ANTI-CD154 ANTIBODIES HAVING IMPAIRED FcR BINDING AND/OR COMPLEMENT BINDING PROPERTIES AND USE IN THERAPY

Abstract

Improved anti-CD154 antibodies are provided herein which have ablated FcR binding and/or complement binding/activation. The use of these antibodies for inducing tolerance and treating immune diseases including autoimmunity, inflammation and allergic disorders is disclosed herein.

1 - 26 . (canceled) 
     
     
         27 . A method of inhibiting the activity of CD154 in a subject in need thereof which comprises the administration of a therapeutically effective amount of a anti-CD154 antibody of the human IgG1 isotype, comprising an Fc region which comprises an E269R mutation and a K322A mutation and which further comprises one additional Fc modification which impairs FcR binding and/or impairs complement binding, wherein said anti-CD154 antibody comprises the V L  complementarity determining regions (CDRs) of SEQ ID NO: 17, 18, and 19 and the V L  CDRs of SEQ ID NO: 24, 25 and 26 wherein the one additional Fc modification which impairs FcR binding is selected from the following mutations: E233P, D265A, D265N, D270N, N297A, S298N, P329A, D270A, K326V, V369R, F405K, L410P, V427R, L234N, G237M, S239F, V262E, V264F, V266T, S267N, N268E, N297R, T299A, R301D, N325L, N325E and L328R, and the additional Fc modification which impairs complement binding is a P331G or P331G/K322A mutation. 
     
     
         28 . An anti-CD154 antibody of the human IgG1 isotype, comprising an Fc region which comprises an E269R mutation and a K322A mutation and which further comprises an additional Fc modification which impairs FcR binding and/or impairs complement binding, wherein said anti-CD154 antibody comprises the Vt, complementarity determining regions (CDRs) of SEQ ID NO: 17, 18, and 19 and the VII CDRs of SEQ ID NO: 24, 25 and 26; wherein the additional Fc modification which impairs FcR binding is selected from the following mutations: E233P, D265A, D265N, D270N, N297A, S298N, P329A, D270A, K326V, V369R, F405K, L410P, V427R, L234N, G237M, S239F, V262E, V264F, V266T, S267N, N268E, N297R, T299A, R301D, N325L, N325E and L328R, and the additional Fc modification which impairs complement binding is a P331G or P331G/K322A mutation. 
     
     
         29 . A composition comprising the anti-CD154 antibody of  claim 28  and a pharmaceutically acceptable carrier. 
     
     
         30 . A nucleic acid encoding an anti-CD154 antibody of the human IgG1 isotype according to  claim 28 , which comprises an Fc region which comprises an E269R mutation and a K322A mutation and which further comprises one additional Fc modification which impairs FcR binding and/or impairs complement binding, wherein said anti-CD154 antibody comprises the V L  complementarity determining regions (CDRs) of SEQ ID NO: 17, 18, and 19 and the V H  CDRs of SEQ ID NO: 24, 25 and 26 wherein the one additional Fc modification which impairs FcR binding is selected from the following mutations: E233P, D265A, D265N, D270N, N297A, S298N, P329A, D270A, K326V, V369R, F405K, L410P, V427R, L234N, G237M, S239F, V262E, V264F, V266T, S267N, N268E, N297R, T299A, R301D, N325L, N325E and L328R, and the additional Fc modification which impairs complement binding comprises a P331G or P331G/K322A mutation. 
     
     
         31 . An expression vector comprising the nucleic acid of  claim 30  operably linked to a promoter. 
     
     
         32 . A recombinant cell comprising the nucleic acid of  claim 30  operably linked to a promoter. 
     
     
         33 . A method of producing an anti-CD154 antibody of the human IgG1 isotype, which comprises an Fc region which comprises an E269R mutation and a K322A mutation, comprising culturing a cell comprising the nucleic acid of  claim 30  operably linked to a promoter under conditions that permit expression of the anti-CD154 antibody encoded by said nucleic acid. 
     
     
         34 . The method of 33, which comprises isolating the expressed anti-CD154 antibody from the cell or culture medium comprising the cell. 
     
     
         35 . A method of obtaining an antibody that comprises an antigen binding region (ABD) which binds to a desired antigen and which comprises impaired FcR binding and complement binding by:
 (i) obtaining an isolated nucleic acid that encodes for said ABD and a nucleic acid encoding an IgG1 Fc region of the human IgG1 isotype comprising an E269R mutation and a K322A mutation, wherein said isolated nucleic acids may be on the same or different nucleic acid constructs;   (ii) introducing said isolated nucleic acids into an isolated host cell;   (iii) culturing said host cell under conditions which provide for the expression of an antibody that comprises an antigen binding region (ABD) which binds to said desired antigen and which comprises an IgG1 Fc region of the human IgG1 isotype comprising an E269R mutation and a K322A mutation that comprises impaired FcR binding and complement binding; and   (iv) optionally isolating the resultant expressed antibody from the host cell or culture containing the host cell.   
     
     
         36 . The method of  claim 35 , wherein the desired antigen is human CD154. 
     
     
         37 . The method of  claim 35 , wherein the encoded IgG1 Fc region further comprises an additional Fc modification which impairs FcR binding and/or an additional Fc modification which impairs complement binding, wherein said additional Fc modification which impairs FcR binding is selected from the following mutations: E233P, D265A, D265N, D270N, N297A, S298N, P329A, D270A, K326V, V369R, F405K, L410P, V427R, L234N, G237M, S239F, V262E, V264F, V266T, S267N, N268E, N297R, T299A, R301D, N325L, N325E and L328R, and wherein said additional Fc modification which impairs complement binding is a P331G or a P331G/K322A mutation.