Relative expression of gammadelta T-cell receptor gene families detected by RT–PCR and capillary electrophoresis
| Authors | |
|---|---|
| Year of publication | 2012 |
| Type | Article in Periodical |
| Magazine / Source | International Journal of immunogenetics |
| MU Faculty or unit | |
| Citation | |
| Web | http://onlinelibrary.wiley.com/doi/10.1111/j.1744-313X.2012.01114.x/full |
| Doi | http://dx.doi.org/10.1111/j.1744-313X.2012.01114.x |
| Field | Genetics and molecular biology |
| Keywords | Reverse transcription PCR; Capillary electrophoresis; gammadelta T cell; TCR; Vgamma and Vdelta gene families |
| Description | Gammadelta 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 gamma variable (Vgamma) and delta variable (Vdelta) gene families. PCR product evaluation is routinely carried out by Southern blot analysis or the third complementarity-determining region (CDR3) spectratyping, but a fast and simple assessment of Vgamma and Vdelta 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 gammadelta T-cell population. This report provides optimized PCR conditions for gammadelta T-cell receptor amplification. Further it describes the utilization of capillary electrophoresis in the Agilent 2100 Bioanalyzer to evaluate the relative expression of Vgamma and Vdelta gene families after their amplification. An application of the methodology to peripheral blood mononuclear cell samples from patients during hemato-oncological treatment is shown. The described methodology is fast and simple to operate and is therefore suitable as a first screening of the gammadelta T-cell population composition in tissues of interest. |
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