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Hexamerization-enhanced CD20 Antibody Mediates Complement-Dependent Cytotoxicity in Serum Genetically Deficient in C9

Hexamerization-enhanced CD20 Antibody Mediates Complement-Dependent Cytotoxicity in Serum Genetically Deficient in C9

Ronald P Taylor, Margaret A Lindorfer, Erika M Cook, Frank J Beurskens, Janine Schuurman, Paul W H I Parren, Clive S Zent, Karl R VanDerMeid, Richard Burack, Masashi Mizuno, B Paul Morgan

Clin Immunol. 2017 Aug;181:24-28.

PMID: 28578024

Abstract:

We examined complement-dependent cytotoxicity (CDC) by hexamer formation-enhanced CD20 mAb Hx-7D8 of patient-derived chronic lymphocytic leukemia (CLL) cells that are relatively resistant to CDC. CDC was analyzed in normal human serum (NHS) and serum from an individual genetically deficient for C9. Hx-7D8 was able to kill up to 80% of CLL cells in complete absence of C9. We conclude that the narrow C5b-8 pores formed without C9 are sufficient for CDC due to efficient antibody-mediated hexamer formation. In the absence of C9, we observed transient intracellular increases of Ca2+ during CDC (as assessed with FLUO-4) that were extended in time. This suggests that small C5b-8 pores allow Ca2+ to enter the cell, while dissipation of the fluorescent signal accompanying cell disintegration is delayed. The Ca2+ signal is retained concomitantly with TOPRO-3 (viability dye) staining, thereby confirming that Ca2+ influx represents the most proximate mediator of cell death by CDC.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP80295596	Complement C9 from human serum		80295-59-6	Price

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