Adenovirus-mediated expression of antisense RNA transcripts complementary to pig alpha(1,3) galactosyltransferase mRNA inhibits expression of Gal alpha(1,3) Gal epitope

AIM: To examine the effects of the expression of antisense RNA transcripts complementary to the pig alpha (1,3) galactosyltransferase [alpha (1,3)GT]m RNA on the expression of Gal cr(1,3) Gal structure (gal epitope) in culture d cell lines. METHODS: Human adenoviral vectors were used to mediate the ex pression of antisense RNA. The expression levels of H blood group antigens and gal epitopes were analyzed by flow cytometry using FITC-UEA-I and FITC- GS-IB4 lectins, respectively. RESULTS: Recombinant adenoviruses, Ad5anti-sG T600 and Ad5anti-sGT1100, which express antisense RNA complementary to diff erent regions of the pig alpha (1,3) GT mRNA, were constructed and used to infect cell line of NIH3T3. The results showed about 30 % reduction in the expression level of gal epitopes on the surface of NIH3T3 cells. In additio n, co-expression of human secretor type alpha (1,2) fucosyltransferase [alp ha (1,2)FT]cDNA and antisense RNA complementary to the pig alpha (1,3) GT m RNA led to a further reduction in the gal epitope level. CONCLUSION: Recomb inant adenoviruses, Ad5anti-sGT600 and Ad5anti-sGT1100, are effective to do wn-regulate the gal epitope expression.