TY - JOUR
T1 - Anti-GD2 with an FC point mutation reduces complement fixation and decreases antibody-induced allodynia
AU - Sorkin, Linda S.
AU - Otto, Mario
AU - Baldwin, William M.
AU - Vail, Emily
AU - Gillies, Stephen D.
AU - Handgretinger, Rupert
AU - Barfield, Raymond C.
AU - Ming Yu, Hui
AU - Yu, Alice L.
N1 - Funding Information:
We would like to thank Damon McCumber for performing the blinded tail vein injections and Julie Nguyen for secretarial support. This work was supported by the Cindy Matters Fund (L.S.S.) and NIH NS048563-04 (L.S.S.), and in part by a grant from FDA, FD-R-002319 (A.L.Y.). None of the authors has any financial arrangements that represent a conflict of interest.
PY - 2010/4
Y1 - 2010/4
N2 - Monoclonal antibodies against GD2 ganglioside, such as ch14.18, the human-mouse chimeric antibody, have been shown to be effective for the treatment of neuroblastoma. However, treatment is associated with generalized, relatively opiate-resistant pain. We investigated if a point mutation in ch14.18 antibody (hu14.18K332A) to limit complement-dependent cytotoxicity (CDC) would ameliorate the pain behavior, while preserving antibody-dependent cellular cytotoxicity (ADCC). In vitro, CDC and ADCC were measured using europium-TDA assay. In vivo, allodynia was evaluated by measuring thresholds to von Frey filaments applied to the hindpaws after injection of either ch14.18 or hu14.18K332 into wild type rats or rats with deficient complement factor 6. Other rats were pretreated with complement factor C5a receptor antagonist and tested following ch14.18 injection. The mutation reduces the antibody's ability to activate complement, while maintaining its ADCC capabilities. Injection of hu14.18K322 (1 or 3 mg/kg) produced faster resolving allodynia than that engendered by ch14.18 (1 mg/kg). Injection of ch14.18 (1 mg/kg) into rats with C6 complement deficiency further reduced antibody-induced allodynia, while pre-treatment with complement factor C5a receptor antagonist completely abolished ch14.18-induced allodynia. These findings showed that mutant hu14.18 K322 elicited less allodynia than ch14.18 and that ch14.18-elicited allodynia is due to activation of the complement cascade: in part, to formation of membrane attack complex, but more importantly to release of complement factor C5a. Development of immunotherapeutic agents with decreased complement-dependent lysis while maintaining cellular cytotoxicity may offer treatment options with reduced adverse side effects, thereby allowing dose escalation of therapeutic antibodies.
AB - Monoclonal antibodies against GD2 ganglioside, such as ch14.18, the human-mouse chimeric antibody, have been shown to be effective for the treatment of neuroblastoma. However, treatment is associated with generalized, relatively opiate-resistant pain. We investigated if a point mutation in ch14.18 antibody (hu14.18K332A) to limit complement-dependent cytotoxicity (CDC) would ameliorate the pain behavior, while preserving antibody-dependent cellular cytotoxicity (ADCC). In vitro, CDC and ADCC were measured using europium-TDA assay. In vivo, allodynia was evaluated by measuring thresholds to von Frey filaments applied to the hindpaws after injection of either ch14.18 or hu14.18K332 into wild type rats or rats with deficient complement factor 6. Other rats were pretreated with complement factor C5a receptor antagonist and tested following ch14.18 injection. The mutation reduces the antibody's ability to activate complement, while maintaining its ADCC capabilities. Injection of hu14.18K322 (1 or 3 mg/kg) produced faster resolving allodynia than that engendered by ch14.18 (1 mg/kg). Injection of ch14.18 (1 mg/kg) into rats with C6 complement deficiency further reduced antibody-induced allodynia, while pre-treatment with complement factor C5a receptor antagonist completely abolished ch14.18-induced allodynia. These findings showed that mutant hu14.18 K322 elicited less allodynia than ch14.18 and that ch14.18-elicited allodynia is due to activation of the complement cascade: in part, to formation of membrane attack complex, but more importantly to release of complement factor C5a. Development of immunotherapeutic agents with decreased complement-dependent lysis while maintaining cellular cytotoxicity may offer treatment options with reduced adverse side effects, thereby allowing dose escalation of therapeutic antibodies.
KW - Anti-GD
KW - Antibody-dependent cellular cytotoxicity
KW - C5a complement factor
KW - Complement-dependent cytotoxicity
KW - Membrane attack complex
KW - Neuroblastoma
KW - Neuroblastoma therapy
KW - Pain
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U2 - 10.1016/j.pain.2010.01.024
DO - 10.1016/j.pain.2010.01.024
M3 - Article
C2 - 20171010
AN - SCOPUS:77349092231
VL - 149
SP - 135
EP - 142
JO - Pain
JF - Pain
SN - 0304-3959
IS - 1
ER -