is possible to use the same methods, or aspects
of the same methods that are herein disclosed for treating AD,
sometimes in combination with other methods.
[0050] Preferred embodiments of the invention are directed toward
the treatment of specific autoimmune disease or condition in a
patient, including those identified herein, and particularly
including RA, SLE, MS, juvenile RA, and ankylosing spondylitis.
[0051] Patients who have received, or who are about to receive, an
allogeneic tissue or organ transplant, such as an allogeneic
kidney, liver, heart, skin, bone marrow, are also patients who are
in need of prophylactic treatment with an autoimmune inhibitor
and/or removal of compound(s) by extracorporeal immunosorption in
accordance with the present invention. The autoimmune inhibitor of
the present invention will minimize or prevent the adaptive and
humoral immune response of the donee from rejecting the allogeneic
tissue or organ of the donor. Likewise, for patients suffering from
graft-versus-host disease treatment with an autoimmune inhibitor in
accordance with the method of the present invention will minimize
or prevent the adaptive and humoral immune response of the
transplanted tissue or organ from rejecting the allogeneic tissue
or organ of the donee.
[0052] Based on standard clinical and laboratory tests and
procedures, an attending diagnostician, physician or other person
skilled in the art, can readily identify those patients who are in
need of treatment with an autoimmune inhibitor. Such an individual
can also determine the compound or compounds to be included in the
autoimmune inhibitor for treatment in accordance with the methods
of the present invention, based upon the increased synthesis of
cytokines typifying the general onset and progression of autoimmune
disease, and on the clinical manifestations of the particular
disease being treated.
[0053] The term "fluid" refers to blood, plasma, plasma containing
leukocytes, serum, serum and leukocytes, peritoneal fluid,
cerebrospinal fluid, synovial fluid, amniotic fluid, or the like,
drawn from the patient in the practice of the present
invention.
[0054] An effective amount of autoimmune inhibitor is that amount
which is effective, upon single or multiple dose administration to
a patient, to bind to, neutralize or inhibit the autoimmunogen(s)
causing (directly or indirectly) or involved with the clinical
manifestation(s) of the autoimmune disease in the patient. In
addition, an effective amount of the autoimmune inhibitor in an
immunosorbent column over which the patient's fluid is passed, is
that amount which removes, neutralizes or inhibits the
autoimmunogen(s) causing (directly or indirectly) or involved with
the clinical manifestation(s) of the autoimmune disease in the
patient. The effect of administering the autoimmune inhibitor
and/or of extracorporeally passing fluid from the patient over
immunosorbent(s) comprising the autoimmune inhibitor in accordance
with the method of the present invention, can be seen as a slowing,
interruption, inhibition, neutralization or prevention of the
adaptive immune response associated with the autoimmune disease,
often displayed as an alleviation of clinical manifestations of the
disease. For example, the immunosuppressive effect of administering
an effective amount of antibody to IFN.gamma. to a patient in need
of such treatment would be the inhibition or prevention of further
expression of IFN.gamma. by the patient, which could be
quantitatively determined in terms of reduced fluid activity level
of one or more of the elevated cytokines, i.e., INF.gamma. or
TNF-.alpha.. The lowering of the cytokine activity level may be
measured directly in the treated patient, or the reduction in
cytokine activity level may be projected from clinical studies in
which dose regimens useful in achieving such reduction are
established.
[0055] An effective amount of autoimmune inhibitor can be readily
determined by the use of known techniques and by observing results
obtained under analogous circumstances. In determining the
effective amount or dose, a number of factors are considered by the
attending diagnostician, including, but not limited to: the species
of mammal; its size, age, and general health; the specific disease
involved; the degree of or involvement or the severity of the
disease; the response of the individual patient; as well as for
purposes of administration, the particular compound being
administered; the mode of administration; the bioavailability
characteristics of the preparation administered; the dose regimen
selected; the use of concomitant medication; and other relevant
circumstances.
[0056] The autoimmune inhibitor of the present invention may
comprise a single compound or anti-cytokine, e.g., anti-IFN.gamma.
antibody administered to the patient or used in extracorporeal
immunosorption, or it may be a combination of anti-cytokines or
compounds, e.g., a combination of antibodies to IFNs, TNFs, and the
like, administered to the patient or used in extracorporeal
immunosorption, and/or antigens such as a target cell, including a
CD4 cell, used in extracorporeal immunosorption. When combined, the
compounds may be used concomitantly in an admixture or as
simultaneous processes, or the compounds may be used sequentially
to provide a combined effect without being in physical combination.
For example, an AIDS patient may be treated by passing his blood,
plasma or the like extracorporeally over an immunosorbent
comprising CD4 cells to remove autoimmune antibodies against his
own CD4 cells, while at the same time, or sequentially,
anti-cytokines may be administered to neutralize, for instance the
interferons and TNFs that have been induced within his body. The
sequential treatments may occur in any order, so long as the
autoimmune inhibitors have the desired anti-autoimmune effect.
[0057] Combined treatments, comprising the use of one or more
autoimmune inhibitors in accordance with a preferred embodiment of
the invention, may be mechanistically advantageous. This is because
although circulating immunogens can be removed extracorporeally by
passing the patient's body fluid over an immunosorbent comprising
the autoimmune inhibitor(s), the administration of suitable
autoimmune inhibitor(s), such as anti-cytokine antibodies, can
effectively neutralize the immunogens, such as cytokines, both in
circulation and on the cell surface. For example, to remove
autoantibodies to CD4 cells, CD4 cells must be placed into an
immunosorbent column. The body fluid from the patient is
extracorporeally exposed to an immunosorbent comprising CD4 cells
or their fragments, then the treated fluid (minus the antibodies
that would otherwise attack his own CD4 cells) is returned to the
patient. An attending diagnostician, physician or other person
skilled in the art, can readily identify those patients who are in
need of administrative treatment with an autoimmune inhibitor, or
those who would benefit from extracorporeal treatment of their body
fluids, or those who would benefit from a combination of the
two.
[0058] The compound(s) comprising the autoimmune inhibitor, e.g.,
antibodies to IFNs, TNFs, and the like, and/or antigens such as a
target cell, including CD4 cells, in accordance with the methods of
the present invention, include cytotoxic amino acid sequence and
glycosylation variants which also are used herein. The terms
likewise cover biologically active functional equivalents,
derivatives, or allelic or species variants of each compound, e.g.,
those differing by one or more amino acids(s) in the overall
sequence. Further, the terms used in this application are intended
to cover substitution, deletion and insertion amino acid variants
of each compound, or post-translational modifications thereof.
[0059] Removal, neutralization and/or inhibition of alpha and gamma
IFNs, TNF, and HLA class II antigen, and the like, and/or their
receptors can be accomplished by the administration to the patient
of one or more antibodies, or by including one or more antibodies
in the immunosorbent over which the patient's body fluid is passed
for extracorporeal treatment. As used herein, the term "antibody"
is intended to include monoclonal or polyclonal antibodies, or a
combination thereof, humanized forms of the monoclonal antibodies
(comprising only human antibody protein), and chimeric monoclonal
antibodies, as well as biologically active fragments, functional
equivalents, derivatives, or allelic or species variants thereof.
Treatment can include polyclonal antibodies from different animal
species.
[0060] The term "biologically active fragment" is intended to mean
a part of the complete molecule which retains all or some of the
catalytic or biological activity possessed by the complete
molecule, especially activity that allows specific binding of the
antibody to an antigenic determinant. "Functional equivalents" of
an antibody include any molecule capable of specifically binding to
the same antigenic determinant as the antibody, thereby
neutralizing the molecule, e.g., antibody-like molecules, such as
single chain antigen binding molecules.
[0061] "Derivative" is intended to include both functional and
chemical derivatives, including fragments, segments, variants or
analogs of a molecule. A molecule is a "chemical derivative" of
another, if it contains additional chemical moieties not normally a
part of the molecule. Such moieties may improve the molecule's
solubility, absorption, biological half life, and the like, or they
may decrease toxicity of the molecule, eliminate or attenuate any
undesirable side effect of the molecule, and the like. Moieties
capable of mediating such effects are disclosed in Remington's
Pharmaceutical Sciences (1980). Procedures for coupling such
moieties to a molecule are well known in the art.
[0062] A "variant" or "allelic or species variant" of a protein
refers to a molecule substantially similar in structure and
biological activity to the protein. Thus, if two molecules possess
a common activity and may substitute for each other, it is intended
that they are "variants," even if the composition or secondary,
tertiary, or quaternary structure of one of the molecules is not
identical to that found in the other, or if the amino acid or
nucleotide sequence is not identical.
[0063] The term "IFN" is intended to refer to any known subtype of
IFN. For example, "IFN.alpha." is broadly intended to include any
of the known 15 subtypes of IFN.alpha., or any that may be
determined in the future. The term "HLA class II antigens" is
intended to mean not only HLA class II antigens, but also where
appropriate, HLA class I or III antigens.
[0064] Any animal (mouse, rabbit, human, etc.) which is known to
produce antibodies can be utilized to produce antibodies with the
desired specificity. Methods for immunization are well known in the
art. Such methods include subcutaneous or interperitoneal injection
of the polypeptide. One skilled in the art will recognize that the
amount of polypeptide used for immunization will vary based on the
animal which is immunized, the antigenicity of the polypeptide and
the site of injection. Chimeric antibodies, generated by recognized
methods can also be used, including antibodies produced by
recombinant methods.
[0065] If the antibody is to be administered intramuscularly or
intravenously into the patient, then it may be preferable to use a
substantially purified monoclonal antibody produced in human
hybridoma. Humanized forms of the antibodies of the present
invention may be generated using one of the procedures known in the
art such as chimerization or CDR grafting. Also monoclonal
antibodies of completely human protein may be applied. Until a
satisfactory partner for human B-cells or activated human B-cells
suitable for fusion become more readily available, a recognized
procedure based upon immortalization of human B-cells with
Epstein-Barr virus has provided a source of human antibodies (see,
Burton, Hospital Practice (August 1992), 67).
[0066] In general, techniques for preparing monoclonal antibodies
are well known in the art (Campbell, A. M., "Monoclonal Antibody
Technology: Laboratory Techniques in Biochemistry and Molecular
Biology," Elsevier Science Publishers, Amsterdam, The Netherlands
(1984); St. Groth et al., J. Immunol. Methods 35:1-21 (1980). For
example, in one embodiment an antibody capable of binding to
IFN.gamma. is generated by immunizing an animal with natural,
synthetic or recombinant IFN.gamma..
[0067] To produce the antibodies of the present invention, a
cytokine or antigen may be modified or administered in an adjuvant
in order to increase the peptide antigenicity. Methods of
increasing the antigenicity of a polypeptide are well known in the
art. Such procedures include coupling the antigen with a
heterologous protein (such as globulin or .beta.-galactosidase) or
through the inclusion of an adjuvant during immunization.
[0068] For monoclonal antibodies, spleen cells from the immunized
animals are removed, fused with myeloma cells, such as SP2/0-Ag14
myeloma cells, and allowed to become monoclonal antibody producing
hybridoma cells. A hybridoma is an immortalized cell line which is
capable of secreting a specific monoclonal antibody. Any one of a
number of methods well known in the art can be used to identify the
hybridoma cell which produces an antibody with the desired
characteristics. These include screening the hybridomas with an
ELISA assay, western blot analysis, or radioimmunoassay (Lutz et
al., Exp. Cell Res. 175:109-124 (1988)). Hybridomas secreting the
desired antibodies are cloned and the class and subclass are
determined using procedures known in the art (See, Monoclonal
Antibody Technology: Laboratory Techniques in Biochemistry and
Molecular Biology, supra).
[0069] For polyclonal antibodies, antibody containing antisera is
isolated from the immunized animal and is screened for the presence
of antibodies with the desired specificity using one of the
above-described procedures. Polyclonal antibodies raised from
animals immunized with specific antigens (IFNs, TNF, etc.) can be
used after the isolation of the active fraction (e.g., IgG) or
isolated Fab fragment.
[0070] The autoimmune inhibitor antibody(ies) also may be produced
and/or isolated from discordant animal species. For example,
porcine or bovine antibodies may be used for the treatment of
humans. To use animal-derived antibodies for a prolonged period,
antibodies from a variety of different animal species must be used,
permitting the source of the antibodies to be changed if the
patient develops a hypersensitivity or deleterious response to a
component of the originally administered antibody, antibody
fragment or polypeptide. In some cases, to prevent allergenic
reaction between injections of antibodies from a discordant
species, immunodepressant drugs, such as steroid hormones or
cyclophosphamide are administered. A preferred compound of the
present invention is derived from a mature compound from
recombinant microbial cell culture, prepared, isolated and
substantially purified in accordance with known techniques. A
combination of monoclonal and polyclonal antibodies can also be
utilized.
[0071] To evaluate the antibody or antibodies, conditions for
incubating the antibody or antibodies with a test sample vary.
Incubating conditions depend on the format employed in the assay,
the detection methods employed, the nature of the test sample, and
the type and nature of the antibody used in the assay. One skilled
in the art will recognize that any one of the commonly available
immunological assay formats (such as, radioimmunoassays,
enzyme-linked immunosorbent assays; diffusion based Ouchterlony, or
rocket immunofluorescent assays, or the like) can readily be
adapted to employ the antibodies of the present invention.
[0072] Autoimmune inhibitor(s) of the present invention include
polypeptides comprising the epitope of the antibody or biologically
active fragment thereof, or polypeptide that is functional in
conferring protection in the individual suffering from autoimmune
disease, or functionally conserved fragments or amino acid variants
thereof. Identification of the epitope is a matter of routine
experimentation. Most typically, one would conduct systematic
substitutional mutagenesis of the compound molecule while observing
for reductions or elimination of cytoprotective or neutralizing
activity. In any case, it will be appreciated that due to the size
of many of the antibodies, most substitutions will have little
effect on binding activity. The great majority of variants will
possess at least some cytoprotective or neutralizing activity,
particularly if the substitution is conservative. Conservative
amino acid substitutions are substitutions from the same class,
defined as acidic (Asp, Glu), hydroxy-like (Cys, Ser, Thr), amides
(Asn, GIn), basic (His, Lys, Arg), aliphatic-like (Met, Ile, Leu,
Val, Gly, Ala, Pro), and aromatic (Phe, Tyr, Trp).
[0073] Homologous antibody or polypeptide sequences generally will
be greater than about 30 percent homologous on an identical amino
acid basis, ignoring for the purposes of determining homology any
insertions or deletions from the selected molecule in relation to
its native sequence. The compounds discussed herein, i.e.,
autoimmune inhibitors for administration to the patient with
autoimmune disease and/or for removal, neutralization or inhibition
of the autoimmunogen(s) by extracorporeal immunosorption in
accordance with the present invention, also include glycosylation
variants as well as unglycosylated forms of the agents, fusions of
the agents with heterologous polypeptides, and biologically active
fragments of the agents, again so long as the variants possess the
requisite neutralizing or cytoprotective activity.
[0074] The autoimmune inhibitor antibody(ies) is also effective
when immobilized on a solid support. Examples of such solid
supports include, but are not limited to, plastics such as
polycarbonate, complex carbohydrates such as agarose and sepharose,
and acrylic resins, such as polyacrylamide and latex beads.
Techniques for coupling antibodies to such solid supports are well
known in the art (Weir et al., "Handbook of Experimental
Immunology" 4th Ed., Blackwell Scientific Publications, Oxford,
England, Chap. 10 (1986); Jacoby et al., Meth. Enzym. 34 Academic
Press, N.Y. (1974).
[0075] Additionally, one or more of the antibodies used in the
above described methods can be detectably labeled prior to use.
Antibodies can be detectably labeled through the use of
radioisotopes, affinity labels (such as, biotin, avidin, etc.),
enzymatic labels (such as, horse radish peroxidase, alkaline
phosphatase, etc.) fluorescent labels (such as, FITC or rhodamine,
etc.), paramagnetic atoms, etc. Procedures for accomplishing such
labeling are well-known in the art, for example see Sternberger et
al., J. Histochem. Cytochem. 18:315 (1970); Bayer et al., Meth
Enzym. 62:308 (1979); Engval et al., Immunol. 109:129 (1972);
Goding, J. Immunol. Meth. 13:215 (1976). The labeled antibodies of
the present invention can be used for in vitro, in vivo, and in
situ assays to identify cells or tissues which express a specific
cytokine or antigenic protein.
[0076] For administration purposes, an effective amount of
autoimmune inhibitor is expected to vary from about 0.1 milligram
per kilogram of body weight per day (mg/kg/day) to about 500
mg/kg/day. Preferred amounts are expected to vary from about 1 to
about 50 mg/kg/day. Humanized monoclonal antibodies can be
administered daily for one or more weeks, depending on need;
whereas polyclonal antibodies can be given for no more than 5-6
days. If antibodies are used from a variety of species, a different
antibody can be given every 5-6 days.
[0077] Cytokines and other pathological agents can also be
neutralized or removed from the patient in accordance with the
methods of the present invention by administering vaccines against
the cytokines or agents. However, vaccines may be dangerous to use
in vivo, unless the antibodies that may be induced by the treatment
can be controlled. Otherwise, such vaccines, although initially
effective, may lead to immunological disaster in the patient.
[0078] In effecting treatment of a patient, an autoimmune inhibitor
can be administered in any form or mode which makes the compound
bioavailable in effective amounts, including oral and parenteral
routes. For example, autoimmune inhibitors can be administered by
inhalation, orally, subcutaneously, intramuscularly, intravenously,
transdermally, intranasally, rectally, and the like. Parenteral
administration is generally preferred. One skilled in the art of
preparing formulations can readily select the proper form and mode
of administration depending upon the particular characteristics of
the compound selected, the disease state to be treated, the stage
of the disease, and other relevant circumstances.
[0079] The autoimmune inhibitor can be administered alone, or in
the form of a pharmaceutical composition in combination with
pharmaceutically acceptable carriers or excipients, the proportion
and nature of which are determined by the solubility and chemical
properties of the compound selected, the chosen route of
administration, and standard pharmaceutical practice. The compounds
of the invention, while effective themselves, may be formulated and
administered in the form of their pharmaceutically acceptable acid
addition salts for purposes of stability, convenience of
crystallization, increased solubility and the like.
[0080] In one embodiment, the present invention provides a method
of treatment in which the autoimmune inhibitor is admixed or
otherwise associated with one or more inert carriers. These
compositions are useful, for example, as assay standards, as
convenient means of making bulk shipments, or as pharmaceutical
compositions. An assayable amount of an autoimmune inhibitor is an
amount which is readily measurable by standard assay procedures and
techniques as are welt known and appreciated by those skilled in
the art. Assayable amounts of the autoimmune inhibitor will
generally vary from about 0.001% to about 75% of the composition by
weight. Inert carriers can be any material which does not degrade
or otherwise covalently react with an autoimmune inhibitor.
Examples of suitable inert carriers include water; aqueous buffers,
such as those which are generally useful in High Performance Liquid
Chromatography (HPLC) analysis; organic solvents, such as
acetonitrile, ethyl acetate, hexane and the like; and
pharmaceutically acceptable carriers or excipients.
[0081] More particularly, in accordance with the present invention,
pharmaceutical compositions are provided comprising an effective
amount of autoimmune inhibitor in admixture or otherwise in
association with one or more pharmaceutically acceptable carriers
or excipients.
[0082] The pharmaceutical compositions are prepared in a manner
well known in the pharmaceutical art. The carrier or excipient may
be a solid, semi-solid, or liquid material which can serve as a
vehicle or medium for the active ingredient. Suitable carriers or
excipients are well known in the art. The pharmaceutical
composition may be adapted for oral, parenteral, or topical use,
and may be administered to the patient in the form of tablets,
powders, granules, capsules, suppositories, solution, suspensions,
or the like.
[0083] The compounds of the present invention may be administered
orally, for example, with an inert diluent or with an edible
carrier. They may be enclosed in gelatin capsules or compressed
into tablets. For the purpose of oral therapeutic administration,
the compounds may be incorporated with excipients and used in the
form of tablets, troches, capsules, elixirs, suspensions, syrups,
wafers, chewing gums and the like. These preparations should
contain a measurable amount of autoimmune inhibitor as the active
ingredient, but the amount may vary depending upon the particular
form and may conveniently be between about 1% to about 90% of the
weight of the pharmaceutical composition. The amount of the
compound present in compositions is such that a suitable dosage
will be obtained. Preferred compositions and preparations according
to the present invention are prepared so that an oral dosage unit
form contains between 5.0-300 milligrams of an autoimmune inhibitor
of the invention. Dosage, in tablet or capsule form, is at a
preferred dose of 1 to 25 mg/kg patient body weight per day. The
dose may be increased or decreased appropriately depending on the
response of the patient, and patient tolerance.
[0084] The tablets, pills, capsules, troches and the like may also
contain one or more of the following adjuvants: binders such as
microcrystalline cellulose, starch paste, gum tragacanth or
gelatin; excipients such as starch or lactose, disintegrating
agents such as alginic acid, corn starch and the like; lubricants
such as magnesium stearate; glidants such as colloidal silicon
dioxide; and sweetening agents such as sucrose or saccharin may be
added or a flavoring agent such as peppermint, methyl salicylate or
orange flavoring, of the types usually used in the manufacture of
medical preparations. When the dosage unit form is a capsule, it
may contain, in addition to materials of the above type, a liquid
carrier such as polyethylene glycol or a fatty oil. Other dosage
unit forms may contain other various materials which modify the
physical form of the dosage unit, for example, as coatings. Thus,
tablets or pills may be coated with sugar, shellac, or other
enteric coating agents.
[0085] For use in oral liquid preparation, the compound(s) may be
prepared as a liquid suspension, emulsion, or syrup, being supplied
either in liquid form or a dried form suitable for hydration in
water or normal saline. A syrup may contain, in addition to the
present compounds, sucrose as a sweetening agent and certain
preservatives, dyes and colorings and flavors.
[0086] Materials used in preparing these various compositions
should be pharmaceutically pure and non-toxic in the amounts used.
As used herein, a protein is said to be "pharmaceutically pure" if
the autoimmune inhibitor comprises no substance that would be
harmful to the patient. A "substantially pure" or "substantially
purified" protein is one in which specific activity cannot be
significantly increased by further purification, and if the
specific activity is greater than that found in whole cell extracts
containing the protein.
[0087] The method of the present invention is also accomplished by
injecting the selected compound(s) in the autoimmune inhibitor,
e.g., intravenously, intramuscularly, or subcutaneously, in the
form of aqueous solutions, suspensions or oily or aqueous
emulsions, such as liposome suspensions. Typically, for parenteral
administration, the extract is formulated as a lipid, e.g.,
triglyceride, or phospholipid suspension, with the extract
components being dissolved in the lipid phase of the suspension.
These preparations should contain at least 0.1% of an autoimmune
inhibitor of the invention, but may be varied to be between 0.1 and
about 50% of the weight thereof. The amount of autoimmune inhibitor
present in such compositions is such that a suitable dosage will be
obtained. Preferred compositions and preparations according to the
present invention are prepared so that a parenteral dosage unit
contains between 5.0 to 100 milligrams of autoimmune inhibitor.
Dosage level may be increased or decreased appropriately, depending
on the conditions of disease, the age of the patient, etc.
[0088] The solutions or suspensions may also include one or more of
the following adjuvants: sterile diluents such as water for
injection, saline solution, fixed oils, polyethylene glycols,
glycerine, propylene glycol or other synthetic solvents;
antibacterial agents such as benzyl alcohol or methyl paraben;
antioxidants such as ascorbic acid or sodium bisulfite; chelating
agents such as ethylene diaminetetraacetic acid; buffers such as
acetates, citrates or phosphates and agents for the adjustment of
tonicity such as sodium chloride or dextrose.
[0089] The parenteral preparation can be enclosed in ampules,
disposable syringes or multiple dose vials made of glass or
plastic.
[0090] Moreover, the invention provides for the treatment of a
patient with autoimmune disease by the use (administration or use
in extracorporeal immunosorbent) of one or more antisense
molecules, which are characterized by the ability to bind to the
autoimmunogen, or a functionally equivalent derivative, or allelic
or species variant thereof. "Antisense sequence," or "antisense
molecule" refers to peptides derived from pseudogenes which are
constructed by reversing the orientation of the gene encoding the
autoimmunogen with regard to its promoter, so that the antisense
strand is transcribed. The term also refers to the antisense strand
of RNA or of cDNA which compliments the strand of DNA encoding the
cytokine, autoimmunogen, protein or peptide of interest.
[0091] When introduced into the patient, the anti-sense molecule
binds to, neutralizes or inhibits the autoimmunogen, much the same
as an antibody. Thus, the present methods can be practiced by means
of one or more antisense molecules. Moreover, when the nucleic acid
sequence encoding the autoimmune anti-sense molecule is introduced
into the cells under the control of a promoter, the anti-sense gene
molecule binds to, neutralizes or inhibits the gene(s) encoding the
autoimmunogen(s), inhibiting or preventing further pathogenesis.
The inhibition appears to depend on the formation of an RNA-RNA or
cDNA-RNA duplex in the nucleus or in the cytoplasm. Thus, if the
antisense gene is stably introduced into a cultured cell, the
normal processing and/or transport is affected if a sense-antisense
duplex forms in the nucleus; or if antisense RNA is introduced into
the cytoplasm of the cell, the expression or translation of the
autoimmunogen is inhibited. Such antisense nucleic acid sequences
may further include modifications which could affect the biological
activity of the antisense molecule, or its manner or rate of
expression. Such modifications may also include, e.g., mutations,
insertions, deletions, or substitutions of one or more nucleotides
that do not affect the function of the antisense molecule, but
which may affect intracellular localization. Also, the nucleic acid
sequence may determine an uninterrupted antisense RNA sequence or
it may include one or more introns.
[0092] In a particular embodiment of the invention, a unique
combination of compounds may be combined to form the autoimmune
inhibitor to be used for the treatment of multiple sclerosis
("MS"), for which there is no other rational treatment. The
administration of beta interferon ("INF.beta.") has been shown to
decrease the rate of exacerbation of the disease in some patients.
This positive effect can be explained by the fact that INF.beta.
decreases the synthesis of IFN.gamma. and TNF (Henniger et al.,
Neurology 46:1633-1639 (1996)). These data both confirm the
negative effect of IFN.gamma. and TNF on the autoimmune process,
and validate the synergic action in MS of anti-cytokine antibodies
(anti-IFN.gamma. antibodies and anti-TNF antibodies) together with
the administration of the cytokine IFN.beta. to decrease the
production of IFN.gamma. and TNF.
[0093] In one embodiment of the invention, treatment comprises
passing the fluid drawn from the patient over immunosorbent
comprising the autoimmune inhibitor, followed by returning the
treated fluid to its source. This method is particularly suited for
treating certain autoimmune conditions in which the autoimmune
inhibitor cannot be administered to the patient. For example, in a
preferred embodiment, the patient's fluid is exposed to an
immunosorbent comprising an effective amount of target cells, CD4
cells, and/or DNA, to remove, neutralize or inhibit the
autoantibodies in the patient's fluid, followed by returning the
treated fluid to the patient. The immunosorbent for extracorporeal
treatment may further comprise one or more antibodies (e.g.,
anti-IFN.alpha. antibodies, antibodies to IFN.alpha. receptor,
anti-IFN.gamma. antibodies, antibodies to IFN.gamma. receptor,
anti-TNF antibodies, antibodies to TNF receptor, antibodies to an
HLA class II antigen or to its receptor, or immunoglobulin E
("IgE").
[0094] To counter transplant rejection, antibodies to IFN.alpha.
and IFN.gamma., or in some cases IFN.gamma. alone, and the antigen
of the transplanted cell or organ are placed in the immunosorbent
column. To treat myocardial infarction or stroke, antibodies to
IFNs and cardiac or brain antigens, respectively, are placed in the
immunosorbent column. Further, the present invention may be used in
combination with immunosuppressive therapy to achieve the desired
results.
[0095] In another preferred embodiment of the invention, the
patient's fluid is extracorporeally exposed to an immunosorbent
comprising target cells. For example, for the treatment of
rheumatoid arthritis, target cell antigens from joints, skin,
collagen, and possibly other target antigens, are used as
immunosorbents, alone or in conjunction with other autoimmune
inhibitors, such as antibodies to IFNs and/or TNF or their
receptors. In addition, for the treatment of rheumatic fever, the
invention provides an immunosorbent comprising antibodies to IFNs
and/or TNF or their receptors and/or other substances, in
conjunction with a second cardiac tissue sorbent for removing
autoantibodies against cardiac tissue. The second sorbent can also
include selected serotypes of Streptococcus (group "A"), because
certain antigens from cardiac tissue and some serotypes of
Streptococcus are antigenically similar. For the treatment of
autoimmune diseases of the central nervous system, target cell
antigens from brain cells, e.g., to nuclear, membrane or cytoplasm
antigens, are used to absorb autoantibodies formed against the
brain cells.
[0096] In yet another preferred embodiment of the invention, the
patient's fluid is extracorporeally exposed to an immunosorbent
comprising DNA. For example, for the treatment of SLE the
immunosorbent comprises DNA to remove, reduce or neutralize the
patient's anti-DNA autoantibodies. For a description of anti-DNA
antibodies as they appear in SLE, see Graninger et at, J.
Rheumatol. 18:1621-1622 (1981).
[0097] In a further preferred embodiment the fluid is
extracorporeally exposed to an immunosorbent comprising antibody to
IgE. For example, for treating certain diseases related to
hypersensitivity of the immediate type, e.g., bronchial asthma,
antibody to IgE is used as an immunosorbent, alone or in
conjunction with other autoimmune inhibitors, such as antibodies to
IFNs and/or TNF or their receptors.
[0098] In an additional preferred embodiment of the invention the
patient's fluid is extracorporeally exposed to an immunosorbent
comprising CD4 cells. For example, for the treatment of AIDS, the
immunosorbent comprises CD4 cells, alone or in conjunction with
other autoimmune inhibitors, such as antibodies to IFNs and/or TNF
and/or HLA class II antigen, or their receptors. The CD4 component
of the immunosorbent comprises lymphocytes, primarily CD4 cells,
from healthy donors to absorb serum autoantibodies which react with
the patient's own CD4 cells.
[0099] For extracorporeal treatment, the pathogenic antibodies
and/or immune lymphocytes can be removed or reduced by passing any
of the previously described fluids over the prepared immunosorbent
column comprising an autoimmune inhibitor. When using whole blood,
plasma, or plasma with leukocytes, one can use a blood cell
separator (e.g., Cobe "Spectra") to which the immunosorbent column
is connected. See, e.g., U.S. Pat. No. 4,362,155, which is
incorporated herein by reference. To remove pathological substances
from joint or spinal fluids or the like, a special extracorporeal
device with a small amount of immunosorbent is used. To neutralize
antibodies to autoimmunogens, such as antibodies to target cells,
including CD4 cells, the cells themselves or that portion of the
cells containing the antigenic determinant(s) for the subject
antibodies, must be placed directly in the immunosorbent
column.
[0100] For the removal of compound(s) by extracorporeal
immunosorption in accordance with the present invention, particles
of sorbent material, such as amorphous silica or Sepharose, can be
readily placed in a container to prepare the immunosorbent for the
extracorporeal procedure. The container can be constructed of any
material which can readily undergo steam, chemical, or
gamma-irradiation sterilization. For instance, glass,
polycarbonate, polystyrene, polymethylmethacrylate, polyolefins
such as polyethylene and polypropylene, are all suitable.
[0101] Various ways of retaining or immobilizing sorbent material
within a container are available. For instance, sorbent material
may be placed between layers of retaining filters, or placed within
a porous solid matrix. The solid matrix immobilizes the sorbent,
while simultaneously permitting flow of blood or other fluids, and
contact with the sorbent. As is readily apparent to one of ordinary
skill in the art, a wide variety of structures are available for
providing suitable fluid/sorbent contact, structures which do not
cause significant hemolysis. Prudent use of additional filters to
retain the sorbent particles in their container is preferred. The
pretreated, immobilized sorbent may be contacted with the fluid in
a variety of ways, e.g., admixture, elution, and the like, which
would be recognized in the art.
[0102] Although a columnar sorbent bed is exemplified in Example 1,
beds of any other shape capable of functioning in the manner
described herein may also be used. The length-to-diameter ratio of
the sorbent bed should be selected so as to minimize any pressure
drop along the bed, and to ensure that shear rates remain below the
known values that correlate with cellular damage or destruction.
The pressure drop along the sorbent bed (and thus the increase in
shear rate) is directly proportional to the length of the bed.
However, mitigating against use of a short bed is the fact that
clearance of a substance from the fluid increases with a longer
bed. The capability of the sorbent to adsorb can be assessed by
experiments in which a test solution (such as whole blood or
plasma) is contacted with the prepared sorbent at a constant
temperature. The data generated from such an experiment can be used
to determine an equilibrium constant (K), according to which the
capacity of the prepared sorbent is determined. An equilibrium
constant (K) is defined in units of (ml solution/g composition).
The capacity of a composition provides a way to estimate the mass
of the prepared sorbent required to remove a certain quantity of
material, such as a cytokine, from solution.
[0103] In one embodiment of the invention, one skilled in the art
will readily recognize that the disclosed autoimmune inhibitor or
immunosorbent comprising the autoimmune inhibitor of the present
invention can readily be incorporated into one of the established
kit formats which are well known in the art. While in yet another
embodiment of the present invention, kits are provided which
contain the necessary reagents to carry out the previously
described methods. For example, in one instance such a kit
comprises a pharmaceutical composition or antibody cocktail
comprising the necessary autoimmune inhibitor, with or without
pharmaceutically acceptable carriers, excipients and the like, in
an amount suitable for administration to a patient suffering from
an autoimmune disease. In another instance, such a kit comprises
the autoimmune inhibitor bound to an immunosorbent that may be used
for the extracorporeal treatment of autoimmune disease in a
patient. In particular, such a kit comprises an effective amount to
extracorporeally remove, reduce or neutralize one or more
autoimmunogens from the fluid of a patient with autoimmune disease
of at least one of the following: anti-IFN.alpha. antibodies,
antibodies to IFN.alpha. receptor, anti-IFN.gamma. antibodies,
antibodies to IFN.gamma. receptor, anti-TNF antibodies, antibodies
to TNF receptor, antibodies to an HLA class II antigen or to its
receptor, and/or antibodies to IgE. Another preferred kit comprises
an effective amount to extracorporeally remove, reduce or
neutralize one or more autoantibodies from the fluid of a patient
with autoimmune disease of at least one of the following: target
cells, CD4 cells, or DNA. While, yet additional kits comprise
components of each of the previously defined kits, to provide the
combined treatments of the present invention.
[0104] All essential publications mentioned herein are hereby
incorporated by reference.
[0105] In order that those skilled in the art can more fully
understand this invention, the following examples are set forth.
These examples are included solely for the purpose of illustration,
and should not be considered as expressing limitations unless so
set forth in the appended claims.
EXAMPLES
[0106] In the following examples and protocols, all commercially
available reagents were utilized in accordance with the
manufactures recommendations. The cell and protein purification
methods utilized in this application are established in the art and
will not be described in detail. Methodologic details may be
readily derived from the cited publications.
Example 1
Preparation of the Immunosorbent Column
[0107] Using a column and tubing made of plastic approved for the
use of blood, a column is prepared of small total volume,
approximately 30-35 ml. The column is filled with immunosorbent,
consisting essentially of one or more antigens or antibodies bound
to Sepharose 4B or another suitable matrix, through a short filling
tube placed at one end of the column. After the column has been
filled, an input tube to introduce the fluid sample, and a return
tube to return the treated sample to its source, are connected to
either end of the column. A filter is interposed between the input
tube and the column, and a second filter is interposed between the
column and the return tube. The two filters prevent the flow of
immunosorbent from the column. Two way stopcocks on the tubes
regulate flow throughout the system.
[0108] Sepharose CL-4B (Pharmacia, Piscattaway, N.J.) (100 ml) is
washed thoroughly with pyrogen free water, then suspended in 300 ml
ice cold 1 M NaCO.sub.3 pH 11.0. 20 gms CNBr in 10 ml acetonitrile
is added to the Sepharose. After 2 minutes this is collected on a
fretted glass funnel. The Sepharose cake is washed with 5 volumes
of ice cold 0.2M Na Bicarbonate buffer, pH 9.5, and 5 volumes of
ice cold 0.5 M Na Bicarbonate buffer, pH 8.5.
[0109] The prepared Sepharose is immediately resuspended in a
solution of the selected antigen or antibody or combination of one
or more antigens and/or antibodies. In this case, the immunosorbent
column is specifically prepared to bind to IFN.alpha., so the
prepared Sepharose is resuspended in a solution of 780 mg
anti-alpha IFN antibody in 200 ml of 0.2 M Bicarbonate buffer, pH
9.3. This is incubated for 20 hours at 4.degree. C. This is then
centrifuged, the supernatant is decanted, and sediment is
resuspended in 100 ml of 0.05 PBS (phosphate buffered saline) and 2
M glycine, pH 8.0, for 12 hours at room temperature. This is then
washed thoroughly with 20 volumes of PBS.
[0110] The column is positioned lower than the source of the fluid
sample, whereupon the fluid drawn from the patient flows into the
column under the influence of gravity. After the fluid perfuses
through the immunosorbent, it is collected in a holding tube from
which it is returned to the source of the fluid.
Example 2
Production of Antibody to Human IFN.gamma.
[0111] Adult rabbits are immunized with purified human IFN.gamma.
(10.sup.5-10.sup.6 unit/mg protein). The interferon is first mixed
with equal volumes of Freund's Complete Adjuvant and 30% Arlacel A
and injected IM or subcutaneously on day 1, 4, 14 and 43 (100
units, 200 units, 200, 200 respectively). Next, 200,000 units of
the interferon is injected per month, for an additional 6 months.
The serum is drawn from the rabbit when the titer has reached 100
units (1 unit of antibody neutralizes 10 units of gamma IFN), after
which IgG is isolated and substantially purified in accordance with
recognized methods.
Example 3
Responses to TNF.alpha., IFN.alpha., and IFN.gamma. Antibodies
Separately and Together, in Patients with Active Rheumatoid
Arthritis and Ankylosing Spondylitis
[0112] Polyclonal antibodies were obtained by immunizing sheep with
natural human IFN.alpha., and goats with recombinant human
IFN.gamma. ("r-Hu-IFN.gamma.") or recombinant human TNF.alpha.
("r-Hu-TNF.alpha."), and isolating the IgG from the animals. Each
milliliter of IgG contained approximately 50 mg of protein, and the
antibodies showed a 1:5 signal to noise ratio at 1:1250
(anti-IFN.alpha. antibodies) and 1:12,500 (anti-IFN.gamma.
antibodies and anti-TNF.alpha. antibodies) dilutions by ELISA
(CytoImmune Sciences, Inc.). After obtaining approval and informed
consent, 20 human patients with very severe rheumatoid arthritis
("RA"), aged 27-64, average disease duration 9 years, were equally
randomized to one of four (4) treatment groups. The patients in
Group A, B and C were given one intramuscular administration of 2-3
ml/day for 5 consecutive days of (Group A) anti-TNF.alpha.
antibodies; (Group B) anti-IFN.alpha. antibodies; or (Group C)
anti-IFN.gamma. antibodies. The patients in Group D were given a
combination of anti-TNF.alpha. antibodies+anti-IFN.alpha.
antibodies+anti-IFN.gamma. antibodies (6 ml/day--2 ml of each
antibody). All patients met the criteria of the American College of
Rheumatology for the diagnosis of RA and had not responded to any
of the standard disease-modifying rheumatoid drugs. Other criteria
for entry into the study included radiographic evidence of bone
erosion, the presence of severe illness as indicated by the
presence of 6 or more swollen joints and 3 of 4 secondary
indications including 45 minutes or more of continuous morning
stiffness, 6 or more painful joints, erythrocyte sedimentation rate
(ESR) of 25 mm/hr or higher, and C-reactive protein of 20 mg/l or
higher. Patients who were pregnant or who had serious illnesses or
conditions such as anemia, leukopenia, marked ankylosis of the
joints were excluded.
[0113] The primary response was determined by the Paulus index
(Paulus et al., Arthritis Rheum. 33:477-484 (1990)), i.e.,
.gtoreq.20% or .gtoreq.50% improvement in .gtoreq.4 of 6 measures
of laboratory and clinical effects (Table 2), which were obtained
through day 28. These include morning stiffness, number of painful
and inflamed joints, ESR, and at least a 2-point improvement on a
5-point scale of disease severity assessed by patient and by
physician. To maintain consistency, the same physician was used to
make all assessments.
[0114] Results
[0115] Signs of inflammation dropped in some patients within each
group on day one. All groups demonstrated marked improvement by day
7, though individual variation appeared in each treatment group.
Table 2 shows the proportion of patients achieving .gtoreq.20%
improvement in the Paulus measures. Based on these 6 measures, the
most positive response for all treatment groups was in the number
of swollen and painful joints. At day 7, the positive responses
using anti-TNF.alpha. antibodies (Group A), and the combined
antibody treatment (antibodies to all three cytokines; Group D),
were the strongest. Three (3) of the five (5) patients receiving
anti-TNF.alpha. antibodies, and two (2) of the five (5) receiving
the combined antibody treatment achieved .gtoreq.20% improvement in
4 or more Paulus measures, and at least one patient in each group
achieved at least 50% improvement.
[0116] In both Group A and D, all patients had at least 20%
improvement in morning stiffness and reduction in the number of
painful and swollen joints. Three (3) of the five (5) patients in
both groups reported at least a 2-point reduction (on a 5-point
scale) in overall disease severity. At day 28, the response to
anti-IFN.gamma. antibodies (Group C) was the strongest, including
one (1) patient reporting at least 50% improvement, and two (2)
others reporting at least 20% improvement in at least 4 of the 6
measures. In Group D (the combined antibody therapy), two (2)
patients reported at least 20% improvement in 4 or more measures.
By comparison, at day 28 only 1 of 4 patients in Group A (the
anti-TNF.alpha.antibody treatment group) reported having at least
20% improvement in 4 of the 6 measures. Comparable results are
achieved by extracorporeal immunosorption as defined above, or by
extracorporeal immunosorption in conjunction with administration of
an autoimmune inhibitor.
[0117] Four (4) of the 20 patients were taken off therapy or
follow-up after a temporary redness appeared at the point of
injection. Two (2) patients receiving anti-IFN.alpha. antibodies
(Group B) and one patient each receiving anti-TNF.alpha. antibodies
(Group A), and the combination therapy (Group D) exhibited such
reactions.
2TABLE 2 Proportion of Patients Achieving .gtoreq.20% Improvement
in Six Measures at Day 7 and Day 28, and Paulus Index by Treatment
Group Anti-TNF.alpha. Combined Anti-IFN.gamma. Ab Anti-IFN.alpha.
Ab Ab d.7 Paulus Measures d.7 d.28 d.7 d.28 d.7 d.28 d.28 Morning
stiffness (min.) 2/5 4/5 3/4 3/3 5/5 3/4 5/5 3/4 No Swollen Joints
4/5 3/5 2/4 2/3 5/5 3/4 5/5 3/4 No Painful Joints 4/5 4/5 2/4 3/3
5/5 4/4 5/5 3/4 Disease Severity 1/5 1/5 0/4 0/3 3/5 1/4 2/5 2/4
(by Physician*) Disease Severity 1/5 2/5 0/4 0/3 3/5 2/4 3/5 1/4
(by Patient*) ESR 2/5 3/5 1/4 2/3 1/5 1/4 1/5 1/4 Paulus Index
.gtoreq.20%** 1/5 2/5 0/4 2/3 3/5 1/4 2/5 2/4 .gtoreq.50%** 0/5 1/5
0/4 0/3 1/5 0/4 1/5 0/4 *.gtoreq.2-point improvement on 5-point
scale as assessed by physician or patient. **Proportion of patients
achieving .gtoreq.20% (or .gtoreq.50%) improvement in .gtoreq.4 of
the 6 measures at day 7 and day 28. .gtoreq.20% includes any
patient achieving .gtoreq.50% improvement.
[0118] One ankylosing spondylitis ("AS") patient, age 22, disease
duration one year, was treated with the combined antibody regimen
(antibodies to IFN.alpha., IFN.gamma., and TNF.alpha.). Improvement
in painful sacroiliac joint disease, diminution of radiating pain,
and normalization of the erythrocyte sedimentation rate was seen on
days 7-8.
[0119] For repeated treatment of human patients with autoimmune
disease, or for treatment of a human patient with a secondary
autoimmune condition, fully humanized monoclonal antibodies must be
used or, as a temporary alternative, chimeric monoclonal or
multi-specied IgG polyclonal antibodies or active antibody fragment
preparations.
[0120] The results indicate that a common mechanism appears to
underlie all AD, with disturbed cytokine production in different
target cells producing the various clinical manifestations.
Moreover, the results establish that each cytokine (e.g.,
IFN.alpha., IFN.gamma., TNF.alpha.) plays its own pathological role
in the mutual induction and activation of other cytokines,
suggesting a single target in treatment. Although other autoimmune
diseases may require treatment with different anti-cytokines,
antibodies or combination of autoimmune inhibitors, neutralization
of such agents, e.g., the exemplified cytokines, appears to break
the chain of pathological reactions typifying AD and normalize the
synthesis of other induced cytokines in AD patients, including AIDS
patients.
Example 4
Long-Term Improvement in Child with Juvenile Rheumatoid Arthritis
in Response to Treatment with IFN.alpha. and TNF.alpha.
Antibodies
[0121] The patient was a seven-year old girl who had been diagnosed
three years earlier (January 1993) as having juvenile rheumatoid
arthritis ("JRA"), polyarticular form, sero-negative, after
presenting with fever, arthralgias, extreme limitation of motion in
the right hip joint, neutrophilia, high ESR, and anemia. The
patient improved slightly on an initial regimen of non-steroidal
anti-inflammatory drugs (NSAID). But within six (6) months (Fall,
1993) exacerbation of her disease necessitated enhancing the
treatment with azathioprine, NSAIDs, and with pulse therapy using
Solumedrol. The patient was maintained on weekly methotrexate from
February 1994 until July 1995, when her disease relapsed. However,
despite increased NSAID therapy, her condition continued to
deteriorate. In light of the ineffectiveness of conventional
therapy, and because the disease had progressed to include hip
joint involvement, which invariably leads to crippling of a child,
this child became a candidate for the combined antibody treatment
of the present invention.
[0122] As described above, and using immunological techniques,
antibodies to IFN.gamma. ("anti-IFN.gamma. antibodies") and
antibodies to TNF.alpha. ("anti-TNF.alpha. antibodies") were
obtained by immunizing goats with r-IFN.gamma. and r-TNF.alpha.,
respectively, and isolating IgG from the immunized animals. Each
milliliter of IgG contained approximately 50 mg of protein, and the
antibodies showed a 1:5 signal to noise ratio at 1:12,500 dilutions
by ELISA (assays performed by CytoImmune Sciences, Inc., College
Park, Md.).
[0123] Two (2) ml/day each of anti-IFN.gamma. antibodies (3 days)
and anti-TNF.alpha. antibodies (5 days) were administered
parenterally to the child. By the second week of observation,
absence of morning stiffness, elimination of hip joint pain, and
considerable increases in the level of physical activity, range of
motion in the affected joints, and grip strength were noted (See,
Table 3). X-rays of the child showed improvement in the appearance
of the femurs and hip joints, and greater delineation of articular
spaces. Repeated testing of the child indicated a significant drop
in disease activity, as shown by clinical and laboratory
parameters, including pain, stiffness, grip strength, C-reactive
protein, and others (See, Table 3). The improvement in clinical
status and the nearly normal range of motion in the child's hip
joints persisted into the fourth month, as shown by x-rays at
regular check-ups. After six months (the most recent data
available), damage to the child's femurs and acetabulae were less
marked as shown on x-rays, and she continued to improve in other
parameters, to the point that on the advice of an orthopedist, her
joints were-allowed to bear greater weight.
3TABLE 3 Dynamics of clinical and laboratory parameters in patient
with JRA, after treatment with anti-IFN.gamma. antibodies and
anti-TNF.alpha. antibodies Before Week Week Week Week Parameter
Treatment 1 2 3 4 Arthralgia score * 4 2 2 0 0 Joint stiffness
(min.) 30 10 0 0 0 Grip strength (mm/Hg) 20 44 72 68 70 Angie of
abduction-rt hip 15 15 20 n/a 30 (degrees) Circumference of right
wrist 12.9 12.7 12.2 11.9 12.0 (cm) ESR 6 3 8 6 6 C reactive
protein (g/l) 0.6 neg neg neg neg * Scale of 0-5 where 5 is most
intense pain. n/a = Not available.
[0124] These data point to a role of cytokines in AD, and again
reinforce the conclusion that a common pathological mechanism
underlies clinically disparate forms of AD. It is the differences
in the target cells affected that result in the varying clinical
manifestations of the autoimmune response in a patient.
[0125] As demonstrated by the results produced in this child,
neutralization of certain cytokines with antibodies can break the
chain of pathological reactions and normalize the synthesis of
other induced cytokines in the patient. Other types of AD can be
treated by the use of anti-cytokines, singly or in combinations, to
counteract autoimmune aggression and inflammation. Good results
have been reported from double-blind placebo controlled trials
using chimeric monoclonal anti-TNF.alpha. antibodies to treat RA
(Elliott et al., Lancet 344: 1105-1110 (1994)). But until the
present invention, there has been no suggestion of treatment of AD
with anti-IFN.gamma. antibodies, nor with a combination of
anti-cytokine antibodies. Nor have the effects of such treatments
been evaluated in clinical trials. Given the striking long-term
results produced by the present method, the combined
anti-cytokines, e.g., anti-TNF.alpha. antibodies in conjunction
with anti-IFN.gamma. antibodies, may even act synergistically.
Example 5
Treatment of Patients with Systemic Lupus Erythematosus
[0126] Human patients with systemic lupus erythematosus (SLE) were
selected after obtaining approval and informed consent, in much the
same manner as set forth in Example 3, and divided into two groups
consisting of at least four (4) patients each. The basis for
selection was the patient's failure to respond to conventional
therapy for SLE. Using polyclonal anti-IFN.gamma. antibodies and
anti-TNF antibodies in accordance with Example 3, one group of
patients was treated with anti-IFN.gamma. antibodies, while the
other group was treated with anti-IFN.gamma. antibodies and
anti-TNF antibodies. The antibodies were administered in accordance
with the schedule and amounts set forth in Example 3 for 5
consecutive days.
[0127] Preliminary results, based upon at least one patient in each
group, indicate that pain and swelling in joints have decreased and
skin lesions have disappeared, further indicating that a common
mechanism underlies all AD, with disturbed cytokine production in
different target cells producing the various clinical
manifestations.
[0128] Comparable results are achieved by extracorporeal
immunosorption as defined above, or by extracorporeal
immunosorption in conjunction with administration of an autoimmune
inhibitor.
Example 6
Treatment of Patients with Multiple Sclerosis
[0129] Human patients with multiple sclerosis (MS) were selected
after obtaining approval and informed consent, in much the same
manner as set forth in Example 3, and divided into three groups
consisting of at least five (5) patients each. The basis for
selection was the presence of active MS and the patient's failure
to respond to conventional therapy for MS. Using polyclonal
anti-IFN.gamma. antibodies and anti-TNF antibodies in accordance
with Example 3, one group of patients was treated with
anti-IFN.gamma. antibodies, one group with anti-TNF antibodies, and
one group with anti-IFN.gamma. antibodies and anti-TNF antibodies.
The antibodies were administered in accordance with the schedule
and amounts set forth in Example 3 for 5 consecutive days, and the
patients were followed for at least two and one half (21/2)
months.
[0130] Results of the treatment were evaluated in terms of measured
neurological deficiencies and general patient function at the end
of the 21/2-month period, as compared with pretreatment
determinations of the same criteria. Determinations were based upon
the Disability Status Scale (DSS) devised by J. F. Kurztke, and the
Functional System Scale (FSS), respectively. Decreasing numbers
indicate improvement on the DSS scale, while increasing numbers
indicate improvement on the FSS scale. Preliminary results indicate
that improvement was most evident in the group treated with
anti-IFN.gamma. antibodies and in the group treated with
anti-IFN.gamma. antibodies and anti-TNF antibodies, as determined
by the two scales.
[0131] Additional studies indicate that the treatment may be
further enhanced by the administration of beta interferon
(IFN.beta.). When eight million international units (IU) of
IFN.beta. were given subcutaneously to patients every other day for
two years, there was a decrease in the rate of exacerbated symptoms
in some patients. Consequently, an optimal treatment of an MS
patient appears to be the use of anti-IFN.gamma. antibodies or a
combination of anti-IFN.gamma. antibodies and anti-TNF antibodies
(by administration or by extracorporeal immunosorption, or both, as
defined above), plus the administration of an effective amount of
IFN.beta..
Example 7
Treatment of AIDS Patients
[0132] A pilot study has been conducted with AIDS patients which
indicated the correlation between a reduction in serum IFN levels
and improved clinical status. In one study, four (4) patients with
very high serum levels of IFN and low levels of CD4 cells
(25/mm.sup.3), when injected with anti-IFN.alpha. antibodies
capable of neutralizing the circulating IFN.alpha., reported an
increased sense of well-being, energy, and appetite, and a
disappearance of skin rashes as the circulating IFN.alpha. was
neutralized and removed. By corollary, when the symptoms returned
in one patient 5 months later, it was determined that circulating
IFN.alpha. was again present in his blood. However, following a
second cycle of treatment with anti-IFN.alpha. antibodies, his
condition improved as the levels of circulating IFN.alpha.
diminished. See, Skurkovich et al., Med. Hypoth. 42:27-35 (1994),
herein incorporated by reference.
[0133] In light of the previously demonstrated effects of reducing
circulating IFN.alpha. in AIDS patients, and the consistently
positive effect that has resulted from the combined neutralization
of IFN.alpha., IFN.gamma. and/or TNF in patients with other
autoimmune diseases, similar effects are seen in AIDS patients when
treated with the combined antibodies of the present invention.
However, greater reduction in the clinical manifestations of AIDS
disease in patients results from-a combined therapy, including the
neutralization or removal of IFN.alpha., IFN.gamma. and/or TNF (by
administration of antibodies to IFN.alpha., IFN.gamma. and/or TNF,
and/or their receptors, and/or by the extracorporeal exposure of
the patient's fluid to an immunosorbent comprising antibodies to
IFN.alpha., IFN.gamma. and/or TNF, and/or their receptors), in
conjunction with inhibition, removal or neutralization of
autoimmune autoantibodies in the patient. This is accomplished by
extracorporeally exposing the patient's fluid to an immunosorbent
comprising CD4 cells and/or target cells in an amount sufficient to
remove, neutralize or inhibit autoantibodies to CD4 cells and/or to
target cells in the patient's fluid, followed by returning the
fluid to the patient, in accordance with the methods disclosed
herein.
[0134] Based on the assumption that a common mechanism underlies
all AD, and that it is the effect of the modified cytokine
production, as well as the production of subsequent components of
the autoimmune cascade, on different target cells that results in
the various clinical manifestations of each specific disease or
condition, the quality of life can be improved, or even extended,
in general in patients with an autoimmune disease or condition.
Consequently, although the present invention has been described
with reference to the presently preferred embodiments and examples,
the skilled artisan will appreciate that various modifications,
substitutions, omissions and changes may be made without departing
from the spirit of the invention.
* * * * *