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• W1981127637 abstract "Although techniques that permit analysis of the clonal composition of T cell populations have been used extensively to provide a better understanding of the mechanisms that influence efficacy of T cell responses in humans and mice, such methods are lacking for other animal species. In this paper we report the establishment and validation of a panel of Vβ subfamily-specific semi-nested PCR assays, and a CDR3β heteroduplex technique for analysing the clonal diversity of bovine αβ T cell responses. Development of these methods was based on available sequence data for 48 functional Vβ genes classified within 17 subfamilies. These techniques were used to determine the clonal composition of parasite-reactive CD8+ T cells obtained from two animals immunised with the protozoan parasite Theileria parva. Analyses of uncloned T cell lines as well as large panels of cloned T cells derived from each of these lines confirmed the specificity and sensitivity of the assays. Specific PCR products were obtained from 96% of the T cell clones examined, indicating that the currently identified Vβ genes represent most of the functional Vβ subfamilies in cattle. Heteroduplex analyses, coupled with sequencing of PCR products, identified over 20 clonal expansions within each of the T cell lines, distributed over a large number of Vβ subfamilies, although a limited number of clonotypes numerically dominated the response in both animals. The development and validation of these methods provides for the first time a generic set of molecular tools that can be used to perform detailed analysis of the TCR diversity and clonal composition of bovine T cell responses." @default.
• W1981127637 created "2016-06-24" @default.
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• W1981127637 creator A5072471787 @default.
• W1981127637 creator A5078926164 @default.
• W1981127637 creator A5086265967 @default.
• W1981127637 date "2008-06-01" @default.
• W1981127637 modified "2023-10-02" @default.
• W1981127637 title "Dissection of the clonal composition of bovine αβ T cell responses using T cell receptor Vβ subfamily-specific PCR and heteroduplex analysis" @default.
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• W1981127637 doi "https://doi.org/10.1016/j.jim.2008.02.015" @default.
• W1981127637 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/18436232" @default.
• W1981127637 hasPublicationYear "2008" @default.
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