Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
, 10 (7), 7947-7955
eCollection

Expression of Toll-Like Receptor 2, 4 and Related Cytokines in Intraperitoneally Inoculated Balb/C Mice With Echinococcus multilocularis

Affiliations

Expression of Toll-Like Receptor 2, 4 and Related Cytokines in Intraperitoneally Inoculated Balb/C Mice With Echinococcus multilocularis

Shadike Apaer et al. Int J Clin Exp Pathol.

Abstract

Immune response pattern between host and Echinococcus multilocularis (E. multilocularis) is considered as a crucial point in development of alveolar echinococcosis (AE). In this study, we are aiming to study the expression patterns of TLR2 and TLR4 with related cytokines and transcription factors in secondary E. multilocularis infected murine model. The murine model of AE was developed by using intraperitoneal inoculation of E. multilocularis protoscolexes and albendazole (E. m+ABZ group) or carboxy methyle cellulose (CMC; E. m+CMC group) administration via gastric tube was initiated in the third month and continued for one month. Mice with CMC administration served as negative controls (C+CMC group). The splenic cells and peritoneal exudates cells (PECs) were prepared and the levels of IFN-γ, IL-10, and IL-5 in splenic cells and PECs culture supernatants were detected using enzyme linked immune-sorbent assay (ELISA). Besides, the mRNA expression levels of TLR2, 4, transcription factors and cytokines were detected by using real-time fluorescent quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The concentration levels of IFN-γ, IL-10, and IL-5 in PECs and splenic cell supernatants were extremely lower, however, significantly elevated after stimulated with Concanavalin A (ConA) for 36 h with higher concentrations in E. m+CMC group comparing to both E. m+ABZ and C+CMC group. The mRNA levels of TLR2, 4 and GATA3, IFN-γ, IL-10 in splenic cells were significantly increased in E. m+CMC group comparing with other groups. Simultaneously, T-bet mRNA expressions were elevated in E. m+ABZ and C+CMC group compared to E. m+CMC group. In addition, T-bet/GATA3 ratios was higher in E. m+ABZ group compared to E. m+CMC group and were higher in C+CMC group than those in E. m+CMC group. TLR2 mRNA expression in splenic cells showed a positive correlation with IL-10 concentration levels in splenic cell culture supernatants. The present study provides evidence on the possible role of TLR2 in the process of immune tolerance during E. multilocularis infection and suggests albendazole treatment might reverse the immune tolerance situation and improve parasite clearance process.

Keywords: Toll-like receptor; albendazole; echinococcus multilocularis; immune modulation.

Conflict of interest statement

None.

Figures

Figure 1
Figure 1
Assessment of parasite load in intraperitoneal cavity of mice infected with E. multilocularis. All mice were sacrificed, and lesion tissues and masses were removed for monitoring parasite burden in the peritoneal cavity (A). Mice treated with ABZ administration resulted in a significantly reduced median parasite weight which was lower when compared to E. m+CMC mice (B).
Figure 2
Figure 2
qRT-PCR analyses of TLR2 and TLR4 mRNA expressions in different groups. The relative expressions of TLR2 and TLR4 were calculated, and results showed TLR2 mRNA expressions were significantly elevated in E. m+CMC group compared to C+CMC and E. m+ABZ group with statistical differences (P<0.05, P<0.01 respectively). Relative mRNA expressions of TLR4 were higher in E. m+CMC group comparing with both E. m+ABZ and C+CMC group, albeit no statistical differences were found between them (P>0.05). P value <0.001 with marker ***; P value <0.01 with marker **; P value <0.05 with marker *.
Figure 3
Figure 3
Relative mRNA expressions of cytokines and transcription factors in splenic cells. Relative cytokines and transcription factors GATA3, IFN-γ, T-bet, and IL-10 mRNA expression levels were calculated by using qRT-PCR technique. GATA3, IFN-γ and IL-10 mRNA expression levels were elevated in E. m+CMC group compared to both E. m+ABZ and C+CMC group. Relative mRNA expressions of T-bet were found to be increased in E. m+ABZ group comparing with both E. m+CMC and C+CMC group. Similarly, T-bet/GATA3 was significantly elevated in E. m+ABZ group comparing with both E. m+CMC and C+CMC group. P value <0.001 with marker ***; P value <0.01 with marker **; P value <0.05 with marker *.
Figure 4
Figure 4
Concentration levels of cytokines in PECs and spleen cell supernatants. Cytokine concentration levels before and after Con A stimulation were detected by using ELISA technique. After stimulating with Con A, concentrations of IFN-γ from both PECs and spleen cell supernatants were similar between all three groups. IL-10 concentration levels were elevated after Con A stimulation and significantly decreased in E. m+ABZ group when comparing with E. m+CMC group. Concentration levels of IL-5 were significantly increased in E. m+CMC and E. m+ABZ group compared to C+CMC group in PECs, however, it was higher than that in Control group in spleen cell supernatants with no statistical differences. P value <0.001 with marker ***; P value <0.01 with marker **; P value <0.05 with marker *.
Figure 5
Figure 5
Correlation analysis between TLR2 mRNA expression levels and IL-10 concentration levels. Spearman correlation coefficients indicated that TLR2 mRNA expressions in splenic cells had a positive correlation with spleen cells IL-10 concentration levels (r=0.4344, P=0.0339). No correlations were found between spleen cell TLR4 mRNA expressions and IL-10 concentrations (r=0.2153, P=0.3124).

Similar articles

See all similar articles

LinkOut - more resources

Feedback