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[Intestinal amebiasis].

Infection produced by Entamoeba histolytica constitutes a clinical problem of great importance due to its frequence and serious complications. Age, diagnosis and clinical manifestations, phisic exploration, lab. procedures, proctosigmoidoscopy, radiology, complications, pathologic anatomy, statistical reports from the Ministry of Health and Social Asistence in addition we emphasize on the importance of an accurate and precocious diagnosis. We expose an effective terapeutic regimen for the different amibiasis states.

Adolescent↗

Intestinal invasive amebiasis: an experimental model in rodents using axenic or monoxenic strains of Entamoeba histolytica.

A new experimental model for the production of early stages of invasive intestinal amebiasis in hamsters or guinea pigs with axenic or monoxenic cultures of Entamoeba histolytica of the HM1:IMSS strain is reported. The model is called the washed-closed cecal loop, because it involves the washing out of the cecum contents of conventionally raised animals and the formation of a closed loop, into which amebas are inoculated. Colonization occurred in all inoculated animals. After 48 hr, macroscopic ulcerations of the cecal mucosa were found in approximately one-half (5/12) of the animals inoculated with axenic amebas and in more than two-thirds (10/12) of those inoculated with monoxenic parasites. The amebic nature of the mucosal ulcerations was confirmed by light and transmission electron microscopy, as well as by scanning electron microscopy (SEM) of fractured samples of cecal mucosa. SEM provided a rapid and reliable technique to assess both the extent and the nature of the lesions. This model is particularly useful for the study of the pathogenesis of early intestinal lesions produced by virulent amebas, and may also be applied to experimental studies on the immunology of invasive intestinal amebiasis.

Amebiasis↗

Amebiasis at an evacuation site on the Thai-Cambodian border.

Symptomatic intestinal amebiasis was highly endemic among the Cambodians living at Green Hill, an evacuation site on the Thai-Cambodian border between June 1987 through May 1989. Monthly incidence rates of intestinal amebiasis were determined to be inversely proportional to cumulative monthly rainfall. The highest incidence of amebic dysentery was 63/1000 in children 12-23 months old. Behavioral risk factors were investigated by conducting a case-control study. A questionnaire was administered to 73 families, each having at least one member with confirmed intestinal amebiasis within the past 3 months, and to 95 randomly selected control families having no individual with diarrhea for at least 3 months. Individuals from families with greater than 4 members were at higher risk for acquiring intestinal amebiasis. No significant differences in behavioral risk factors were identified between case and control families. Eighty-six percent of 51 water samples drawn from wells where amebiasis patients obtained their drinking water had greater than 10 coliforms/100 ml. The main route of transmission of E. histolytica was not identified, but was most likely via the fecal-oral route.

Adolescent↗

Ultrastructure of experimental intestinal invasive amebiasis.

The morphological features of early intestinal ulcerations induced in rodents with axenic cultures of Entamoeba histolytica were studied by scanning and transmission electron microscopy. Amebas did not attach to the luminal surface of the mucosa except at interglandular regions, where parasites penetrated apparently through pseudopodial movement. Once in the lamina propria, trophozoites multiplied and destroyed mucosal components. Damage extended laterally through the mucosa, but progression to deeper layers of the intestinal wall was prevented by the muscularis mucosae, which acted as a partial barrier. This was eventually breached at focal points where amebas invaded the submucosa. Electron microscopy clearly showed the lysis of abundant polymorphonuclear neutrophil leukocytes (PMNs) by the amebas at the periphery of intestinal ulcerations as well as the lack of bacteria at these sites. This study demonstrates that recruitment and destruction of inflammatory cells following intestinal amebic invasion may take place in the absence of bacterial multiplication. The observations provide histological support to the hypothesis that lysis of PMNs by trophozoites participates in the genesis of amebic intestinal lesions.

Animals↗

Differentiation of pathogenic Entamoeba histolytica infections from nonpathogenic infections by detection of galactose-inhibitable adherence protein antigen in sera and feces.

We determined whether epitope-specific monoclonal antibodies to the galactose-inhibitable adherence protein (GIAP) of Entamoeba histolytica could be used in an enzyme-linked immunosorbent assay (ELISA) to detect antigen in serum and feces and differentiate between nonpathogenic zymodemes and the potentially invasive pathogenic organisms that require treatment. Overall, 57% of subjects from Cairo, Egypt, with symptomatic intestinal amebiasis and 42% with asymptomatic infection possessed GIAP antigen in their sera, whereas 4% of uninfected controls or subjects with other parasitic infections possessed GIAP antigen in their sera (P < 0.001). In subjects from Durban, South Africa, only 6% of uninfected controls or those with nonpathogenic E. histolytica infection were positive for GIAP in serum, whereas 3 of 4 with asymptomatic pathogenic intestinal infection and 75% with amebic liver abscess were positive for GIAP in serum. Fifteen stool samples from patients with intestinal amebiasis were available for study; all had a positive ELISA result for fecal GIAP antigen. Epitope-specific monoclonal antibodies identified 8 of 15 subjects with fecal antigen from pathogenic strains. Seven of those eight subjects had adherence protein antigen in their sera, whereas none of seven with apparent nonpathogenic E. histolytica infection had adherence protein antigen in their sera. In summary, we were able to detect E. histolytica adherence protein antigen directly in serum and fecal samples by ELISA. The presence of amebic antigen in serum demonstrated 94% specificity for pathogenic E. histolytica infection, and amebic antigen is present during asymptomatic intestinal infection. In conjunction with antibody detection, this method should be very useful in the diagnosis and management of intestinal amebiasis.

Animals↗

Serology of amebiasis using the FIAX-TM system.

A new fluorescent immunoassay system (FIAX TM) has permitted development of a promising serologic test for amebiasis. The test is similar but far superior to the soluble antigen fluorescent antibody test. The main differences are the unique StiQ TM sampler and the flourometer itself. Quantitative results can be obtained from a single standard dilution of serum. The procedure is simple to perform, gives objective readings, and uses the proven principle of immunofluorescence. It is possible to test for different immunoglobulin classes and to use purified antigen prepared from axenically cultured Entamoeba histolytica. Conservation of reagents is comparable to microplate systems, and results are available in less than two hours. FIAX TM for amebiasis has demonstrated excellent discriminating power between amebic and nonamebic sera. Sensitivity is 100 percent for extra-intestinal amebiasis (33 sera), 90 percent for symptomatic intestinal amebiasis (20 sera), and 43 percent for asymptomatic amebiasis (seven sera). Specificity is 97 percent (158 sera).

Amebiasis↗

Cloning and characterization of Entamoeba histolytica antigens recognized by human secretory IgA antibodies.

To identify the Entamoeba histolytica antigens capable of inducing secretory IgA (sIgA) responses in humans, a cDNA library from the strain HM1:IMSS was immuno-screened with saliva from patients with intestinal amebiasis or amebic liver abscess. Clones isolated with sIgA antibodies from patients with intestinal amebiasis corresponded to the known serine-rich protein isoform, a 29 kDa cysteine-rich protein and 1-alpha elongation factor. Clones corresponding to enolase, cyclophilin, ribosomal protein L23a, and an Hsp70 family protein were isolated with sIgA from a patient with amebic liver abscess. A glutamic acid-rich peptide (EhGARP) positive with sIgA from a patient with amebic liver abscess was also isolated; for EhGARP, no homologs were found in the protein databases. The antigens isolated are potentially useful in the development of an oral vaccine or new diagnostic tools for amebiasis.

Amino Acid Sequence↗

Experimental amebiasis: immunohistochemical study of immune cell populations.

Distribution of immune cell populations was studied in a C3H/HeJ mouse model of intestinal amebiasis from 5 to 60 days post-inoculation with Entamoeba histolytica, using immunoperoxidase techniques. At various time intervals, the ceca from mice were fixed in 10% formalin, dehydrated, embedded and sectioned at 5 microm. Sections were incubated with conjugated peroxidase-labelled antibodies to mouse IgA, IgM, and IgG. Color was developed with 3, 3'-diaminobenzidine tetrahydrochloride (DAB)/H2O2 solution. CD3, CD4, and CD8 cells, as well as neutrophils were detected by reacting with biotin-conjugated anti-mouse CD3, CD4, CD8, and CD11 monoclonal antibodies, followed by their incubation with avidin-peroxidase and color development with DAB/H2O2 solution. Erythrocin B and toluidine blue were used to stain eosinophils and Mast cells, respectively. It was observed that the IgA+ plasma cell was the dominating immune cell present in the mucosa, although eosinophils, neutrophils, CD3+, CD4+, CD8+, IgM+, IgG+ cells and Mast cells were also seen. Results of this study suggest that infiltration of immune cells at the mucosal surface during intestinal amebiasis might be important in the defense against this parasite.

Animals↗