A novel EGFP-CEM-NKr flow cytometric method for measuring antibody dependent cell mediated-cytotoxicity (ADCC) activity in HIV-1 infected individuals

Kantakamalakul, Wannee; Pattanapanyasat, Kovit; Jongrakthaitae, Surat; Assawadarachai, Vatcharain; Ampol, Silawun; Sutthent, Ruengpung

doi:10.1016/j.jim.2006.06.005

Cited by 18 publications

(13 citation statements)

References 24 publications

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“…Our assay measures the number of target cells that are killed. 10 Identification of potential ADCC antibody epitopes using a matrix format led to the finding that the spectrum of peptide specific reactivity varied for each serum tested. All tested sera showed reactivity against a total of 1-37, and 1-18 peptides for Env and Gag proteins, respectively (Tables 2, 3).…”

Section: Discussionmentioning

confidence: 99%

“…The actual percent lysis of target cells at each serum dilution was calculated as previously reported. 10 The cut-off for positive results using gp120-and peptide pool-pulsed target cells are 7.21% lysis and 5.46% lysis, respectively. Identification of HIV-1 CRF01_AE gp160-and Gag-epitopes that are the target of ADCC reactive antibodies Twenty sera with relatively high ADCC antibody activity were further examined against each of the seven peptide pools:E1-E7 of the HIV-1 CRF01_AE Env peptide set and each of the five peptide pools:G1-G5 of HIV-1 subtype A Gag formulated as described in the methods section.…”

Section: Egfp-adcc Flow Cytometric Assaymentioning

confidence: 99%

“…10 Cells were later thawed and overlaid overnight at 1 × 10 6 cells/ml in growth medium at 37 °C with 5% CO2. The cells were then re-suspended in growth media at a concentration of 2.5 × 10 6 cells/ml and used as effector cells.…”

Section: Effector Cellsmentioning

confidence: 99%

“…10 Lysis of target cells by ADCC activity was measured and analyzed with a FACScan flow cytometer (Becton Dickinson Biosciences {BDB}, San Jose, CA, USA). The actual percent lysis of target cells at each serum dilution was calculated as previously reported.…”

Section: Egfp-adcc Flow Cytometric Assaymentioning

confidence: 99%

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Common antibody dependent cell mediated cytotoxicity (ADCC) antibody epitopes of HIV-1 CRF01_AE Env and Gag in early HIV-1 infected individuals

Sangjan

Ampol

Tabprasit

et al. 2018

Asian Pac J Allergy Immunol

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Section: Discussionmentioning

confidence: 99%

Section: Egfp-adcc Flow Cytometric Assaymentioning

confidence: 99%

Section: Effector Cellsmentioning

confidence: 99%

Section: Egfp-adcc Flow Cytometric Assaymentioning

confidence: 99%

See 2 more Smart Citations

Common antibody dependent cell mediated cytotoxicity (ADCC) antibody epitopes of HIV-1 CRF01_AE Env and Gag in early HIV-1 infected individuals

Sangjan

Ampol

Tabprasit

et al. 2018

Asian Pac J Allergy Immunol

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“…The RFADCC assay has also been used to evaluate RV144 vaccine induced ADCC antibodies showing immune selection of viruses by these Abs [160]. In a modification of RFADCC approach EGFP was expressed in the CEM-NK R cell line with dual staining by propidium iodide marking killed target cells [161].…”

Section: Cell Based Functional Assays For Fcr Activating Antibodiesmentioning

confidence: 99%

Antibody functional assays as measures of Fc receptor-mediated immunity to HIV - new technologies and their impact on the HIV vaccine field.

Wines¹,

Billings²,

Mcleand³

et al. 2017

CHR

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Background:There is now intense interest in the role of HIV-specific antibodies and the engagement of FcγR functions in the control and prevention of HIV infection. The analyses of the RV144 vaccine trial, natural progression cohorts, and macaque models all point to a role for Fc-dependent effector functions, such as cytotoxicity (ADCC) or phagocytosis (ADCP), in the control of HIV. However, reliable assays that can be reproducibly used across different laboratories to measure Fcdependent functions, such as antibody dependent cellular cytotoxicity (ADCC) are limited. Method: This brief review highlights the importance of Fc properties for immunity to HIV, particularly via FcγR diversity and function. We discuss assays used to study FcR mediated functions of HIV-specific Ab, including our recently developed novel cell-free ELISA using homo-dimeric FcγR ectodomains to detect functionally relevant viral antigen-specific antibodies. Results: The binding of these dimeric FcγR ectodomains, to closely spaced pairs of IgG Fc, mimics the engagement and cross-linking of Fc receptors by IgG opsonized virions or infected cells as the essential prerequisite to the induction of Ab-dependent effector functions. The dimeric FcγR ELISA reliably correlates with ADCC in patient responses to influenza. The assay is amenable to high throughput and could be standardized across laboratories. Conclusion: We propose the assay has broader implications for the evaluation of the quality of antibody responses in viral infections and for the rapid evaluation of responses in vaccine development campaigns for HIV and other viral infections.

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A novel assay for detecting virus-specific antibodies triggering activation of Fcγ receptors

Corrales-Aguilar

Trilling

Reinhard

et al. 2013

Journal of Immunological Methods

119

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A novel EGFP-CEM-NKr flow cytometric method for measuring antibody dependent cell mediated-cytotoxicity (ADCC) activity in HIV-1 infected individuals

Cited by 18 publications

References 24 publications

Common antibody dependent cell mediated cytotoxicity (ADCC) antibody epitopes of HIV-1 CRF01_AE Env and Gag in early HIV-1 infected individuals

Common antibody dependent cell mediated cytotoxicity (ADCC) antibody epitopes of HIV-1 CRF01_AE Env and Gag in early HIV-1 infected individuals

Antibody functional assays as measures of Fc receptor-mediated immunity to HIV - new technologies and their impact on the HIV vaccine field.

A novel assay for detecting virus-specific antibodies triggering activation of Fcγ receptors

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