Role of monocytes in mediating HIV-specific antibody-dependent cellular cytotoxicity

M Kramski; A Schorcht; APR Johnston; GF Lichtfuss; S Jegaskanda; R De Rose; I Stratov; AD Kelleher; MA French; RJ Center; A Jaworowski; SJ Kent

Journal article

Role of monocytes in mediating HIV-specific antibody-dependent cellular cytotoxicity

M Kramski, A Schorcht, APR Johnston, GF Lichtfuss, S Jegaskanda, R De Rose, I Stratov, AD Kelleher, MA French, RJ Center, A Jaworowski, SJ Kent

Journal of Immunological Methods | Published : 2012

DOI: 10.1016/j.jim.2012.07.006

Abstract

Antibodies (Abs) that mediate antibody-dependent cellular cytotoxicity (ADCC) activity against HIV-1 are of major interest. A widely used method to measure ADCC Abs is the rapid and fluorometric antibody-dependent cellular cytotoxicity (RFADCC) assay. Antibody-dependent killing of a labelled target cell line by PBMC is assessed by loss of intracellular CFSE but retention of membrane dye PKH26 (CFSE-PKH26. +). Cells of this phenotype are assumed to be derived from CFSE. +. PKH26. + target cells killed by NK cells.We assessed the effector cells that mediate ADCC in this assay. Backgating analysis and phenotyping of CFSE-PKH26. + revealed that the RFADCC assay's readout mainly represents CD3-CD..

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University of Melbourne Researchers

Rob De Rose Author

Stephen Kent Author

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Grants

Awarded by National Institutes of Health

Funding Acknowledgements

This work was supported by Australian NHMRC award 510448 and NIH award R21AI081541. We are grateful to all the subjects who kindly provided blood samples and their carers. We also thank Damian Purcell for helpful discussions.