Research on molecular chimeric mice pre-T cells infusion to reduce response to allogeneic T cells stimulation / 国际生物医学工程杂志

ABSTRACT

Objective To research the effect of molecular chimeric mice pre-T cells on proliferation ability of allogeneic mouse T cells.Methods The MHC-Ⅰ gene (H-2Kband H-2Db gene) were extracted and amplified by RT-PCR,the identified pre-T cells were transfected by the constructed eukaryotic expression vector of C57BL/6mouse MHC-I (pIRES-H-2Db and pIRES-H-2Kb),non-transfected group and sham pIRES-transfected control group were set.The molecular chimeric cells were transfused back to BALB/c mouse.After 7 days,T lymphocyte cells of each group were extracted,the ability of molecular chimeric cells inducing spleen T lymphocyte response to allogeneic T cells was observed through mixed lymphocyte culture (MLC).Results Sequencing of the plasmid we have constructed showed that insertion sequence contained C57BL/6 mice H-2Kb and H-2Db series which could be retrieved from GenBank.The result of flow cytometry analysis indicated that H-2Kb and H-2Db protein had an increased expression in pre-T cells,the difference with other two groups was statistically significant (P＜0.05).The result of MLC demonstrated that the stimulation index (SI) of T lymphocyte in co-injection transfected pIRES-H-2Kb and pIRES-H-2Db pre-T cells group (0.764±0.074) were significantly decreased compared with non-transfected group(0.983±0.081)and sham pIRES-transfected group(0.994±0.142) (P＜0.05).Conclusions The molecular chimeric pre-T cells infusion can reduce spleen T lymphocyte response to allogeneic T cells and it may induce immune tolerance in vivo.