RESUMO
Murine hybridoma monoclonal antibodies (MAbs) were produced against the capsular polysaccharide (CPs) of serogroups A, C, W135 and Y meningococci (MenA, MenC, MenW, MenY) in order to develop immunological reagents for the identification of meningococcal polysaccharides. Each serogroup-specific MAb reacted with the CPs from its homologous serogroup only and did not react with CPs from the other three serogroups. The affinity constant (Ka) of the four MAbs measured by non-competitive ELISA was 6.62 × 10(9), 2.76 × 10(9), 1.48 × 10(9) and 3.8 × 10(9) M(-1) for MenA, MenC, MenW and MenY MAbs respectively. The application of these MAbs for identity tests was demonstrated by their abilities to correctly identify the CPs from serogroups A, C, W135 and Y in meningococcal CPs-based vaccines through ELISA. The MAbs obtained in this work are a very valuable set of tools for study meningococcal polysaccharides vaccines.
Assuntos
Anticorpos Antibacterianos , Anticorpos Monoclonais Murinos , Cápsulas Bacterianas , Neisseria meningitidis Sorogrupo A , Neisseria meningitidis Sorogrupo C , Neisseria meningitidis Sorogrupo W-135 , Neisseria meningitidis Sorogrupo Y , Polissacarídeos Bacterianos , Animais , Anticorpos Antibacterianos/química , Anticorpos Antibacterianos/imunologia , Anticorpos Monoclonais Murinos/química , Anticorpos Monoclonais Murinos/imunologia , Especificidade de Anticorpos , Cápsulas Bacterianas/química , Cápsulas Bacterianas/metabolismo , Ensaio de Imunoadsorção Enzimática/métodos , Camundongos , Neisseria meningitidis Sorogrupo A/química , Neisseria meningitidis Sorogrupo A/imunologia , Neisseria meningitidis Sorogrupo C/química , Neisseria meningitidis Sorogrupo C/imunologia , Neisseria meningitidis Sorogrupo W-135/química , Neisseria meningitidis Sorogrupo W-135/imunologia , Neisseria meningitidis Sorogrupo Y/química , Neisseria meningitidis Sorogrupo Y/imunologia , Polissacarídeos Bacterianos/química , Polissacarídeos Bacterianos/imunologiaRESUMO
BACKGROUND: Dengue is the most important human viral disease transmitted by arthropod vectors. The availability of random peptide libraries (RPL) displayed on phage has provided a powerful tool for selecting sequences that mimic epitopes from microorganisms that are useful for diagnostic and vaccine development purposes. In this paper, we describe peptides that resemble the antigenic structure of B-cell epitopes of dengue virus identified from a phage-peptide library using human sera containing polyclonal antibodies against dengue virus. MATERIALS AND METHODS: Eighteen phage clones were isolated from the phage-display peptide library, J404, by affinity selection using human antisera against dengue virus type 3. These clones were tested for reactivity by ELISA with a panel of hyperimmune ascitic fluids (HAFs) containing antibodies either against all four dengue serotypes, West Nile virus (WNV) or Eastern equine encephalitis virus (EEEV) with control ascitic fluid (NAF) used as a negative control. RESULTS: Eight clones were recognized by HAFs against the four dengue serotypes, of which four significantly inhibited binding of anti-dengue antibodies to the virus. Two peptides with similar sequences to regions of NS3 and NS4B non-structural dengue virus proteins were identified. CONCLUSION: Our results suggest that these peptides could be used for the development of diagnostic tools for the detection of dengue virus infection and for a potential vaccine against this pathogen.
RESUMO
Se describieron los diferentes comportamientos de anticuerpos monoclonales anti-dengue serotipo 2 (H3/6, 4G3) y anti-proteína recombinante de la envoltura del virus dengue cuando fueron enfrentados a diferentes cepas de los serotipos 2 y 4 de este virus (D2 Nueva Guinea C, A15 cepa cubana y A15 propagada 53 veces en cultivo de riñón de perro Beagly y D4 H-2412 cepa prototipo) en un ensayo de inmunoamplificación dependiente de anticuerpos. Los anticuerpos monoclonales han demostrado ser una herramienta eficaz para explicar que la neutralización y el incremento de la multiplicidad viral pueden realizarse como funciones biológicas separadas. Solamente el AcM H3/6 fue capaz de producir el fenómeno amplificación dependiente de anticuerpos frente a la cepa A15 con un incremento significativo en la multiplicación viral. Los AcMs 4G3 y 4B6 no fueron capaces de inmunoamplificar la multiplicación viral de las cepas estudiadas.
The different behaviors of antidengue monoclonal antibodies serotype 2 (H3/6, 4G3) and antiprotein recombinant from the dengue virus sheath were described when they faced diverse strains from the serotypes 2 and 4 of this virus (D2 New Guinea C, A15 Cuban strain and A15 propagated 53 times in culture of Beagley dog kidney and D4 H-22412 prototype strain) in an immunoamplification assay dependent of antibodies. Themonoclonal antibodies have prove to be an efficient tool to explain that the neutralizatrion and increase of viral multiplicity may be carried out as separate biological functions. Only the H3/6 monoclonal antibdoy was capable of producing the amplification assay dependent phenomenon against the A15 strain with a significant rise in the viral multiplication. The 4G3 and 4B6 monoclonal antibodies were not capable of immunoamplifying the viral multiplication of the studied strains.
RESUMO
Se describen los resultados del estudio de Enterovirus como agentes productores de meningoencefalitis viral, desde 1990 hasta 1994. Fueron estudiados en este periodo 546 muestras de heces fecales, 95 liquidos cefalorraquideos y 1 058 sueros pareados, obtenidos de 1 388 pacientes diagnosticados clinicamente con esta enfermedad. Las muestras para aislamiento viral se inocularon en 2 sistemas celulares diferentes, el mayor numero de aislamiento se encontro en celulas diploides de fibroblasto humano. Las determinaciones de anticuerpos se realizaron por prueba de neutralizacion (micrometodo) con 11 antigenos de Enterovirus (Echo 4,6,9,11 y 30 y Coxsackie B1,2,3,4,5 y 6) y en periodos epidemicos con el virus aislado. En los anos estudiados se produjeron 2 brotes epidemicos; uno por Coxsackie A9 (1990-1991) y otro por Echo 30 (1994). En los sueros pareados se encontro mayor positividad a los Echo 6 y 11