The Role Of Thymic Stromal Lymphopoietin In Polarization Helper T Cell In Lung Cancer

Objective:We investigated the effect of thymic stromal lymphopoietin (TSLP) on tumor immunity Thl/Th2 shift, further explored the mechanism of Th cell differentiation and also explored new treatment in immune therapy through dendritic cells (DCs) models in vitro. To observe thymic stromal lymphopoietin expression in lung cancer patients, and used rhTSLP to stimulate DC, then test phenotype of DC and cytokines, further investigated the effection of TSLP-DC on Th cell differentiation, to explore the immune mechanism of Thl/Th2 shift in lung cancer.Methods:30 cases of lung cancer tumors, normal lung tissue, tumor infiltrating lymph nodes and negative lymph nodes were collected, immunohistochemistry assay was used to test the expression of TSLP in cells to analyze the differences between tissues. Then we collect peripheral blood of lung cancer patients, and establish DC model in vitro, moreover DCs were induced to mature by LPS, rhTSLP and rhTSLP +LPS, tested their phenotypes using flow cytometry, finally we detect cytokines by euzymelinked immunosorbent assay(ELISA). We futher isolated CD4+T cell using magnetic beads, and was co-culture with imDCs,DCs/LPS,DCs/TSLP in order to induce Th cell differentiation, then IFN-γand IL-4 enzyme-linked immunosorbent spot test ELISPOT was used to determine the differentiation direction of CD4+T cells.Result:Immunohistochemistry showed that, TSLP was not or very lowly expressed in normal lung tissue, the expression score of TSLP in lung cancer tissue and positive lymph nodes were significantly higher than negative lymph nodes (P<0.05), the expression in lung cancer tissue was higher than the metastatic lymph nodes (P＜0.05). Tested DC phenotype by flow cytometry showed that:there were no significant difference between TSLP-DC group and the LPS-DC group of cell markers (including CD83, CD86, CDllc, CD14, OX40L, HLA-DR), howerever CD83 and CD86 in TSLP-DC group and the LPS-DC group was significantly higher than the im-DC group. ELISA tested DC culture supernatants showed:IL-6, IL-12, TNF-α, IL-10 in LPS-DC group are significantly higher than im-DC group and TSLP-DC group (P＜0.05), and there is no difference between the latter two; TSLP-DC group of TGF-βwere significantly higher than LPS-DC group and the im-DC group (P＜0.05), no difference between the latter two; and IFN-γno difference among the groups (P=0.19). DC cells and CD4+T cells were incubated,ELISPOT results indicated:IL-4 spots in TSLP-DC group are significantly more than im-DC group and the LPS-DC group, there was no significant difference between the latter two; And IFN-γspots in TSLP-DC group was significantly fewer than im-DC group and the LPS-DC group, there was no significant difference between the latter two;Conclusion:(1) The expression of TSLP in tumor tissue and positive lymph nodes were significantly higher than in negative lymph nodes or normal lung tissue; (2) TSLP can induce DC differentiation and maturation, and compared with im-DC, The expression rates of CD83, CD86 were upregulated,after DCs were treated with TSLP or LPS, but the expression of CDllc, CD14, OX40L, and HLA-DR have no significant difference between all groups. (3) DCs treated with TSLP expressed lowly IFN-γ, IL-12, TNF-α, IL-10 and IL-6, but secreted a large amount of TGF-β1. (4) DCs treated with TSLP can induce differentiation of CD4+T to Th2 cells.