Anti-GD(2) with an FC point mutation reduces complement fixation and decreases antibody-induced allodynia.
Monoclonal antibodies against GD(2) 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 3mg/kg) produced faster resolving allodynia than that engendered by ch14.18 (1mg/kg). Injection of ch14.18 (1mg/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.
Sorkin
Linda S
LS
Department of Anesthesiology, University of California, San Diego School of Medicine, La Jolla, CA 92093, USA Department of Oncology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA Division of Immunology, Cleveland Clinic, Cleveland, Ohio 44195, USA Provenance Biopharmaceuticals Corp. Waltham, MA 01821, USA Department of Hematology/Oncology, Children's University Hospital, Tuebingen 72076, Germany Division of Pediatric Hematology and Oncology, Duke University, Durham, NC 27708, USA Department of Oncology, University of California San Diego, La Jolla, CA 92093, USA Genomics Research Center, Academia Sinica, Taipei, Taiwan.
Otto
Mario
M
Baldwin
William M
WM
Vail
Emily
E
Gillies
Stephen D
SD
Handgretinger
Rupert
R
Barfield
Raymond C
RC
Yu
Hui Ming
HM
Yu
Alice L
AL
D000818:Animals; D000906:Antibodies / Q000009:adverse effects / Q000235:genetics* / Q000276:immunology*; D005732:Gangliosides / Q000276:immunology*; D006930:Hyperalgesia / Q000139:chemically induced* / Q000235:genetics*; D007141:Immunoglobulin Fc Fragments / Q000009:adverse effects / Q000235:genetics; D008297:Male; D017354:Point Mutation / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley
D000906:Antibodies; D005732:Gangliosides; D007141:Immunoglobulin Fc Fragments; C019403:ganglioside, GD2