Development of IFN-γ secretory ELISPOT based assay for screening of ADCC responses

Shete, Ashwini; Suryawanshi, Poonam; Chavan, Chetan; Kulkarni, A. G.; Godbole, Sheela; Ghate, Manisha; Thakar, Madhuri

doi:10.1016/j.jim.2016.12.001

Cited by 4 publications

(3 citation statements)

References 41 publications

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“…Furthermore, in vivo protection in monkey correlated with P1-specific IgG capable of mediating ADCC ( 19 ). We thus used HIV-1 envelope subunit P1 derived from HIV-1 Clade A (P1-A), Clade B (P1-B), and Clade C (P1-C) to coat CEM-NK r cells as an alternative target cell model, a procedure commonly used in ADCC studies ( 9 , 19 , 33 – 35 ).…”

Section: Resultsmentioning

confidence: 99%

“…Here, we demonstrate for the first time that 2F5-IgA can trigger ADCC of target cells present at different steps of HIV infection ( 39 ). Hence, HIV-1 envelope subunit-coated target cells, an experimental model frequently used in ADCC studies by us ( 9 , 19 , 30 – 32 ) and others ( 33 – 35 ), could recapitulate the initial step in infection, when viruses bind to target cells exposing specific epitopes poorly accessible at the free virus surface ( 40 ). Alternatively, HIV-1-infected cells represent targets once infection is established, although in this case, CD4 downregulation and Tetherin/BST-2 antagonism could reduce HIV envelope ADCC-mediating epitope exposure ( 41 ).…”

Section: Discussionmentioning

confidence: 99%

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IgA Targeting Human Immunodeficiency Virus-1 Envelope gp41 Triggers Antibody-Dependent Cellular Cytotoxicity Cross-Clade and Cooperates with gp41-Specific IgG to Increase Cell Lysis

et al. 2018

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The protective efficacy of human immunodeficiency virus-1 (HIV-1) antibodies (Abs) remains mostly correlated with their in vitro neutralizing activity engaging their Fab region. However, anti-HIV-1 Abs also mediate a broad array of Fc-mediated effector functions including Ab-dependent cellular cytotoxicity (ADCC), which depend primarily on the Ab isotype. While ADCC is commonly associated with HIV-1 gp120 envelope-specific IgGs, whether IgAs, especially those targeting the HIV-1 gp41 envelope, also mediate ADCC remains elusive. Therefore, to assess the capacity of IgA specific for HIV-1 to induce Fcα-mediated ADCC, we used the gp41 envelope-specific IgA transformed from the broadly neutralizing 2F5-IgG we have previously reported to induce ADCC. We demonstrate that 2F5-IgA engages FcαRI (CD89), expressed on human monocytes used as effector cells, to induce the lysis of HIV-1 Clade A- and B-infected target cells by ADCC. Furthermore, the 2F5-IgA and 2F5-IgG cooperate to enhance target cells lysis by ADCC. Cooperation in ADCC is also observed between 2F5-IgA and the broadly neutralizing 10E8-IgG. These results provide a new perspective for IgA in protection against HIV-1 acquisition or reservoir eradication and suggest that inducing IgA by vaccination, in particular when targeting gp41, in combination with IgG could strengthen protection by complementary and cooperative activities with IgG.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

IgA Targeting Human Immunodeficiency Virus-1 Envelope gp41 Triggers Antibody-Dependent Cellular Cytotoxicity Cross-Clade and Cooperates with gp41-Specific IgG to Increase Cell Lysis

et al. 2018

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“…Detection of single-cell secreted proteins is primarily divided into Elispot direct detection and flow cytometry indirect detection. [8,[35][36][37][38][39] Indirect detection is accomplished through coupling flow cytometry with droplet microfluidics, [40][41][42] of which the narrow nanoliter space improves detection sensitivity but reduces detection throughput due to spectral overlap. [39,[43][44][45][46][47] Direct detection is achieved using a microfluidic chip, [20,43,46] for instance, a single-cell barcode chip that overcomes spectral overlap and increases detection throughput by using spatially distributed antibody barcodes.…”

Section: Introductionmentioning

confidence: 99%

High‐Throughput, Living Single‐Cell, Multiple Secreted Biomarker Profiling Using Microfluidic Chip and Machine Learning for Tumor Cell Classification

Wang

et al. 2022

Adv Healthcare Materials

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Secreted proteins provide abundant functional information on living cells and can beused as important tumor diagnostic markers, of which profiling at the single-cell level is helpful for accurate tumor cell classification. Currently, achieving living single-cell multi-index, high-sensitivity, and quantitative secretion biomarker profiling remains a great challenge. Here, a high-throughput living single-cell multi-index secreted biomarker profiling platform is proposed, combined with machine learning, to achieve accurate tumor cell classification. A single-cell culture microfluidic chip with self-assembled graphene oxide quantum dots (GOQDs) enables high-activity single-cell culture, ensuring normal secretion of biomarkers and high-throughput single-cell separation, providing sufficient statistical data for machine learning. At the same time, the antibody barcode chip with self-assembled GOQDs performs multi-index, highly sensitive, and quantitative detection of secreted biomarkers, in which each cell culture chamber covers a whole barcode array. Importantly, by combining the K-means strategy with machine learning, thousands of single tumor cell secretion data are analyzed, enabling tumor cell classification with a recognition accuracy of 95.0%. In addition, further profiling of the grouping results reveals the unique secretion characteristics of subgroups. This work provides an intelligent platform for high-throughput living single-cell multiple secretion biomarker profiling, which has broad implications for cancer investigation and biomedical research.

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An improved method to quantify human NK cell-mediated antibody-dependent cell-mediated cytotoxicity (ADCC) per IgG FcR-positive NK cell without purification of NK cells

Sung

Tang

Guglielmo

et al. 2018

Journal of Immunological Methods

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Development of IFN-γ secretory ELISPOT based assay for screening of ADCC responses

Cited by 4 publications

References 41 publications

IgA Targeting Human Immunodeficiency Virus-1 Envelope gp41 Triggers Antibody-Dependent Cellular Cytotoxicity Cross-Clade and Cooperates with gp41-Specific IgG to Increase Cell Lysis

IgA Targeting Human Immunodeficiency Virus-1 Envelope gp41 Triggers Antibody-Dependent Cellular Cytotoxicity Cross-Clade and Cooperates with gp41-Specific IgG to Increase Cell Lysis

High‐Throughput, Living Single‐Cell, Multiple Secreted Biomarker Profiling Using Microfluidic Chip and Machine Learning for Tumor Cell Classification

An improved method to quantify human NK cell-mediated antibody-dependent cell-mediated cytotoxicity (ADCC) per IgG FcR-positive NK cell without purification of NK cells

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