IE922028A1 - Antibody composition acting synergistically - Google Patents

Abstract

The invention disclosed concerns a synergistic antibody composition which produces an enhanced immunosuppressive effect, the composition containing (a) at least one monoclonal anti-CD4 antibody which, by itself, has a strong inhibitory action and (b) at least one monoclonal anti-IL2R alpha or anti-IL2R beta antibody which, by itself, has a strong inhibitory action. The invention also concerns a drug based on this antibody composition.

Description

The present invention concerns an antibody composition acting synergistically for improving immunosuppression which contains two antibodies against different T cell surface markers.

Major advances have been made in the last ten years in transplantation surgery. In particular there has been a considerable improvement in the therapeutic measures which are intended to ensure the survival time of the transplanted organ. Despite this positive development, the established measures of today do not guarantee a one hundred percent chance of transplantation success even for a limited time period. This is partly due to the quality of the transplanted organ and the operation technique but, the main reason is that the donor organs are derived from a genetically different individual.

This genetic non-compatibility between recipient and donor (allogenic transplant), causes an immune reaction in the recipient against major histocompatibility complex (MHC)-coded surface antigens of the transplant. In order to prevent this rejection reaction which can take an acute as well as a chronic course, the immunolgical reactivity of the recipient has to be specifically suppressed by an immunosuppressive therapy. However, all immunosuppressive agents used today in clinical practice cause an unspecific immunosuppression to a greater or lesser extent. The side effects caused by this are sometimes serious.

,E 922028 Thus the purine analogue azathioprin (Drug Evaluation, 6th Edition American Medical Association 1151 (1986)) acts as an antimetabolite on all proliferating cells. Drawbacks of this substance are, however, its potential liver toxicity and induction of bone marrow depression. This results in a decrease of all blood cells.

Glucocorticoids are examples of non-cytotoxic immunosuppresive agents. However, in particular when the therapy is over a longer period, they lead to intolerable side effects (disturbances of growth, hypertension, cardiac insufficiency) which compel the medication to be discontinued.

Antilymphocyte globulin (ALG) and cyclosporin have a higher specificity than the aforementioned substances but they act on the whole immune system and this general immunosuppression increases the risk of viral and bacterial infections. Further disadvantages are that ALG can only be administered over a limited time period because of its potential sensitizing effect. Treatment with cyclosporin can lead to an increase in the incidence of tumours.

Acute rejection crises in organ transplantations have also been successfully treated with anti-T cell antibodies (Cosimi AB: Transplant. Proc. 15 (1983), p. 1889; Kreis, H. et al., Transplant. Proc. 17 (1985), p. 1315; Krikma, R.L. et al., Transplantation 36 (1983), p. 620). T cells (T lymphocytes) are divided into CD4positive and CD8-positive cells. CD4 and CD8 are surface antigens on these cells. CD8-positive T cells interact with the T cell receptor and this is associated with MHC class I molecules (killer cells). CD4-positive T cells interact with the T cell receptor and this is associated - 3 with MHC class II molecules on corresponding recipient cells (helper T cells, suppressor T cells).

It has been shown that the rejection reaction in organ transplantations can be prevented by anti-CD4 antibodies as well as by anti-CD8 antibodies (cf. e.g. Benjamin, R. J. and Waldmann, H.: Induction of tolerance by monoclonal antibody therapy, Nature 320 (1986), p. 449; Benjamin, R.J., Cobbold, S.P., Clark, M.P. and Waldmann, H.: Tolerance to rat monoclonal antibodies. Implications for serotherapy. J. Exp. Med. 163 (1986), p. 1539; Cobbold, S.P., Martin, G., Quin S.X. and Waldmann, H.: Monoclonal antibodies to promote marrow engraftment and tissue graft tolerance. Nature 323 (1986), p. 164; Quin S. X, Cobbold, S.P., Tighe, H., Benjamin, R. and Waldmann, H.: CD4 Mab pairs for immunosuppression and tolerance induction. Eur. J. Immunol. 18 (1987) p. 495).

The disadvantage of immunosuppressive treatment with such antibodies is, however, that they have to be used in a relatively large amount. This can lead to massive immune reactions even when using chimerized antibodies.

EP-A 0 240 344 discloses a combination of anti-CD4 MAB's with anti-IL2Ra (CD25) MAB's in example 5 (column 8) to prevent transplant rejections. A complete inhibition is, however, only achieved when this is combined with a third MAB against CD8. The antibody YCTLD45.1 used in the combination as anti-IL2Ra MAB by itself only causes a negligible inhibition of the immune reaction. However, it is not disclosed which properties the antibodies used in the combination have to possess in order that in general a complete inhibition can be expected. - 4 An object of the present invention is thus to provide an immunosuppressive agent which can reliably suppress the rejection reaction against transplants and which only has to be administered in small doses.

This object is achieved by an antibody composition acting synergistically for improving immunosuppression which is characterized in that it contains (a) at least one monoclonal anti-CD4 antibody which by itself is already strongly inhibitory and (b) at least one monoclonal anti-IL2Ra or anti-IL2RB antibody which by itself is already strongly inhibitory, whereby a strongly inhibitory antibody at a concentration of 10000 ng/ml inhibits allogen-induced lymphocyte proliferation by at least 40 % in the absence of other antibodies.

If the antibody (b) is an anti-IL2Ra antibody, then the molar ratio of the antibodies (a) and (b) is preferably between 1:10 and 10:1. If in contrast the antibody (b) is an anti-IL2RB antibody then the molar ratio of the antibodies (a) and (b) is preferably between 1:1000 and 10:1. The combination of anti-CD4 MAB and anti-IL2Ra MAB is preferred according to the present invention.

A prerequisite for the synergistic effect in the combination of monoclonal anti-CD4 or anti-IL2 receptor antibodies is that each of these antibodies must already by itself have an immunosuppressive action i.e. it must strongly inhibit the lymphocyte proliferation. This strongly inhibitory action within the meaning of the present invention is manifested by the fact that the antibody concerned already alone inhibits the allogeninduced lymphocyte proliferation (MLR) by at least 40 % - 5 at a concentration of 10000 ng/ml. Antibodies which at a concentration of 10000 ng/ml do not demonstrate an inhibitory effect on the MLR or have an inhibitory effect of less than 40 % are therefore in general not suitable as components of a composition according to the present invention.

It was found that surprisingly the combination of a strongly inhibitory monoclonal antibody against the CD4 structure with a strongly inhibitory monoclonal antibody against the a chain of the IL2 receptor (anti-IL2Ra antibody) or the β chain of the IL2 receptor (anti-IL2RB antibody) synergistically increases the inhibition of the proliferation of T helper cells so that in this way transplant rejections are avoided. It turns out that the simultaneous addition of two monoclonal antibodies (anti-CD4, anti-IL2 receptor) synergistically inhibits the lymphocyte proliferation in an in vitro system which is usually used to test the immunological histocompatibility of two individuals (Mixed Lymphocyte Reaction, MLR, Selected Methods in Cellular Immunology, Mishell, B.B., Shiigi S.M. eds. WH Freeman and Company, San Francisco 1980). The effective dose of this antibody combination is thus surprisingly much less than the effective dose of the antibodies when they are used singly. In addition it was surprisingly found that an inhibition of the MLR of up to 96 % can be achieved by using the combination of anti-IL2Ra antibodies with anti-CD4 antibodies. In contrast when anti-IL2Ra antibodies or anti-CD4 antibodies are used alone the inhibition is not greater than 70 % even at concentrations of 30 gg/ml. Surprisingly the use of the combination of anti-IL2RB antibodies with anti-CD4 antibodies can also lead to up to 96 % inhibition of the MLR. - 6 The synergism according to the present invention which is surprising can only be demonstrated when an antiIL2Ra or anti-IL2RB MAB which by itself is already strongly inhibitory is used in combination with an antiCD4 MAB. If in contrast an anti-IL2Ra MAB is for example used which is non-inhibitory or only weakly inhibitory then when it is combined with an anti-CD4 MAB which by itself is already strongly inhibitory then no increased suppressive action can be determined compared to the use of the anti-CD4 MAB alone.

In a combination of anti-CD4 MAB and anti-IL2Ra MAB, the two antibodies (a) and (b) are preferably used in a molar ratio of 1:10 to 10:1 to achieve an optimal synergistic effect. The molar ratio of the antibodies (a) and (b) is particularly preferably 1:5 to 5:1, most preferred is a ratio of 3:10 to 3:1. When the combination anti-CD4 MAB and anti-IL2RB MAB is used, a molar ratio of the antibodies (a) and (b) of 1:1000 to 10:1 is preferably selected. The antibody composition according to the present invention can contain one as well as several anti-CD4 antibodies and one as well as several anti-IL2Ra or anti-IL2RB antibodies whereby when using several antibodies with one specificity the molar ratio always in each case refers to the total of all antibodies of one specificity.

In addition it was found that for the inventive use both antibodies have to be used in combination to demonstrate the synergistic effect but this does not necessarily have to take place at the same time. The term used in combination within the meaning of the present invention is understood to mean that a certain time lag between the administration of the two antibodies can be tolerated. However, it is necessary that the activity of the antibody administered first should not have significantly decreased before the second antibody is administered. In practice such a time lag can be up to about 12 hours.

A further advantage of the antibody combination according to the present invention is its high specificity. Since anti-IL2Ra or anti-IL2RB antibodies bind especially to the activated T lymphocytes and only the anti-CD4 antibodies bind to the whole population of T helper cells, the combination of these antibodies according to the present invention in the small amounts according to the present invention only negligibly suppresses the general immune reaction.

Antibodies which are suitable as components of the antibody combination according to the present invention can be murine, human, chimerized or humanized antibodies or antibody fragments. One preferably uses human, chimerized or humanized antibodies or such antibody fragments since by this means the chances of an immune reaction against the administered antibodies can be kept as small as possible. Chimerized or humanized antibodies within the sense of the present invention are non-human antibodies, e.g. of murine origin, in which murine sequences have been replaced by human sequences by means of known genetic engineering methods. In these chimerized antibodies only the constant region of the antibody is replaced by a corresponding human region whereas in the humanized antibodies the CDR regions (complementary-determining regions) of human V regions for the light and the heavy chain of an antibody can be substituted by the CDR regions of a murine or other rodent antibody so that the humanized antibody corresponds to a human antibody apart from the CDR IE 922°28 regions. Antibody fragments such as Fab or F(ab)2 antibody fragments which can be obtained according to standard methods are also suitable for the process according to the present invention.

An exact desription of suitable anti-IL2Ra antibodies is given in PCT/EP91/01737. Anti-CD4 antibodies are disclosed in Eur. J. Immunol. 18 (1987) 495-502.

Suitable anti-IL2RB antibodies are described in Takeshita, R. , Goto, Y., Tada, K., Nagata, K. Asao, H., Sugamura, K.: J. Exp. Med. 169 (1989) 1323-1332; Tsudo, M., Kitamura, F., Mijasaka, M.: Proc. Natl. Acad. Sci. USA 86 (1989) 1982-1986 and Niguma, T., Sakagami, K., Kawamura, T., Haisa, M. , Fujiwara, T., Kusaka, S., Uda, M., Orita, K.: Transplantation 52 (1991) 296-302.

A combination of anti-CD4 and anti-IL2Ra or anti-IL2RB antibodies is particularly suitable, for which the sequences of the variable regions of the light and heavy chains are described in the attached sequence protocols. SEQ ID NO. 1 and 2 show nucleotide and amino acid sequences of the anti-CD4 antibody MT 15.1, SEQ ID NO. 3 and 4 show nucleotide and amino acid sequences of the anti-CD4 antibody MT 3.10, SEQ ID NO. 5 and 6 show nucleotide and amino acid sequences of the anti-IL2Ra antibody MAB 179. SEQ ID NO. 9 and 10 show nucleotide and amino acid sequences of the anti-ILRB antibody A41. Whereas the aforementioned antibodies are suitable for producing the composition according to the present invention, it was found that the anti-ILRa antibody M-215 (SEQ ID NO. 7 and 8), which by itself only has a weak inhibitory effect, does not have a synergistic effect when combined with a strongly inhibitory anti-CD4 antibody. Suitable constant regions (murine or human) for these antibodies are described in: Sequences of - 9 proteins of immunological interest; E. Rabat, T. Wu, M. Reid-Miller, H. Perry and K. Gottesman, U.S. Department of Health and Human Services, 1987, p. 282-325. The fusion of the genes for the constant regions with the genes for the variable regions can be carried out by means of standard cloning techniques used in molecular biology.

The present invention in addition concerns a pharmaceutical agent which comprises components (1) and (2), whereby both components have to be administered in combination but can be formulated separately, which is characterized in that the component (1) contains a monoclonal anti-CD4 antibody (a) which by itself is strongly inhibitory as the active substance and component (2) contains a monoclonal anti-IL2Ra or antiIL2RB antibody (b) which by itself is strongly inhibitory as the active substance, if desired, together with the usual pharmaceutical auxiliary, dilution, carrier and filling agents. If an anti-IL2Ra antibody is used as the active substance of component (2), then the molar ratio of the two antibodies of components (1) and (2) is preferably 1:10 to 10:1 in the pharmaceutical agent. The molar ratio of the two antibodies is particularly preferably 1:5 to 5:1, most preferred is a ratio of 3:10 to 3:1. If on the other hand an anti-IL2RB antibody is used as the active substance of component (2) then the ratio of the two antibodies of components (1) and (2) is preferably 1:1000 to 10:1.

As already set forth above, the two components of the pharmaceutical agent according to the present invention have to be administered in combination whereby the term in combination does not, however, necessarily mean at the same time. It is therefore obvious that the two - 10 components of the pharmaceutical agent according to the present invention can be formulated separately. The formulation of the antibodies is carried out by means of standard methods, e.g. in physiological solutions which can be administered intravenously.

The present invention also concerns the use of the antibody composition according to the present invention or of the pharmaceutical agent according to the present invention in an immunosuppressive therapy in particular in a therapy to suppress an immune reaction after organ or tissue transplantations or in autoimmune diseases.

In such a method of treatment a pharmaceutical agent according to the present invention is administered which contains the combination of an anti-CD4 antibody and an anti-IL2Ra or anti-IL2RB antibody. In such a therapeutic procedure it is surprising that it is possible to reduce the amount of antibodies used by a factor of 10 compared to the separate use of the MAB's. In the various in vivo models for transplantation, MAB's against surface structures are used at a concentration of 1 to 5 mg/kg body weight (daily for 10 to 14 days). With a combination of anti-CD4 plus anti-IL2Ra or anti-CD4 plus anti-IL2RB according to the present invention the effective dose can be reduced to 100 to 200 Mg/kg .

The following antibodies were deposited at the ECACC, Public Health Laboratory Service, Porton Down, Salisbury, Wiltshire SP 5 OJG, Great Britain: Anti CD4 MAB MT 15.1 designated clone 15-1/P3/14 (ECACC 90090705) Anti CD4 MAB MT 3.10 designated clone 3.101/5B10 (ECACC 90090702) and Anti IL2Rct MAB 179 designated 3G10/179 (ECACC 90071905). - 11 The monoclonal anti-IL2RB antibody A41 designated MAKM-A23A41 was deposited at the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSM), Mascheroder Weg lb, D-3300 Braunschweig, on 30.07.1991 under the number DSM ACC 2015.

The following examples in conjunction with the sequence protocols SEQ ID NO. 1 to NO. 10 are intended to elucidate the invention in more detail.

SEQ ID N0.1 SEQ ID NO.2 SEQ ID NO.3 SEQ ID NO.4 shows the nucleotide and amino acid sequence of the light chain of the anti CD4 antibody MT 15.1, whereby the amino acids 1-95 correspond to the V region and the amino acids 96-107 correspond to the J2 region, shows the nucleotide and amino acid sequence of the heavy chain of the antiCD4 antibody MT 15.1 whereby the amino acids 1-107 correspond to the V region and the amino acids 108-120 correspond to the J4 region, shows the nucleotide and amino acid sequence of the light chain of the antiCD4 antibody MT 3.10 whereby the amino acids 1-100 correspond to the V region and the amino acids 101-111 correspond to the J1 region, shows the nucleotide and amino acid sequence of the heavy chain of the antiCD4 antibody MT 3.10 whereby the amino acids 1-106 correspond to the V region and the amino acids 107-118 correspond to the J3 region, - 12 SEQ ID NO.5 SEQ ID NO.6 SEQ ID NO.7 SEQ ID NO.8 SEQ ID NO.9 shows the nucleotide and amino acid sequence of the light chain of the antiIL2Ra antibody MAB 179 whereby the amino acids 1-95 correspond to the V region and the amino acids 96-107 correspond to the J1 region, shows the nucleotide and amino acid sequence of the heavy chain of the antiIL2Ra antibody MAB 179 whereby the amino acids 1-98 correspond to the V region, the amino acids 99-102 correspond to the D region and the amino acids 103-113 correspond to the J3 region, shows the nucleotide and amino acid sequence of the light chain of the antiIL2Ra antibody MAB M-215 whereby the amino acids 1-94 correspond to the V region, the amino acids 95-106 correspond to the J4 region and the amino acids 107-123 correspond to the beginning of the C region, shows the nucleotide and amino acid sequence of the heavy chain of the antiIL2Ra antibody MAB M-215 whereby the amino acids 1-97 correspond to the V region, the amino acids 98-104 correspond to the D region, the amino acids 105-119 correspond to the J3 region and the amino acids 120164 correspond to the beginning of the C region, shows the nucleotide and amino acid sequence of the light chain of the antiIL2RB antibody MAB A41 whereby the amino acids 1-96 correspond to the V region, the amino acids 97-107 correspond to the J region and the nucleotide 322 corresponds - 13 to the first base pair of the C region and SEQ ID NO.10 shows the nucleotide and amino acid sequence of the heavy chain of the antiIL2RB antibody MAB A41 whereby the amino acids 1-98 correspond to the V region, the amino acids 99-104 correspond to the D region, the amino acids 105-118 correspond to the J region and the nucleotide 355 corresponds to the first base pair of the C region.

Example 1 Measurement of the inhibition of the mixed lymphocyte reaction (MLR) by antibodies The mixed lymphocyte reaction (mixed lymphocyte culture) is based on the property of T lymphocytes to increase their proliferation in vitro after recognizing foreign antigens. Foreign antigens can be bacteria or viruses and even transplantation antigens on foreign tissues or cells. The human lymphocyte line RPMI 1788 (ATCC CLL156) was used to induce the T cell proliferation of peripheral blood lymphocytes (PPBL) isolated by plasmaphoresis. The cell proliferation of this cell line was completely blocked by treatment with mitomycin C. Thus in the MLR only the T lymphocytes of the PPBL proliferate. The activation of the T lymphocytes can be completely inhibited by addition of immunosuppressive substances .

The MLR is carried out according to Selected Methods in Celular Immunology Mishell B.B., Shiigi S.M. eds., W.H. Freeman and Company, San Francisco (1980).

Medium (complete RPMI 1640) 440 ml RPMI 1640 (Boehringer Mannheim, catalogue No. 209 945) ml foetal calf serum (FCS) (Boehringer Mannheim, catalogue No. 210 471) ml glutamine solution, 200 mmol/1 (Boehringer Mannheim, catalogue No. 210 277) ml vitamin solution (1 %) (Boehringer Mannheim, catalogue No. 210 307) ml penicillin (50000) and streptomycin (50 mg) (Boehringer Mannheim, catalogue No. 210 404) in RPMI 1640 Vitamin solution (Boehringer Mannheim, catalogue No. 210 307) : mg/100 ml mg/100 ml Ca D(+)-pantothenate 10.0 nicotinamide 10 choline chloride 10.0 pyridoxal · HC1 10 folic acid 10.0 riboflavin 1 meso-inositol 20.0 thiamine · HCl 10 PPBL cells: The lymphocytes were isolated by density centrifugation from lymphocyte concentrate (obtained by plasmaphoresis, Bavarian Red Cross, Munich) using lymphocyte separation medium (Boehringer Mannheim, catalogue No. 295 949). After washing the cells twice in complete RPMI 1640 the cell titre was adjusted to 10^/ml.

RPMI 1788 cells: Human lymphocyte line (ATCC CCL 156; secreting IgM-lambda chains) RPMI 1788 cells (ATCC CCL 156) are treated with mitomycin C. For this, 50 Mg mitomycin (dissolved in 100 μ! complete RPMI 1640) is added to 10? cells/ml and incubated for 45 minutes at 37°C, 5 % CO2. Afterwards the cells are centrifuged down and washed twice with RPMI 1640 (complete).

The cell titre is adjusted to 1 x 105/ml.

In order to carry out the MLR, 100 μΐ PPBL (105 cells) in complete RPMI 1640 medium, 100 μΐ RPMI 1788 cells (105 cells) in complete RPMI 1640 medium and 20 μΐ sample substance (MAB 179 and/or MAB M 15.1) at the concentrations stated in tables 1 to 2b are placed in flat-bottomed tissue culture plates (96 wells, Nunc Company). They are incubated in an incubator for four days at 37°C and 5 % CO2. The extent of the proliferation is quantified by incorporation of radioactive thymidine into the DNA. For this 0.5 Mci/well methyl-3H-thymidine (specific activity 25 Ci/mmol, TRK 120, Amersham-Buchler, Braunschweig) in 25 Ml medium is added. The cells are incubated for a further 24 hours in an incubator at 37°C and 5 % CO2.

Subsequently the cells are harvested with an Inotech harvester (Inotech Company, Wohlen, Switzerland) on glass-fibre filter discs. The radioactivity of the filter discs is determined with the filter counting system INB-384 (Inotech). The inhibition in percent which is given in tables 1, 2a and 2b is determined from - 16 the ratio of the measured radioactivity for a test mixture with sample substance to that for a test mixture without sample substance. (The concentrations stated in the tables are to be understood as amounts per ml test volume.) The following Table 1 shows the dose-effect curve when the monoclonal antibody MAB 179 (anti-IL2Ra antibody) or MT 3.10 or MT 15.1 (anti-CD4 antibody) are each used alone in the allogen-induced induced lymphocyte proliferation (MLR). It can be seen from the values that the inhibitory effects of the individual antibodies reach a maximum value between 60 and 70 % at a concentration of ca. 300 ng/ml. Further addition of the same antibody up to a concentration of 30000 ng/ml does not further improve the inhibitory effect.

Table 1: Inhibition of the MLR in percent compared to a control Concentration ng/ml MAB 179 MT 3.10 MT 15.1 1 4 9 5 3 11 18 2 10 28 26 24 30 48 57 52 100 58 58 54 300 61 67 67 1000 61 68 67 3000 60 62 65 10000 61 68 70 30000 61 70 70 - 17 The following Table 2a shows the inhibition of the allogen-induced lymphocyte proliferation (MLR) by a combination of MAB 179 and MT 3.10. Surprisingly it was found that the combined use of both antibodies leads to a substantial improvement in the inhibitory effect. Even when only 100 ng/ml of each of the two antibodies are used it can be seen that there is an almost complete inhibition of the allogen-induced lymphocyte proliferation.

Table 2a: Concentration in ng/ml % Inhibition MAB 179 MT 3.10 1 - 0 10 - 23 100 - 53 — 1 0 - 10 19 - 100 43 1 1 0 10 1 14 100 1 46 1 10 29 10 10 45 100 10 74 1 100 39 10 100 69 100 100 92 - 18 The following Table 2b shows the inhibition of the allogen-induced lymphocyte proliferation (MLR) by a combination of MAB 179 and MT 15.1. One finds that already the use of 100 ng/ml of each of the two antibodies leads to an almost complete inhibition of the allogen-induced lymphocyte proliferation.

Table 2b: Concentration in ng/ml % Inhibition MAB 179 MT 15.1 1 a> - 0 10 - 23 100 - 53 - 1 1 - 10 18 - 100 53 1 1 0 10 1 25 100 1 62 1 10 16 10 10 48 100 10 86 1 100 28 10 100 64 100 100 96 - 19 Comparative example The effect of combining an anti-IL2Ra antibody (M-215), which alone is only weakly inhibitory, with the strongly inhibitory anti-CD4 antibody 3.10 or 15.1 was tested.

The test procedure was carried out as described in example 1. The results are shown in tables 3a and 3b.

Table 3a: Concentration in ng/ml % Inhibition M-215 MT 3.10 10000 - 20 - 100 43 10000 1 18 10000 10 31 10000 100 40 Table 3b: Concentration M-215 in MT ng/ml 15.1 % Inhibition 10000 - 20 - 100 44 10000 1 19 10000 10 26 10000 100 45 ,Ε 922028 It can be seen from tables 3a and 3b that a combination of the antibodies M-215 and MT 3.10 or M-215 and MT 15.1 does not show any synergistic effect even at extremely high concentrations.

Example 2 The dose effect curve of the anti-IL2R5 antibody A41, which alone is strongly inhibitory, was tested alone and in combination with the strongly inhibitory anti-CD4 antibodies MT 3.10 and MT 15.1 which were already examined in example 1. The test procedure was carried out according to example 1. The results are shown in the tables 4a and 4b. ®922028 - 21 Table 4a Concentration in ng/ml % Inhibition MAB A 41 MT 3.10 100 - 11 1000 - 26 10000 - 50 — 1 4 - 10 31 - 100 60 100 1 8 1000 1 32 10000 1 60 100 10 48 1000 10 70 10000 10 85 100 100 64 1000 100 76 10000 100 96 - 22 Table 4b Concentration in ng/ml % Inhibition MAB A 41 MT 15.1 100 - 11 1000 - 26 10000 - 50 - 1 0 - 10 34 - 100 55 100 1 27 1000 1 39 10000 1 49 100 10 48 1000 10 70 10000 10 80 100 100 59 1000 100 78 10000 100 92 It can be seen from tables 4a and 4b that a combination of the antibodies A41 and MT 3.10 or A41 and MT 15.1 shows a synergistic effect compared to the single administration of the antibodies.

SEQUENCE LISTING (1) GENERAL INFORMATION: (iii) NUMBER OF SEQUENCES: 10 (2) INFORMATION FOR SEQ ID NO:1: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 381 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA to mRNA (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..381 (ix) FEATURE: (A) NAME/KEY: matjoeptide (B) LOCATION: 61..381 (ix) FEATURE: (A) NAME/KEY: sig_peptide (B) LOCATION: 1..60 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1: ATC ATS Met Met -20 TOC Ser TCT Ser GCT CAG Ada Gin -15 TTC CTT GCT CTC CTG TTC CTC TCT TTT CAA 48 Phe Leu Gly Leu Leu -10 Leu Leu Cys Phe Gin -5 GCT ACC AGA TCT GAT ATC CAG ATC ACA CAG ACT AIA TCC TCC CTC TCT 96 Gly Thr Arg Cys Asd He Gin Met Thr Gin Thr He Ser Ser Leu Ser Ϊ 5 10 GCC TCT CTG GGA GAC AGA GTC ACC ATC AGT TOC AGG GCA AGT CAG GAC 144 Ala Ser Leu Gly Asp Arg Val Thr lie Ser Cys Arg Ala Ser Gin Asp 15 20 25 ATT AAC AAT TAT TTA AGC TGG TAT CAG CAG AAA CCA GAT GGA ACT CTT 192 lie Asn Asn Tyr Leu Ser Trp Tyr Gin Gin Lys Pro Asp Gly Thr Val 30 35 40 AAA CTO CTG ATC TAG TAC ACA TCA AGA TTA CAT TCA GGA GTC CCA TCA 240 Lys Leu Leu lie Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser 45 50 55 60 AGG TTC ACT GGC AGT TCT GGA ACA GAT TAT TCT CTC ACC ATT ACC 288 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr He Thr 70 75 AAC Asn CTC GAG CAA GAA GAT GTT GCC ACT TAG ΤΤΓ TGC CAA CAG GGT AAT Leu Glu Gin Glu 80 Asp Val Ale. Thr 85 Tyr Phe Cys Gin Gin 90 Gly Asn ACG CTT CCG TAG ACG TTC GGA ACC AAG CTC GAA ATA AAA Thr Leu Pro Tyr Thr Phe C-ly Gly Gly Thr Lys Leu Glu lie Lys 100 105 336 381 - 25 (2) INFORMATION FDR SEQ ID NO: 2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 417 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA to mRNA (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..417 (ix) FEATURE: (A) NAME/KEY: mat_peptide (B) LOCATION: 58..417 (ix) FEATURE: (A) NAME/KEY: sig_peptide (B) LOCATION: 1..57 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: ATG GCT TGG GTG TGG ACC TTG CTT TIC CTG ATG GCA GCT GCC CAA ACT 48 Met Ala Trp Val Trp Thr Leu Leu Phe Leu Met Ala Ala Ala Gin Ser -19 -15 -10 -5 ATC CAA GCA CAG ATC CAG TTG CTG CAG TOT GGA CCT GAG CTG AAG ACG 96 lie Gin Ala Gin lie Gin Leu Val Gin Ser Gly Pro Glu Leu Lys Thr CCT GGA GAG ACA CTC AAG ATC TOC TOC AAG GCT TOT GCT TAT ACC TTC 144 Pro Gly Glu Thr Val Lys lie Ser Cys Lys Ala Ser Gly Tyr Thr Phe 25 ACA GAC TAT TCA ATA CAC TOG GTG AAG CAG GCT CCA GGG AAG GAT TEA 192 Thr Asp Tyr Ser lie His Trp Val Lys Gin Ala Pro Gly Lys Asp Leu 40 45 AAG TGG ATG GGC TGG ATA AAC ACT GAG ACT GCT GAG CCA ACA TAT GCA 240 Lys Trp Met Gly Trp lie Asn Thr Glu Thr Gly Glu Pro Thr Tyr Ala 55 60 GAT GAC TIC ACG GGA CGG ΊΤΓ GCC TTC TOT TTG GAA ACC TOT GCC AGC 288 Asp Asp Phe Thr Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser 70 75 ACT CTC TAT TTG CAG ATC AAC AAC CTC AAA AAT GAG GAC ACG TCT ACA 336 Thr Val Tyr Leu Gin lie Asn Asn Leu Lys Asn Glu Asp Thr Ser Thr 85 90 TAT TIC TGT GCT ATT CAT TAC TAC GCC TAC GGcj GAT CCT TTG GAC TAC 284 Tyr Phe Cvs Ala lie His Tyr Tvr Ala Tyr Gly Aso Pro Leu Asp Tyr 95 100 105 TGG GGT CAA GGA ACC TCA GTC ACC GTC TCC TCA 417 Trp Gly Gin Gly Thr Ser Val Thr Val Ser Ser 110 115 120 (2) INFORMATION FOR SEQ ID NO: 3: (i) SEQUENCE CHARACTERISTICS: (A) TENCTH: 393 base pai rs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA to mRNA (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..393 (ix) FEATURE: (A) NAME/KEY: matjoeptide (B) LOCATION: 61..393 (ix) FEATURE: (A) NAME/KEY: sig_peptide (B) LOCATION: 1..60 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: ATS Met -20 GAG ACA GAC Asp ACA ATC CTG CTA TGG GIG CTG CTO CTC TGG GTT CCA 48 Glu Thr Thr lie Leu -15 Leu Trp Val Leu Leu Leu Trp Val Pro -10 -5 GGC TCC ACT GCT GAC ATT CTG CTG ACC CAA TOT CCA GCT TOT TIG CCT 96 Gly Ser Thr Gly Asp lie Val 1 Leu Thr 5 Gin Ser Pro Ala Ser Leu Pro 10 AIG TOT CTA GGG CAG AGG GCC ACC ATC TCC TGC AAG GCC AGC CAA ACT 144 Met Ser Leu 15 Gly Gin Arg Ala Thr 20 lie Ser Cys Lys Ala Ser Gin Ser 25 CTT GAT TAT GAT GCT GAT ACT TAT ATG AAC TGG TAC CAA CAG AAA CCA 192 Leu Asp 30 Tyr Asp Gly Asp Ser 35 Tyr Met Asn Trp Tyr Gin Gin Lys Pro 40 GGA CAG CCA CCC AAA CTC CTC ATC TAT GCT GCA TCC AAT CTA GAA TCT 240 Gly 45 Gin Pro Pro Lys Leu Leu 50 He Tyr Ala Ala 55 Ser Asn Leu Glu Ser 60 GGG ATC CCA GCC AGA ΊΤΤ ACT GGC ACT GGG TCC GGG ACA GAC TIG ACC 288 Gly lie Pro Ala Arg Phe Ser 65 Gly Ser Gly Ser 70 Gly Thr Asp Phe Thr 75 CTC AAC ATC CAT CCT GTG GAG GAG GAG GAT GCT GCA ACC TAT TAC TCT 336 Leu Asn lie His 80 Pro Val Glu Glu Glu 85 Asp Ala Ala Thr Tyr Tyr Cys 90 CAG CAA ACT ACT GAG GAT CCT CCG ACG TTC GCT GGA GGC ACC AAG CTG Gin Gin Ser Ser Glu Asp Pro Pro Thr Phe Gly Gly Glv Thr Lys Leu 100 105 584 GAA ATC AAA Glu lie Lvs 110 393 - 29 (2) INFORMATION FOR SEQ ID NO:4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 408 base pairs (B) TYPE: nucleic acid (C) STRANDEDiESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA. to mRNA (lx) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..408 (ix) FEATURE: (A) NAME/KEY: mat_peptide (B) LOCATION: 55..408 (ix) FEATURE: (A) NAME/KEY: sig_peptide (B) LOCATION: 1..54 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: ATC GAA TGG AGG ATC ΤΊΤ CTC TTC ATC CTG TCA GGA ACT GCA GCT GTC Met -18 Glu Trp Arg -15 lie Phe Leu Phe He -10 Leu Ser Gly Thr Ala -5 Gly Val CAC TCC CAG GTT CAC CTG CAG CAG TCT GGA CCT GAG CTG GTG AAG CCT His Ser Gin 1 Val His Leu Gin 5 Gin Ser Gly Pro Glu 10 Leu Val Lys Pro GGG CCT TCA GTC AAG ATC TCC TCC AAG GCT TCT GGA TAC ACA TTC ACT Gly 15 Pro Ser Val Lys Met 20 Ser Cys Lys Ala Ser Gly 25 Tyr Thr Phe Thr 30 GAC TAT GTT CTA ACT TGG ATC CAA CAG AGA ACT GGA CAG GTC CTT GAG Asp Tyr Val Val Ser 35 Trp Met Gin Gin Arg 40 Thr Gly Gin Val Leu 45 Glu TOG ATT GGA GAG ATT TAT CCT GGA ACT GCT AGT GCT TAT TAC AAT GAA Trp He Gly Glu 50 He Tyr Pro Gly Ser 55 Gly Ser Ala Tyr Tyr 60 Asn Glu AAA TIC AAG GGC AAG GCC ATA CTG ACT GCA GAC AAA TCC TCC AGC ACA Lys Phe Lys 65 Gly Lys Ala He Leu 70 Thr Ala Asp Lys Ser 75 Ser Ser Thr GCC TAC ATC GAG TIC AGC AGC CTG ACA TCT GAG GAC TCT GCG CTC TIT Ala Tyr 80 Met Glu Phe Ser Ser 85 Leu Thr Ser Glu Asp 90 Ser Ala Val Phe 144 192 240 288 336 TTC TGT GGA AGA CGG xxjxj GAT GGT TCC CTG GXX. ITT GCT CAC «TV'’''*' Lax. 384 Phe Cys Ala Arg Arg Gly Asp Gly Ser Leu Glv Phe Ala His Trp Glv 95 100 105 110 CAA GGG rhCT CTG GIG ACT GIG GCT 408 Gin Gly Thr Leu Val Thr Val Ala 115 ,Ε 922028 - 31 (2) INFORMATION FOR SEQ ID NO:5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 381 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA to mRNA (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..381 (ix) FEATURE: (A) NAME/KEY: irat_peptide (B) LOCATION: 61..381 (ix) FEATURE: (A) NAME/KEY: sig_peptide (B) LOCATION: 1..60 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5: ATG Met -20 ATG GTC CTT Leu GCT CAG ΤΊΤ CTT GCA TIC TTG TTG CTT TGG ITT CCA 48 Met Val Ala Gin Phe -15 Leu Ala Phe Leu Leu Leu Trp Phe Pro -10 -5 GGT GCA AGA TCT GAC ATC CTC ATG ACC CAA TCT CCA TCC TCC ATG TCT 96 Gly Ala Arg Cys Asp lie Leu Met Thr Gin Ser Pro Ser Ser Met Ser 1 5 10 CTA TOT CTG GGA GAC ACA GTC AGC ATC ACT TGC CAT GCA ACT CAG GGC 144 Val Ser Leu Gly Asp Thr Val Ser He Thr Cys His Ala Ser Gin Gly 15 20 25 ATC AGA ACT AAT ATA CTG TGG TIG CAG CAG AAA CCA G3G AAA TCA TIT 192 lie Arg Ser Asn He Val Trp Leu Gin Gin Lys Pro Gly Lys Ser Phe 30 35 40 AGG GGC CTG ATC TAT CAT GGA ACC AAG TTG GAA GAT GGA CTT CCA TCA 240 Arg Gly Leu lie Tyr His Gly Thr Lys Leu Glu Asp Gly Val Pro Ser 45 50 55 60 AGG TTC ACT GGC ACT GGA TCT GGA GCA GAT TAT TCT CTC ACC ATC AGC 288 Arg Phe Ser Gly Ser Gly Ser Gly Ala Asp Tyr Ser Leu Thr He Ser 65 70 75 AGC CTG GAA TCT GAA GAT TIT GCA GAC TAT TAT TCT CTA CAG TAT GCT 336 Ser Leu Glu Ser Glu Asp Phe Ala Asp Tyr Tyr Cys Val Gin Tyr Ala 80 85 90 - 32 CAG TIT CCT CGG ACG TTC GGT GGA GGC ACC AAG CTG GAA ATC AAA Gin Phe Pro Arg Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lvs 95 100 105 381 - 33 (2) INFORMATION FOR SEQ ID NO:6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 396 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA to mRNA (lx) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..396 (ix) FEATURE: (A) NAME/KEY: mat__peptide (B) LOCATION: 58..396 (ix) FEATURE: (A) NAME/KEY: sig_peptide (B) LOCATION: 1..57 (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:6: ATC GAC TCC AGG CTC AAT TTA GTT TTC CTT CTC CTT ATI TEA AAA GCT Phe Leu Val Leu lie Leu Lys Gly 48 Met Asp -19 Ser Arg Leu -15 Asn Leu Val -10 -5 CTC CAG TCT GAT CTG CAG CTG CTG GAG TCT GGG GGA GGC TIA GTG CAG 96 Val Gin Cys Asp Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin 1 5 10 CCT GGA GGG TCC CGG AAA CTC TCC TCT GTT GCC TCT GGA TIC ACT TTC 144 Pro Gly Gly Ser Arg Lys Leu Ser Cys Val Ala Ser Gly Phe Thr Phe 15 20 25 ACT ACC ΊΤΤ GGA ATC CAC TCG CTT CCT CAG GCT CCA GAG AAG GGG CTG 192 Ser Thr Phe Gly Met His Trp Val Arg Gin Ala Pro Glu Lys Gly Leu 30 35 40 45 GAG TGG CTC GCA TAC ATT ACT ACT GGC ACT GCT ACC ATC TAC TAT GCA 240 Glu Trp Val Ala Tyr He Ser Ser Gly Ser Gly Thr He Tyr Tyr Ala 50 55 60 GAC ACA CTG AAG GGC CGA TIC ACC ATC TCC AGA GAC AAT CCC AAG AAT 288 Asp Thr Val Lys Gly Arg Phe Thr He Ser Arg Asp Asn Pro Lys Asn 65 70 75 ACC CTC TTC CTC CAA ATC ACC ACT CTA AGG TCT GAG GAC ACG GCC ATC 336 Thr Leu Phe Leu Gin Met Thr Ser Leu Arg Ser Glu Asp Thr Ala Met 80 85 90 TAT TAG TCT GCA AGA GAT TGG ATG AAC TGG GGC CAA GGG ACT CTC CTC Tyr Tyr Cys Ala Arg Asp Trp Met Asn Trp Gly Gin Gly Thr Leu Val 100 105 ACT CTC TCT GCA Thr Val Ser Ala 110 334 396 - 35 (2) INFORMATION FOR SEQ ID NO:7: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 435 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA to mRNA (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..435 (ix) FEATURE: (A) NAME/KEY: nat_j5epride (B) LOCATION: 67..435 (ix) FEATURE: (A) NAME/KEY: sig_peptide (B) LOCATION: 1..66 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7: ATC GAT ITT CAA GTC CAG Val Gin ATT lie TIC AGC TTC CTC CTA ATC ACT GCT TCA 48 Met Asp Phe Gin Phe Ser -15 Phe Leu Leu He Ser Ala Ser -10 -22 -20 GIG ATA ATC TCC AGA GGC AAA ATT CTT CTC TCC CAG TCT CCA GCA ATC 96 Val He Met Ser Arg Gly Lys He Val Leu Ser Gin Ser Pro Ala He -5 1 5 10 CTC TCT GCA TCT CCA GGG GAG AAG CTC ACA. ATC ACT TCC AGG GCC AGC 144 Leu Ser Ala Ser Pro Gly Glu Lys Val Thr Met Thr Cys Arg Ala Ser 15 20 25 TCA ACT ATA ACT TAC ATC CAC TCG TAC CAG CAG AAG CCA GGA TCC TCC 192 Ser Ser lie Ser Tyr Met His Trp Tyr Gin Gin Lys Pro Gly Ser Ser 30 35 40 ccc AAA CCC TCG ATT CAA GCC AGA TCC AAC CTC GCT ITT GGA CTC CCT 240 Pro Lys Pro Trp lie Gin Ala Thr Ser Asn Leu Ala Phe Gly Val Pro 45 50 55 TGT CGC TTC ACT GGC ACT GGG TCT GGG ACC TCT TAC TCT CTC ACA ATC 288 Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr He 60 65 70 AGC AGA GIG GAG GCT GAA GAT GCT GCC ACT TAT TAC TCC CAG CAG TCG 336 Ser Arg Val Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gin Gin Ttp 80 85 90 ACT ACT Ser Ser AAC CCA TIC Asn Pro Phe 95 ACG Thr TTC GGC TCG Phe Gly Ser GGG ACA AAG TTC GAA ATC AAA 384 Gly 100 Thr Lys Leu Glu Met 105 Lys CGG GCT GAT GCT GCA CCA ACT GLA TCC ATC TTC CCA CCA TCC ACT GAG 432 Arg Ala Asp Ada Ala Pro Thr Val Ser lie Phe Pro Pro Ser Ser Glu 110 115 120 CAG 435 Gin - 37 (2) INFORMATION FOR SEQ ID NO: 8: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 549 base pai rs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA to mRNA (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..549 (ix) FEATURE: (A) NAME/KEY: mat_peptide (B) LOCATION: 58.7549 (ix) FEATURE: (A) NAME/KEY: sig_peptide (B) LOCATION: 1..57 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8: ATC GOT GIG CTC GGG CTC CTT CTC TOO CTG CTG ACT TIG CCA AGC TCT Cys Leu Val Thr Phe Pro Ser Cys 48 Met Ala -19 Val Leu Gly -15 Leu Leu Leu -10 -5 GTC CCG TCC CAG GIG CAG CTG AAG GAG TCA GGG CCT GGC CTC GTG GCG 96 Val Pro Ser Gin 1 Val Gin Leu Lys 5 Glu Ser Gly Pro Gly Leu Val Ala 10 CCC TCA CAG AGC CTC TCC ATC ACA TOO ACC GTC TCA GGG TTC TCA TEA 144 Pro Ser 15 Gin Ser Leu Ser lie 20 Thr Cys Thr Val Ser Gly Phe Ser Leu 25 ACT ACC TAT AGT CTA TAC TGG GIT CGC CAG CCT CCA GGA AAG. GGE CTC 192 Ser Thr 30 Tyr Ser Val Tyr Trp 35 Val Arg Gin Pro 40 Pro Gly Lys Gly Leu 45 GAG TGG CTG GGA GTG ATA TGG AGE GAT GGA AGO ACA ACC TAT AAT TCA 240 Glu Trp Leu Gly Val 50 lie Trp Ser Asp Gly Ser 55 Thr Thr Tyr Asn Ser 60 ACT orc AAA TCC AGA CTC ACC ATC AGO AAG GAC AAC TCC AAG ACT CAA 288 Thr Leu Lys Ser 65 Arg Leu Thr lie Ser Lys Asp 70 Asn Ser Lys Ser Gin 75 CTE TTC TEA AAA GIG AAC AGT CTC CAA ACT GAT GAC ACA GCC ATC TAC 336 Val Phe Leu Lys 80 Val Asn Ser Leu 85 Gin Thr Asp Asp Thr Ala Met Tyr 90 TAC TCT GCC AGA ACC TAT GCT TAT GAC GGG TCC TGG CTT GCT TAC TGG 384 Tyr Cys Ala Arg Thr Tyr Gly Tyr Asp Gly Ser Tro Leu Ada Tyr Trp 95 100 105 GGC CAA GGG ACT CTG CTC ACT GTC TCT GCA GCC AAA ACA ACA CCC CCA 432 Gly Gin Gly Thr Leu Val Thr Val Ser Ala Ala Lys Thr Thr Pro Pro 110 115 120 125 TCA GTC TAT CCA CTG GCC CCT GGG TCT GGA GAT ACA ACT GCT TCC TCC 480 Ser Val Tyr Pro Leu Ala Pro Gly Cys Gly Asp Thr Thr Gly Ser Ser 130 135 140 GTG ACT CTG GGA TGC CTG CTC AAG GGC TAC ITC CCT GAG TCA GTG ACT 528 Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Ser Val Thr 145 150 155 GTG ACT TGG AAC TCT GGA TCC Val Thr Trp Asn Ser Gly Ser 160 549 - 39 (2) INFORMATION FOR SEQ ID NO:9: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 322 base pairs (B) TYPE: nucleic acid (C) STRANDEDiESS: single (D) TOPOLOGY: linear (ii) iOLECULE TYPE: cDNA to mRNA (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..321 (ix) FEATURE: (A) NAME/KEY: mat_peptide (B) LOCATION: 1..321 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: GAC GTC TTC CTC ACT CAG TCT CCA GCC ATC CTC TCC GTC ACT CCA GGA 48 Asp 1 Val Leu Leu Thr 5 Gin Ser Pro Ala lie 10 Leu Ser Val Ser Pro 15 Gly GAA AGA GIC ACT TTC ICC TCT AGG GCC ACT CAG AGC ATT GGC ACA AGC 96 Glu Arg Val Ser 20 Phe Ser Cys Arg Ala 25 Ser Gin Ser He Gly 30 Thr Ser ATA CAC TGG TAT CAG CAA AGA ACA AAT GCT CCT CCA AGG CTT CTC ATA 144 lie His Trp Tyr 35 Gin Gin Arg Thr 40 Asn Gly Pro Pro Arg 45 Leu Leu lie AAG TAT GCG TCT GAG TCA ATC TCT GGG ATC CCT TCC AGG TIT ACT GGC 192 Lys Tyr 50 Ala Ser Glu Ser He Ser 55 Gly He Pro Ser 60 Arg Phe Ser Gly AGT GGA TCA GGG ACA GAT TIT ACT CTT AGC ATC AGC AGT CTG GAG TCT 240 Ser 65 Gly Ser Gly Thr Asp 70 Phe Thr Leu Ser lie 75 Ser Ser Val Glu Ser 80 GAA GAT ATT GCA GAT TAT TAC TCT CAA CAA ACT AAT AGC TGG CCA ACC 288 Glu Asp He Ala Asp 85 Tyr Tyr Cys Gin Gin 90 Thr Asn Ser Trp Pro 95 Thr ACG TIC GGA GGG GGG ACC AAG CTG GAA AIT AAA C Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lys 100 105 322 - 40 (2) INFORMATION FOR SEQ ID NO:10: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 355 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA to mRNA (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..354 (ix) FEATURE: (A) NAME/KEY: mat_peptide (B) LOCATION: 1..354 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10: GAG GTC CAG CTG CAA Glu Val Gin Leu Gin CAG TTT GGA GCT GAA TTC GTC AAG CCT GGG ACT Gin Phe Gly Ala Glu Leu Val Lys Pro Gly Thr 1 5 10 15 TCG CTG AAG ATA TCC TCC AAG GCT TCT GGC TAC ATT TTC ACT GAC TAC Ser Val Lys lie Ser Cys Lys Ala Ser Gly Tyr He Phe Thr Asp Tyr 20 25 30 AAC ATC GAC TGG GTC AAG CAG AGC CAT GGA AAG AGC CTT GAG TCG AIT Asn Met Asp Trp Val Lys Gin Ser His Gly Lys Ser Leu Glu Trp He 35 40 45 GGA GAT ATT GAT CCT AAC TTT GAT ACT TCC ACT TAC AAC CAG AAG TTC Gly Asd He Asp Pro Asn Phe Asp Ser Ser Ser Tyr Asn Gin Lys Phe 50 55 60 AAG GGA AAG GCC ACA TTC ACT GTA GAC AAG TCC TCC AAC ACA GCC TAC Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Asn Thr Ala Tyr 65 70 75 80 ATC GAG CTC CGC AGC CTC ACA TCT GAG GAC ACT GCA CTC TAT TAC TCT Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 GCA AGA GGG GGA TTC CCC TAT GCT ATG GAC TAC TOG GCT CAA GGA ACC Ala Arg Gly Gly Phe Pro Tyr Gly Met Asp Tyr Trp Gly Gin Gly Thr 100 105 110 144 192 240 288 336 TCA CTC ACC GTC TCC TCA G Ser Val Thr Val Ser Ser 115 355

Claims (28)

Claims

1. Antibody composition acting synergistically for improving immunosuppression, wherein it contains (a) at least one monoclonal anti-CD4 antibody which by itself is already strongly inhibitory and (b) at least one monoclonal anti-IL2Ra or anti-IL2RB antibody which by itself is already strongly inhibitory, whereby a strongly inhibitory antibody at a concentration of 10000 ng/ml inhibits allogeninduced lymphocyte proliferation by at least 40 % in the absence of other antibodies.

2. Antibody composition as claimed in claim 1, wherein it contains at least one anti-IL2Ra antibody as the antibody (b) whereby the molar ratio of antibodies (a) and (b) is from 1:10 to 10:1.

3. Antibody composition as claimed in claim 2, wherein the molar ratio of the antibodies (a) and (b) is from 1:5 to 5:1.

4. Antibody composition as claimed in claim 2, wherein the molar ratio of the antibodies (a) and (b) is from 3:10 to 3:1. - 42

5. Antibody composition as claimed in one of the claims 1-4, wherein the antibody (a) has the amino acid sequences shown in SEQ ID NO. 1 and 2 or in SEQ ID NO. 3 and 4 as the variable regions of the light or heavy chain.

6. Antibody composition as claimed in one of the claims 1-5, wherein the antibody (b) has the amino acid sequences shown in SEQ ID NO. 5 and 6 as the variable regions of the light or heavy chain.

7. Antibody composition as claimed in claim 1, wherein it contains at least one anti-IL2RB antibody as antibody (b) whereby the molar ratio of the antibodies (a) and (b) is from 1:1000 to 10:1.

8. Antibody composition as claimed in claim 7, wherein the antibody (b) has the amino acid sequences shown in SEQ ID NO. 9 and 10 as the variable regions of the light or heavy chain.

9. Antibody composition as claimed in one of the previous claims, wherein the antibodies (a) and (b) are murine, human, chimerized or humanized antibodies or antibody fragments. - 43

10. Pharmaceutical agent comprising components (1) and (2) whereby the two components are to be administered in combination but can be formulated separately, wherein the component (1) contains a monoclonal anti-CD4 antibody (a) which by itself is strongly inhibitory as the active substance and component (2) contains a monoclonal anti-IL2Ra or anti-IL2RB antibody (b) which by itself is strongly inhibitory as the active substance, if desired, together with the usual pharmaceutical auxiliary, dilution, carrier and filling agents.

11. Pharmaceutical agent as claimed in claim 10, wherein the component (2) contains a monoclonal anti-IL2Ra antibody (b) which by itself is strongly inhibitory as the active substance, whereby the molar ratio of the two antibodies of the components (1) and (2) is from 1:10 to 10:1 in the pharmaceutical agent.

12. Pharmaceutical agent as claimed in claim 11, wherein the molar ratio of the two antibodies is from 1:5 to 5:1.

13. Pharmaceutical agent as claimed in claim 11, wherein the molar ratio of the two antibodies is from 3:10 to 3:1.

14. Pharmaceutical agent as claimed in one of the claims 10 to 13, wherein the antibody (a) has the amino acid sequences shown in SEQ ID NO. 1 and 2 or the amino acid sequences shown in SEQ ID NO. 3 and 4 as the variable regions of the light or heavy chain.

15. Pharmaceutical agent as claimed in one of the claims 10 to 14, wherein the antibody (b) has the amino acid sequences shown in SEQ ID NO. 5 and 6 as the variable regions of the light or heavy chain.

16. Pharmaceutical agent as claimed in claim 10, wherein the component (2) contains a monoclonal anti-IL2RB antibody (b) which by itself is strongly inhibitory as the active substance, whereby the molar ratio of the two antibodies of the components (1) and (2) is from 1:1000 to 10:1 in the pharmaceutical agent.

17. Pharmaceutical agent as claimed in claim 16, wherein the antibody (b) has the amino acid sequences shown in SEQ ID NO. 9 and 10 as the variable regions of the light or heavy chain. - 45

18. Pharmaceutical agent as claimed in one of the claims 10 to 17, wherein the antibodies (a) and (b) are murine, human, chimerized or humanized antibodies or antibody fragments.

19. Use of an antibody composition as claimed in one of the claims 1 to 9 or of a pharmaceutical agent as claimed in one of the claims 10 to 18 for immunosuppressive therapy, in particular for therapy after organ or tissue transplantations.

20. Process for the production of a pharmaceutical agent for an immunosuppressive therapy, in particular for therapy after organ and tissue transplantations in which a pharmaceutical agent comprising components (1) and (2) is provided the two components of which are to be administered in combination but can be formulated separately, wherein the component (1) contains a strongly inhibitory monoclonal anti-CD4 antibody (a) as the active substance and the component (2) contains a strongly inhibitory monoclonal anti-IL2Ra or anti-IL2Re antibody (b) as the active substance, if desired, with the usual pharmaceutical auxiliary, dilution, carrier and filling substances.

21. Process as claimed in claim 20, wherein an anti-IL2Ra antibody is used as the antibody (b) whereby the molar ratio of the two antibodies of the components (1) and (2) is from 1:10 to 10:1.

22. Process as claimed in claim 20, wherein an anti-IL2RB antibody is used as the antibody (b) whereby the molar ratio of the two antibodies of the components (1) and (2) is from 1:1000 to 10:1.

23. Process for immunosuppressive therapy, in particular for therapy after organ and tissue transplantations, wherein a pharmaceutical agent is administered as claimed in one of the claims 10 to 18.

24. An antibody composition according to claim 1, substantially as hereinbefore described.

25. A pharmaceutical agent according to claim 10, substantially as hereinbefore described.

26. Use according to claim 19, substantially as hereinbefore described.

27. A process for the production of a pharmaceutical agent according to claim 10, substantially as hereinbefore described and exemplified.

28. A pharmaceutical agent according to claim 10, whenever produced by a process claimed in any one of claims 20-22 or 27.