Characteristics of gamma-delta T cells as antigen-presenting cells during transplantation rejection / 中国组织工程研究

ABSTRACT

BACKGROUND: A latest research indicates that γ~δ T cells following touching with microbe products show characteristics as dendritic cells and antigen-presenting cells (APC) and induce intensive immune response of CD4~+ CD8~+ γ~δ T cells.OBJECTIVE: To verify APC-like functions of γ~δ T cells during transplantation rejection, investigate a simple and effective method to amplify γ~δ T cells in vitro, and to infect γ~δ T cells with FasL retrovirus system.DESIGN, TIME AND SETTING: An animal experiment was performed at Central Laboratory of the First Affiliated Hospital of the Fourth Military Medical University of Chinese PLA from August 2007 to August 2008. MATERIALS A total of 60 healthy adult Wistar rats of clean grade and weighing 200-320 g were used to establish donor and receptor models with segmental heterotopic small intestine transplantation.METHODS: Models of segmental heterotopic small intestine transplantation were established using three sleevelet vascular anastomosis. γ~δ T cells were obtained using flow cytometry and its function was demonstrated. Mononuclear cells were routinely separated and amplified with Mtb-Ag. MAIN OUTCOME MEASURES: Cell proliferation was observed; percentage of γ~δ T cells for lymphocytes was detected using TCRγ~δ magnetic beads kit; γ~δ T cells following transfection were detected using pLXSN FasL retrovirus system. RESULTS: Activated γ~δ T cells showed dendritic cell-like adhesion function and APC-like functions during transplantation rejection. γ~δ T cells accounted 4.5% for mononuclear cells, the purification of γ~δ T cells was up to 72.2% on the 10~(th) day after activated by Mtb-Ag, and the purification of γ~δ T cells was up to 99.1% through positive magnetic sorting. 285 bp FasL fragment demonstrated that the gene integration was observed in PA317 cells. The ration of FasL+ cells was 97.3% after infected by pLXSN FasL retrovirus system.CONCLUSION: This study demonstrated APC-like functions of γ~δ T cells during transplantation rejection. γ~δ T cells were successfully amplified in vitro. PA317/ pLXSN2FasL+ retrovirus system was successfully constructed and γ~δ T cells were modified by FasL retrovirus system.