Found programs: Open Project of Henan Key Laboratory of Immunology and Targeted Drugs (No. HNKLITD2024001); Open Project of State Key Laboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases in Central Asia (No. SKL-HIDCA-2024- GX2); Innovative Research Project of Xinjiang Uygur Autonomous Region Health and Wellness, Science and Technology Program (No. 2025001CXKYXM650030870)
Authors：Wang Peng1 ，Gong Xiaoyun1 ，Zhang Manli1 ，Zhang Yunian1 ，La Xiaolin1,2,3,4
Keywords：follicular helper T cells; embryo implantation disorder; C-X-C Chemokine Receptor Type 5 protein; B-cell lymphoma 6 protein; cytokines; endometrial receptivity
DOI：专辑：医药卫生科技
〔Abstract〕 To detect the proportion of splenic follicular helper T (Tfh) cells and their functionally related cytokine expression levels in the incomplete embryo implantation disorder (EID) model mice, and to explore the immunological mechanism of Tfh in infertility caused by embryo implantation disorder. Methods Sixteen female Kunming mice were randomly divided into two groups, with eight mice in each group. On day 4 of pregnancy, an incomplete EID mouse model was established by oral gavage of mifepristone suspension, while an equal volume of saline was administered to the control group. On day 8 of pregnancy, the mice were euthanized. Flow cytometry was used to detect the levels of Tfh cells in the spleen lymphocytes of both incomplete EID mice and normal control mice. qRT-PCR was performed to measure the mRNA levels of B-cell lymphoma 6 (Bcl-6) and chemokine receptor 5 (CXCR5) in the spleen lymphocytes of both groups. Western blotting was employed to assess the protein expression levels of Bcl-6 and CXCR5 in the spleen lymphocytes of both groups. Serum levels of interleukin-4 (IL-4), IL-6, and IL-21 were measured by ELISA. Immunohistochemistry (IHC) was used to observe the expression levels of progesterone receptor (PR), Bcl-6, and CXCR5 proteins in the uterine endometrial tissue of mice in both groups. Results Incomplete-type EID mice had a reduced number of embryo implantation points and reduced endometrial PR receptor expression. Flow assay results showed that the proportion of CD4+CXCR5+Tfh cells in splenic lymphocytes of incomplete-type EID mice was significantly higher than that of normal controls (P<0.05).Compared with the normal control group, Bcl-6 and CXCR5 mRNA levels and protein levels were elevated in splenic lymphocytes of incomplete EID mice, with statistically significant differences(P<0.05); serum IL-4, IL-6, and IL-21 levels were elevated in incomplete EID mice, and Bcl-6 and CXCR5 proteins in the endometrium were significantly elevated (P<0.05). Conclusion The increase of Tfh cells and their associated cytokines Bcl-6 and CXCR5 is associated with the development of incomplete EID, and may be involved in the development of female immune infertility.