Immunogenic compositions for induction of anti-tumor immunity

Abstract

The invention relates to the use of an immunogen selected from the group consisting of (i) an anti-p53 mAb; (ii) a fragment of an anti-p53 mAb; (iii) a peptide based on a CDR of the heavy or light chain of an anti-p53 mAb, which peptide is capable of eliciting antibodies to p53; and (iv) a DNA molecule coding for the variable (V) region of an anti-p53 mAb in a suitable gene delivery vehicle, for the preparation of a pharmaceutical composition useful for induction of anti-tumor immunity in mammals, for activating an enhanced immune response to a p53 molecule in mammals, and/or for induction of immune responses to mutated and wild-type forms of a p53 in mammals. The use of anti-p53 mAbs and novel peptides based on the CDR2 and CDR3 of the heavy chains and CDR3 of the light chains of different anti-p53 mAbs are disclosed.

1 . A method for inducing anti-tumor immunity in a mammal which comprises administering to the mammal an effective amount of at least one immunogen selected from the group consisting of:
 (i) an anti-p53 mAb;   (ii) a fragment of an anti-p53 mAb; and   (iii) a DNA molecule coding for the variable (V) region of an anti-p53 mAb in a suitable gene delivery vehicle.   
     
     
         2 . The method according to  claim 1 , wherein the method is used for inducing immune responses to mutated and wild-type forms of p53 in the mammal. 
     
     
         3 . The method according to  claim 1 , wherein the immunogen is selected from the group consisting of:
 (a) a murine anti-p53 selected from the group consisting of: mAb 240, mAb 246 and mAb 421;   (b) a humanized anti-p53 mAb derived from a murine mAb of (a);   (c) a human anti-p53 mAb; and   (d) a fragment of an anti-p53 mAb selected from antigen-binding fragments (Fab), F(ab′) 2  and single chain Fv fragments.   
     
     
         4 . The method of  claim 1 , which comprises administering to a patient effective amounts of two or more different anti-p53 mAbs, or fragments thereof, either concomitantly or sequentially. 
     
     
         5 . A method for activating an enhanced immune response to p53 in a mammal which comprises immunizing the mammal with an effective amount of at least one immunogen selected from the group consisting of:
 (i) an anti-p53 mAb;   (ii) a fragment of an anti-p53 mAb; and   (iii) a DNA molecule coding for the variable (V) region of an anti-p53 mAb in a suitable gene delivery vehicle.   
     
     
         6 . The method according to  claim 5 , wherein the method is used for inducing immune responses to mutated and wild-type forms of p53 in the mammal 
     
     
         7 . The method according to  claim 1 , wherein the immunogen is selected from the group consisting of:
 (a) a murine anti-p53 selected from the group consisting of: mAb 240, mAb 246 and mAb 421;   (b) a humanized anti-p53 mAb derived from a murine mAb of (a);   (c) a human anti-p53 mAb; and   (d) a fragment of an anti-p53 mAb selected from antigen-binding fragments (Fab), F(ab′) 2  and single chain Fv fragments.   
     
     
         8 . The method of  claim 1 , which comprises administering to a patient effective amounts of two or more different anti-p53 mAbs, or fragments thereof, either concomitantly or sequentially. 
     
     
         9 . A method for activating an enhanced anti-tumor immune response in a mammal which comprises:
 determining at least one immunogen that can elicit antibodies to p53 wherein the immunogen is selected from the group consisting of:   (i) an anti-p53 mAb;   (ii) a fragment of an anti-p53 mAb; and   (iii) a DNA molecule coding for the variable (V) region of an anti-p53 mAb in a suitable gene delivery vehicle; and   administering an effective amount of the at least one immunogen to a mammal having a tumor characterized by mutation, over-expression or accumulation of p53.   
     
     
         10 . The method according to  claim 9 , wherein the method is used for inducing immune responses to mutated and wild-type forms of p53 in the mammal. 
     
     
         11 . The method according to  claim 9 , wherein the anti-p53 mAb is selected from mAb 240, mAb 246 and mAb 421. 
     
     
         12 . The method according to  claim 9 , wherein the anti-p53 mAb contains at least one sequence selected from the group consisting of:
 (i) sequences herein designated la-lb, based on the CDR2 and CDR3, respectively, of the heavy chain (240VH), and sequence Ic based on the CDR3 of the light chain (240V f). of the anti-p53 mAb 240, of the sequences:   
       (Ia) Glu-Ile-Asp-Pro-Ser-Asp-Ser-Tyr-Thr-Asn-Tyr-Asn-Gln-Asn-Phe-Lys-Asp (SEQ ID NO:9), (Ib) Leu-Leu-Arg-Tyr-Phe-Ala-Met-Asp-Tyr (SEQ ID NO: 10), or (Ic) Gln-His-Ile-Arg-Glu-Leu-Thr-Arg (SEQ ID NO:11);
 (ii) sequences herein designated IIa-IIb, based on the CDR2 and CDR3, respectively, of the heavy chain (246VH), and sequence IIc based on the CDR3 of the light chain (246VL), of the anti-p53 mAb 246, of the sequences: 
 
       (IIa) Asp-IIe-Asn-Pro-Asn-Asn-Gly-Tyr-Thr-Ile-Tyr-Asn-Gln-Lys-Val-Lys-Gly (SEQ ID NO:12), (IIb) Gly-Gly-Gly-Leu-Lys-Gly-Tyr-Pro-Phe-Val-Tyr (SEQ ID NO:13), or (IIc) Gln-Gln-Arg-Ser-Ser-Phe-Pro-Phe-Thr (SEQ ID NO:14); and
 (iii) sequences herein designated IVa-IVb, based on the CDR2 and CDR3, respectively, of the heavy chain (421VH), and sequence IVc based on the CDR3 of the light chain (421VL), of the anti-p53 mAb 421, of the sequences: 
 
       (IVa) Trp-Ile-Asp-Pro-Glu-Asn-Gly-Asp-Thr-Glu-Tyr-Ala-Pro-Lys-Phe-Gln-Gly (SEQ ID NO:18), (IVb)Tyr-Gly-Asp-Ala-Leu-Asp-Tyr (SEQ ID NO:19), or (IVc) Trp-Gln-Gly-Thr-His-Ser-Pro-Leu-Thr (SEQ ID NO:20). 
     
     
         13 . A method for activating an enhanced immune response to p53 in a mammal that is afflicted with a tumor characterized by mutation, overexpression or accumulation of p53 which comprises:
 determining at least one immunogen that can elicit antibodies to p53 wherein the immunogen is selected from the group consisting of:   (i) an anti-p53 mAb;   (ii) a fragment of an anti-p53 mAb; and   (iii) a DNA molecule coding for the variable (V) region of an anti-p53mAb in a suitable gene deliver vehicle; and   immunizing the mammal by administering an effective amount of the at least one immunogen.   
     
     
         14 . The method according to  claim 13 , wherein the method is used for inducing immune responses to mutated and wild-type forms of p53 in the mammal. 
     
     
         15 . The method according to  claim 13 , wherein the anti-p53 mAb is selected from mAb 240, mAb 246 and mAb 421. 
     
     
         16 . The method according to  claim 13 , wherein the anti-p53 mAb contains at least one sequence selected from the group consisting of:
 (i) sequences herein designated Ia-Ib, based on the CDR2 and CDR3, respectively, of the heavy chain (240VH), and sequence Ic based on the CDR3 of the light chain (240VL), of the anti-p53 mAb 240, of the sequences:   
       (Ia) Glu-Ile-Asp-Pro-Ser-Asp-Ser-Tyr-Thr-Asn-Tyr-Asn-Gln-Asn-Phe-Lys-Asp (SEQ ID NO:9), (Ib) Leu-Leu-Arg-Tyr-Phe-Ala-Met-Asp-Tyr (SEQ ID NO: 10), or (Ic) Gln-His-Ile-Arg-Glu-Leu-Thr-Arg (SEQ ID NO:11);
 (ii) sequences herein designated IIa-IIb, based on the CDR2 and CDR3, respectively, of the heavy chain (246VH), and sequence He based on the CDR3 of the light chain (246VL), of the anti-p53 mAb 246, of the sequences: 
 
       (IIa) Asp-Ile-Asn-Pro-Asn-Asn-Gly-Tyr-Thr-Ile-Tyr-Asn-Gln-Lys-Val-Lys-Gly (SEQ ID NO:12), (IIb) Gly-Gly-Gly-Leu-Lys-Gly-Tyr-Pro-Phe-Val-Tyr (SEQ ID NO:13), or (IIc) Gln-Gln-Arg-Ser-Ser-Phe-Pro-Phe-Thr (SEQ ID NO:14); and
 (iii) sequences herein designated IVa-IVb, based on the CDR2 and CDR3, respectively, of the heavy chain (421VH), and sequence IVc based on the CDR3 of the light chain (421VL), of the anti-p53 mAb 421, of the sequences: 
 
       (IVa) Trp-Ile-Asp-Pro-Glu-Asn-Gly-Asp-Thr-Glu-Tyr-Ala-Pro-Lys-Phe-Gln-Gly (SEQ ID NO:18), (IVb)Tyr-Gly-Asp-Ala-Leu-Asp-Tyr (SEQ ID NO: 19), or (IVc) Trp-Gln-Gly-Thr-His-Ser-Pro-Leu-Thr (SEQ ID NO:20). 
     
     
         17 . A method for the generation of sequence-specific anti-DNA antibodies which comprises immunizing a mammal with a mAb directed to a domain containing a DNA-binding site of a DNA-binding protein, and recovering the thus elicited sequence-specific anti-DNA antibodies, wherein the DNA-binding protein is p53. 
     
     
         18 . The method of  claim 16 , wherein the mAb is directed against the central DNA-binding domain of the p53 molecule. 
     
     
         19 . The method of  claim 17 , wherein the anti-p53 mAb is the mAb 246.