[Transduction of antisense RNA of MHC-II gene into CD3+ cord blood stem/progenitor cells inhibits the expression of HLA-DR antigen].
OBJECTIVE
To explore whether the antisense RNA of HLA-DR gene could modulate the expression of the gene and reduce the immunogenicity of cord blood (CB) cells.


METHODS
A retroviral vector encoding antisense RNA of HLA-DR gene was constructed and transducted into packaging cell line PA317 by lipofectin. CD34+ hematopoietic stem cells/progenitors were sorted and enriched by MACS.


RESULTS
A high titer (1 x 1095) cfu/ml) helper-free,virus producing cell PA317 was obtained. The vector was identified by blot hybridization and mapping of restriction points. The CD3+ CB cells were infected by the supernatant of the viral cultures. The G418-resistant clone was demonstrated to express the antisense RNA of HLA-DR gene by RT-PCR technique. The expression rate of HLA-DR antigen on CB cells was 28%, which was apparently lower than that of the control (45%) with an inhibition rate of 38.25%.


CONCLUSIONS
The introduction of antisense RNA of HLA-DR gene into CB hematopoietic stem cells can successfully decrease the expression of HLA-DR antigen. These studies provide evidences for the prevention and treatment of GVHR in clinical CB transplantation.
Deng
Y
Y
University of Medical Sciences, Guangzhou 510089.
Li
S
S
Huang
S
S
D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism*; D002460:Cell Line; D005312:Fetal Blood / Q000166:cytology*; D015870:Gene Expression; D005802:Genes, MHC Class II / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D006684:HLA-DR Antigens / Q000235:genetics*; D006801:Humans; D016372:RNA, Antisense / Q000235:genetics*; D012190:Retroviridae / Q000235:genetics; D013234:Stem Cells / Q000378:metabolism*; D014161:Transduction, Genetic
D018952:Antigens, CD34; D006684:HLA-DR Antigens; D016372:RNA, Antisense