PubMed HealthSearch

Biomedical subjects

J Kumate

Publications and source records attributed to J Kumate.

At least 19 recordsLinked to original sources

Active protection of mice against Salmonella typhi by immunization with strain-specific porins.

NIH mice were immunized with between 2.5 and 30 micrograms of two highly purified porins, 34 kDa and 36 kDa, isolated from the virulent strain Salmonella typhi 9,12, Vi:d. Of mice immunized with 10 micrograms of porins, 90% were protected against a challenge with up to 500 LD50 (50% lethal doses) of S. typhi 9,12,Vi:d and only 30% protection was observed in mice immunized with the same dose of porins but challenged with the heterologous strain Salmonella typhimurium. These results demonstrate the utility of porins for the induction of a protective status against S. typhi in mice.

Animals

Non-pathogenic Entamoeba histolytica: functional and biochemical characterization of a monoxenic strain.

We have cultured under monoxenic conditions and characterized an Entamoeba histolytica clone, MAV-I CINVESTAV (MAV-I), obtained from feces from an asymptomatic carrier. The clone shows the non-pathogenic E. histolytica zymodeme type I, which did not change through the process of monoxenization. Clone MAV-I was non-pathogenic in both in vivo and in vitro tests, and it did not have a functional 112-kDa adhesin. As far as we know, this is the first non-pathogenic monoxenic strain reported. Clone A (strain HM1:IMSS), a highly virulent clone with pathogenic zymodeme type II, and which has the 112-kDa adhesin, was used as a control. Protein patterns from both clones were almost identical in one-dimensional gels. In two-dimensional gels, differences in high-molecular-weight proteins were detected. Clone MAV-I adhered and phagocytosed only 12% of the red blood cells adhered and phagocytosed by clone A. MAV-I trophozoites did not destroy cell culture monolayers and did not produce hepatic abscesses in hamsters. They also showed deficiency in protease activity. The absence of virulence in clone MAV-I correlated directly with the absence of a functional 112-kDa adhesion, supporting the role that this protein plays in virulence.

Animals

[Obtaining monoclonal antibodies against outer membrane glycoproteins of Entamoeba histolytica].

The goal of this paper was the production of monoclonal antibodies capable of detecting relevant antigens from the surface of Entamoeba histolytica trophozoites, with the purpose of using them as a diagnostic test. The cellular fusion for obtaining the monoclonal antibodies (mAb) was done with spleen cells from BALB/c mice, previously immunized with glycoproteins from the membrane, as well as Sp2/0 cells. The hybridoma supernatants were tested with ELISA, using glycoproteins and lipopeptide phosphoglycans (LPPG) as antigens. Seven hybridomas producing mAb against the glycoproteins were found. Among these, three recognize LPPG. The ability of reacting with the mAb against two molecules disappeared for all the LPPG positive ones when were treated with meta-periodate, and only three reacted against the glycoproteins. All of the mAb were of the Ig M isotypes. They were characterized by Dot blot and Western blot assays. From the results, one may deduce that some mAb recognize as epitopes the polysaccharide portion, and thus infer that they are directed of against the surface and therefore, in the future, could be used with a diagnostic purpose.

Animals

[Seroepidemiology of amebiasis in the northern region of the Republic of Mexico].

The object of paper has been the study of amebic seroepidemiology in the northern region of Mexico, by means of an immunoenzymatic assay in solid phase (ELISA). The proteic antigen was obtained from axenic culture of E. histolytica's trophozoites of strain HM1:IMSS, by means of solubilization with phenoxy polyethoxyethanol (Triton X-100). Standardization of the method was obtained by analysis of 80 sera from patients with both a clinical and radiologic diagnosis of amebic hepatic abscess and 80 sera from healthy patients. The sensitivity reached 95% and the specificity was 90.7%. 15,457 sera from eight Mexican northern states were tested as part of the National Seroepidemiologic Survey. A semiautomatic BIOMEK 1,000 (Beckman Ins.) equipment was used. The positive ratio varied between 2.0% in Southern Baja California and 4.92% in Chihuahua, with an average ratio for the whole group of 3.34%. The first three places, in descending order, are Chihuahua (4.92%), Sinaloa (4.01%) and Northern Baja California (3.78%). The positive man: woman ratio was 0.87:1 and fluctuated between 0.38:1 in Coahuila and 1.76:1 in Nuevo Leon. In the whole sample the positive ratio of men was 3.00% and in women was 3.43%. The antibody profile related to age, is similar to diseases reported to be highly endemic; the percentage of positive sera rose progressively up to the age of 20 years (4.05%), at which a plateau occurred with a slight descending tendency up to 60 years of age (3.54%), after which a clear descent in the positive index is seen. No important variations were observed when comparing groups of ages and sexes. The antibody level seems to show an inverse relationship with the incidence rate of amebiasis.

Adolescent

[Immunochemical differences in the surface polysaccharides obtained from Entamoeba histolytica strain HM1:IMSS and its virulent (C-A) and non-virulent (L-6) clones].

There probably exist many different surface molecular polysaccharides with different compositions of their constituent sugars, in the different zymodemes. This fact might have a repercussion upon the resistance to the lysis of the complement. Seeking to prove this hypothesis, the ideal would be to obtain polysaccharide molecules of type LPFG of pathogenic and non-pathogenic zymodemes, demonstrating that there is a difference in the chemical composition of their polysaccharides. If this were true, a serologic classification, of the different zymodemes, would be possible in a fashion similar to gram-negative bacteria. Our results suggest the existence of structural differences between the polysaccharides of a virulent clone (C-A) and a non-virulent one (L-6).

Animals

[Vaccination against measles. The situation in Mexico and America. Advances in the method of aerosol immunization].

We present general comments on the epidemiology of measles considering the pre-vaccine era as well as the post-vaccine period in which some changes can be observed: the decrease in morbidity and mortality, the extension of the inter-epidemic interval, the increase in the mean age of infection, etc. We make some estimations about the vaccine coverage and the ideal age of immunization for the goal of eradication (assuming a lifelong immunity for the vaccinees). The technical problems in measles immunization are also revised explaining why no continental country has been able to eliminate the disease. We describe the epidemiological situation in North America, Mexico and Latin American countries. Lastly we present the Mexican experience with the inhaled aerosolised vaccine: the studies in Monterrey (Sabin et al, 1982), other investigation in Mexico, D.F. and in the State of Jalisco, as well as the mass campaigns in Aguascalientes in 1988 and in Coahuila and Nuevo León in 1989. We propose it as an effective, harmless, simple, inexpensive and practical method.

Adolescent

Immunoblot detection of class-specific humoral immune response to outer membrane proteins isolated from Salmonella typhi in humans with typhoid fever.

The studies reported here were undertaken to assess the ability of the outer membrane proteins (OMPs) of Salmonella typhi to induce a humoral immune response in humans with typhoid fever. OMPs were isolated with the nonionic detergent Triton X-100 and were found to be contaminated with approximately 4% lipopolysaccharide. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns showed protein bands with molecular size ranges from 17 to 70 kilodaltons; the major groups of proteins were those that correspond to the porins and OmpA of gram-negative bacteria. Rabbit antiserum to OMPs or to S. typhi recognized OMPs after absorption with lipopolysaccharide. Sera from patients with typhoid fever contained immunoglobulin M antibodies which reacted with a protein of 28 kilodaltons and immunoglobulin G antibodies which reacted mainly with the porins, as determined by immunoblotting. These results indicate that the porins are the major immunogenic OMPs from S. typhi and that the immune response induced in the infection could be related to the protective status.

Adult

[Serology of typhoid fever in children. I. Capillary flocculation- agglutination. Its specificity in an asymptomatic population and its utility at a pediatric emergency service].

Using a capillary flocculation technique we evaluated serum samples of asymptomatic children for typhoid fever serology. Thirty one (5.16%) of the 600 serum samples tested were positive for a specificity of 95%. To evaluate the sensitivity of the test, serum samples from 36 children with proven typhoid fever and a similar number of control patients were evaluated. The sensitivity of the capillary flocculation test in this group was 100%. The test compared favorably with the Ruiz Castañeda serologic test for typhoid fever.

Adolescent

[Comparative serology in typhoid fever in children. II. Comparison of a microhemagglutination plate technic with Ruiz-Castañeda surface fixation].

Two lipopolysaccharide antigens that differ by the technique of preparation from S. typhi, one named crude is easily extracted, the other one is water-phenol extracted. Both were adsorbed by papain and cysteine to lamb erythrocyte surfaces, fixed then by glutaraldehyde and used in a plate microhemagglutination test (MHA). The MHA test was done with 30 sera from children with S. typhi blood isolates. The same number of control sera were tested for comparison. The crude LPS antigen showed better sensitivity: 73% versus 53% of the water-phenol extracted one despite a high index correlation (r = 0.88). The latest one was evaluated in a prospective study, using it during a six-month period in an emergency room with S. typhi blood isolated sera. The MHA test was compared with the surface fixation test. Both assays showed similar sensitivity and a specificity of 91 and 100%, respectively.

Adolescent

Protection against Salmonella typhi infection in mice after immunization with outer membrane proteins isolated from Salmonella typhi 9,12,d, Vi.

The current studies were undertaken to assess the ability of the outer membrane proteins (OMPs) of Salmonella typhi to induce protection against challenge with the bacteria in mucin. OMPs were isolated as described by Schnaitman (J. Bacteriol. 108:553-556, 1971) and were found to be contaminated with approximately 4% lipopolysaccharide (LPS). Immunization with as little as 30 micrograms of OMPs conferred 100% protection to mice challenged with up to 1,000 50% lethal doses (LD50) of two strains of S. typhi (9,12,d, Vi and Ty2). In addition, 30% protection against challenge with up to 500 LD50 of Salmonella typhimurium was achieved. Immunization with LPS at doses equivalent to those found in the OMPs was considerably inferior to the OMPs in the induction of an immune status. Moreover, LPS was effective only when the challenge was performed with S. typhi 9,12,d, Vi (40% protection to 100 LD50). An antiserum raised in rabbits reacted mainly against the bands of the molecular weights corresponding to the so-called porins contained in the OMP preparation as shown by Western blotting (immunoblotting). This rabbit antiserum protected 100% of mice against challenge with 100 LD50 of either strain of S. typhi and 80% of mice against challenge with the same LD50 of S. typhimurium. These results indicate the usefulness of OMPs in the induction of active immunity against S. typhi in mice.

Animals