12 research outputs found
Comparison of in house and commercial conjugates by rapid fluorescent focus inhibition test (RFFIT) in rabies antibodies evaluation
Submitted by Alexandre Sousa ([email protected]) on 2015-12-01T15:45:44Z
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Previous issue date: 2010Fundação Oswaldo Cruz. Centro de Criação de Animais de Laboratório. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Controle de Qualidade em Saúde. Departamento de Imunologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos. Rio de Janeiro, RJ, Brasil.A raiva é uma zoonose letal transmitida ao homem pela inoculação do vírus rábico, principalmente pela mordedura de animais infectados. Em 2005, o Ministério da Saúde brasileiro gastou cerca de R 66 millions in epidemiological surveillance actions, in order to carry out vaccination campaigns and acquisition of immunobiologicals. The serological evaluation is the basic requirement for surveillance of individuals vaccinated. Ninety one sera, from 34 vaccinees, were selected to evaluate the antibody titration by rapid fluorescent focus inhibition test adapted to 96-well microplates, to compare an in house produced conjugate and a commercial one. Seventy four sera (82.2%) had titers of ≥0.5 IU/mL and 12 sera (13.3%) had titers of <0.5 IU/mL. Theses results showed that the rapid fluorescent focus inhibition test using the in house conjugate was as sensitive as the commercial one. The difference between the results using both conjugates was not significant; the antibody titers were highly correlated (r=0.94)
Lymphocyte subset analyses in healthy adults vaccinated with yellow fever 17DD virus
Submitted by Sandra Infurna ([email protected]) on 2019-11-28T13:17:55Z
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Previous issue date: 2005Fundação Oswaldo Cruz. Bio-Manguinhos. Laboratório de Tecnologia Imunológica. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Imunologia. Laboratório de Imunoparasitologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Bio-Manguinhos. Laboratório de Tecnologia Imunológica. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Imunologia. Laboratório de Imunoparasitologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Bio-Manguinhos. Laboratório de Tecnologia Imunológica. Rio de Janeiro, RJ, Brasil.In this study the kinetics of humoral and cellular immune responses in first-time vaccinees and re-vaccinees with the yellow fever 17DD vaccine virus was analyzed. Flow cytometric analyses were used to determine percentual values of T and B cells in parallel to the yellow fever neutralizing antibody production. All lymphocyte subsets analyzed were augmented around the 30th post vaccination day, both for first-time vaccinees and re-vaccinees. CD3+ T cells increased from 30.8% (SE +/- 4%) to 61.15% (SE +/- 4.2%), CD4+ T cells from 22.4% (SE +/- 3.6%) to 39.17% (SE +/- 2%) with 43% of these cells corresponding to CD4+CD45RO+ T cells, CD8+ T cells from 15.2% (SE +/- 2.9%) to 27% (SE +/- 3%) with 70% corresponding to CD8+CD45RO+ T cells in first-time vaccinees. In re-vaccinees, the CD3+ T cells increased from 50.7% (SE +/- 3%) to 80% (SE +/- 2.3%), CD4+ T cells from 24.9% (SE +/- 1.4%) to 40% (SE +/- 3%) presenting a percentage of 95% CD4+CD45RO+ T cells, CD8+ T cells from 19.7% (SE +/- 1.8%) to 25% (SE +/- 2%). Among CD8+CD38+ T cells there could be observed an increase from 15 to 41.6% in first-time vaccinees and 20.7 to 62.6% in re-vaccinees. Regarding neutralizing antibodies, the re-vaccinees presented high titers even before re-vaccination. The levels of neutralizing antibodies of first-time vaccinees were similar to those presented by re-vaccinees at day 30 after vaccination, indicating the success of primary vaccination. Our data provide a basis for further studies on immunological behavior of the YF 17DD vaccine
The sample processing time interval as an influential factor in flow cytometry analysis of lymphocyte subsets
The objective of this paper is to propose a protocol to analyze blood
samples in yellow fever 17DD vaccinated which developed serious adverse
events. We investigated whether or not the time between sample
collection and sample processing could interfere in lymphocyte subset
percentage, for it is often impossible to analyze blood samples
immediately after collection due to transport delay from collection
places to the flow cytometry facility. CD4+CD38+ T, CD8+CD38+ T, CD3+
T, CD19+ B lymphocyte subsets were analyzed by flow cytometry in nine
healthy volunteers immediately after blood collection and after
intervals of 24 and 48 h. The whole blood lysis method and gradient
sedimentation by Histopaque were applied to isolate peripheral blood
mononuclear cells for flow cytometry analyses. With the lysis method,
there was no significant change in lymphocyte subset percentage between
the two time intervals (24 and 48 h). In contrast, when blood samples
were processed by Histopaque gradient sedimentation, time intervals for
sample processing influenced the percentage in T lymphocyte subsets but
not in B cells. From the results obtained, we could conclude that the
whole blood lysis method is more appropriate than gradient
sedimentation by Histopaque for immunophenotyping of blood samples
collected after serious adverse events, due to less variation in the
lymphocyte subset levels with respect to the time factor
Lymphocyte subset analyses in healthy adults vaccinated with yellow fever 17DD virus
In this study the kinetics of humoral and cellular immune responses in first-time vaccinees and re-vaccinees with the yellow fever 17DD vaccine virus was analyzed. Flow cytometric analyses were used to determine percentual values of T and B cells in parallel to the yellow fever neutralizing antibody production. All lymphocyte subsets analyzed were augmented around the 30th post vaccination day, both for first-time vaccinees and re-vaccinees. CD3+ T cells increased from 30.8% (SE ± 4%) to 61.15% (SE ± 4.2%), CD4+ T cells from 22.4% (SE ± 3.6%) to 39.17% (SE ± 2%) with 43% of these cells corresponding to CD4+CD45RO+ T cells, CD8+ T cells from 15.2% (SE ± 2.9%) to 27% (SE ± 3%) with 70% corresponding to CD8+CD45RO+ T cells in first-time vaccinees. In re-vaccinees, the CD3+ T cells increased from 50.7% (SE ± 3%) to 80% (SE ± 2.3%), CD4+ T cells from 24.9% (SE ± 1.4%) to 40% (SE ± 3%) presenting a percentage of 95% CD4+CD45RO+ T cells, CD8+ T cells from 19.7% (SE ± 1.8%) to 25% (SE ± 2%). Among CD8+CD38+ T cells there could be observed an increase from 15 to 41.6% in first-time vaccinees and 20.7 to 62.6% in re-vaccinees. Regarding neutralizing antibodies, the re-vaccinees presented high titers even before re-vaccination. The levels of neutralizing antibodies of first-time vaccinees were similar to those presented by re-vaccinees at day 30 after vaccination, indicating the success of primary vaccination. Our data provide a basis for further studies on immunological behavior of the YF 17DD vaccine
Comparative analysis of the intracerebral mouse protection test and serological method for potency assays of pertussis component in DTP vaccine
The aim of this study was to compare the PSPT standardized in-house as an alternative to MPT for potency assays of pertussis component. Statistical analyses have showed similar pertussis potency values when PSPT was compared to MPT. Significant correlation between the potency results obtained by in vivo and in vitro assays was also been observed. Results by PSPT have demonstrated reproducibility and accuracy for potency pertussis control and this approach has been considered promising for use at least during the steps of production
Imunogenicidade da cepa avirulenta RV194-2 do vírus rábico em camundongos
O vírus rábico RV194-2, uma variante avirulenta da cepa CVS (Challenge Vírus Standard), produz uma infecção inaparente quando inoculado intracerebralmente em camundongos adultos. Sugerindo que a resposta imunológica do hospedeiro permite a eliminação do vírus do sistema nervoso central. Por esta razão foram estudadas a indução de interferon e a resposta imune humoral em camundongos BALB/c inoculados com vírus RV194-2. Durante a infecção, estes camundongos apresentaram elevados níveis de interferon no plasma e no cérebro com altos títulos de anticorpos neutralizantes anti-rábicos. A 2-5A sintetase. um marcador da ação dos interferons,foi também analisada no cérebro destes animais. Sua atividade, aumentou, paralelamente, á produção de interferon, demonstrando que este interferon é bioquímicamente ativo. O vírus RV194-2 também induziu, 45 dias após sua inoculação, proteção aos animais quando desafiados com a cepa virulenta CVS. Estes resultados demonstram que a cepa RV194-2possui um alto nível imunogênico.<br>RV194-2 rabies virus, an avirulent mutant of CVS strain, induces an inapparent infection limited to the central nervous system (CNS) in adult mice inoculated intracerebrally. This fact suggest that immune response of the host is able to eliminate the virus in CNS. For this reason, we have studied the induction of interferon and the humoral immune responses in BALB/c mice after RV194-2 inoculation. These mice presented high levels of interferon in the plasma and in the brain, with elevated levels of neutralizing antirabies antibodies. The 2-5A synthetase, an enzyme marker of interferon action, was analyzed in the brain of inoculated animals. Its enhancement in parallel to the interferon production in the brain, showed biochemical evidence that this interferon is active. Forty five days after RV194-2 virus inoculation, mice were protected against a challenge with the CVS virulent strain. The results presented herein show that RV194-2 strain has a bigh level of immunogenicity
Immunogenicity test of tetanus component in adsorbed vaccines by toxin binding inhibition test
Samples from 20 lots of diphtheria-tetanus (adult use dT) vaccine and from 20 lots of diphtheria-tetanus-pertussis (DTP) vaccine were used to standardize and validate the in vitro toxin binding inhibition (ToBI) test for the immunogenicity test of the tetanus component. The levels of tetanus antitoxin obtained by ToBI test were compared to those obtained using the toxin neutralization (TN) test in mice routinely employed to perform the quality control of the tetanus component in adsorbed vaccines. The results ranged from 1.8 to 3.5 IU/ml for dT and 2 to 4 IU/ml for DTP by ToBI test and 1.4 to 3 IU/ml for dT and 1.8 to 3.5 IU/ml for DTP by TN in mice. These results were significantly correlated. From this study, it is concluded that the ToBI test is an alternative to the in vivo neutralization procedure in the immunogenicity test of the tetanus component in adsorbed vaccines. A substantial refinement and a reduction in use of animals can be achieved