The study of prolongation of rat renal allograft survival by CD4+ CD25- T cells induced by recipient dendritic cells transfected with dominant negative form of I.KAPPA.B kinases 2

Abstract

Objective To observe whether CD4 + CD25-T cells induced by recipient-derived immature dendritic cells (imDC) transfected with dominant negative form of IκB kinases 2 (IKK2dn) gene and loaded with donor antigen can prolong the survival time of kidney transplantation in rats and investigate its probably.Methods DC were cultured from recipient rats (Lewis) bone marrow,transfected with AdvIKK2dn and loaded with donor antigen,then cultured with Lewis rats lymphocytes in primary mixed lymphocyte reaction (MLR).The CD4 + CD25-T cells were separated by immune magnetic beads method after 72 hours.Male Brown Norway rats and Lewis rats were used as donors and recipients respectively,Four groups were set up(naive T cells group,Adv0-CD4 + T cells group,CD4 + CD25-T cells group and rejection group),receiving 1 × 107 naive CD4 + T-cell,Adv-0-DC-generated CD4 + CD25-T-cell,Adv-IKK2dnDC-generated CD4 + CD25 T-cell and equal volume of normal saline,respectively 7 days before transplantation.In the third party donor group,wistar rats as donors were treated the same as CD4 + CD25 T cells group before transplantation.After transplantation,the survival time of recipients were observed; the T lymphocyte proliferation in recipients was measured; the levels of serum interleukin (IL)-2,IL-10,interferon-γ(IFN-γ),transforming growth factor-β (TGF-β) were detected; serum creatinine levels were monitored ; pathological changes were examined to identify the grade of rejection,Results Compared with other groups,CD4 + CD25-T cells group markedly prolonged the survival time of renal allografts [(28.50 ± 2.36) d,P ＜0.01],and had higher expression of IL-10 and TGF-β (P ＜0.05).Compared with Adv0CD4 + T cells group,rejection group,the third party donor group,expression levels of IL-2 and IFN-γwere lower(P ＜0.05).In contrast to other groups,CD4 + CD25-T cells group elicited markedly lower proliferative responses (P ＜ 0.05) and lower grade as estimated by pathological examination.Conclusion These findings suggested that CD4 + CD25 T cells induced by recipient-derived immature DC transfected by IKK2dn and loaded with donor antigen could prolong renal allograft survival in a donor-specific manner,and it maybe by expressing high levels of IL-10 and TGF-β. Key words: Dendritic cells ; Dominant negative form of IκB kinases 2 ; CD4+ CD25-T cells; Kidney allotransplantation ; Rat