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 humanized anti-CD4 antibody which is a humanized version of a mouse monoclonal anti-CD4 antibody, wherein the mouse monoclonal anti-CD4 antibody comprises a Vh region having SEQ ID NO: 9 and a Vk region having SEQ ID NO: 10, and wherein the humanized anti-CD4 antibody further comprises an IgG1 constant domain.
2. A humanized anti-CD4 antibody which comprises an L chain V domain having the complementarity-determining regions of SEQ ID NO: 2 shown in FIG. 3 and an H chain V domain having the complementarity-determining regions of SEQ ID NO: 1 shown in FIG. 4 , wherein the H chain V domain further comprises a Leucine residue at the position 37, and wherein the humanized anti-CD4 antibody further comprises an IgG1 constant domain.
3. A humanized anti-CD4 antibody according to claim 2 which comprises an H chain V domain having SEQ ID NO: 1 and an L chain V domain having SEQ ID NO: 2, and wherein the humanized anti-CD4 antibody further comprises an IgG1 constant domain.
4. A therapeutic composition comprising the humanized anti-CD4 antibody of claim 1 .
5. A method of making a humanized anti-CD4 antibody according to claim 3 comprising the steps of:
culturing a host cell comprising a polynucleotide encoding SEQ ID NO: 1 and a polynucleotide encoding SEQ ID NO: 2, under conditions suitable for expression thereof, and
recovering the expressed humanized anti-CD4 antibody.
6. A method according to claim 5 wherein the host cell is a mammalian cell.
7. A method according to claim 6 wherein the mammalian cell is a CHO cell.
8. A method according to claim 6 wherein the mammalian cell is a Sp2/0 cell.
9. A method according to claim 5 further comprising a step of purifying the expressed humanized anti-CD4 antibody by affinity chromatography using protein A Sepharose.
10. A method according to claim 7 further comprising a step of purifying the expressed humanized anti-CD4 antibody by affinity chromatography using protein A Sepharose.
11. A method according to claim 8 further comprising a step of purifying the expressed humanized anti-CD4 antibody by affinity chromatography using protein A Sepharose.
12. A method of treating a subject suffering from an autoimmune disease comprising administering to said subject an antibody according to claim 1 .
13. A polynucleotide comprising a DNA molecule encoding the antibody of claim 1 .
14. A method of using the humanized antibody of claim 1 to activate CD25+CD4+ regulatory T cells comprising contacting the humanized antibody with CD25 + CD4 + regulatory T cells in vitro.
15. A method of preventing graft rejection in a subject comprising administering to said subject the therapeutic composition of claim 4 .