César R. González

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.

Immunogenicity of a Salmonella typhi CVD 908 candidate vaccine strain expressing the major surface protein gp63 of Leishmania mexicana mexicana

Attenuated Salmonella typhi are attractive for use as live vector vaccines to express protozoal antigens and deliver them to the human immune system. The gene encoding the mature form of Leishmania mexicana mexicana gp63 under control of tac promoter was integrated into the delta aroC locus of the chromosome of attenuated delta aroC, delta aroD S. typhi strain CVD 908. After oral immunization of BALB/c mice with two 1 x 10(9) colony forming unit doses given 21 days apart, CVD 908 omega (delta aroC::Ptac-gp63) elicited a broad T cell-mediated immune response against L. m. mexicana gp63 as demonstrated by: (1) lymphoproliferative response to fixed whole L. m. mexicana promastigotes; (2) activation of IL-2 (but not IL-4)-producing lymphocytes; (3) appearance of cytotoxic T cells against mouse mastocytoma cells expressing gp63. This T-cell mediated immune response was associated with significant protection in F1 (BALB/cXC57Bl/6) mice challenged in their footpads with a wild type strain of L. m. mexicana.

Human Cell Mediated Immunity to Porins from Salmonella typhi

The current studies were undertaken to assess the role of the porins and outer membrane proteins (OMP) in the human immune response to Salmonella typhi 9, 12 Vi:d. Experiments were performed to determinate the lymphocyte activation response to porins in individuals who had been vaccinated against typhoid fever. 10 healthy volunteers were studied before and 10 days after oral or subcutaneous immunisation. Five patients with typhoid fever were also studied. Lymphocyte activation was measured by the 3H thymidine incorporation assay. Individuals with typhoid fever as well as those immunised with oral vaccine responded well to porins and outer membrane proteins, as opposed to those immunised with the subcutaneous vaccine. These results suggest that the porins and OMP play a role in the cellular immune response against Salmonella typhi.

Lymphocytic Proliferative Response to Outer‐Membrane Proteins Isolated from Salmonella

Porins isolated from Salmonella typhi have been demonstrated to protect against the challenge with this bacteria in mice. The mechanism has not been clarified, but could be associated with activation of both humoral and cellular immunity. In order to evaluate the induction of specific T cell responses, the lymphocytic proliferation to porins isolated from Salmonella typhimurium, Salmonella typhi and Escherichia coli was examined by 3H‐thymidine incorporation assay in mice immunized with three different antigens: acetone‐killed S. typhimurium, its porins, or outer‐membrane proteins (OMPs) isolated from S. typhi. Higher proliferative responses were observed in mice immunized with porins and OMPs compared with those which received the acetone‐killed bacteria. Although cross‐reactivity was observed between porins, they were not mitogenic. Moreover, porins were able to activate T lymphocytes isolated from mice immunized with S. typhi OMPs. These results suggest that T cell activation, through the release of lymphokines, may play a role in the induction of protective immunity with porins.

Prevalence of antibodies against Entamoeba histolytica in Mexico measured by ELISA

The prevalence of antibodies against Entamoeba histolytica was studied in the Mexican population using an immunoenzyme assay in solid phase (ELISA) and semiautomatic equipment. The antigen was a mixture of membrane proteins obtained by Triton X-100 extraction from an axenic culture of Entamoeba histolytica HM1-IMSS. The method was standardized by comparing serum samples from amoebic liver abscess patients with healthy volunteers. From the 60,538 samples supplied by the National Seroepidemiology Survey, antibodies were found in 4.49% (4.32-4.65% at 95% confidence limit). More significant titres occurred in the central region of the country. The ratio female to male was 1.25:1. The population living in metropolitan areas had probably been infected at a younger age than those living in the country. Important differences were found in the seroprevalence obtained by ELISA compared with a study which used indirect haemagglutination (IHA) in the same sample frame.

The Role of Outer Membrane Proteins From Gram-Negative Bacteria as VACCINES with Special Emphasis in Typhoid Fever: Monoclonal antibodies against S. typhi porins

Gram negative bacteria contain in their outer membrane a group of proteins that have been classified into major and minor according to their level of expression (1). The major members of the outer membrane proteins (OMP) include the Braun’s lipoprotein, the porins (ompC, ompF, ompD, phoE, etc.) and the heat-modifiable protein (Omp A) (2). The porins are a group of proteins with molecular weights (Mr) ranging from 36 to 38 Kd per monomeric unit (3). In the membrane, porins are expressed as non-covalently linked homotrimers. These proteins function in the transmembrane transport of nutrients and ions (4). Moreover, some members of the porin family (lamB) function as receptors for phagi (5). Biochemical and molecular genetic studies have indicated that there is extensive homology among the porins of Gram negative bacteria, although some species-specific sequences have been demonstrated (6,7).