In vitro fertilization-embryo transfer (IVF-ET) can be used by infertile couples to assist with reproduction; however, failure of the embryo to implant into the endometrial lining results in failure of the IVF treatment. The present study investigated the expression of chemokine receptor 7 (CCR7)(lo) programmed death-1(PD-1)(hi) chemokine receptor type 5 (CXCR5)+ cluster of differentiation 4 (CD4)+ T cells and associated factors in patients with repeated implantation failure (RIF). A total of 30 females with RIF and 30 healthy females were enrolled in the current study. Flow cytometry was used to detect the proportion of CCR7(lo)PD-1(hi) CXCR5+ CD4+ T cells in the peripheral blood. Cytokine bead arrays were performed to detect the levels of interleukin (IL)-6, −4 and −2 in the serum. ELISAs were used to detect the level of IL-21 in the serum. Quantitative real time polymerase chain reaction analysis and immunohistochemistry were used to investigate the expression of B-cell lymphoma 6 (Bcl-6), chemokine receptor type 5 (CXCR5) and IL-21 in the endometrium. The results revealed that the percentage of CCR7(lo)PD-1(hi) CXCR5+ CD4+ T cells was increased in the RIF group compared with the control group during the mid luteal phase. The mRNA and protein levels of Bcl-6, IL-21 and CXCR5 in the endometrium and the concentrations of IL-21 and IL-6 in the serum were significantly increased in the RIF group; however, no significant difference was observed between the two groups in regards to the expression of IL-4 and IL-2. Furthermore, a significant positive correlation was identified between the percentage of CCR7(lo)PD-1(hi) CXCR5+ CD4+ T cells and IL-21 and IL-6 levels. The expression of IL-21 also had a positive correlation with Bcl-6 and CXCR5 expression in the RIF group. These results suggest that increased levels of CCR7(lo)PD-1(hi) CXCR5+ CD4+ T cells and associated factors contribute to RIF and could therefore be a potential therapeutic target.

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Background: In vitro cultivation of Echinococcus is essential for vaccine development to prevent transmission of echinococcosis to dogs. We optimized the conditions for Echinococcusprotoscolex evagination and adult worm development in vitro, including those of water, bile, bile salt, trypsin, and serum in the culture system. Results: Short stimulation (2 × 20 s) with water significantly increased the evagination of protoscoleces (pre-worms) of both E. granulosus and E. multilocularis. However, medium containing fetal calf serum (FCS) invaginated 92% of these evaginated protoscoleces. Preculture of the evaginated protoscoleces in no-serum RPMI1640 medium containing dog bile or bile salt for three days maintained 80.5% of the evaginated protoscoleces. Dog serum gel-base maintained 79.8% of the evaginated protoscoleces developing adult worms, which was higher than newborn bovine serum gel-base. The rapidly developing worms had 3–4 progloids after 56 days of culture. E. granulosus worms were longer and wider in size than E. multilocularis after five weeks of in vitro culture. Conclusions: Brief stimulation with water for proscolex evagination and pre-worm maintenance in no-serum medium are crucial for in vitro worm development of E. granulosus and E. multilocularis. Dog serum gel-base and bile salts are important for long-term tapeworm development. This study was supported by the National Natural Science Foundation of China (81830066, U1803282)

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