ArticleJournal of translational medicine2008
Relative quantification of TCR Vbeta-chain families by real time PCR for identification of clonal T-cell populations.
Article in Journal of translational medicine, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed.
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- Signatures of T cell immunity revealed using sequence similarity with TCRDivER algorithm.Communications biology · 2023Article
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- Emergence of a STAT3 mutated NK clone in LGL leukemia.Leukemia research reports · 2015Article
- Profiling the repertoire of T-cell receptor beta-chain variable genes in peripheral blood lymphocytes from subjects who have recovered from acute hepatitis B virus infection.Cellular & molecular immunology · 2014Article
- Developing an in vitro model of T cell type of large granular lymphocyte leukemia.Leukemia research · 2013Article
- Molecular characterization of T cell receptor beta variable in the peripheral blood T cell repertoire in subjects with active tuberculosis or latent tuberculosis infection.BMC infectious diseases · 2013Article
- Wilms' tumor protein 1 (WT1) peptide vaccination in AML patients: predominant TCR CDR3β sequence associated with remission in one patient is detectable in other vaccinated patients.Cancer immunology, immunotherapy : CII · 2012Article
- Quantitative analysis of T cell receptor diversity in clinical samples of human peripheral blood.Journal of immunological methods · 2012Article
- Molecular features of the complementarity determining region 3 motif of the T cell population and subsets in the blood of patients with chronic severe hepatitis B.Journal of translational medicine · 2011Article
- Avidity of CD1d-ligand-receptor ternary complex contributes to T-helper 1 (Th1) polarization and anticancer efficacy.Proceedings of the National Academy of Sciences of the United States of America · 2011Article
- High-scatter T cells: a reliable biomarker for malignant T cells in cutaneous T-cell lymphoma.Blood · 2011Article
- Comparison of T-cell receptor repertoire restriction in blood and tumor tissue of colorectal cancer patients.Journal of translational medicine · 2010Article
- A quantitative real time PCR method to analyze T cell receptor Vbeta subgroup expansion by staphylococcal superantigens.Journal of translational medicine · 2010Article
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Authors and funding
8 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundQuantification of T-cell receptor (TCR) chain families can be utilized for detection of clonal T-cell populations. Besides southern blotting and antibody-based approaches, quantitative real time PCR (qRT PCR) has been more widely applied in this context during the last years. Here, the heterogeneity of sequences within single families is the most challenging problem for exact quantification.
methodVbeta-families were quantified using a universal reverse primer and family-specific forward primers with TaqMan technology on a light cycler instrument. Relative concentrations were calculated considering slopes and crossing points of each PCR reaction. Total expression of alpha/beta TCR was assessed by quantification of the constant alpha-chain as a further control.
resultsThe method was tested by serial dilutions of clonal T-cells in mononuclear cells from healthy volunteers. Calculated percentages were in good correspondence with qRT PCR results demonstrating high reliability. Duplicates showed excellent technical reproducibility. We analyzed blood samples of 20 healthy volunteers for determination of mean and standard deviation for each family. The method was applied both to tissue and blood samples from patients with carcinomas and hematological disorders.
conclusionWe introduce a versatile method for the relative quantification of Vbeta-families by real time PCR. The experimental strategy described allows the identification of alterations in the Vbeta-family repertoire.
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