Detection of CDR3 skewing of TCR beta gene repertoire in peripheral blood of HIV infected people

Objective To detect the CDR3 skewing of TCR beta gene repertoire in the peripheral blood of HIV infected people by using real-time fluorescence quantitative reverse transcription polymerase chain reaction (FQ-PCR) with DNA melting curve analysis. Methods The total RNA of peripheral blood mononuclear cell (PBMC) from 4 healthy donors and 6 HIV infected people were transcribed reversely into cDNA. The cDNA of 26 TRBV gene families CDR3 was amplified by FQ-PCR, The monoclonal/oligoclonal/polyclonal CDR3 spectratyping were analyzed with DNA melting curve. Results The 26 TRBV families CDR3 showed different frequency in healthy donors and patients. The CDR3 spectratyping of 26 TRBV families showed polyclonal peak (Gaussian distribution) in healthy donors but showed different monoclonal/oligoclonal/polyclonal peak in HIV infected people with DNA melting curve analysis. Conclusion The technique of FQ-PCR with DNA melting curve analysis can be used for detecting the CDR3 skewing of TCR beta gene repertoire in HIV infected people.