Definitions

• CD4 is a non-polymorphic surface glycoprotein receptor with partial sequence identity to immunoglobulins. CD4 receptors define distinct subsets of mature peripheral T cells. In general, CD4 T cells expressing helper or regulatory functions interact with B cells in immune responses, while T cells expressing the
• CD8 surface antigen function as cytotoxic T cells and have regulatory effects on immune responses. Since T-cell receptors are the pathway through which stimuli augment or modulate T-c ⁇ ll responses, they present a potential target for i munological intervention.
• CD4+ T cells with antigen presenting cells lies at the root of the immune response. Many aspects of the autoimmune response are essentially similar to that of normal immune responses.
• CD4+ autoantigen reactive T cells are resti ulated by APC expressing class II with autoantigen peptides in the binding groove.
• rheumatoid arthritis in rheumatoid arthritis (RA) , in vivo activated T cells recognizing collagen type II have been isolated from joints of an RA patient in three consecutive operations during the course of three years (Londei, M. et al. r Proc. Natl. Acad. Sci. 86: 636- 640 (1989)).
• CD4+ T cells from the blood have been cloned, including CD4+ T cells recognizing the acetylcholine receptor in myasthenia gravis (Hohlfeld, R. ⁇ t al.. Nature 310: 224-246 (1984)); myelin basic protein in multiple sclerosis (Hafler, D.A.
• TNF ⁇ tumor necrosis factor- ⁇
• cachectin cy o ine tumor necrosis factor- ⁇
• TNF ⁇ is a protein secreted primarily by monocytes and acrophages in response to endotoxin or other stimuli as a soluble homotrimer of 17 kD protein SUbunits (Smith, R.A. e al.. J. Biol. Chem. 262: 6951- 6954 (1987)).
• a membrane-bound 26 kD precursor form of TNF has also been described (Kriegler, M. et al.. Cell 53: 45-53 (1988).
• TNF is also produced by CD4+ and CD8+ peripheral blood T lymphocytes, and by various cultured T and B cell lines (Cuturi, M.C. £__j_L_. J. EXP. Med. 165: (1581 (1987); Sung, S.-S.J. ⁇ ____ , J. Exp. Med. 168: 1539 (1988); Turner, M. __ ai ⁇ ., Eur. J. Immunol. 17: 1807-1814 (1987)). Recent evidence implicates TNF in the autoimmune pathologies and graft versus host pathology (Piguet, P.-F. *t flit. J. EXP. Med. 166: 1280 (1987).
• CD4+ T cell inhibiting agents include agents which block, diminish, inhibit, or interfere with the activation of CD4+ T cells or the interaction of CD4+ T cells with antigen presenting cells (APC) , such as antibodies to T cells or to their receptors; antibodies to APC or to their receptors; and other appropriate pep ides or small molecules.
• APC antigen presenting cells
• TNF antagonists include agents which block, diminish, inhibit, or interfere with TNF activity, TNF receptors, or TNF synthesis, such as anti-TNF antibodies; soluble TNF receptors; and other appropriate peptides or small molecules.
• anti-CD4 antibodies are administered in conjunction (either simultaneously or sequentially) with anti-TNF antibodies.
• anti-CD4 antibodies are administered in conjunction with soluble TNF receptor, such as a TNF receptor/TgG fusion protein.
• cyclosporin is administered in conjunction with anti-TNF antibody.
• the combination therapy can utilize any CD4+ T cell inhibiting agent in conjunction with any TNF antagonist, including multiple CD4+ T cell inhibiting agents in conjunction with multiple TNF antagonists.
• Combination therapy can also utilize inflammatory mediators other than TNF antagonists, in conjunction with CD4+ T cell inhibiting agents.
• the CD4+ T cell inhibiting agent and TNF antagonist can be administered together with a pharmaceutically acceptable vehicle; administration can be in the form of a Bingle dose, or a series of doses separated by intervals of days or weeks.
• the benefits of combination therapy with CD4+ T cell inhibiting agents and TNF antagonists include improved results in comparison with the effects of treatment with each therapeutic modality separately.
• lower dosages can be used to provide the same reduction of the immune and inflammatory response, thus increasing the therapeutic window between a therapeutic and a toxic effect.
• Lower doses may also result in lower financial costs to the patient, and potentially fewer side effects.
• Figure 1 contains a set of graphs, individually labelled as Fig. 1A and Fig. IB, from an experiment which illustrates the suppression of arthritis as assessed by clinical score (Fig. 1A) and pawswelling measurements (Fig. IB) after the administration of 50 ⁇ g anti-TNF (hamster TN3.19.2) and 200 ⁇ g anti-CD4 to DBA/1 male mice.
• Figure 2 contains a set of graphs, individually labelled as Fig. 2A, Fig. 2B, Fig. 2C, and Fig.
• FIG. 2D from a second experiment which illustrates the potentiation of anti-CD4 with low dose (50 ⁇ g) anti-TNF or high dose (300 ⁇ g) anti-TNF on clinical score and pawswelling measurements.
• Fig. 2A clinical score with low-dose anti-TNF
• Fig. 2B clinical score with high-dose anti- TNF
• Fig. 2D pawswelling with high-dose anti-TNF.
• Figure 3 is a graph illustrating the suppression of arthritis as assessed by pawswelling measurements after the administration of 250 ⁇ g cyclosporin A, 50 ⁇ g anti-TNF antibody, and a combination of 250 ⁇ g cyclosporin A and 50 ⁇ g anti-TNF antibody to DBA/1 mice.
• the present invention concerns the treatment of autoimmune or inflammatory diseases, such as rheumatoid arthritis, through the administration of a CD4+ T cell inhibiting agent in conjunction with a TNF antagonist.
• the invention also encompasses the use of multiple CD4+ T cell inhibiting agents in conjunction with multiple TNF antagonists.
• CD4+ T cell inhibiting agent refers to an agent which blocks, diminishes, inhibits, or interferes with the activation of CD4+ T cells or the interaction of CD4+ T cells with antigen presenting cells (APC) .
• CD4+ T cell inhibiting agents include antibodies to T cells or to their receptors, such as anti-CD4, anti-CD28, anti-CD52 (e.g., CAMPATH-1H) and anti-IL-2R; antibodies to APC or to their receptors, such as anti-class II, anti-ICAM-1, anti-LFA-3, and anti-LFA-1; peptides and small molecules blocking the T cell/APC interaction, including those which block the HLA class II groove, or block signal transduction in T-cell activation, such as cyclosporins, particularly cyclosporin A, or FK- 506; and antibodies to B cells including CD5+ B cells, such as CD19, 20, 21, 23 and BB/7 or B , ligands for CD28, B cells including CD5+ B cells are considered to be an important type of APC in disease processes (Plater-Zyberk, C. SLU. _-_L_ . Ann. N.Y. Acad. Sci. 651: 540-555 (1992)),
• TNF antagonist refers to an agent which blocks, diminishes, inhibits, or interferes with TNF activity, TNF synthesis, or TNF receptors, such as anti-TNF antibody; soluble TNF receptor (monomeric receptor and/or fusion proteins comprising the receptor, such as receptor/IgG fusion proteins, etc.); and other appropriate peptides or small molecules, such as pentoxyfilline or other phosphodiesterase inhibitors, and thalidomide.
• Inflammatory mediators other than TNF antagonists can also be used instead of or in addition to TNF antagonists in the current invention.
• In rheumatoid joint cell cultures Brennan e£ j ⁇ f ancet 11. 244-247 (1989) have shown that blocking TNF results in down-regulation of IL-1 production, and down-regulation of the pro-inflammatory cytokine GM-CSF (Haworth ⁇ £ ____ E.J.I. 21:2575-2579
• inflammatory mediators include agents which block, diminish, inhibit, or interfere with IL-1 activity, synthesis, or receptor signalling, such as anti-IL-1 antibody, soluble IL-1R, IL- 1 receptor antagonist, or other appropriate peptides and small molecules; agents which block, diminish, inhibit, or interfere with IL-6 activity, synthesis, or receptor signalling, such as anti-IL-6 antibody, anti-gp 130, or other appropriate peptides and small molecules; modalities which block, diminish, inhibit, or interfere with the activity, synthesis, or receptor signalling of other inflammatory mediators, such as GM-CSF and members of the ch ⁇ mokin ⁇ (IL-8) family; and cytokine ⁇ with anti- inflammatory properties, such as IL-4, IL-10, and TGF/S.
• agents which block, diminish, inhibit, or interfere with IL-1 activity, synthesis, or receptor signalling such as anti-IL-1 antibody, soluble IL-1R, IL- 1 receptor antagonist, or other appropriate peptides and small molecules
• anti-CD4 antibody is used in conjunction with anti-TNF antibody.
• the term antibody is intended to encompass both polyclonal and monoclonal antibodies.
• the term antibody is also intended to encompass mixtures of more than one antibody reactive with CD4 or with TNF (e.g., a cocktail of different types of monoclonal antibodies reactive with CD4 or with TNF) .
• the term antibody is further intended to encompass whole antibodies, biologically functional fragments thereof, bifunctional antibodies, and chimeric antibodies comprising portions from more than one species.
• Biologically functional antibody fragments which can be used are those fragments sufficient for binding of the antibody fragment to CD4 or to TNF.
• the chi eric antibodies can comprise portions derived from two different species (e.g., human constant region and urine variable or binding region) .
• the portions derived from two different species can be joined together chemically by conventional techniques or can be prepared as single contiguous proteins using genetic engineering techniques.
• DNA encoding the proteins of both the light chain and heavy chain portions of the chimeric antibody can be expressed as contiguous proteins.
• Monoclonal antibodies reactive with CD4 or with TNF can be produced using somatic cell hybridization techniques (Kohler and Milstein, Nature 256: 495-497 (1975)) or other techniques.
• somatic cell hybridization techniques Kohler and Milstein, Nature 256: 495-497 (1975)
• a crude or purified protein or peptide comprising at least a portion of CD4 or of TNF can be used as the immunogen.
• An animal is vaccinated with the immunogen to obtain anti-CD4 or anti-TNF antibody- producing spleen cells.
• the species of animal immunized will vary depending on the species of monoclonal antibody desired.
• the antibody producing cell is fused with an immortalizing cell (e.g., myeloma cell) to create a hybridoma capable of secreting anti-CD4 or anti-TNF antibodies.
• the unfused residual antibody-producing cells and immortalizing cells are eliminated.
• Hybridomas producing desired antibodies are selected using conventional techniques and the selected
• Polyclonal antibodies can be prepared by immunizing an animal with a crude or puri ied protein or peptide comprising at least a portion of CD4 or of TNF. The animal is maintained under conditions whereby antibodies reactive with either CD4 or TNF are produced. Blood is collected from the animal upon reaching a desired titre of antibodies. The serum containing the polyclonal antibodies (antisera) is separated from the other blood components. The polyclonal antibody-containing serum can optionally be further separated into fractions of particular types of antibodies (e.g., IgG, IgM) .
• Antibodies specific for CD4 have been used in treatment of a wide range of both experimentally-induced and spontaneously-occurring autoimmune diseases.
• a more detailed description of anti-CD4 antibodies and their use in treatment of disease is contained in the following references, the teachings of which are hence incorporated by reference: U.S. Application NO. 07/867,100, filed June 25, 1992; Grayheb, J. £__ al. ⁇ J. of Autoi_ranunl_v 2:627-642 (1989); Ranges, G.E. e al. J. Exp. Med. 162: 1105-1110 (1985) ; Horn, J.T. et al.. gur. J. Immunol. 18: 881-888 (1988); Wooley, P.H.
• the CD4 + T cell inhibiting agent and TNF antagonist can be administered by various routes , including subcutaneously, intravenously, intramuscularly, topically, orally, rectally, nasally, buccally, vaginally, by inhalation spray, or via an implanted reservoir in dosage formulations containing conventional non-toxic pharmaceutical ly-acceptable carriers, adjuvants and vehicles .
• the form in which the agents are administered e . g. , capsule, tablet, solution, emulsion
• a therapeutically effective amount of the combination of anti-CD4 agent and anti-TNF agent is that amount necessary to significantly reduce or eliminate symptoms associated with a particular autoimmune or inflammatory disorder.
• the therapeutically effective amount will be determined on an individual basis and will be based, at least in part, on consideration of particular agents used, the individual ' s size, the severity of symptoms to be treated, the result sought, etc.
• the preferred therapeutically effective amount of anti-CD4 antibody administer e in conjunction with anti- TNF antibody is in the range of 0.1 - 10 mg/kg/dose of each antibody.
• the therapeutically effective amount can be determined by one of ordinary skill in the art employing such factors and using no more than routine experimentation.
• the therapeutically effective amount can be administered in the form of a single dose, or a series of doses separated by intervals of days or weeks .
• a maintenance amount of anti-CD4 agent, of anti-TNF agent, or of a combination of anti-CD4 agent and anti-TNF agent can be administered.
• a maintenance amount is the amount of anti-CD4 agent, anti-TNF agent, or combination of anti- CD4 agent and anti-TNF agent necessary to maintain the reduction or elimination of symptoms achieved by the therapeutically effective dose.
• the maintenance amount can be administered in the form of a single dose, or a series or doses separated by intervals of days or weeks. Like the therapeutically effective amount, the maintenance amount will be determined on an individual basis.
• the combination therapy of the current invention is thus useful for the treatment of many autoimmune or inflammatory diseases of humans and of animals.
• diseases for which the therapy is appropriate include rheumatoid arthritis (RA) and juvenile chronic arthritis (JCA) .
• other diseases and conditions or which combination therapy is appropriate include spondyloarthropathies, such as ankylosing spondylitis, psoriatic arthritis, or arthritis associated with inflammatory bowel disease; vasculitidee, such as polyarteritis nodosa, Wegener's granulomatosis, giant cell arteritis, Henoch-Schoenlein purpura, and microscopic vasculiti ⁇ of the kidneys; Sjogren's syndrome; systemic lupus erythematosus; inflammatory bowel disease, including Crohn' ⁇ disease and ulc ⁇ rative colitis; chronic active hepatitis; primary biliary cirrhosis; cryptogenic fibrosing alveolitis and other fibrotic
• EXAMPLE 1 Treatment of Induced Arthritis in a Murine Model usin ⁇ Anti-CD4 Antibody and Anti-TNF Antibody
• the murine model of collagen type II induced arthritis has similarities to rheumatoid arthritis (RA) in its marked MHC class II predisposition, as well as in histology, im unohistology, and erosions of cartilage and bone. Furthermore, there is a good correlation of therapeutic response with human rheumatoid arthritis.
• anti-TNF antibody has beneficial effects (Williams, R.O. fit ⁇ L__, _____ 12:9784-9788 (1992); El? >tt, M. J.
• mice Male DBA/1 mice were immunized intradermally at 8-12 weeks of age with 100 ⁇ g of bovine type II collagen emulsified in complete Freund's adjuvant (Difco Laboratories, East Molsey, UK), and 21 days later with 100 ⁇ g of collagen intra-peritoneally (i.p.). Immediately after the onset of clinically evident arthritis (redness and/or swelling in one or more limbs) , which was about 35 days after the initial injection, mice were injected i.p. with anti-CD4; anti-TNF; anti-CD4 and anti-TNF; or isotype controls. Arthritis was monitored for clinical score and paw-swelling for 10 days. Antibody treatment was administered on day l (onset) , day 4 and day 7.
• mice Male DBA/1 mice were immunized intradermally at 8-12 weeks of age with 100 ⁇ g type II collagen emulsified in Freund's complete adjuvant (Difco Laboratories, East Molsey, UK) .
• Day one of arthritis was considered to be the day that erythema and/or swelling was first observed in one or more limbs. Arthritis became clinically evident around 30 days after immunization with type II collagen.
• treatment was started on the first day that arthritis was observed and continued over a 10 day period, after which the mice were sacrificed and joints were processed for histology.
• Monoclonal antibody (mAb) treatment was administered on days 1, 4, and 7.
• TN3-19.12 a neutralizing hamster IgGl anti- TNF ⁇ /£ monoclonal antibody (mAb)
• mAb monoclonal antibody
• the isotype control was L2.
• the anti-TNF antibody and the isotype control were provided by R. Schreiber, Washington University Medical School (St. Louis, O, USA) , in conjunction with Celltech (Slough, UK).
• the cell- depleting anti-CD4 monoclonal antibody (rat IgG2b) consisted of a 1:1 mixture of YTS 191.1.2 and YTA 3.1.2, provided by H. Wald ann (University of Cambridge, UK)
• anti-TNF/anti- CD4 treatment also referred to herein as anti-CD4/TNF treatment
• anti-CD4/TNF treatment produced a significant reduction in paw ⁇ swelling relative to anti-CD4 alone, and anti-TNF alone (P ⁇ 0.05) .
• mice After 10 days, the mice were sacrificed; the first limb that had shown clinical evidence of arthritis was removed from each mouse, formalin-fixed, decalcified, and wax-embedded before sectioning and staining with hae atcxylon and eosin.
• a sagittal section of the proximal interphalangeal (PIP) joint of the middle digit was studied in a blind fashion for the presence or absence of erosions in either cartilage or bone (defined as demarcated defects in cartilage or bone filled with inflammatory tissue) . The comparisons were made only between the same joints, and the arthritis was of identical duration.
• PIP proximal interphalangeal
• CD4+ T cells The possible persistence of CD4+ T cells in the joint despite virtual elimination of peripheral CD4+ T cells was next investigated by immunohistochemical analysis of sections from treated arthritic mice. Wax-embedded sections were de-waxed, trypsin digested, then incubated with anti-CD4 mAb (YTS 191.1.2/YTA 3.1.2). To confirm the T cell identity of the CD4+ T cells, sequential sections were stained with anti-Thy-1 mAb (YTS 154.7) (Cobbold, S.P. et al.. Nature 312:548-551 (1984)). Control sections were incubated with HRPNi ⁇ /l2a.
• Serum anti-collagen IgG levels were measured by enzyme-linked immunosorbent assay (ELISA) .
• ELISA enzyme-linked immunosorbent assay
• Microtitre plates were coated with bovine type II collagen (2 ⁇ g/ml) , blocked, then incubated with test sera in serial dilution steps. Detection of bound IgG was by incubation with alkaline phosphata ⁇ e-conjugated goat anti-mouse IgG, followed by substrate (dinitrophenol phosphate) . Optical densities were read at 405 nm. A reference sample, consisting of affinity-purified mouse anti-type II collagen antibody, was included on each plate. Results are shown in Table 3.
• Serum levels of anti-type II collagen IgG were not significantly altered within the 10 day treatment period by anti-CD4 alone, anti-TNF alone, or anti-CD4 plus anti- TNF.
• IgM anti-TN4-19.12 levels on day 10 were compared. At this time, an IgG anti-TN3-19.12 response was not detected.
• Microtitre plates were coated with TN3-19.12 (5 ⁇ g/ml), blocked, then incubated with serially diluted test sera. Bound IgM was detected by goat anti-mouse IgM-alkaline phosphatase conjugate, followed by substrate. The results demonstrated that anti-CD4 was highly effective in preventing the development of an anti-TN3-19.12 antibody response (Table -24-
• mice Male DBA/1 mice were immunized intradermally with 100 ⁇ g of bovine type II collagen emulsified in complete Freund's adjuvant (Difco Laboratories, East Molsey, UK). The mean day of onset of arthritis was approximately one month after immunization. After the onset of clinically evident arthritis (erythema and/or swelling) , mice were injected intraperitoneally with therapeutic agents. Arthritis was monitored for clinical score and paw swelling (measured with calipers) for 10 days, after which the mice were sacrificed and joints were processed for histology. Sera were collected for analysis on day 10. Therapeutic agents were administered on day 1 (onset) , day 4 and day 7.
• the therapeutic agents included TNF receptor/IgG fusion protein (p55-sf2), anti-TNF antibody, anti-CD4 antibody, and methylprednisolone acetate.
• __ Comparison of Treatment with TNF Receptor/IgG Fusion Protein.
• mice were subjected to treatment with TNF receptor/IgG protein (2 ⁇ g) (18 mice) , TNF receptor/IgG protein (20 ⁇ g) (18 mice) , TNF receptor/IgG protein (100 ⁇ g) (12 mice), anti-TNF monoclonal antibody (mAb) (300 ⁇ g) (17 mice), methylprednisolone acetate (6 mice), an irrelevant human IgGl monoclonal antibody (mAb) (6 mice), or saline (control).
• the TNF receptor/IgG fusion protein herein referred to as p55-sf2, (Butler et al., Cvtokine (in press): (1994)), was provided by Centocor, Inc.
• the anti-TNF antibody was TN3-19.12, a neutralizing hamster IgGl anti-TNF ⁇ /,3 monoclonal antibody (Sheehan, K. C. et al., J. Immunology 142:3884-3893 (1989)), and _was provided by R. Schreiber, Washington University Medical School (St. Louis, MO, USA), in conjunction with Celltech (Slough, UK) .
• Neutralizing titres were defined as the concentration of TNF ⁇ neutralizing agent required to cause 50% inhibition of killing of WEHI 164 cells by trimeric recombinant murine TNF ⁇ ; the neutralizing titre of p55-sf2 was 0.6 ng/ml, compared with 62.0 ng/ l for anti-TNF mAb (TN3-19.12), using 60 pg/ml mouse TNF ⁇ .
• Paw-Swelling Treatment with p55-sf2 resulted in a dose-dependent reduction in paw-swelling over the treatment period, with the doses of 20 ⁇ g and 100 ⁇ g giving statistically significant reductions in paw-swelling relative to mice given saline (P ⁇ 0.05).
• the group of mice given an irrelevant human IgGl mAb as a control did not show any deviation from the saline-treated group (data not shown) , indicating that the therapeutic effects of p55-sf2 were attributable to the TNF receptor rather than the human IgGl constant region.
• a reduction in paw-swelling was observed in the ethylprednisolone acetate treated group that was comparable in magnitude to the reductions given p55-sf2 at 100 ⁇ g or anti-TNF mAb at 300 ⁇ g.
• mice After 10 days, the mice were sacrificed; the first limb to show clinical evidence of arthritis was removed from each mouse, fixed, decalcified, wax-embedded, and sectioned and stained with hae atoxylon and eosin. Sagittal sections of the proximal lnterphalangeal (PIP) joint of the middle digit of each mouse were studied in a blind fashion and classified according to the presence or absence of erosions, as defined above. Comparisons were thus made between identical joints, and the arthritis was of equal duration. Results are shown in Table 6.
• PIP proximal lnterphalangeal
• Anti-collagen IgG levels on day 10 were measured by ELISA as described (Williams, R.O. et al., PNAS 89: 9784- 9788 (1992)). Microtitre plates were sensitized with type II collagen, then incubated with serially-diluted test sera. Bound IgG was detected using alkaline pho ⁇ phatase- conjugated goat anti-mouse IgG, followed by substrate (dinitrophenol phosphate) . Optical densities were read at 405 nm. No differences between any of the treatment groups were detected (data not shown) . this suggests that the therapeutic effect of p55-sf2 is not due to a generalized im unosuppressive effect.
• the cell-depleting anti-CD4 mAb (rat IgG2b) consisted of a 1:1 mixture of YTS 191.1.2 and YTA 3.1.2, provided by H. Wald ann (University of Cambridge, UK) (Galfre, G. et al.. Nature 277: 131-133 (1979); Cobbold, S.P. et al. , Nature 3___: 548-551 (1984); Qin, S. et al. , European J. Immunology 17:1159-1165 (1987)).
• p55-Sf2 is described abov . Paw-Swelllire
• Microtitre plates were coated with recombinant murine TNF- ⁇ (Genentech Inc., San Francisco, CA) , blocked, then incubated with test sera. Goat anti-human IgG-alkaline phosphatase conjugate was then applied followed by substrate. Quantitation was by reference to a sample of known concentration of p55-sf2.
• the murine model of collagen type II induced arthritis was used to investigate the efficacy of the CD4+ T cell inhibiting agent cyclosporin A in conjunction with anti-TNF monoclonal antibody (mAb) , for the ability to modulate the severity of joint disease in collagen-induced arthritis.
• mAb monoclonal antibody
• mice Male DBA/1 mice were immunized intradermally with 100 ⁇ g of bovine type II collagen emulsified in complete Freund's adjuvant (Difco Laboratories, East Molsey, UK). The mean day of onset of arthritis was approximately one month after immunization.
• mice 11 mice each were subjected to treatment with one of the following therapies: 50 ⁇ g (2 mg/kg) L2 (the isotype control or anti-TNF antibody) , intraperitoneally once every three days (days 1, 4 and 7); 250 ⁇ g (10 mg/kg) cyclosporin A intraperitoneally daily; 50 ⁇ g (2 mg/kg) anti-TNF mAb TN3-19.12, intraperitoneally once every three days (days l, 4 and 7); or 250 ⁇ g cyclosporin A intraperitoneally daily in conjunction with 50 ⁇ g anti-TNF mAb intraperitoneally once every three days. Arthritis was monitored for paw swelling (measured with calipers) for 10 days, after which the mice were sacrificed and joints were processed for histology.
• Paw-swellin ⁇ Treatment with cyclosporin A in conjunction with anti-TNF mAb resulted in a reduction in paw-swelling over the treatment period, relative to mice treated with control antibody. Results are shown in Figure 3.

Applications Claiming Priority (3)

Publications (1)

Family

ID=34803520

Family Applications (1)

Country Status (2)

Citations (1)

• 1994
  • 1994-03-10 AU AU61495/94A patent/AU6149594A/en not_active Abandoned
  • 1994-03-10 EP EP94908462A patent/EP0765171A1/de not_active Withdrawn

Patent Citations (1)

Non-Patent Citations (1)

Also Published As

Similar Documents

Legal Events