Prevalence of Cryptosporidium species isolated from diarrheic children in Makkah, Saudi Arabia.

Cryptosporidiosis has been identified as a significant underlying cause of morbidity and mortality worldwide. Studies in high and low income countries have recognized the importance of Cryptosporidium as a cause of diarrhea. The objectives of the current study were to determine the prevalence rate and genotypes of Cryptosporidiumin in diarrheic children in Makkah Region. A total of 1,380 fecal samples were collected from children aged up to 14 years attending 3 major hospitals of Makkah between March 2015 and March 2016. Stool collected were subjected to direct microscopic examination and crypto antigen detection using ImmunoCard STAT, Cryptosporidium/Giardia rapid test. Part of each positive stool sample was kept frozen at -20ºC for molecular characterization. Initial screening by immunochromatographic detection kit revealed 23 positive cases. PCR was performed for positive cases by amplification of a piece of the gene encoding the small (18S) subunit of rRNA producing a 435-438 bp product. Cryptosporidium genotyping was performed by RFLP analysis of PCR products. Genotyping revealed 18 cases C. hominis genotype, 4 cases C. parvaum genotype and one sample failed to be amplified. The data revealed a higher incidence of the common human species C. hominis (81.8%). The detection of both C. hominis and C. parvaum genotypes point to the possibility of both anthroponotic and zoonotic transmission routes occurring in Makkah region. Further studies are needed to verify the subgenotypes of Cryptosporidium to elucidate the real transmission modes and hence plan for effective control strategies.

Helminth infections: therapeutic potential in autoimmune disorders.

Knowledge of immunity enables us to predict that the reactions set in response to infection with helminth would prevent concomitant disease driven by an opposing spectrum of immune events. In another way, the immune response generated to combat the helminth infection could counteract the immunopathological reactions that drive autoimmune diseases. Rodent model systems recapitulate many aspects of human autoimmune diseases and have been enormously useful in defining mechanisms of immunopathology after infection. From this theoretical perspective, many researchers have proved that infection with a variety of helminth can ameliorate disease in murine model systems. Thus, helminth-evoked Th2 events were shown to improve disorders in which Th1 events predominated. This raised the question, 'Can this information be translated into therapies for autoimmune diseases in humans via actual infection, cell delivery or drug intervention?' In this review, we will present some experimental trails to treat autoimmune disorders through establishment of some parasitic infections.

Secreted adhesion molecules of Strongyloides venezuelensis are produced by oesophageal glands and are components of the wall of tunnels constructed by adult worms in the host intestinal mucosa.

The parasitic female of Strongyloides venezuelensis keeps invading the epithelial layer of the host intestinal mucosa. Upon invasion, it adheres to the surface of the intestinal epithelial cells with adhesion molecules secreted from the mouth. It has been demonstrated that S. venezuelensis are expelled from the intestine because mucosal mast cells inhibit the attachment of adult worms to the mucosal surface. In the present study, we generated specific antibodies against secreted adhesion molecules to investigate their function in vivo, because these molecules have been demonstrated only in vitro in spite of the importance in the infection processes. A mouse monoclonal antibody specific to S. venezuelensis adhesion molecules inhibited the attachment of adult worms to plastic dishes and the binding of adhesion molecules to rat intestinal epithelial cells. Immunohistochemical study revealed that adhesion molecules were produced by oesophageal glands and were continuously secreted in vivo to line the wall of the tunnels formed by adult worms in the intestinal mucosa. Our findings indicate that adhesion molecules play essential roles in the infection processes of S. venezuelensis in the host intestine.

Vaccination with calpain induces a Th1-biased protective immune response against Schistosoma japonicum.

A large subunit of calpain, a calcium-activated neutral proteinase, from Schistosoma japonicum was cloned and expressed in Escherichia coli. When BALB/c mice were immunized with purified recombinant calpain (r-calpain) emulsified in complete Freund's adjuvant, a significant reduction in the number of recovered worms and also in egg production per female worm was observed (P<0.01). Spleen cells of the immunized mice showed enhanced production of gamma interferon (IFN-gamma) by activated CD4(+) T cells. Considering our observation of elevated expression of inducible nitric oxide synthase mRNA in immunized mice, r-calpain-induced IFN-gamma seemed to upregulate the production of nitric oxide by macrophages and subsequently mediated the killing of schistosomulae in the lung. On the other hand, spleen cells of immunized mice showed only faint interleukin-4 production in response to r-calpain in vitro, suggesting that immunization with r-calpain alters the Th1-Th2 balance in murine hosts even during a Th2-promoting S. japonicum infection. Furthermore, histopathological study of the livers of immunized mice showed that granulomas formed around eggs were diminished in both size and number. Egg production by female worms was clearly decreased in immunized mice, suggesting that r-calpain also has antifecundity effects. Taken together, these results point to S. japonicum calpain as a potential vaccine candidate for both worm killing and disease prevention, possibly through the induction of a strong Th1-dominant environment in immunized mice.

Strongyloides venezuelensis: heparin-binding adhesion substances in immunologically damaged adult worms.

Immunologically damaged Strongyloides venezuelensis adult worms were examined for their mucosal invasion ability and secretion of heparin-binding adhesion substances. S. venezuelensis was expelled from male Wistar rats 4 to 5 weeks after infection. Four-week-old adult worms were smaller and had fewer eggs than 1-week-old adult worms. One-week-old, 4-week-old, and 5-week-old adult worms equally established in the recipient mouse intestine when surgically implanted. Adult worms of 4 and 5 weeks of age secreted adhesion substances as much as 1-week-old adult worms. There was no difference in the heparin-binding activities and the lectin-binding profile of adhesion substances among adult worms of different ages. The rate of secretion of adhesion substances from the mouth was also identical. Heparin-binding activities were detected in crude adult worm proteins; however, proteins of 5-week-old adult worms had weaker heparin-binding activities than those of 1-week-old adult worms. Western blotting revealed that a number of heparin-binding proteins were lost in 5-week-old adult worms. A heparin-binding protein of 42. 0 kDa, which was consistently expressed in adult worms, was a possible component of heparin-binding adhesion substances which are secreted from the mouth.