Relative expression of γδ T‐cell receptor gene families detected by RT–PCR and capillary electrophoresis
Identifieur interne : 001D98 ( Main/Exploration ); précédent : 001D97; suivant : 001D99Relative expression of γδ T‐cell receptor gene families detected by RT–PCR and capillary electrophoresis
Auteurs : L. Tesarova [République tchèque] ; I. Koutna [République tchèque] ; M. Dvorakova [République tchèque] ; M. Klabusay [République tchèque]Source :
- International Journal of Immunogenetics [ 1744-3121 ] ; 2012-10.
English descriptors
- Teeft :
- Agilent, Allogeneic transplantation, Annealing temperature, Autoimmune disease, Autoimmune diseases, Bioanalyzer, Bioanalyzer analysis, Bioanalyzer software, Biomedical image analysis, Biophysical research communications, Blackwell publishing, Blood samples, Bone marrow transplantation, Brewer ericson, Cancer immunology, Capillary electrophoresis, Cdr3 spectratyping, Cell carcinoma, Cell harvest, Cell receptor, Cell receptor gamma, Cell response, Cell transplantation, Chip electrophoresis, Current study, Cytomegalovirus infection, Czech republic, Ethidium bromidestained agarose, European journal, Experimental immunology, Experimental medicine, Family expression, Fragment sizing, Gamma delta, Gammadelta, Gene, Gene families, Gene family, Gene family expression, Gene primers, Gene repertoire, Gene segments, Haematological malignancies, Healthy donors, Immunogenetics, Immunology, Individual gene families, Initial chemotherapy, International journal, Isopentenyl pyrophosphate, Masaryk university, Methodology, Microchip device, Mobilization regimen, Monoclonal antibodies, Multiple myeloma, Oligonucleotide microarrays, Pbmc samples, Peripheral blood, Peripheral blood mobilization, Peripheral blood samples, Primer, Product concentrations, Receptor, Receptor gene families, Relative expression, Same pcrs, Smith hayday, Southern blot analysis, Statistical analysis, Transplantation, Tumor cells, Tumour, Tumour cells, Zoledronic acid.
Abstract
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.
Url:
DOI: 10.1111/j.1744-313X.2012.01114.x
Affiliations:
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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.</div></front></TEI><affiliations><list><country><li>République tchèque</li></country><region><li>Moravie</li></region><settlement><li>Brno</li></settlement></list><tree><country name="République tchèque"><region name="Moravie"><name sortKey="Tesarova, L" sort="Tesarova, L" uniqKey="Tesarova L" first="L." last="Tesarova">L. Tesarova</name></region><name sortKey="Dvorakova, M" sort="Dvorakova, M" uniqKey="Dvorakova M" first="M." last="Dvorakova">M. Dvorakova</name><name sortKey="Klabusay, M" sort="Klabusay, M" uniqKey="Klabusay M" first="M." last="Klabusay">M. Klabusay</name><name sortKey="Koutna, I" sort="Koutna, I" uniqKey="Koutna I" first="I." last="Koutna">I. Koutna</name><name sortKey="Koutna, I" sort="Koutna, I" uniqKey="Koutna I" first="I." last="Koutna">I. Koutna</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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