A flow cytometry based assay that simultaneously measures cytotoxicity and monocyte mediated antibody dependent effector activity

Journal of Immunological Methods
Aljawharah AlrubayyiMichael A Eller

Abstract

Antibody effector functions such as antibody dependent cellular cytotoxicity (ADCC) and antibody dependent cellular phagocytosis (ADCP) are considered important immunologic parameters following results from the RV144 clinical trial where a reduced risk of infection was associated with non-neutralizing antibody against the V1/V2 region of HIV envelope. The rapid and fluorometric ADCC (RFADCC) assay has been widely used to measure ADCC, however, the mechanism behind the activity measured remains unclear. Here, we demonstrate that monocytes acquire the PKH26 dye used in the RFADCC assay and that the commonly used RFADCC readout correlates with phagocytosis. The RFADCC assay was combined with an amine reactive dye staining to confirm target cell killing. Interestingly, the majority of RFADCC and amine indices were mutually exclusive. In fact, the amine reactive assay results correlated with results from another assays that directly measure NK cell antibody effector functions not associated with phagocytosis. Together, this combined assay offers the opportunity to discriminate monocytes and NK cell antibody effector functions simultaneously.

References

Dec 1, 1989·AIDS Research and Human Retroviruses·D S TylerK J Weinhold
Feb 28, 2001·Journal of Immunological Methods·M E SheehyD F Nixon
Feb 9, 2005·The Journal of Immunology : Official Journal of the American Association of Immunologists·V Raúl Gómez-RománMarjorie Robert-Guroff
Dec 14, 2005·Journal of Immunological Methods·V Raúl Gómez-RománMarjorie Robert-Guroff
Dec 8, 2007·Nature Reviews. Immunology·Falk Nimmerjahn, Jeffrey V Ravetch
Jan 7, 2009·The Journal of Immunology : Official Journal of the American Association of Immunologists·Amy W ChungIvan Stratov
Jan 5, 2011·Journal of Immunological Methods·Margaret E AckermanGalit Alter
Apr 20, 2011·Proceedings of the National Academy of Sciences of the United States of America·Amy W ChungIvan Stratov
Jul 8, 2011·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Justin PollaraGuido Ferrari
Apr 6, 2012·The New England Journal of Medicine·Barton F HaynesJerome H Kim
Jul 31, 2012·Journal of Immunological Methods·M KramskiS J Kent
Dec 4, 2016·The Journal of Immunology : Official Journal of the American Association of Immunologists·Yunda HuangMargaret E Ackerman
Jan 18, 2017·JCI Insight·Rui André Saraiva RaposoDouglas F Nixon
Jun 16, 2017·The Journal of Immunology : Official Journal of the American Association of Immunologists·Milla R McLeanStephen J Kent
Mar 3, 2018·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Justin PollaraGuido Ferrari
Apr 2, 2018·Journal of Immunological Methods·Matthew J WorleyAmy W Chung

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Citations

Jul 18, 2020·Current Opinion in HIV and AIDS·Margaret C Carpenter, Margaret E Ackerman
Jan 10, 2019·Cell & Bioscience·Yongjun Guan
Dec 9, 2020·PLoS Pathogens·Thembi MdluliMorgane Rolland
Sep 3, 2021·Proceedings of the National Academy of Sciences of the United States of America·Hannah A D KingDiane L Bolton

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