Anti-GD2 with an FC point mutation reduces complement fixation and decreases antibody-induced allodynia

Linda S. Sorkin, Mario Otto, William M. Baldwin, Emily Vail, Stephen D. Gillies, Rupert Handgretinger, Raymond C. Barfield, Hui Ming Yu, Alice L. Yu

Research output: Contribution to journalArticle

77 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)135-142
Number of pages8
JournalPain
Volume149
Issue number1
DOIs
StatePublished - Apr 2010
Externally publishedYes

Keywords

  • Anti-GD
  • Antibody-dependent cellular cytotoxicity
  • C5a complement factor
  • Complement-dependent cytotoxicity
  • Membrane attack complex
  • Neuroblastoma
  • Neuroblastoma therapy
  • Pain

ASJC Scopus subject areas

  • Neurology
  • Clinical Neurology
  • Anesthesiology and Pain Medicine

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