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Benaroya Research Institute at Virginia Mason

2 ARTICLES PUBLISHED IN JoVE

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Biology

Visualizing Antigen Specific CD4+ T Cells using MHC Class II Tetramers
Eddie A. James 1, Rebecca LaFond 2, Ivana Durinovic-Bello 3, William Kwok 2
1Tetramer Core Laboratory, Benaroya Research Institute, 2Kwok Laboratory, Benaroya Research Institute, 3Nepom Laboratory, Benaroya Research Institute

This procedure demonstrates the purification and in vitro expansion of antigen specific CD4+ T cells from whole peripheral blood and their visualization using MHC class II tetramers. Tetramers permit the direct visualization of T cells with a single antigen specificity and defined MHC class II restriction.

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Immunology and Infection

Streamlined Single Cell TCR Isolation and Generation of Retroviral Vectors for In Vitro and In Vivo Expression of Human TCRs
Maran L. Sprouse 1, Gabriele Blahnik 2, Thomas Lee 1, Natalie Tully 1, Pinaki Benarjee 3, Eddie A. James 2, Maria J. Redondo 4, Matthew L. Bettini 1, Maria Bettini 1
1Department of Pediatrics, Section of Diabetes and Endocrinology, McNair Medical Institute, Baylor College of Medicine, Texas Children's Hospital, 2Benaroya Research Institute at Virginia Mason, 3Center for Human Immunobiology, Baylor College of Medicine, Texas Children's Hospital, 4Department of Pediatrics, Section of Diabetes and Endocrinology, Baylor College of Medicine, Texas Children's Hospital

The current protocol combines single cell paired human TCR alpha and beta chain sequencing with streamlined generation of retroviral vectors compatible with in vitro and in vivo TCR expression.

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