Frequency of clonally expanded T cells evaluated by PCR from a single cell.
In analyses of antigen-specific immune responses, it is essential to estimate the frequency of individual T cell clonotypes. This frequency has been estimated, however, only indirectly by the frequency of T cell receptor (TCR) mRNA. We have developed a method to determine T cell frequency directly by cell count using reverse transcription polymerase chain reaction (RT-PCR) amplification of TCR beta genes from single cell-derived cDNA (single cell PCR). In a study of clinical samples, the frequency of clonally expanded T cells estimated by TCR frequency analysis was found to be higher than that by single cell PCR. Single cell PCR can estimate T cell frequency accurately, as it is not affected by skewed PCR amplification or different TCR mRNA expressions in individual T cells.
Kurokawa
M
M
Rheumatology, Immunology and Genetic Program, Institute of Medical Science, St. Marianna University School of Medicine, Kawasaki, Japan. manae@mail-1.marianna-u.ac.jp
Furukawa
H
H
Yabe
T
T
Matsui
T
T
Toda
M
M
Hamada
C
C
Kasukawa
R
R
Yamamoto
K
K
Nishioka
K
K
Kato
T
T
D001172:Arthritis, Rheumatoid / Q000235:genetics / Q000276:immunology*; D006801:Humans; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D013601:T-Lymphocytes / Q000276:immunology*
D016693:Receptors, Antigen, T-Cell, alpha-beta