Investigation of alpha nascent polypeptide-associated complex functions in a human CD8(+) T cell ex vivo expansion model using antisense oligonucleotides.

In order to determine molecules involved in the differentiation and proliferation of human CD8(+) cells, two ex vivo expansion models were established: coculture of freshly purified human CD8(+) cells with irradiated autologous feeders (AF) or stimulation with anti-CD3. Two different proliferation kinetics of CD8(+) cells and expression patterns of CD57 were observed between these conditions. Differential display reverse transcriptase-polymerase chain reaction was applied to investigate the differential expression of mRNA species between CD8(+) CD57(+) and CD8(+) CD57(-) populations. A differentially expressed RNA species called alpha nascent polypeptide associated complex (alpha NAC) was found at a higher level in CD8(+) CD57(-) cells than in CD8(+) CD57(+) cells. In the presence of AF, the expression of alpha NAC was reduced on culturing whilst proliferation increased. Similarly, in cultures stimulated with anti-CD3, alpha NAC reverted to its inactive form and differentiation and proliferation increased. Using a phosphorothioate-modified oligodeoxynucleotide antisense directed specifically against alpha NAC mRNA, protein expression was inhibited and increased CD8(+) cell proliferation and CD25 expression were observed irrespective of the culture conditions. This suggests that alpha NAC protein is antiproliferative molecule. This is the first description of the function of the alpha NAC protein in human CD8(+) T cells.