SHIOKAWA Satoshi Medical Institute of Bioregulation, Kyushu University, Assistant Professor, 生体防御医学研究所, 助手 (20215940)
EZAKI Ichiko Medical Institute of Bioregulation, Kyushu University, Research Associate, 生体防御医学研究所, 助手 (40037453)
|Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1997: ¥2,100,000 (Direct Cost: ¥2,100,000)
To identity the existence of antigen-specific T cell responses and to follow the changes of these reactions, it is considered useful to evaluate whether certain T cells clonally accumulate in the lymphocyte population. For this purpose, we have established a novel method to analyze T cell clonality using a combination of reverse transcriptase-polymerase chain reaction of T cell receptor beta chain transcripts and single-strand conformation polymorphism (SSCP). Using this method, we obtained a smear-like pattern of electrophoresed DNA from the heterogeneous T cell population. On the other hand, a single T cell clone exhibits a band in appropriate Vbeta amplification and an accumulatedT cell clone in a heterogeneous lymphocyte population is identified as a band in the background smear pattern. If a lymphocyte population was stimulated by an antigen either in vitro or in vivo, several distinct bands were found to be generated in the background smear. Thus, the dynamic changes of T cell clonal responses could be monitored with this method. Analyzes of several immunological disorders, including autoimmune diseases, malignant disorders, andtransplantation, revealed the involvement of antigen-spesific T cell immune responses in these disorders. Furthermore, taking advantege of the reproduciblemobility of a band of SSCP gel, we are now able tocompare identities of the accumulated T cell Clones in different samples without the need for nucleoptidesequencing of each clone. Such information can thus elucidate the occurrence of uniform or stable immunological reactions in the host and also suggests that these reactions play an important role in vivo. Therefore, taken togeter, the above described novel T cell clonality analysis is considered to be useful in studying the T cell immune responses in various fields of immunology.