AGENT FOR THE TREATMENT AND/OR PROPHYLAXIS OF AN AUTOIMMUNE DISEASE AND FOR THE FORMATION OF REGULATORY T CELLS
The present invention relates to an agent for the treatment and/or prophylaxis of an autoimmune disease, an agent for the formation of regulatory T cells (T Reg ) in an organism and various methods in which the agents according to the invention are used.
1 . Method for the preparation of a medicament for the treatment and/or prophylaxis of an autoimmune disease, comprising the following steps:
(1) providing a mutein of human interleukin-2 (hIL-2 mutein) or of a fragment thereof, which is numbered in accordance with the hIL-2 wild type and has an amino acid substitution in at least one of the positions 20, 88 or 126,
(2) formulating said mutein of fragment into a pharmaceutically acceptable carrier.
2 . Method according to claim 1 , wherein through the substitution at position 88 an asparagine is exchanged for an amino acid which is selected from the group consisting of:
arginine (hIL-2-N88R), glycine (hIL-2-N88G), or isoleucine (hIL-2-N88I).
3 . Method according to claim 1 , wherein through the substitution at position 20 an aspartic acid is exchanged for an amino acid which is selected from the group consisting of: histidine (hIL-2-D2OH), isoleucine (hIL-2-D2OH), or tyrosine (hIL-2-D20Y).
4 . Method according to claim 1 , wherein through the substitution at position 126, a glutamine is exchanged for a leucine (hIL-2-Q126L).
5 . Method according to claim 1 , wherein the hIL-2 mutein or the fragment thereof has at least one further amino acid substitution in any position except the positions 20, 88 or 126, so that the thus further substituted hIL-2 mutein or the thus further substituted fragment thereof has an amino acid sequence which is at least 80% identical with the amino acid sequence of the hIL-2 mutein or of the section thereof, which is not further substituted, compared to the hIL-2 wild type, other than in at least one of the positions 20, 88 or 126.
6 . Method according to claim 5 , wherein the further substitution in at least any position, except the positions 20, 88 or 126, is a conservative amino acid substitution.
7 . Method according to claim 1 , wherein the medicament in addition contains an immunosuppressant.
8 . Method according to claim 7 , wherein the immunosuppressant is selected from the group consisting of: glucocorticoid, including decortin, prednisol; azathioprine; cyclosporin A; mycophenolate mofetil; tacrolimus; anti-T lymphocyte globulin, anti-CD3 antibodies, including muromonab; anti-CD25 antibodies, including basiliximab and daclizumab; anti-TNF-α antibodies, including infliximab and adalimumab; azathioprine; methotrexate; cyclosporin; sirolimus; everolimus; fingolimod; CellCept®; myfortic; and cyclophosphamide.
9 . Method for the treatment and/or prophylaxis of an autoimmune disease in an organism, which comprises the following steps:
(a) providing a mutein of human interleukin-2 (hIL-2 mutein) or of a fragment thereof, which is numbered in accordance with the hIL-2 wild type and has an amino acid substitution in at least one of the positions 20, 88 or 126,
(b) administering said hIL-2 mutein or of said fragment thereof to an organism, and
(c) if necessary repeating the steps (a) and (b).
10 . Method according to claim 9 , wherein through the substitution at position 88 an asparagine is exchanged for an amino acid which is selected from the group consisting of: arginine (hIL-2-N88R), glycine (hIL-2-N88G), or isoleucine (hIL-2-N88I).
11 . Method according to claim 9 , wherein through the substitution at position 20 an aspartic acid is exchanged for an amino acid which is selected from the group consisting of: histidine (hIL-2-D2OH), isoleucine (hIL-2-D2OH), or tyrosine (hIL-2-D20Y).
12 . Method according to claim 9 , wherein through the substitution at position 126, a glutamine is exchanged for a leucine (hIL-2-Q126L).
13 . Method according to claim 10 , wherein the hIL-2 mutein or the fragment thereof has at least one further amino acid substitution in any position except the positions 20, 88 or 126, so that the thus further substituted hIL-2 mutein or the thus further substituted fragment thereof has an amino acid sequence which is at least 80% identical with the amino acid sequence of the hIL-2 mutein or of the section thereof, which is not further substituted, compared to the hIL-2 wild type, other than in at least one of the positions 20, 88 or 126.
14 . Method according to claim 13 , wherein the further substitution in at least any position, except the positions 20, 88 or 126, is a conservative amino acid substitution.
15 . Method according to claim 9 , wherein the medicament in addition contains an immunosuppressant.
16 . Method according to claim 15 , wherein the immunosuppressant is selected from the group consisting of: glucocorticoid, including decortin, prednisol; azathioprine; cyclosporin A; mycophenolate mofetil; tacrolimus; anti-T lymphocyte globulin, anti-CD3 antibodies, including muromonab; anti-CD25 antibodies, including basiliximab and daclizumab; anti-TNF-α antibodies, including infliximab and adalimumab; azathioprine; methotrexate; cyclosporin; sirolimus; everolimus; fingolimod; CellCept®; myfortic; and cyclophosphamide.
17 . Method for the treatment and/or prophylaxis of an autoimmune disease in an organism, which comprises the following steps:
(a) providing a mutein of human interleukin-2 (hIL-2 mutein) or of a fragment thereof, which is numbered in accordance with the hIL-2 wild type and has an amino acid substitution in at least one of the positions 20, 88 or 126,
(b) contacting said hIL-2 mutein or of said fragment thereof with peripheral mononuclear blood cells (PBMCs) deriving from a first organism,
(c) incubating said hIL-2 mutein or of the fragment thereof with the PBMCs to obtain a cell population which contains regulatory T cells (T Reg ), and
(d) introducing the cell population into a second organism.
18 . Method according to claim 17 , wherein the first and the second organisms are identical organisms.