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 needed 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 and was hybridized with 28 different TCR beta variable (TCRBV) genes fixed on nylon membranes, and the percentage of each TCRBV gene was measured spectrophotometrically.


RESULTS
The specificity and linearity of the hybridization technique and the constancy of the TCRBV percentages over a wide range of sample amounts were demonstrated by control experiments. Analysis of RAITs of biopsy specimens from four patients showed broad or skewed TCRBV usage, indicating the presence of polyclonal and oligoclonal RAIT populations, respectively. In one patient who received OKT3 immunosuppressive treatment, the TCRBV skewness was dramatically reduced after the treatment.


CONCLUSIONS
We have established a powerful method for analyzing RAIT clonality, which is especially useful for monitoring RAIT dynamics after immunosuppression therapy.
Obata
F
F
Department of Immunology, Kitasato University School of Allied Health Sciences, Sagamihara, Japan.
Kumano
K
K
Endo
T
T
Takeuchi
Y
Y
Ohkubo
M
M
Tsunoda-Iizuka
M
M
D000208:Acute Disease; D001706:Biopsy / Q000379:methods*; D002465:Cell Movement; D002908:Chronic Disease; D002999:Clone Cells / Q000737:chemistry; D005784:Gene Amplification; D006084:Graft Rejection / Q000235:genetics; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D016030:Kidney Transplantation / Q000276:immunology / Q000473:pathology*; D008832:Microchemistry; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics / Q000276:immunology*; D013601:T-Lymphocytes / Q000473:pathology*
D007135:Immunoglobulin Variable Region; D016693:Receptors, Antigen, T-Cell, alpha-beta