During pregnancy, there are important changes in hormone levels such as the huge production of human chorionic gonadotropin (hCG), which is supposed to influence the immune system. The aim of this study was to investigate the effect of hCG on immune response against Leishmania, through the evaluation of the functions of human macrophages infected with L. tropica. This study demonstrated that hCG significantly increased the NO production by rHu-IFNγ-primed macrophages then infected with L. tropica, which was correlated with decrease in the number of infected macrophages as well as the number of amastigotes per macrophage in a dose-dependent manner; however, the greatest effect was shown with the 250 U/mL concentration. The addition of the same concentration of hCG to rHu-IFNγ-primed macrophages caused also a major increase in both IL-6 and IL-12p40 production. In conclusion, hCG enhances different macrophage functions involved in immunity against L. tropica.
In Leishmania species, protein disulfide isomerase (PDI) is an essential enzyme that catalyzes thiol-disulfide interchange. The present work describes the isolation, cloning, sequencing and expression of the pdI-2 gene. Initially, the gene was amplified from L. tropica genomic DNA by PCR using specific primers before cloning into the expression vector pET-15b. The construct pET/pdI-2 was transformed into BL21(DE3) cells and induced for the protein expression. SDS-PAGE and western blot analysis showed that the expressed protein is about 51 kDa. Cloned gene sequence analysis revealed that the deduced amino acid sequence showed significant homology with those of several parasites PDIs. Finally, recombinant protein was purified with a metal-chelating affinity column. The putative protein was confirmed as a thiol - disulfide oxidoreductase by detecting its activity in an oxidoreductase assay. Assay result of assay suggested that the PDI-2 protein is required for both oxidation and reduction of disulfide bonds in vitro. Antibodies reactive with this 51 kDa protein were detected by Western blot analysis in sera from human infected with L. tropica. This work describes for the first time the enzymatic activity of recombinant L. tropica PDI-2 protein and suggests a role for this protein as an antigen for the detection of leishmaniasis infection.
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