The sequence of a highly homologous gene to hgl2* of Entamoeba histolytica that is present and expressed in E.dispar.
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Animals↗
Biomedical subjects
J I Ravdin
Publications and source records attributed to J I Ravdin.
Humoral and mucosal IgA antibody response to a recombinant 52-kDa cysteine-rich portion of the Entamoeba histolytica galactose-inhibitable lectin correlates with detection of native 170-kDa lectin antigen in serum of patients with amebic colitis.
Humoral and mucosal IgA responses to a recombinant cysteine-rich portion (designated LC3) of the Entamoeba histolytica galactose-inhibitable lectin's 170-kDa subunit were determined in patients with amebic colitis. All patients had 170-kDa amebic antigen in serum, compared with 1 of 50 cyst passers and 1 of 31 controls (P < .01). Seven days after treatment, serum and fecal 170-kDa antigen became undetectable in 12 of the 13 patients (P < .001). Serum anti-LC3 IgA was found in 83.8% of colitis patients, compared with 2% of controls and 12% of asymptomatic cyst passers (P < .001). Salivary and fecal anti-LC3 IgA levels were higher in patients than in cyst passers (P < .001). In conclusion, in amebic colitis, development of humoral and mucosal IgA responses to the recombinant LC3-encoded protein correlates with detection of amebic 170-kDa antigen in serum and feces.
Animals↗
Oral immunization with a recombinant cysteine-rich section of the Entamoeba histolytica galactose-inhibitable lectin elicits an intestinal secretory immunoglobulin A response that has in vitro adherence inhibition activity.
The LC3-encoded 52-kDa recombinant protein includes amino acids 758 to 1134 of the 170-kDa subunit of the galactose-inhibitable lectin. Oral immunization of BALB/c mice with the LC3-encoded protein and cholera holotoxin induced an intestinal secretory immunoglobulin A (IgA) response (P < 0.01 compared with the control). There was a negative correlation (P = 0.001) between intestinal anti-LC3 IgA and serum IgA and IgG antibody responses. Intestinal secretions from immunized mice completely inhibited the galactose-specific adherence of axenic trophozoites ot Chinese hamster ovary cells (P < 0.01).
Adhesiveness↗
Galactose-specific adhesion mechanisms of Entamoeba histolytica: model for study of enteric pathogens.
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Animals↗
Interdisciplinary integration for quality improvement: the Cleveland Veterans Affairs Medical Center Firm System.
BACKGROUND: Many of the characteristics of Firm Systems lend themselves to the application of principles of continuous quality improvement (CQI). A Firm System is defined as two or more parallel practices organized on the principle of continuity of relationships between patients and an interdisciplinary group of health care professionals and trainees. Firm Systems are organized around the care of the patient or customer and emphasize access, continuity, and quality of care. CASE STUDY: The Firm System was implemented at the Cleveland Veterans Affairs Medical Center (VAMC) not as a CQI initiative per se, but as an effort to coordinate the processes involved in the delivery of patient care. The primary goals of this implementation were to improve the quality of patient care, medical education, and health care research. The main strategy to deal with problems caused by uncoordinated care were to move from a departmental approach to an integrated interdisciplinary approach. This approach represented a paradigm shift within the organization that extended to planning, documentation, and the general work environment. Most important, the institution had leaders who were committed to the Firm System and willing to authorize resources to ensure its success. CONCLUSION: VA hospitals are ideal settings for Firm Systems because they provide longitudinal, comprehensive care with a centralized, prepaid payment mechanism, and they have well-developed information systems that allow the random assignment of patients to Firms. Recommendations to others interested in implementing Firm Systems include creation of a written plan that can gain general support; identification of resources needed for successful implementation; remembering that the patient is the most important customer, as well as that complex systems have many customers; monitoring of performance; and the importance of randomizing patients and providers.
Continuity of Patient Care↗
A recombinant cysteine-rich section of the Entamoeba histolytica galactose-inhibitable lectin is efficacious as a subunit vaccine in the gerbil model of amebic liver abscess.
The 170-kDa subunit of the galactose-inhibitable adherence lectin of Entamoeba histolytica mediates attachment to colonic mucins and host cells. The DNA fragment encoding the 170-kDa subunit was produced by polymerase chain reaction (PCR) and divided into four sections by restriction endonucleases. The third section (designated LC3, base pairs 2273-3397) encodes a cysteine-rich fusion protein that was recognized by adherence-inhibitory anti-lectin monoclonal antibodies and serum antibodies from 95% of subjects with amebic liver abscess. Immunization of gerbils with purified recombinant LC3-encoded protein (10 micrograms) with Titermax adjuvant elicited a high-titer serum anti-LC3 IgG antibody response and protective immunity against intrahepatic challenge with 0.5 x 10(6) virulent axenic trophozoites (strain HM1:IMSS; 71% vaccine efficacy, P < .01). In summary, a recombinant cysteine-rich portion of the 170-kDa lectin subunit was highly antigenic, immunogenic, and effective as a subunit vaccine in an experimental animal model of amebic liver abscess.
Animals↗
Protection of gerbils from amebic liver abscess by immunization with recombinant Entamoeba histolytica 29-kilodalton antigen.
The goal of our study was to obtain a highly conserved Entamoeba histolytica recombinant antigen for study as a subunit amebiasis vaccine. We screened a Uni-Zap cDNA library of E. histolytica (strain HM1:IMSS) with human immune sera and isolated a dominant 804-bp cDNA clone. A 33-kDa fusion protein expressed from the cDNA clone was determined by monoclonal antibody binding, DNA hybridization, and nucleotide sequence to be the complete E. histolytica 29-kDa antigen. Serum antibodies to the recombinant protein were detected by enzyme-linked immunosorbent assay in 80% of subjects from Egypt and South Africa with amebic liver abscess. Similar results were found with the native 29-kDa protein. Native and recombinant 29-kDa antigens induced proliferation of lymphocytes harvested from patients with amebic liver abscess (P < 0.01 compared with controls). Intraperitoneal immunization of gerbils with the recombinant fusion protein (10 micrograms) with Titermax adjuvant elicited an antigen-specific serum immunoglobulin G antibody response and was partially protective (54%) against intrahepatic challenge with 5 x 10(5) virulent axenic trophozoites (strain HM1:IMSS). In summary, the recombinant form of the E. histolytica 29-kDa antigen demonstrated serologic specificity for amebic liver abscess, exhibited conserved T-cell epitopes, and was effective as a subunit vaccine in an experimental animal model of amebic liver abscess.
Animals↗
Immunization of rats with the 260-kilodalton Entamoeba histolytica galactose-inhibitable lectin elicits an intestinal secretory immunoglobulin A response that has in vitro adherence-inhibitory activity.
The 260-kDa galactose-inhibitable lectin of Entamoeba histolytica mediates binding of amebic trophozoites to purified colonic mucins and intestinal epithelial cells. Parenteral immunization of Lewis rats with immuno-affinity-purified lectin with Freund's adjuvant and then intra-Peyer's patch inoculation of lectin with cholera toxin B subunit as adjuvant elicited a significant anti-lectin secretory immunoglobulin A response in the bowel lumen. Purified intestinal secretory immunoglobulin A from three or four immunized animals studied possessed inhibitory activity (P < 0.02) in an in vitro assay of trophozoite galactose-specific adherence to Chinese hamster ovary cells.
Animals↗
Development of amebicidal cell-mediated immunity in gerbils (Meriones unguiculatus) immunized with the galactose-inhibitable adherence lectin of Entamoeba histolytica.
The galactose-inhibitable adherence lectin of Entamoeba histolytica is a protective antigen in the gerbil model of amebic liver abscess. To further understand the mechanisms of vaccine efficacy, we studied the cell-mediated immune response to the lectin in gerbils. Splenocytes harvested from immunized gerbils demonstrated in vitro proliferation and production of interleukin-2 and gamma-interferon in response to purified adherence lectin (P < 0.05 for each compared to control splenocytes). Splenocytes from immunized gerbils developed direct amebicidal activity (P = 0.014) following in vitro stimulation with the adherence lectin. Splenocytes harvested from immunized gerbils following intrahepatic challenge with viable E. histolytica adherence lectin. Splenocytes harvested from immunized gerbils following intrahepatic challenge with viable E. histolytica trophozoites demonstrated proliferative and amebicidal responses (P < 0.05). In conclusion, immunization with the E. histolytica galactose-inhibitable adherence lectin elicits an amebicidal cell-mediated response that is enhanced by parasite challenge.
Animals↗
Diagnosis of invasive amoebiasis--time to end the morphology era.
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Animals↗
Secretory immunoglobulin A antibodies to the galactose-inhibitable adherence protein in the saliva of patients with amebic liver disease.
Monoclonal antibodies directed against the 260-kD galactose-inhibitable adherence protein (GIAP) of Entamoeba histolytica inhibit binding of amebic trophozoites to purified colonic mucins, suggesting that anti-GIAP secretory immunoglobulin A (sIgA) may have a role in host defense in invasive amebiasis. We determined by enzyme-linked immunosorbent assay (ELISA) whether a salivary anti-GIAP sIgA response was present in patients from the Republic of South Africa with invasive E. histolytica infection. In 13 patients with amebic liver abscess (ALA), salivary anti-GIAP sIgA was significantly higher (mean +/- SD optical density [OD] = 0.448 +/- 0.258) than that determined for seven South African adult patients hospitalized with nonamebic illness (0.084 +/- 0.072; P = 0.002), seven healthy South African Adults (0.194 +/- 0.119: P = 0.025), and seven healthy adults from Charlottesville, Virginia (0.036 +/- 0.023; P = 0.004). Of the patients with ALA, nine had acute disease, and four had been cured of amebiasis 2-8 months previously. There was no significant difference between these two groups in the anti-GIAP sIgA levels. All ALA patients had a high titer serum anti-amebic antibody response, and there was no direct correlation between the level of anti-GIAP salivary IgA and anti-GIAP serum antibodies (R = 0.187). These findings demonstrate that the E. histolytica GIAP is a mucosal antigen in naturally occurring invasive E. histolytica infection.
Adult↗
Amebiasis: human infection with Entamoeba histolytica.
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Animals↗
Role of mucosal secretory immunity in the development of an amebiasis vaccine.
Invasive colonic infection by the enteric protozoan Entamoeba histolytica elicits a mucosal anti-amebic IgA antibody response. The E. histolytica galactose-inhibitable adherence protein (GIAP) mediates parasite binding to colonic mucins and epithelial cells. Anti-GIAP secretory IgA antibodies are found during invasive amebiasis and can be elicited by immunization of rats with the native protein. Entamoeba histolytica contains potent IgA degradative activities due to its cysteine-specific proteases. Research is needed to further understand the role of secretory antibodies in natural or vaccine-induced immunity to E. histolytica.
Animals↗
Antigenicity, immunogenicity and vaccine efficacy of the galactose-specific adherence protein of Entamoeba histolytica.
Entamoeba histolytica is an enteric protozoan that causes amoebic colitis and liver abscess. Human immunity to E. histolytica is apparently mediated by a serum antibody response and amoebicidal cellular mechanisms. The galactose-specific adherence protein of E. histolytica is a 260 kDa glycoprotein which mediates amoebic in vitro adherence to human colonic mucins, epithelium, and inflammatory cells. Amoebic lysis of cells is dependent upon binding by this adherence protein. Serum IgG and salivary IgA antibodies from greater than 90% of subjects with invasive amoebiasis recognize the adherence protein's 170 kDa heavy subunit. Incubation of peripheral blood mononuclear cells from antibody-positive subjects with the purified galactose-specific adherence protein induces in vitro T lymphocyte proliferation, IL-2 and gamma interferon production, and direct lymphocyte amoebicidal activity. The rationale for an adherence protein vaccine includes sIgA blockage of amoebic binding to colonic mucins, humoral IgG prevention of parasitic adherence in tissues, and development of amoebicidal cell-mediated immunity. Immunization of gerbils with purified adherence protein in Freund's adjuvant provides protection against intrahepatic challenge with the trophozoites. In summary, the galactose specific adherence protein of E. histolytica contains highly conserved B- and T-cell epitopes, and has a high degree of vaccine efficacy in the gerbil model of amoebic liver abscess.
Animals↗
Degradation of human IgA by Entamoeba histolytica.
To determine whether the virulent enteric pathogen Entamoeba histolytica degrades human IgA molecules, serum and secretory IgA was exposed to viable axenic trophozoites (strain HM1:IMSS), a parasite sonicate, and medium conditioned by incubation with live trophozoites. IgA was completely degraded under all conditions, proteinase activity was maximal at a neutral pH, and there was a four- to eightfold enrichment of amebic IgA proteolytic activity in a soluble fraction of amebic sonicate. Degradation of serum IgA by amebic sonicate was completely inhibited by the cysteine proteinase inhibitors trans-epoxysuccinyl-L-leucyl-amino(4-guanidino)butane (E-64, 100 microM) and benzyloxycarbonyl-phenyl-alanyl-alanyl-fluoromethyl (Z-Phe-Ala-CH2F, 12.5 microM). Secretion of degradative activity, the optimal pH, and the inhibition by E-64 and Z-Phe-Ala-CH2F indicates that cysteine proteinase activity is predominantly responsible for the degradation of human IgA by E. histolytica.
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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↗
Serologic reactivity to purified recombinant and native 29-kilodalton peripheral membrane protein of pathogenic Entamoeba histolytica.
The 29-kDa peripheral membrane protein of Entamoeba histolytica has recently been demonstrated to have epitopes on pathogenic clinical isolates which were not detected by monoclonal antibodies on nonpathogenic isolates. To analyze the serological response to this protein, we tested 93 serum specimens (from 33 patients with amebic liver abscess, 7 patients with colitis, 2 patients with ameboma, 18 individuals harboring a nonpathogenic zymodeme strain, 10 healthy Mexican migrant workers, and 23 healthy controls) by enzyme-linked immunosorbent assay (ELISA) using immunoaffinity-purified native or recombinant protein. When tested by ELISA with the native antigen, 79% (26 of 33) of the serum specimens from patients with amebic liver abscess, 4 of 9 serum specimens from symptomatic patients with colitis or ameboma, and serum from one migrant worker were positive. None of the 18 subjects harboring a nonpathogenic strain or 23 control individuals were seropositive to the native antigen (sensitivity, 71%; specificity, 98%). Of 30 serum specimens from patients with amebic liver abscess tested with recombinant antigen, 27 were seropositive (90%). In addition, six patients with colitis or ameboma and two individuals who harbored a nonpathogenic strain were seropositive to the recombinant antigen. One healthy Mexican migrant worker tested positive by both ELISAs (sensitivity, 87%; specificity, 94%). Immunoblotting of 51 serum specimens to sodium dodecyl sulfate-denatured native 29-kDa protein was less sensitive (65%) than ELISA in detecting serum antibodies to the antigen. These results suggest a similar antibody response to native and recombinant antigens (r = 0.86) and support the potential utility of a quantitative assay with defined recombinant antigen for the serodiagnosis of invasive amebiasis in nonendemic areas in conjunction with other diagnostic tools.
Amebiasis↗