Detection, sorting, and immortalization of dual expresser lymphocytes from human peripheral blood samples

Rizwan Ahmed; Kusuma Ananth; Zahra Omidian; Neha Majety; Hao Zhang; Abdel Rahim A. Hamad

doi:10.1016/j.xpro.2021.100925

Detection, sorting, and immortalization of dual expresser lymphocytes from human peripheral blood samples

Rizwan Ahmed, Kusuma Ananth, Zahra Omidian, Neha Majety, Hao Zhang, Abdel Rahim A. Hamad

School of Medicine

Research output: Contribution to journal › Article › peer-review

Abstract

This protocol describes how to identify Dual Expressers (DEs), a rare type of lymphocyte that co-expresses B-cell receptors and T-cell receptors, by flow cytometry using a cocktail of four antibodies. It also shows the subsequent gating strategy for detecting and sorting DEs and the generation of EBV-immortalized DE lymphoblastoid cell lines and clones for antibody production and cloning antigen receptors. Use of this protocol maximizes detection of DEs and minimizes inclusion of doublets. For complete details on the use and execution of this protocol, please refer to Ahmed et al. (2019).

Original language	English (US)
Article number	100925
Journal	STAR Protocols
Volume	2
Issue number	4
DOIs	https://doi.org/10.1016/j.xpro.2021.100925
State	Published - Dec 17 2021

Keywords

Antibody
Cell Biology
Cell culture
Cell isolation
Flow Cytometry/Mass Cytometry
Immunology

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology
General Neuroscience
General Immunology and Microbiology
General Medicine

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10.1016/j.xpro.2021.100925

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title = "Detection, sorting, and immortalization of dual expresser lymphocytes from human peripheral blood samples",

abstract = "This protocol describes how to identify Dual Expressers (DEs), a rare type of lymphocyte that co-expresses B-cell receptors and T-cell receptors, by flow cytometry using a cocktail of four antibodies. It also shows the subsequent gating strategy for detecting and sorting DEs and the generation of EBV-immortalized DE lymphoblastoid cell lines and clones for antibody production and cloning antigen receptors. Use of this protocol maximizes detection of DEs and minimizes inclusion of doublets. For complete details on the use and execution of this protocol, please refer to Ahmed et al. (2019).",

keywords = "Antibody, Cell Biology, Cell culture, Cell isolation, Flow Cytometry/Mass Cytometry, Immunology",

author = "Rizwan Ahmed and Kusuma Ananth and Zahra Omidian and Neha Majety and Hao Zhang and Hamad, {Abdel Rahim A.}",

note = "Funding Information: We thank participants enrolled in this study and their families for blood donation. This work was supported by NIH , United States grant R0 AI099027 (A.R.A.H.); the Norman Raab Foundation ; W.M. Keck foundation . R.A. is partly supported by Diabetes Research Connection grant. Publisher Copyright: {\textcopyright} 2021",

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doi = "10.1016/j.xpro.2021.100925",

language = "English (US)",

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AU - Ahmed, Rizwan

AU - Ananth, Kusuma

AU - Omidian, Zahra

AU - Majety, Neha

AU - Zhang, Hao

AU - Hamad, Abdel Rahim A.

N1 - Funding Information: We thank participants enrolled in this study and their families for blood donation. This work was supported by NIH , United States grant R0 AI099027 (A.R.A.H.); the Norman Raab Foundation ; W.M. Keck foundation . R.A. is partly supported by Diabetes Research Connection grant. Publisher Copyright: © 2021

PY - 2021/12/17

Y1 - 2021/12/17

N2 - This protocol describes how to identify Dual Expressers (DEs), a rare type of lymphocyte that co-expresses B-cell receptors and T-cell receptors, by flow cytometry using a cocktail of four antibodies. It also shows the subsequent gating strategy for detecting and sorting DEs and the generation of EBV-immortalized DE lymphoblastoid cell lines and clones for antibody production and cloning antigen receptors. Use of this protocol maximizes detection of DEs and minimizes inclusion of doublets. For complete details on the use and execution of this protocol, please refer to Ahmed et al. (2019).

AB - This protocol describes how to identify Dual Expressers (DEs), a rare type of lymphocyte that co-expresses B-cell receptors and T-cell receptors, by flow cytometry using a cocktail of four antibodies. It also shows the subsequent gating strategy for detecting and sorting DEs and the generation of EBV-immortalized DE lymphoblastoid cell lines and clones for antibody production and cloning antigen receptors. Use of this protocol maximizes detection of DEs and minimizes inclusion of doublets. For complete details on the use and execution of this protocol, please refer to Ahmed et al. (2019).

KW - Antibody

KW - Cell Biology

KW - Cell culture

KW - Cell isolation

KW - Flow Cytometry/Mass Cytometry

KW - Immunology

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ER -

Detection, sorting, and immortalization of dual expresser lymphocytes from human peripheral blood samples

Abstract

Keywords

ASJC Scopus subject areas

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