Humanized anti-CD4 antibody with immunosuppressive properties
A humanized antibody derived from mouse monoclonal anti-CD4 antibody B-F5 is able to activate CD25+CD4+ regulatory T cells and is useful for preparing immunosuppressive compositions.
1. A polynucleotide encoding a V domain of an antibody, wherein the antibody is a humanized hB-F5antibody, wherein said hB-F5 antibody comprises V domains comprising:
H chain V domain comprising the sequence:
(SEQ ID NO: 1)
EEQLVESGGGLVKPGGSLRLSCAASGFSFSDCRMYWLRQAPGKGLEWIGV
ISVKSENYGANYAESVRGRFTISRDDSKNTVYLQMNSLKTEDTAVYYCSA
SYYRYDVGAWFAYWGQGTLVTVSS; and
L chain V domain comprising the sequence:
(SEQ ID NO: 2)
DIVMTQSPDSLAVSLGERATINCRASKSVSTSGYSYIYWYQQKPGQPPKL
LIYLASILESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQHSRELPW
TFGQGTKVEIK,
wherein said hB-F5 antibody is able to activate a subset of CD4 + T cells comprising CD4 + CD25 + regulatory T cells.
2. A polynucleotide selected from the group consisting of
(a) a polynucleotide comprising a sequence encoding for SEQ ID No: 1; and
(b) a polynucleotide comprising a sequence encoding for SEQ ID No: 2.
3. A polynucleotide according to claim 1 or claim 2 , selected from the group consisting of:
(a) a polynucleotide comprising the sequence SEQ ID No: 3; and
(b) a polynucleotide comprising the sequence SEQ ID No: 4.
4. A polynucleotide encoding a V domain of an antibody, wherein the antibody is a humanized anti-CD4 antibody, wherein said humanized anti-CD4 antibody comprises a sequence comprising the complementarity-determining regions (CDRs) of SEQ ID NOs: 10 and 9, displayed in FIGS. 3 and 4 , and wherein the humanized anti-CD4 antibody is capable of activating CD4 + CD25 + regulatory T cells such that the activated CD4 + CD25 + regulatory T cells are able to suppress CD4 + CD45RA+CD25-T cell proliferation in vitro in an allogenic mixed lymphocyte reaction in which the activated CD4 + CD25 + regulatory T cells are cocultured with allogenic CD4 + CD45RA+CD25-T cell in the presence of CD2-depleted peripheral blood mononuclear cells that are syngenic for the CD4 + CD25 + regulatory T cells.
5. A polynucleotide according to claim 4 wherein the humanized anti-CD4 antibody is obtainable from the mouse monoclonal anti-CD4 antibody having V h and V k regions defined by SEQ ID No: 5 and SEQ ID No: 6.
6. An expression vector comprising at least one polynucleotide according to claim 1 .
7. A host cell comprising the expression vector according to claim 6 .
8. A host cell according to claim 7 wherein the cell is a prokaryotic cell.
9. A host cell according to claim 7 wherein the cell is a eukaryotic cell.
10. A host cell according to claim 9 wherein the eukaryotic cell is selected from the group consisting of: plant cell, insect cell, yeast cell or mammalian cell.
11. A polynucleotide according to claim 1 , wherein said subset of CD4+ T cells comprising CD4+CD25+ regulatory T cells are able to suppress CD4 + CD45RA+CD25-T cell proliferation in vitro in an allogenic mixed lymphocyte reaction in which the activated CD4 + CD25 + regulatory T cells are cocultured with allogenic CD4 + CD45RA+CD25-T cell in the presence of CD2-depleted peripheral blood mononuclear cells that are syngenic for the CD4 + CD25 + regulatory T cells.