Identification of Genes Involved in Positive Selection of CD4⁺8⁺ Thymocytes: Expanding the Inventory

Abstract

Positive selection of cortical CD4⁺8⁺ thymocytes represents crucial and mysterious process in T cell development whereby short-lived precursors are rescued from programmed cell death and induced to differentiate towards long-lived CD4 and CD8 T cells. One reason that this process is not fully understood is that the inventory of genes changing their expression in positively selected CD4⁺8⁺ thymocytes is not yet complete. In this work Affymetrics GeneChip cDNA microarrays and cDNA-Representational Difference Analysis were used to search for unknown and known genes that were not identified before as being involved in positive selection. Comparison of transcriptosome of nonstimulated with transcriptosome of PMA/ionomycin stimulated thymoma cell line resembling CD4⁺8⁺ thymocytes and subtraction of cDNA of extrathymic tissues from cDNA of purified CD4⁺8⁺ thymocytes resulted in identification of 36 genes, which have not been previously reported to change their expression during positive selection. One of them represents a novel, third evolutionarily conserved gene within RAG locus.

Keywords:

• CD4⁺8⁺ thymocytes
• positive selection
• gene expression