Streamlined protocol for TCR sequencing, cloning, and in vivo functional analysis of islet antigen reactive human T cells (BA11P.131)
| Autor: | Maran Sprouse, Maria Redondo, Matthew Bettini, Maria Bettini |
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| Rok vydání: | 2015 |
| Předmět: | |
| Zdroj: | The Journal of Immunology. 194:184.13-184.13 |
| ISSN: | 1550-6606 0022-1767 |
| Popis: | Type 1 diabetes (T1D) is an autoimmune disease characterized by the selective destruction of insulin secreting beta cells present in the islets of Langerhans. A large body of evidence has implicated T cells as the predominant mediators of beta cell destruction leading to T1D development. The goal of our research is to develop a system for isolation and characterization of diabetogenic CD4+ T cells from the peripheral blood of early onset T1D patients. We were able to obtain a blood sample from a treatment-naïve T1D patient at the time of diagnosis. Through a novel multiplex-nest PCR protocol, we cloned corresponding alpha and beta TCR chains from individual islet antigen-specific T cells and tested them in vitro for antigenic specificity and HLA restriction. Utilizing a humanized TCR/HLA retrogenic mouse system, we are testing human islet antigen reactive TCRs for their in vivo functionality and diabetogenic potential. |
| Databáze: | OpenAIRE |
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