Co-stimulate Molecules PD-L1 Regulate B Cell Differentiation And Relationship With Tregs And Biological Significance

Part I:The determination of CD19 + Bregs cells and biological significance in Breast ductal carcinomaObjectives: To explore Bregs expression in breast tumor tissue and the correlation with invasive breast carcinoma and their clinical significance.Methods: This study using immunohistochemical technique analysis the expression of CD19,IL-10 in 134 cases of invasive breast cancer and the relationship.Using flow cytometry technology study the CD19 + IL-10 + B cells expressing in breast tumor tissue and the relationship with CD24, CD38Results:(1): In invasive breast carcinoma, IL- 10 mainly locate in tumor infiltrating lymphocytes cell plasma, the expression is associated with tumor differentiation degree(P=0.048).(2): Tumor infiltrating CD19 + B cells and IL-10 exist co-expression(P=0.001), Part of IL-10 can be seen in the cytoplasm of CD19 + cells, show that infiltrating CD19 + B cells may have ability to secrete IL-10.(3): The percentage of CD19 + IL-10 + was higher than in peripheral blood of invasive carcinoma than fibroma(P = 0.046) with high expression of CD24, CD38,Also CD19+ CD24+ CD38+ B cells express high IL-10 + B cells.(4): CD19+ B, Bregs cells express higher in invasive breast cancer tissues than fibroma.Conclusion: This study confirmed CD19+ IL-10 +B cells is higher in breast invasive cancer, and with CD24, CD38. To further research on B cells and the correlation with breast cancer cells, and the correlation with tumor immune lay the foundation.Part II:The relationship between Bregs and Tregs and its biological significanceObjectives: To study the Bregs, Tregs expression in breast cancer, explore the relationship and their biological significance.Methods: Using flow cytometry to detect the expression of Bregs,Tregs in healthy people, fibroma and breast invasive carcinoma, the relationship with clinical pathological parametersResults:(1):The peripheral blood of CD19+ B improve the proportion of Tregs in CD4+ T cell not only in breast invasive cancer patients, but also in healthy people.(2): The proportion of Bregs in peripheral blood is higher in invasive breast cancer, than healthy people(P < 0.001) and fibroma(P < 0.001), The expression of CD19+ B cells in tumor diameter less than 3 cm group is increased, than in diameter greater than 3 cm group(P = 0.019).(3) The proportion of Tregs in peripheral blood is higher in invasive breast cancer, than healthy people(P < 0.001) and fibroma(P< 0.001),CD4+ T cells expressed in 49 years group less than the group of older than 49 years old(P=0.047).(4): In the peripheral blood of invasive breast cancer,the expression of B is associated with T cells(≤0.001,r=0.397),and Bregs and Tregs expression had the significant positive correlation(P=0.001,r=0.361).Conclusion: The peripheral blood of invasive breast carcinoma expressed higher levels of Bregs, Tregs, and had positive correlations. how tumor cells escape immune through Bregs, Tregs still needs further research.Part III:PD-L1 regulate Bregs and biological significanceObjectives: Through PD-L1 molecules expressed on the surface of breast cancer cell, to study the tumor cells improve role and function of Bregs, provides a new therapy strategy for PD-L1 express on tumor cells.Methods: Using activated CD19+ B cells, in culture with MCF-7 and MDA-MB-231 breast cancer cell lines, analysis the expression of Bregs, and in culture with CD4+ T cells tests the expression of Tregs. Si RNA-PD-L1, in culture with CD19+ B, CD4+ T cells,detection Bregs, Tregs expressionResults:(1): The surface of PD-L1 molecules expressed on tumor cell inductd CD19+ Bregs amplification, which facilitate Tregs differentiation.(2): Si RNA PD-L1 expression,the expression of Bregs,Tregs decreased.Conclusion: PD-L1 can induce CD19+ B cells differentiate to Bregs, thereby inhibiting the immune response, so as to evade the host immune attack; This provide evidence for immunotherapy for PD-L1.