Establishment of method of cloning alloantigen-specific T lymphocyte / 安徽医科大学学报

ABSTRACT

Objective　To establish the method of alloantigen-specific T lymphocyte clones. Methods　PBMC of the recipient was stimulated by donor splenocytes for twice and the stimulated T lymphocytes were cloned by limited dilution. Results　Three clones of T lymphocytes were obtained. By analysis of immunohistochemistry, two of the three clones were CD3+CD4-CD8+, another one was CD3+CD4-CD8-. The latter was analyzed by flowcytometry and CD3+ was 98.87%, CD4- 97.05%, CD8- 97.14%. Antigen specificity of the cloned T lymphocytes was determined by stimulation of donor splenocytes and non-related individual splenocytes. When stimulated by donor splenocytes, the cloned T lymphocytes proliferated and did not show proliferative role when stimulated by non-related individual splenocytes. Conclusion　Our method of cloning alloantigen-specific T lymphocytes is successful.