Abstract
Background Antibodies (Abs) that mediate antibody-dependent cellular cytotoxicity (ADCC) activity against HIV-1 are of major interest. Considerable evidence supports a role for ADCC activity in the control of HIV-1 infection and in the context of vaccination. One method widely used to assess the role of ADCC is the rapid and fluorometric antibody-dependent cellular cytotoxicity (RFADCC) assay. In the RFADCC assay specific killing of target cells by PBMC is assessed by loss of intracellular CFSE but retention of membrane dye PKH26 (CFSE-PKH26 +), which is assumed to be derived from CFSE+PKH26+ target cells killed by NK cells. We have revisited this assay to assess the role of effector cells in mediating ADCC.
Highlights
Antibodies (Abs) that mediate antibody-dependent cellular cytotoxicity (ADCC) activity against HIV-1 are of major interest
One method widely used to assess the role of ADCC is the rapid and fluorometric antibody-dependent cellular cytotoxicity (RFADCC) assay
In the RFADCC assay specific killing of target cells by PBMC is assessed by loss of intracellular CFSE but retention of membrane dye PKH26 (CFSE-PKH26 +), which is assumed to be derived from CFSE+PKH26+ target cells killed by NK cells
Summary
Antibodies (Abs) that mediate antibody-dependent cellular cytotoxicity (ADCC) activity against HIV-1 are of major interest. One method widely used to assess the role of ADCC is the rapid and fluorometric antibody-dependent cellular cytotoxicity (RFADCC) assay. In the RFADCC assay specific killing of target cells by PBMC is assessed by loss of intracellular CFSE but retention of membrane dye PKH26 (CFSE-PKH26 +), which is assumed to be derived from CFSE+PKH26+ target cells killed by NK cells. We have revisited this assay to assess the role of effector cells in mediating ADCC
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