Syphacia obvelata: A New Hope to Induction of Intestinal Immunological Tolerance in C57BL/6 Mice

The ability of nematodes to manipulate the immune system of their host towards a Th2 and T regulatory responses has been proposed to suppress the inflammatory response. Clinical trials have proposed a useful effect of helminth infections on improvement of inflammatory disorders. In this study, we investigated the immunomodulatory effect of Syphacia obvelata infection to induce intestinal tolerance in C57BL/6 mice. Mice were infected through the cagemates with self-infected BALB/c mice. Four weeks post-infection, expression levels of IFN-γ, TNF-α, IL-17, and IL-10 were assessed in the supernatant of mesenteric lymph node (MLN) culture. Foxp3⁺Treg were measured in MLN cells by flow cytometry. In the S. obvelata-infected group, the percentage of Tregs (5.2±0.4) was significantly higher than the control (3.6±0.5) (P<0.05). The levels of IL-10 (55.3±2.2 vs 35.2±3.2), IL-17 (52.9±3.8 vs 41±1.8), IFN-γ (44.8±4.8 vs 22.3±2.3) and TNF-α (71.1±5.8 vs 60.1±3.3) were significantly increased in infected mice compared to the control group (P<0.05). The above results showed the potential effects of S. obvelata to induce intestinal tolerance. Therefore, it seems that S. obvelata may increase the immunological suppressive function in the intestinal tract.

experimental model of colitis induced by dextran sulfate sodium from acute progresses to chronicity in C57BL/6: correlation between conditions of mice and the environment

Aim: To induce acute colitis progresses to chronicity in C57BL/6 mice by dextran sulfate sodium

Background: Murine models are essential tools to understand IBD pathogenesis. Among different types of chemically induced colitis models, the dextran sulfate sodium [DSS]-induced colitis model is the most common model of IBD, due to its simplicity

Patients and methods: Male C57BL/6 mice 6-8 weeks old, were collected and matched by age with controls. C57BL/6 mice treated with 2 cycles of 3.5% DSS for 4 days and 4 days of pure water between each cycle. After that, mice were sacrificed and the entire colon was removed. Small sections of the colon were fixed in formaldehyde, embedded in paraffin and sectioned with a microtome. Sections were stained with hematoxylin eosin to analyses the degree of inflammation

Results: After the first cycle oral administration of DSS, mice with severe and visible rectal bleeding and diarrhea entered into the acute phase. After day 4-5, bleeding and diarrhea were improved and mice entered into the chronic phase with peak levels of weight loss. Macroscopically, the inflammation was predominantly located in the distal colon. Microscopically, examination of the distal colon sections showed a decrease number of goblet cells, loss of crypts, signs of surface epithelial regeneration and moderate to severe infiltration of inflammatory cells in the mucosa

Conclusion: In order to achieve an experimental colitis model, our protocol is recommended for future therapies in IBD experimental modeling

Application of multiplex PCR for detection and differentiation of entamoeba histolytica, entamoeba dispar and entamoeba moshkovskii

Entamoeba moshkovskii and E. dispar are impossible to differentiate microscopically from the pathogenic species E. histolytica. Multiplex polymerase chain reaction [Multiplex PCR] is a widespread molecular biology technique for amplification of multiple targets in a single PCR experiment. For detection and differentiation of the three-microscopy indistinguishable Entamoeba species in human, multiplex PCR assay using different DNA extraction methods was studied. A conserved forward primer was derived from the middle of the small-subunit rRNA gene, and reverse primers were designed from signature sequences specific to each of these three Entamoeba species. A 166-bp PCR product with E. histolytica DNA, a 580-bp product with E. moshkovskii DNA and a 752-bp product with E. dispar DNA were generated in a single-round and multiplex PCR reaction. We recommend this PCR assay as an accurate, rapid, and effective diagnostic method for the detection and discrimination of these three Entamoeba species in both routine diagnosis of amoebiasis and epidemiological surveys

Potential treatment of inflammatory bowel disease: a review of helminths therapy

An inflammatory bowel disease [IBD] is most common in highly industrialized Western countries but uncommon in less developed areas of the world where helminths are frequent. The hygiene hypothesis proposes that the recent increase in allergic and autoimmune diseases is due to modern highly hygienic life styles and medical conditions. Loss of routine exposure to parasitic helminths, as a result of increasing lifestyle-associated factors, may be one factor leading to the increased disease prevalence. In animal models and clinical trials of IBD, gastrointestinal nematodes colonization suppresses intestinal inflammation through multiple mechanisms including induction of innate and adaptive regulatory circuits. Studies using helminths like Trichuris suis or Necator americanus showed that these helminths are safe and may be effective therapeutic approaches for the control of IBD and other immune diseases. The aim of present review was to exploring the therapeutic use of helminths for the control of IBD

[Expression of a recombinant serine-rich Entamoeba Histolytica protein [SREHP]]

Entamoeba histolytica antigenic markers such as Serine-Rich E. histolytica protein [SREHP] have recently been used for vaccine preparation, genetic diversity studies of Entamoeba histolytica isolates and for differentiation between E. histolytica and E. dispar species. This study was carried out with the aim of expression of a recombinant Serine Rich E. histolytica protein in the laboratory to use it in the ELISA kit. In this study which is an exploration method, an Iranian isolate of Serine-Rich E. histolytica gene which had previously been cloned in bluescript plasmid [pBSc], was cut using BamHI restriction enzyme. After extracting and purification from gel, the SREHP gene was sub cloned into pET32a expression vector. The inserted gene was confirmed with Rosconis solution, PCR and sequencing methods. PCR was performed with the SREHP specific primers as well as pET T7 promoter primer. The cloned gene was also digested with HindIII and BamHI restriction enzymes. Recombinant plasmid was conveyed to competent cell BL21 [DE3]. A colony of the plasmid including SREHP gene was cultivated and induced with IPTG. The result of expressed protein was observed on the SDS-PAGE gel. The SREHP gene was sub cloned into pET32a expression vector. A recombinant plasmid including an inserted SREHP gene was screened and confirmed with quick check method using Ruscoins solution, as well as PCR by special primers [SREHP and universal pET primer], digested with BamHI and HindIII restriction enzymes. Finally an open reading frame of 666 nucleotides from inserted SREHP gene was obtained with the sequencing method. The recombinant protein of Serine-Rich E. histolytica in presence of IPTG was expressed in five hours and the result of expressed protein in the length of 44 KDa was observed on SDS-PAGE gel. SREHP protein was successfully cloned and expressed in this study. However additional studies are recommended for preparation and purification of the SREHP in a large quantity and the using it for the ELISA test