T-cell receptor variable gene analysis of renal allograft-infiltrating cells in biopsy specimens using a nonradioisotopic micromethod

Background. A sensitive micromethod for T-cell receptor (TCR) analysis is n eeded for clonality analysis of renal allograft-infiltrating T cells (RAITs ) obtained by needle biopsy. Methods. TCR cDNA was amplified by the anchored polymerase chain reaction a nd was hybridized with 28 different TCR beta variable (TCRBV) genes fixed o n nylon membranes, and the percentage of each TCRBV gene was measured spect rophotometrically. Results. The specificity and linearity of the hybridization technique and t he constancy of the TCRBV percentages over a wide range of sample amounts w ere demonstrated by control experiments. Analysis of RAITs of biopsy specim ens from four patients showed broad or skewed TCRBV usage, indicating the p resence of polyclonal and oligoclonal RAIT populations, respectively. In on e patient who received OKT3 immunosuppressive treatment, the TCRBV skewness was dramatically reduced after the treatment. Conclusion. We have established a powerful method for analyzing BAIT clonal ity, which is especially useful for monitoring BAIT dynamics after immunosu ppression therapy.