| Background: CD3~+ CD56~+ NKT like cell is a group of heterogeneous active immune cells that can express T cell and NK cell surface markers simultaneously, also has powerful tumoricidal activity of T lymphocyte and the non MHC restriction tumoricidal characteristic of NK cell. It is considered as the fastest rate of propagation, the strongest tumoricidal activity and the most widely withstand disease spectrum. NKT like cell is also called cytokine induced killer cell which can be acquired by Coculture of various cytokines in vitro. It is extensively investigated in peripheral hemogram of the tumour patients in vitro and in adoptive immunotherapy. NKT like cells are not only efficient immune cells which can kill tumor cells directly in vitro, but also immune-suppressing regulatory cells, which play important role in immune homeostasis of organism, anti-infection, anti-tumor and transplantation immunity. In order to reveal its biological effect in autoimmune diseases, the goal of this project was to clarify NKT like cell's sensibility to different stimuli in vitro and its potential role the regulation of inflammation, therefore to provide experimental evidence for its biological function in autoimmune diseases.Objective: To investigate the immunoregulation mode and biological features of NKT like cells by detecting the multiplication capacity of CD3~+CD56~+ and the expression level of IFN-γof Th1 cytokines and IL-4 of Th2 cytokines in peripheral blood of patients with systemic lupus erythematosus. Methods: Thirty cases of SLE patients from the Rheumatology Department of the First Affiliated Hospital of Anhui Medical University and 20 cases of age- and gender- matched health controls from Physical Check-Ups were collected during March to July in the year of 2010. Meanwhile SLE patients were divided into stable and active group according to disease activity. Peripheral blood mononuclear cell (PBMC) was purified, flow cytometry analysis was used to detect the expression level and multiplication capacity of CD3~+CD56~+, IFN-γof Th1 cytokines and IL-4 of Th2 cytokines in vitro after nonspecific stimulation by PMA , Ionomycin and specific stimulation of CD3mAb and IL-2.Results:1. The expression level of CD3~+ CD56~+ NKT like cells in SLE patients decreased mar- kedly than that in health controls (1.45±0.37 vs 2.81±0.52,P <0.01). However, the expression level of CD3~+ CD56~+ NKT like cells in active group has no statistical significance with stable patients (1.53±0.44 vs 1.41±0.33,P=0.405).2. The expression level of CD3~+ CD56~+ NKT like cells were enhanced signifiantly in SLE patients and health controls by nonspecific stimulation of PMA , Ionomycin and specific stimulation of CD3mAb , IL-2(P <0.01). The multiplication capacity of CD3~+ CD56~+ NKT like cells by the activation of CD3mAb and IL-2 enhanced significantly than by nonspecific stimulation of PMA and Ionomycin. Moreover, the multiplication capacity of CD3~+CD56~+ NKT like cells by the activation of CD3mAb and IL-2 enhanced significantly in health controls than that in SLE patients (16.73±2.44 times vs 13.78±1.87 times,P <0.05).3. The expression level of IFN-γwas increased markedly in SLE patients and in health controls after nonspecific stimulation of PMA, Ionomycin and specific stimulation of CD3mAb, IL-2. However, whether by the specific stimulation or by the nonspecific stimulation, the expression level of IFN-γin SLE patients was higher than that in health controls. Moreover,the expression level of IFN-γby the specific stimulation of CD3mAb and IL-2 was increased markedly than nonspecific stimulation of PMA and Ionomycin( P <0.01). However, the IL-4 of Th2 cytokines is scarcely expressed by two different stimulating methods.Conclusion:1. The expression level of CD3~+ CD56~+ NKT like cells in SLE patients decreased markedly than that in health controls, furthermore, its multiplication capacity is inferior to the health controls, the results may show that NKT like cells are closely related to the formation of SLE, moreover, its dysfunction may associated with the progression of SLE.2. CD3~+CD56~+ NKT like cells can be activated by specific stimulation of CD3mAb, IL-2 in vitro, then can proliferate greatly, which may be applied to the treatment of SLE patients whose NKT like cells are deficient.3. The high expression Th1 cytokines IFN-γand the low expression Th2 cytokines IL-4 was found by the in vitro activation in SLE patients , illustrating that the change of NKT like cells' function may promote the immune response to humoral immunity by Th1, so CD3~+ CD56~+ NKT like cells regulate the inflammatory immune response by IFN-γof Th1 cytokines and inhibit Th2 response.
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