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

被引：19

作者：

Alrubayyi, Aljawharah ^{[1
,2
]}

Schuetz, Alexandra ^{[1
,2
,3
]}

Lal, Kerri G. ^{[1
,2
]}

Jongrakthaitae, Surat ^{[3
]}

Paolino, Kristopher M. ^{[4
]}

Ake, Julie A. ^{[1
]}

Robb, Merlin L. ^{[1
,2
]}

de Souza, Mark S. ^{[5
]}

Michael, Nelson L. ^{[1
]}

Paquin-Proulx, Dominic ^{[1
,2
]}

Eller, Michael A. ^{[1
,2
]}

机构：

[1] US Mil HIV Res Program, Walter Reed Army Inst Res, Silver Spring, MD 20910 USA

[2] Henry M Jackson Fdn Adv Mil Med Inc, Bethesda, MD USA

[3] Armed Forces Res Inst Med Sci, Dept Retrovirol, Bangkok, Thailand

[4] Walter Reed Army Inst Res, Clin Trials Ctr, Translat Med Branch, Silver Spring, MD USA

[5] Thai Red Cross AIDS Res Ctr, SEARCH, Bangkok, Thailand

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 2018年 / 462卷

关键词：

HIV antibodies; ADCC; ADCP; Cytotoxicity; Monocytes; NK cells; HIV-SPECIFIC ANTIBODIES; CELLULAR CYTOTOXICITY; FLUOROMETRIC ASSESSMENT; PROTECTIVE EFFICACY; RHESUS-MONKEYS; VACCINE; ADCC; NEUTRALIZATION; INVOLVEMENT; CHALLENGES;

D O I：

10.1016/j.jim.2018.08.012

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：74 / 82

页数：9

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