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Relative expression of γδ T‐cell receptor gene families detected by RT–PCR and capillary electrophoresis
Author(s) -
Tesarova L.,
Koutna I.,
Dvorakova M.,
Klabusay M.
Publication year - 2012
Publication title -
international journal of immunogenetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.41
H-Index - 47
eISSN - 1744-313X
pISSN - 1744-3121
DOI - 10.1111/j.1744-313x.2012.01114.x
Subject(s) - capillary electrophoresis , microbiology and biotechnology , gene , receptor , biology , real time polymerase chain reaction , cell , chemistry , genetics
Summary γδ T cells are intensively studied because their function in infection, allergy, autoimmune disease, cancer and post‐transplant period is not yet fully understood. PCR‐based techniques were established to study the γ variable (Vγ) and δ variable (Vδ) gene families. PCR product evaluation is routinely carried out by Southern blot analysis or the third complementarity‐determining region spectratyping, but a fast and simple assessment of Vγ and Vδ gene family expression is missing. The aim of our study was to test capillary electrophoresis as a potential method for evaluating the composition of the γδ T‐cell population. This report provides optimized PCR conditions for γδ T‐cell receptor amplification. Further, it describes the utilization of capillary electrophoresis in the Agilent 2100 Bioanalyzer to evaluate the relative expression of Vγ and Vδ gene families after their amplification. An application of the methodology to peripheral blood mononuclear cell samples from patients during haemato‐oncological treatment is shown. The described methodology is fast and simple to operate and is therefore suitable as a first screening of the γδ T‐cell population composition in tissues of interest.