Benaroya Research Institute at Virginia Mason

2 ARTICLES PUBLISHED IN JoVE

Biology

Visualizing Antigen Specific CD4+ T Cells using MHC Class II Tetramers

Eddie A. James¹, Rebecca LaFond², Ivana Durinovic-Bello³, William Kwok²

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.

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. Sprouse¹, Gabriele Blahnik², Thomas Lee¹, Natalie Tully¹, Pinaki Benarjee³, Eddie A. James², Maria J. Redondo⁴, Matthew L. Bettini¹, Maria Bettini¹

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.