PMID: 8601715Mar 28, 1996Paper

A one step PCR procedure for analysis of tumor specific T lymphocyte responses

Journal of Immunological Methods
M M Abu-hadidA K Sood

Abstract

In an effort to develop optimal conditions for analysis of tumor specific T lymphocyte responses, we have studied the effect of changes in the concentration of oligonucleotide primers on the synthesis of TCR cDNAs in one step PCR procedure using Vbeta10 gene subfamily as a model. It was found that synthesis of the TCR cDNAs increases in a roughly linear fashion at primer concentrations between 0.005-0.05 muM. Evaluation of the use of low concentration (0.005 muM) of primers showed these conditions to be adequate for the analysis of TCR Vbeta subfamilies in the spleen of BALB/c mice, but not in the peritoneal exudate cells (PECs), the latter requiring ten-fold higher concentrations of the variable region primers to compensate for the overall low frequency of T lymphocytes in the PECs in comparison to the spleen. Use of these optimal conditions to detect L1210 tumor specific T lymphocyte responses showed that, in the immunized mice, L1210 specific T lymphocyte responses are detectable in the PECs, but not in the spleen cells from these mice. Thus, upon i.p. immunization of DBA/2 mice with irradiated L1210 lymphoma cells, followed by analysis of the PECs by RT/PCR, three TCR Vbeta subfamilies, including Vbeta8.2, Vbeta15 and Vbeta...Continue Reading

References

Oct 1, 1992·European Journal of Immunology·M CochetP Kourilsky
Nov 17, 1989·Cell·S Lewis, M Gellert
Aug 4, 1988·Nature·M M Davis, P J Bjorkman
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May 28, 1993·International Journal of Cancer. Journal International Du Cancer·E WeidmannT L Whiteside

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Citations

Aug 15, 2002·Journal of Environmental Quality·Mitchell J HerbelThomas D Bullen

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