1.Expression of Tim-3 in early stages of Echinococcus granulosus infection in mice
Fangjie XU ; Shuanghong YIN ; Jun HOU ; Hairui FANG ; Hongqun JIANG ; Xiangwei WU ; Xueling CHEN
Chinese Journal of Immunology 2014;(12):1616-1621,1626
Objective:To understand the expression levels of Tim-3,a new proinflammatory factor in the early stages of Echinococcus granulosus infection in mice.Methods: BALB/c mice were infected with E.granulosus.Peritoneal macrophages and spleen cells were collected at 1,5,9 and 13 days post-infection.At different time points ,the levels of Tim-3 in peritoneal macrophages and spleen CD3+lymphocyte subsets were detected by FCM , and the relative expression of TLR 4 mRNA was detected by qRT-PCR.Results:There was no significant difference in the expression levels of Tim-3 of CD3+spleen lymphocyte subsets between E.granulosus group and control group (P>0.05).The expression levels of Tim-3 of spleen macrophages (9,13 days) and peritoneal macrophage (5,9,13 days) were much higher in E.granulosus infected group than those in control group with statistical significance (P<0.05).The numbers of macrophages were no change.Compared with control groups,the relative expression of TLR4 mRNA at 1 day post-infection was statistically higher in E.granulosus infected group ( P<0.05 ).Conclusion:During early stage of E.granulosus infection in mice,the levels of Tim-3 expression are upregulated,while the expression of TLR4 are downregulated,which may inhibit the function of macrophages resulting in host-immunity-defensive-system inhibition and immune tolerance of E.granulosus to host.
2.Effect of protoscolex on T subsets in mice spleen cells in vitro
Hairui FANG ; Hongqun JIANG ; Fangjie XU ; Jun HOU ; Dan DONG ; Congzhe CHEN ; Xiangwei WU ; Xueling CHEN
Chinese Journal of Immunology 2016;(2):174-177
Objective:To observe the effect of protoscolex on Th subsets and correlative cytokine in mice spleen cells in vitro.Methods:Co-culture spleen cells from BALB/c mice with protoscolices,then IL-4,IFN-γand TGF-βproduction in cell culture supernatants were analyzed by ELISA.The percentage of Th subsets were detected by Flow Cytometry analysis.Results:Secretion levels of IL-4 and TGF-βwere significantly increased in spleen cells at different time point in co-culture system with protoscolices.Ratios of Th2 and Treg cells were also significantly increased in co-culture system at different time points than the control groups.However,there was no statistical significance for ratio of Th1 cells at different time points.Conclusion:The protoscolex can increase the ratios of Th2 cells and Treg cells from spleen cells.Secretion levels of IL-4 and TGF-βwere also increased in spleen cells co-cultured with protosco-lices.The results suggest that these Th cell subsets play a role in the immune escape of the hydatid disease.
3.Effect of inhibition of TGF-β1 receptors on lymphocytes during infection of Echi-nococcus granulosus
Shuanghong YIN ; Junbo ZHANG ; Xiaolin CHEN ; Fangjie XU ; Jun HOU ; Xiangwei WU ; Xueling CHEN ; Yuanyong YAO ; Mingming XING ; Wencai YANG
Chinese Journal of Immunology 2015;(5):607-612
Objective:To investigate the effects of TGF-β1 on T lymphocytes of BALB/c mice infected with Echinococcus granulosus( E.granulosus ) in vitro.Methods: The inhibitor group:the spleen cells of BALB/c mouse were co-cultured with E.granulosus and SB525334.The control group:the spleen cells of BALB/c mouse were co-cultured with E.granulosus and PBS.The blank group:the spleen cells of BALB/c mouse were co-cultured with RPMI-1640 medium and SB525334.The lymphocytes were collected at 48 h post-infection.The T lymphocyte subsets, the number of CD4+CD25+T cells, the number of NK cells, and the expression of NKG2D receptor were detected by flow cytometry.The NK cell activity was determined with the lactate dehydrogenase leakage assay(LDH).Results:The inhibition the TGF-β1 receptors resulted in the increase of in the number of CD4+T cells,the decrease in the number of CD8+T cells,the increase of in the ratio CD4+/CD8+T cells,the decrease of in the number of CD4+CD25+T cells,the increase in the expression of the NKG2D receptors,the increase in the lysis rate of Yac-1 cells by NK cells,and a positive cor-relation between the expression of activity receptor NKG2D and killing activity of NK, which were mediated by E.granulosus.Conclusion: The inhibition of TGF-β1 receptors can enhance the immune response of T lymphocytes against E.granulosus infection in vitro.
4.Identification Study on Cryptotympana pustulata Ecdysis Based on 3D Depth of Field Synthesis Technology
Lixiao GUO ; Zijing XUE ; Fangjie HOU ; Baohui SUN ; Yuguang ZHENG ; Qian ZHENG
China Pharmacy 2020;31(4):447-451
OBJECTIVE:To establish the rapid field identification method of Cryptotympana pustulata ecdysis. METHODS : 3D depth of field synthesis technology was used to identify 50 batches of C. pustulata ecdysis and its adulterants from the length of beak ,size and protrusion degree of upper labial base ,the protrusion degree of the lower labial base ecdysis and the color of its upper transverse groove ,the number and shape of main and lateral spines on the foot ,significance of abdominal valves ,the number of webs ,the number and shape of side plates ,the number of tergum rings ,terminaliae,etc. RESULTS :Among 50 batches of samples ,S1-S5,S26-S30,S36-S50 were C. pustulata ecdysis;S21-S25 was adulterants of C. pustulata ecdysis after weight gain ;S31-S35 was adulterants of C. pustulata ecdysis after extraction ;S6-S20 were ecdysis from Tibicen flammatus ,C. flammatta,Lyristes pekinensis ,all of which were adulterants. The main distinguishing feature of C. pustulata ecdysis and its adulterants was that abdomen and ventral surface of C. pustulata ecdysis were triangular ,and the abdomen and ventral surface of other species was nearly parallel ;the valve of C. flammatta ecdysis was obvious ,but those of other varieties were not obvious ;the lateral appearance of terminaliae of C. flammatta ecdysis was sharper than those of other species ;there was an acute angle between the front foot accessory thorns and the end thorns of the T. flammatus ecdysis,and an obtuse angle between the front foot accessory thorns and the end thorns of the L. pekinensis ecdysis. CONCLUSIONS :The method is simple ,reliable and suitable for rapid field identification of C. pustulata ecdysis and its adulterants.