Development and characterization of murine monoclonal antibody specific for the P1,4 Por A proteins from strain B: 4: P1,(7b),4, of Neisseria meningitidis / s. t

Neisseria meningitidis isolates are conventionally classified by serosubtyping that characterizes the reactivities of the PorA outer membrane protein variable-region epitopes with monoclonal antibodies. Porins are outer membrane proteins (OMPs) of N. meningitidis serogroup B and have attracted study principally for two reasons: their use in the classification of meningococcal isolates into serotype and subtype and as potential components of vaccines against this important pathogen. New murine hybridomas, secreting specific monoclonal antibodies against PorA serotype P1.4 of N. meningitidis serogroup B, were generated using conventional hybridoma procedures. The monoclonal antibodies obtained were characterized by Western blot and whole cell ELISA, using reference strains from different N. meningitidis serotypes and subtypes. All monoclonal antibodies belong to isotype IgG1. Others hybridomas producing MAbs against PorB and FrpB were also obtained(AU)

Los aislamientos de Neisseria meningitidis se clasifican convencionalmente por serosubtipos. Su reactividad se realiza entre el epítope de la región variable de la proteína de membrana externa PorA con anticuerpos monoclonales. Las porinas, proteínas de membrana externa de N. meningitidis del serogrupo B, son atractivas para su estudio principalmente por la clasificación en serotipo y subtipo de los aislamientos del meningococo y como posibles componentes de vacunas contra este importante agente patógeno. Se generaron nuevos hibridomas murinos secretores de anticuerpos monoclonales específicos contra la proteína PorA subtipo P1.4 de N. meningitidis del serogrupo B, mediante los procedimientos convencionales de hibridomas. Los anticuerpos monoclonales, pertenecientes al isotipo IgG1, fueron caracterizados mediante Western blot y ELISA de células enteras. Se utilizaron cepas de referencia de diferentes serotipos y subtipos de N. meningitidis y se obtuvieron hibridomas productores de anticuerpos monoclonales contra otras proteínas como PorB y FrpB(AU)

Clonal Distribution of Disease-Associated and Healthy Carrier Isolates of Neisseria meningitidis between 1983 and 2005 in Cuba

In response to epidemic levels of serogroup B meningococcal disease in Cuba during the 1980s, the VA-MENGOC-BC vaccine was developed and introduced into the National Infant Immunization Program in 1991. Since then the incidence of meningococcal disease in Cuba has returned to the low levels recorded before the epidemic. A total of 420 Neisseria meningitidis strains collected between 1983 and 2005 in Cuba were analyzed by multilocus sequence typing (MLST). The set of strains comprised 167 isolated from disease cases and 253 obtained from healthy carriers. By MLST analysis, 63 sequence types (STs) were identified, and 32 of these were reported to be a new ST. The Cuban isolates were associated with 12 clonal complexes; and the most common were ST-32 (246 isolates), ST-53 (86 isolates), and ST-41/44 (36 isolates). This study also showed that the application of VA-MENGOC-BC, the Cuban serogroup B and C vaccine, reduced the frequency and diversity of hypervirulent clonal complexes ST-32 (vaccine serogroup B type-strain) and ST-41/44 and also affected other lineages. Lineages ST-8 and ST-11 were no longer found during the postvaccination period. The vaccine also affected the genetic composition of the carrier-associated meningococcal isolates. The number of carrier isolates belonging to hypervirulent lineages decreased significantly after vaccination, and ST-53, a sequence type common in carriers, became the predominant ST(AU)

En respuesta a niveles de epidemia de meningitis meningocócica B en Cuba durante la década de 1980, la vacuna VA-MENGOC-BC se ha desarrollado e introducido en el Programa Nacional de Vacunación Infantil en 1991. Desde entonces, la incidencia de enfermedad meningocócica en Cuba ha vuelto a los bajos niveles registrados antes de la epidemia. Un total de 420 cepas de Neisseria meningitidis recogidos entre 1983 y 2005 en Cuba se analizaron escribiendo la secuencia multilocus (MLST). El conjunto compuesto por 167 de las cepas aisladas de casos de la enfermedad y 253 obtenidos a partir de portadores sanos. Mediante el análisis MLST, 63 tipos de secuencias (ST) fueron identificados, y 32 de estos fueron reportados a ser un nuevo ST. Se aislaron y se asociaron con 12 complejos clonales, y los más frecuentes fueron (36 aislamientos) ST-32 (246 aislamientos), ST-53 (86 aislamientos), y ST-41/44. Este estudio también mostró que la aplicación de VA-MENGOC-BC, el cubano al serogrupo B y la vacuna C, la reducción de la frecuencia y la diversidad de hipervirulenta complejos clonales ST-32 (la vacuna contra el serogrupo B de tipo tensión) y ST-41/44 y también afectados otros linajes. Linajes ST-8 y ST 11-ya no se encuentran en el período post-vacunación. La vacuna también se ve afectada por composición genética de la compañía meningocócica asociada a los aislados. El número de la compañía aislados pertenecientes a linajes hipervirulenta disminuyeron significativamente después de la vacunación, y ST-53, un tipo de secuencia común en las compañías, se convirtió en el ST predominante(AU)

The genetic structure of Neisseria meningitidis populations in Cuba before and after the introduction of a serogroup BC vaccine.

We investigated the population genetics in collections of meningococci sampled in Cuba during the period 1983-2005, thereby covering a period before and after the introduction of an antimeningococcal B-C vaccine. A total of 163 case isolates and 210 isolates from healthy carriers were characterized by multilocus sequence typing (MLST) and sequence determination of porA, porB and fetA genes. A total of 56 sequence types (STs) including 28 new STs were identified among these isolates. The analysis of surface antigens revealed variants 3-1 and 3-8 to be prevalent for porB; variant F5-1 was the most common FetA epitope, and variants 19 and 15 corresponded to the prevalent variable regions 1 (VR1) and VR2 PorA epitopes, respectively. The strongest associations between specific surface protein variants and clonal complexes were detected in lineages ST-32 and ST-53. All ST-32 complex isolates possessed porB3 alleles, and the most frequent antigen combination among ST-32 complex isolates was P1.19,15;F5-1. Variants PorB3-64 at PorB and P1.30 at PorA VR2, in combination with the PorA VR1 variants P1.12-1, P1.7 and P1.7-2 as well as the FetA variants F1-2 and F1-7, dominated the ST-53 complex organisms. Furthermore, we observed a statistically significant association between the most frequent porA, porB and fetA alleles and strain invasiveness. Finally, this study showed that the application of VA-MENGOC-BC((R)), the Cuban antimeningococcal vaccine, reduced the number and frequency of the hypervirulent Clonal Complexes ST-32 and ST-41/44, and also impacted on other lineages. The vaccine also affected the genetic composition of the carrier-associated meningococcal isolates. The number of carrier isolates belonging to hypervirulent lineages decreased significantly after vaccination, and ST-53, a sequence type common in carriers, became the predominant ST.

Clonal distribution of disease-associated and healthy carrier isolates of Neisseria meningitidis between 1983 and 2005 in Cuba.

In response to epidemic levels of serogroup B meningococcal disease in Cuba during the 1980s, the VA-MENGOC-BC vaccine was developed and introduced into the National Infant Immunization Program in 1991. Since then the incidence of meningococcal disease in Cuba has returned to the low levels recorded before the epidemic. A total of 420 Neisseria meningitidis strains collected between 1983 and 2005 in Cuba were analyzed by multilocus sequence typing (MLST). The set of strains comprised 167 isolated from disease cases and 253 obtained from healthy carriers. By MLST analysis, 63 sequence types (STs) were identified, and 32 of these were reported to be a new ST. The Cuban isolates were associated with 12 clonal complexes; and the most common were ST-32 (246 isolates), ST-53 (86 isolates), and ST-41/44 (36 isolates). This study also showed that the application of VA-MENGOC-BC, the Cuban serogroup B and C vaccine, reduced the frequency and diversity of hypervirulent clonal complexes ST-32 (vaccine serogroup B type-strain) and ST-41/44 and also affected other lineages. Lineages ST-8 and ST-11 were no longer found during the postvaccination period. The vaccine also affected the genetic composition of the carrier-associated meningococcal isolates. The number of carrier isolates belonging to hypervirulent lineages decreased significantly after vaccination, and ST-53, a sequence type common in carriers, became the predominant ST.

Proteomic study via a non-gel based approach of meningococcal outer membrane vesicle vaccine obtained from strain CU385: a road map for discovering new antigens.

This work presents the results from a study of the protein composition of outer membrane vesicles from VA-MENGOC-BC (Finlay Institute, Cuba), an available vaccine against serogroup B Neisseria meningitidis. Proteins were identified by means of SCAPE, a 2DE-free method for proteome studies. More than one hundred proteins were detected by tandem liquid chromatographymass spectrometry analysis of fractions enriched in peptides devoid of histidine or arginine residues, providing a detailed description of the vaccine. A bioinformatic analysis of the identified components resulted in the identification of 31 outer membrane proteins and three conserved hypothetical proteins, allowing the cloning, expression, purification and immunological study of two of them (NMB0088 and NMB1796) as new antigens.

Comparison of phenotypically indistinguishable but geographically distinct Neisseria meningitidis Group B isolates in a serum bactericidal antibody assay.

The "gold standard" assay for measuring serologic protection against Neisseria meningitidis group B (MenB) is the serum bactericidal antibody (SBA) assay. Of vital importance to the outcome of the SBA assay is the choice of the target strain(s), which is often chosen on the basis of phenotype or genotype. We therefore investigated the effect on the results produced by the SBA assay of using phenotypically indistinguishable but geographically distinct MenB isolates. Nine PorA P1.19,15 and 11 PorA P1.7-2,4 MenB isolates were incorporated into the SBA assay using human complement and were assayed against sera obtained either before or after outer membrane vesicle vaccination. Large differences in the results produced by the isolates in the SBA assay were demonstrated. These included differences as great as 5.8-fold in SBA geometric mean titers and in the proportions of subjects with SBA titers of >/=4. Ranges of as many as 9 SBA titers were achieved by individual sera across the panels of isolates. To determine the reasons for the differences observed, investigations into the expression of capsular polysaccharide, PorA, PorB, Opc, and lipooligosaccharide (LOS) and into LOS sialylation were completed. However, minor differences were found between strains, indicating similar expression and no antigen masking. These results have implications for the choice of MenB target strains for inclusion in future studies of MenB vaccines and highlight the requirement for standardization of target strains between laboratories.

New meningococcal C polysaccharide-tetanus toxoid conjugate Physico-chemical and immunological characterization.

The polysaccharides (Ps) are thymus-independent 2 (TI-2) antigens and poor immunogens in infants and young children; as a result of this delayed response to Ps antigens during ontogeny, infants and young children are highly susceptible to infections caused by encapsulated bacteria. Meningococcal group C polysaccharide (PsC)-proteins conjugate vaccines have been reported to induce significant serum IgG antibodies and immunologic memory in infants resulting in very effective vaccines. We describe here the obtainment, by a new method, of a neoglycoconjugate intended to immunize against Neisseria meningitidis serogroup C, its characterization by physico-chemical methods, including (1)H NMR and fluorescence spectroscopy methods, as well as the characterization of the immune response induced in mice by such conjugate. Amine groups generated by basic hydrolysis in the PsC were successfully conjugated to carboxyl groups of tetanus toxoid (TT), using carbodiimide-mediated coupling. The specific anti-Ps IgG and anti-Ps IgG subclasses (IgG1 and IgG2a) were measured by ELISA methods, the bactericidal activity in sera and the cytokines response (IFNgamma or IL5) in spleen cell of mice immunized with conjugated and native Ps were evaluated. The (1)H NMR spectra and the result obtained by the fluorescence spectroscopy method showed that the PsC and TT maintained structural identity after conjugation process. Conjugated PsC elicited an increase of anti-PsC IgG responses, anti-PsC IgG subclass (IgG1, IgG2a), an eight-fold increase in bactericidal activity in sera of mice immunized with conjugate compared with native PsC, was also observed. Higher titres of IFNgamma were observed in mice immunized with conjugated Ps. These results indicated that, the PsC and TT maintained its chemical and antigenic structure after the conjugation process. A change in the immunological pattern of responses of PsC, from TI-2 to a thymus-dependent (TD) pattern, was also demonstrated.

Immunogenicity and safety of three doses of a bivalent (B:4:p1.19,15 and B:4:p1.7-2,4) meningococcal outer membrane vesicle vaccine in healthy adolescents.

An experimental bivalent meningococcal outer membrane vesicle (OMV) vaccine (B:4:P1.19,15 and B:4:P1.7-2,4) has been developed to provide wide vaccine coverage particularly of the circulating strains in Europe. A randomized, controlled phase II study (study identification number, 710158/002; ClinicalTrials.gov identifier number, NCT00137917) to evaluate the immunogenicity and safety of three doses of the OMV vaccine when given to healthy 12- to 18-year-olds on a 0-2-4 month (n = 162) or 0-1-6 month schedule (n = 159). A control group received two doses of hepatitis A and one of conjugated meningococcal serogroup C vaccine on a 0-1-6 month schedule (n = 157). Immune response, defined as a fourfold increase in serum bactericidal titer using a range of vaccine-homologous or PorA-related and heterologous strains, was determined for samples taken before and 1 month after vaccination; assays were performed at two laboratories. As measured at the GlaxoSmithKline (GSK) laboratory, the OMV vaccine induced an immune response against homologous or PorA-related strains (in at least 51% of subjects against strains of serosubtype P1.19,15 and at least 66% against strains of serosubtype P1.7-2,4) and against a set of three heterologous strains (in 28% to 46% of subjects). Both laboratories showed consistent results for immune response rates. The OMV vaccine had a similar reactogenicity profile for each schedule. Pain preventing normal activities occurred in approximately one-fifth of the subjects; this was significantly higher than in the control group. The immune responses induced by the bivalent OMV vaccine demonstrated the induction of bactericidal antibodies against the vaccine-homologous/PorA-related strains but also against heterologous strains, indicating the presence of protective antigens in OMVs and confirming the potential of clinical cross-protection.

Cuban Meningococcal BC Vaccine: Experiences & Contributions from 20 Years of Application.

This paper reviews 20 years of experience and scientific contributions of the Cuban meningococcal BC vaccine (VA-MENGOC-BC®) obtained by the Finlay Institute in Havana, Cuba. The vaccine is the first of its type in the world that is safe, effective, and commercially available for preventing meningococcal disease caused by serogroup B meningococcus; it is also effective against serogroup C. VA-MENGOC-BC® has shown satisfactory results, with no serious adverse events, after application of approximately 55 million doses in some 15 countries. Also included is background information on meningococcal disease, as well as the main characteristics of VA-MENGOC-BC®, the strategy used for controlling meningococcal disease and its prevention in Cuba, and a summary of the main scientific results obtained in basic research, development, clinical evaluation, and post-marketing results (safety, efficacy-effectiveness, post-vaccination adverse events, etc.) in Cuba and elsewhere.

Respuesta de anticuerpos inducidos por la vacuna antimeningocócica cubana VA-MENGOC-BC frente a la cepa de Neisseria meningitidisB: 4: P1. 19, 15 en adolescentes después de 12 años de inmunizados / Response of antibodies induced by the Cuban meningococcal vaccine VA-MENGOC-BC against the

Se estudió la respuesta de anticuerpos inducida por la vacuna antimeningocócica cubana VA-MENGOC-BC® contra la cepa de Neisseria meningitidis B:4:P1.19,15 mediante el Ensayo Bactericida del Suero (EBS) y ELISA de tipo indirecto, para medir anticuerpos contra vesículas de membrana externa (VME) de N. meningitidis B a 184 adolescentes de un politécnico de Ciego de Ávila que fueron inmunizados en campañas masivas 12 años antes. Se realizaron extracciones de sangre antes de aplicar la primera dosis (T0), 4 semanas después de ésta (T1) y 4 semanas después de la segunda dosis (T2). Transcurridos 12 años de esta vacunación el 42por ciento de los adolescentes presentó títulos bactericidas ≥ 1:4 frente a la cepa homóloga (B:4:P1.19,15) y el 98por ciento mostró anticuerpos detectables contra las VME. En el EBS; el porcentaje de seroconversión T1/T0 fue del 57por ciento y T2/T0 del 60por ciento. Mediante ELISA la seroconversión fue del 59por ciento y 54por ciento, respectivamente, por lo que se demostró que la aplicación de una sola dosis después de 12 años indujo una respuesta inmune importante que puede sugerir una respuesta anamnésica(AU)

Mucosal immune responses to meningococcal C polysaccharide-protein conjugate in mice.

Mucosal delivery of vaccines represents an attractive approach because this is a region of first contact point for inhaled antigens. We have obtained a meningococcal group C polysaccharide-tetanus toxoid conjugate (MGCP-TT) and evaluated it for intranasal route in mice. The conjugate was obtained by a developed method in our laboratory. The specific IgA in saliva and specific IgA and IgG in serum were measured by ELISA methods and bactericidal antibodies in sera against a meningococcal group C strain were measured. The conjugated elicited a significant increase in anti-MGCP salivary IgA and serum IgG and bactericidal antibodies concentrations, while specific serum IgA was not observed. These results indicated that after conjugation, there was a change in the responses for MGCP from thymus-independent to thymus-dependent and that it was effective by intranasal route.

Outer membrane vesicles of the VA-MENGOC-BC vaccine against serogroup B of Neisseria meningitidis: Analysis of protein components by two-dimensional gel electrophoresis and mass spectrometry.

Neisseria meningitidis is a Gram-negative bacterium responsible for significant mortality worldwide. While effective polysaccharides-based vaccines exist against serogroups A, C, W135, and Y, no similar vaccine is suitable for children under 4 years against disease caused by serogroup B strains. Therefore, major vaccine efforts against this serogroup are based on outer membrane vesicles (OMVs), containing major outer membrane proteins. The OMV-based vaccine produced by the Finlay Institute in Cuba (VA-MENGOC-BC) contributed to the rapid decline of the epidemic in this Caribbean island. While the content of major proteins in this vaccine has been discussed, no detailed work of an outer membrane proteomic map of this, or any other, commercially available OMV-derived product has been published so far. Since OMVs exhibit a large bias toward a few major proteins and usually contain a high content of lipids, establishing the adequate conditions for high resolution, 2-DE of this kind of preparation was definitely a technical challenge. In this work, 2-DE and MS have been used to generate a proteomic map of this product, detailing the presence of 31 different proteins, and it allows the identification of new putative protective protein components it contains.

Inmunogenicidad inducida por la vacuna antimeningocócica VA-MENGOC-BC® contra la cepa de N. meningitidis ATCC C11 en adolescentes después de 12 años de vacunados / Immunogenecity induced by the VA-MENGOC-BC® antimeningococcal vaccine against the ATCC C11 N. meningitidis strain in adolescents 12 years after being vaccinated

Se estudió la respuesta de anticuerpos inducida por la vacuna antimeningocócica cubana VA-MENGOC-BC® contra la cepa ATCC C11 mediante Ensayo Bactericida del Suero y ELISA, a 184 adolescentes de un Politécnico de Ciego de Ávila, que habían sido inmunizados en campañas masivas 12 años antes. Se realizaron extracciones de sangre antes de aplicar la primera dosis (T0), 4 semanas después de esta (T1) y 4 semanas después de la segunda dosis (T2). Después de 12 años de la vacunación, 25 por ciento de los adolescentes presentó títulos bactericidas ≥ 1:8 frente a la cepa evaluada. Por ELISA, 78 por ciento mostró una concentración de anticuerpos superior al límite de detección contra el polisacárido capsular del meningococo C. Los porcentajes de seroconversión posterior a la primera dosis fueron 59 por Ensayo Bactericida del Suero y 82 por ELISA. No hubo diferencias significativas (p> 0,05) entre los resultados obtenidos después de la primera y segunda dosis por ambos ensayos. La reinmunización con 2 dosis de la vacuna no provocó hiporrespuesta frente a la cepa ATCC C11 en este grupo de edad(AU)

Inmunogenicidad inducida por la vacuna antimeningocócica VA-MENGOC-BC® contra la cepa de N. meningitidis ATCC C11 en adolescentes después de 12 años de vacunados / Immunogenecity induced by the VA-MENGOC-BC® antimeningococcal vaccine against the ATCC C11 N. meningitidis strain in adolescents 12 years after being vaccinated

Se estudió la respuesta de anticuerpos inducida por la vacuna antimeningocócica cubana VA-MENGOC-BC® contra la cepa ATCC C11 mediante Ensayo Bactericida del Suero y ELISA, a 184 adolescentes de un Politécnico de Ciego de Ávila, que habían sido inmunizados en campañas masivas 12 años antes. Se realizaron extracciones de sangre antes de aplicar la primera dosis (T0), 4 semanas después de esta (T1) y 4 semanas después de la segunda dosis (T2). Después de 12 años de la vacunación, 25 por ciento de los adolescentes presentó títulos bactericidas ≥ 1:8 frente a la cepa evaluada. Por ELISA, 78 por ciento mostró una concentración de anticuerpos superior al límite de detección contra el polisacárido capsular del meningococo C. Los porcentajes de seroconversión posterior a la primera dosis fueron 59 por Ensayo Bactericida del Suero y 82 por ELISA. No hubo diferencias significativas (p> 0,05) entre los resultados obtenidos después de la primera y segunda dosis por ambos ensayos. La reinmunización con 2 dosis de la vacuna no provocó hiporrespuesta frente a la cepa ATCC C11 en este grupo de edad

Sistema de Lotes de Siembra de la cepa vacunal Vibrio cholerae 638

En la producción de biológicos, especialmente en aquellos obtenidos a partir de microorganismos, resulta imprescindible contar con un suministro de células caracterizado y estable, como punto de partida para la producción. El concepto de Sistema de Lotes de Siembra en dos niveles, en el cual el Lote de Siembra de Referencia es usado para generar el Lote de Siembra de Trabajo, es considerado el enfoque más práctico para lograr este propósito. En este trabajo se elaboraron los Lotes de Siembra de Referencia y de Trabajo de la cepa vacunal Vibrio cholerae 638, primera etapa de la fabricación de una vacuna contra el cólera, cumpliendo con los requerimientos de Buenas Prácticas de Producción (BPP), lo que constituye una exigencia de las autoridadesregulatorias nacionales e internacionales. La caracterización de los lotes demostró que se lograron adecuados niveles de viabilidad y se conservaron inalterables las características de la cepa original en cuanto a identidad, pureza, atenuación de su virulencia, capacidad de colonizar el intestino delgado de ratones lactantes y de inducir una respuesta inmune(AU)

[Immunogenecity induced by the VA-MENGOC-BC antimeningococcal vaccine against the ATCC C11. N. meningitidis strain in adolescents 12 years after being vaccinated]. / Inmunogenicidad inducida por la vacuna antimeningocócica VA-MENGOC-BC contra la cepa de N. meningitidis ATCC C11 en adolescentes después de 12 años de vacunados.

The antibodies' response induced by the VA-MENGOC-BC Cuban antimeningococcal vaccine against the ATCC C11 strain was studied by Bactericidal Serum Trial and ELISA among 184 adolescents from a polytechnic, in Ciego de Avila, that had been immnunized in mass campaings 12 years before. Blood samples were taken before administering the first dose (T0), 4 weeks later (T1), and 4 weeks after the second dose (T2). 12 years after vaccination, 25% of the adolescents presented bactericidal titers > or =1:8 against the evaluated strain. 78% showed a concentration of antibodies over the limit of detection against the meningococcus C capsular polyssacharide. The percentages of seroconversion after the first dose were 59 by Bactericidal Serum Trial and 82 by ELISA. There were no significant differences (p > 0.05) between the results obtained after the first and second dose by boths trials. The reimmnunization with 2 doses of the vaccine did not cause hyporesponse against the ATCC C11 strain in this age group.

Portadores de Neisseria meningitidis en niños de una escuela primaria

Se realizó, con la autorización de la Dirección Municipal de Educación, Dirección Municipal de Salud y el consentimiento informado de los padres, un estudio transversal descriptivo en 318 niños de la Escuela "Mártires del Corynthia"; con el propósito de conocer la prevalencia de portadores de meningococo en niños de edad escolar, determinar los marcadores epidemiológicos de las cepas aisladas y establecer la posible relación existente entre el portador y las variables como edad, sexo, antecedente de infección respiratoria aguda, hacinamiento, amigdalectomía, efecto inhibitorio de la flora acompañante y el estado secretor de antígenos ABH en la saliva. A todos, se les tomó exudado nasofaríngeo y una muestra de saliva. Además, los padres llenaron una encuesta donde se indagó sobre los factores de riesgo a investigar. Se detectó 6,9 % de portadores de meningococo y predominaron las cepas NA:NT:P1.NST:L3,7,9. Los factores de riesgo que dieron resultados estadísticamente significativos respecto a la condición de portador de Neisseria meningitidis fueron: edad, antecedente de infección respiratoria aguda y la presencia de Streptococcus pneumoniae y Neisseria lactamica de la flora bacteriana acompañante en la nasofaringe de los niños investigados.

A cross-sectional and descriptive study was conducted among 318 children from the "Mártires del Corynthia" Primary School under the authorization of the Municipal Division of Education and the informed consent of their parents aimed at knowing the prevalence of meningoccoco carriers in school children, determining the epidemiological markers of the isolated strains and establishing the possible relation existing between the carrier and variables, such as age, sex, acute respiratory infection history, hacinamiento, amigdalectomy, inhibitory effect of of the accompanying flora and the secretory state of ABH antigens in saliva. All of them underwent nasopharyngeal exudate and a saliva sample was taken. In adition, the paents were surveyed about the risks factors to be investigated. 6.9 % of meningoccoco carriers were found and the NA:NT:P1:NST:L3,7,9 strains predominated. The risk factors with statistically significant results regarding the condition of carrier Neisseria meningitidis carrier were age, acute respiratory infection history, and the presence of Streptococcus pneumoniae and Neisseria lactamica of the accompanying bacterial flora in the nasopharynx of the children under study.

[Carriers of Neisseria meningitidis among children from a primary school]. / Portadores de Neisseria meningitidis en niños de una escuela primaria.

A cross-sectional and descriptive study was conducted among 318 children from the "Mártires del Corynthia" Primary School under the authorization of the Municipal Division of Education and the informed consent of their parents aimed at knowing the prevalence of meningoccoco carriers in school children, determining the epidemiological markers of the isolated strains and establishing the possible relation existing between the carrier and variables, such as age, sex, acute respiratory infection history, hacinamiento, amigdalectomy, inhibitory effect of of the accompanying flora and the secretory state of ABH antigens in saliva. All of them underwent nasopharyngeal exudate and a saliva sample was taken. In adition, the paents were surveyed about the risks factors to be investigated. 6.9 % of meningoccoco carriers were found and the NA:NT:P1:NST:L3,7,9 strains predominated. The risk factors with statistically significant results regarding the condition of carrier Neisseria meningitidis carrier were age, acute respiratory infection history, and the presence of Streptococcus pneumoniae and Neisseria lactamica of the accompanying bacterial flora in the nasopharynx of the children under study.

Restauración en el Inmunoblotting de proteínas de / Neisseria meningitidis dañadas por calor y agentes / reductores / Restoration in Immunoblotting of Neisseria meningitidis proteins denatured by reducing agents and heat

En este trabajo se utilizaron cinco detergentes para restaurar las proteínas de membrana externas (PME) del meningococo dañadas por el efecto del calor y de agentes reductores utilizados en el Inmunoblotting. La acción de los detergentes fue evaluada en la solución de lavado, en el diluente de la muestra y del conjugado. Las bandas de proteínas, reconocidas por la IgG del suero, fueron identificadas usando un conjugado anti IgGhumana peroxidasa. Los antígenos reconocidos por el control positivo se corresponden con las proteínas P1, P3,P4 y P5 atendiendo a su peso molecular. Además, fueron reconocidas bandas de 80, 70, 24 kDa y otra con pesomayor a 150 kDa. En general el reconocimiento de todas las PME, excepto esta última de alto peso molecular(APM), se vieron favorecidas con la utilización del Tween 20, con el que se logró un incremento del número y la intensidad de las bandas así como la disminución de los fondos con respecto al resto de detergentes evaluados(Empigen BB, Triton X-100, Nonidet NP-40 y CHAPS). El reconocimiento de la proteína de APM (>150 kDa) se vio afectado por la presencia de detergente como el Tween 20 y Empigen BB. Los lavados con Tween 20 constituyeron los pasos más importantes en la renaturalización de los sitios de unión de la IgG a las PME(AU)

Guía para la estandarización de técnicas inmuno-enzimáticas / en ensayos de vacunas / Guide for the standardization of enzyme immunoassays for use in vaccine trials

Se realizó una revisión sobre la estandarización de técnicas inmunoenzimáticas para ensayos preclínicos y clínicos de vacunas, elaborándose una guía práctica. Se analizan los principios a usar y los pasos a seguir para la optimización de un ensayo, incluyendo la preparación de estándares y controles. Se exponen nuestras experiencias(AU)