A comparative study of the immunoantigenicity of eight Paracoccidioides brasiliensis isolates

Para se detectar diferenças imuno-antigênicas entre 8 amostras de P. brasiliensis isoladas de diferentes áreas endêmicas (Botucatu: Pb 1, 2 e 3; São Paulo: Pb: 18, 192 e 265; Venezuela: Pb 9 e 73), esutdaram-se: 1. A reatividade antigênica de cada amostra nas reações de imunofluorescência indireta (II) e de imunodifusão dupla em gel de agar (ID) contra painel de 20 soros controles positivos para paracoccidioidomicose; 2. A capacidade de induzir resposta imune humoral (medida por imunodifusão) e celular (medida pelo teste de coxim plantar) em camundongos imunizados com an-tígenos de cada amostra. Observamos: 1. As amostras Pb 265 e Pb 9 mostraram-se mais reativas na II; 2. Os antígenos das amostras Pb 192 e Pb 73 foram significativamente mais reativas na ID; 3. Estes dados demonstram diferenças de antigenicidade entre estas amostras; 4. A amostra Pb 18 mostrou baixo poder indutor de resposta imune celular e alta capacidade de indução de resposta imune humoral em camundongos imunizados, revelando dissociação de sua imunogenicidade. Estas diferenças podem indicar a existência de cepas distintas do fungo ou refletir modificações do parasita no hospedeiro ou du rante seu cultivo.

Fatores associados à vacinação contra influenza em idosos em município do Sudeste do Brasil

OBJETIVO: Mesmo gratuita e disponível no Brasil desde 1999, a cobertura vacinal contra a influenza ainda é inadequada em diversos municípios do País. O objetivo da pesquisa foi estimar a cobertura vacinal e identificar fatores relacionados à vacinação contra a influenza em idosos. MÉTODOS: Realizou-se inquérito domiciliar em amostra aleatória sistemática (N=365) da população urbana maior de 60 anos em Botucatu, São Paulo. Foi aplicado modelo de regressão logística múltipla, cuja variável dependente foi ter sido vacinado em 2002. Foram testadas no modelo as covariáveis: sexo, idade, socioeconômicas (renda per capita, número de pessoas por cômodo, escolaridade, estado civil, ocupação, tempo de moradia), antecedentes mórbidos, de internação, hábito de fumar, sintomas respiratórios nos últimos 15 dias e atividades comunitárias (trabalho voluntário, atividades no bairro, igreja). RESULTADOS: Registrou-se cobertura vacinal de 63,2% (IC 95%: 58,3-68,2). Foi observado menor percentual de vacinados entre os idosos na faixa etária de 60 a 64 anos. As variáveis que se mostraram associadas à vacinação e permaneceram no modelo final foram: idade (OR=1,09 por ano; IC 95%: 1,06-1,13); hipertensão arterial (OR=1,92; IC 95%: 1,18-3,13); inserção em atividades na comunidade (OR=1,63; IC 95%: 1,01-2,65). A vacinação em portadores de doenças crônicas não atingiu níveis adequados conforme esperado para este grupo de risco, com exceção dos hipertensos. A participação em atividades comunitárias e sociais foi relacionada com o estado vacinal. CONCLUSÕES: Condições socioeconômicas, hábitos e idade não restringiram o acesso à campanha vacinal. Por outro lado, campanhas específicas, endereçadas a indivíduos da faixa de 60 a 64 anos, podem ampliar a cobertura da vacinação.

Evaluation of Toxoplasma gondii placental transmission in BALB/c mice model

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efficiency of Matricaria chamomilla CH12 and number of doses of rabies vaccine on the humoral immune response in cattle

This study evaluated the effect of Matricaria chamomilla and vaccination frequency on cattle immunization against rabies. Four groups (n = 15/group) were treated with or without Matricaria chamomilla CH12 and vaccinated with one or two doses of rabies vaccine (30 day interval). No effect of chamomile was found on cattle immunization against rabies; however, antibody titers were protective in cattle vaccinated twice, while 93.3% of cattle vaccinated only once had titers under 0.5 UI/ml after 60 days. In conclusion, the use of chamomile did not alter the Immoral immune response in cattle, and two vaccine doses are suggested for achieving protective antibody titers.

Chicken embryo related (CER) cell line for quantification of rabies neutralizing antibody by fluorescent focus inhibition test

Levels of rabies virus neutralization antibody in sera from vaccinated dogs and cattle were either measured by mouse neutralization test (MNT) or by rapid fluorescent focus inhibition test (RFFIT), performed on CER monolayers. The two tests were compared for their ability to detect the 0.5 International Units/ml (I.U.) recommended by the World Health Organization (WHO) as the minimum response for proof of rabies immunization. A significant correlation was found between the two tests (n = 211; r = 0.9949 in dogs and 0.9307 in cows, p < 0.001), good sensitivity (87.5%), specificity (94.7%) and agreement (96.6%) as well. RFFIT method standardized on CER cell system for neutralizing antibodies detection turns the diagnosis easier and less expensive, specially when a great number of samples must be tested from endemic areas as commonly found in Brazil. (c) 2005 the International Association for Biologicals. Published by Elsevier Ltd. All rights reserved.

Rabies neutralizing antibody detection by indirect immunperoxidase serum neutralization assay performed on chicken embryo related cell line

The aim of this study was to evaluate the indirect immunoperoxidase virus neutralization (IPVN) and mouse neutralization test (MNT) to detect antibodies against rabies virus from vaccinated dogs and cattle. The IPVN was set up for the ability to measure 0.5 International Units/ml (IU) of antibody required by the World Health Organization and the Office International des Epizooties as the minimum response for proof of rabies immunization. IPVN was developed and standardized in chicken embryo related (CER) cell line when 141 dog and 110 cattle sera were applied by serial five-fold dilutions (1:5, 1:25, 1:125) as well as the positive and negative reference controls, all added in four adjacent wells, of 96-well microplates. A 50 µl amount of CVS32 strain dilution containing 50-200 TCID50/ml was mixed to each serum dilution, and after 90 min 50 µl of 3 x 10(5) cells/mlcell suspension added to each well. After five days of incubation, the monolayers were fixed and the IPVN test performed. The correlation coefficient between the MNT and IPVN performed in CER cells was r = 0.9949 for dog sera (n = 100) and r = 0.9307 for cattle sera (n = 99), as well as good specificity (94.7%), sensitivity (87.5%), and agreement (96.6%) were also obtained. IPVN technique can adequately identify vaccinated and unvaccinated animals, even from low-responding vaccinated animals, with the advantage of low cost and faster then MNT standard test.

Characterization of S3Pvac Anti-Cysticercosis Vaccine Components: Implications for the Development of an Anti-Cestodiasis Vaccine

Background: Cysticercosis and hydatidosis seriously affect human health and are responsible for considerable economic loss in animal husbandry in non-developed and developed countries. S3Pvac and EG95 are the only field trial-tested vaccine candidates against cysticercosis and hydatidosis, respectively. S3Pvac is composed of three peptides (KETc1, GK1 and KETc12), originally identified in a Taenia crassiceps cDNA library. S3Pvac synthetically and recombinantly expressed is effective against experimentally and naturally acquired cysticercosis.Methodology/ Principal Findings: In this study, the homologous sequences of two of the S3Pvac peptides, GK1 and KETc1, were identified and further characterized in Taenia crassiceps WFU, Taenia solium, Taenia saginata, Echinococcus granulosus and Echinococcus multilocularis. Comparisons of the nucleotide and amino acid sequences coding for KETc1 and GK1 revealed significant homologies in these species. The predicted secondary structure of GK1 is almost identical between the species, while some differences were observed in the C terminal region of KETc1 according to 3D modeling. A KETc1 variant with a deletion of three C-terminal amino acids protected to the same extent against experimental murine cysticercosis as the entire peptide. on the contrary, immunization with the truncated GK1 failed to induce protection. Immunolocalization studies revealed the non stage-specificity of the two S3Pvac epitopes and their persistence in the larval tegument of all species and in Taenia adult tapeworms.Conclusions/ Significance: These results indicate that GK1 and KETc1 may be considered candidates to be included in the formulation of a multivalent and multistage vaccine against these cestodiases because of their enhancing effects on other available vaccine candidates.

Modulatory effects of tuberculosis vaccines on experimental autoimmune encephalomyelitis

Although BCG is the only accepted vaccine against tuberculosis (TB), its protective ability is very limited. Therefore, many new vaccines are being evaluated. Our group has been working on DNAhsp65 - a genetic construction containing the hsp65 gene from Mycobacterium leprae. In previous experimental works, we demonstrated that both DNAhsp65 alone or associated with BCG, in a prime-boost regimen, were effective to control TB. A possible deleterious effect related to autoimmunity needed to be tested because hsp65 is highly homologous to the correspondent mammalian protein. In this investigation we tested the effect of a previous immunization with DNAhsp65 alone or associated with BCG in a rat model of multiple sclerosis. Female Lewis rats were immunized with three doses of DNAhsp65 or primed with BCG followed by two DNAhsp65 boosters. The animals were, then, immunized with myelin associated with complete Freund's adjuvant to develop experimental autoimmune encephalomyelitis (EAE). The following parameters were evaluated: weight loss, clinical score, central nervous system (CNS) inflammation and anti-myelin immune response. No deleterious effect was associated with these immunizations schedules. Immunized animals equally lost weight, the clinical scores were similar and CNS inflammation did not increase. Interestingly, both procedures determined decreased inflammation in the brain and lumbar spinal cord. This was concurrent with a modulatory effect over cytokine production by peripheral lymphoid organs. Cell cultures from spleen and lymph nodes in vitro stimulated with myelin produced less IFN-gamma and IL-10, respectively. This phenomenon was more clear in rats immunized with the genetic vaccine alone than with the prime-boost strategy. Together the results suggest that these strategies for TB prophylaxis would not accelerate or aggravate multiple sclerosis, being therefore, safe in this aspect. In addition, they indicate that these vaccination regimens have a potential anti-inflammatory activity that could be better explored in the future.

The effects of propolis on antibody production by laying hens

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Exposure to Mycobacterium avium Decreases the Protective Effect of the DNA Vaccine pVAXhsp65 Against Mycobacterium tuberculosis-Induced Inflammation of the Pulmonary Parenchyma

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

An essential role for mast cells as modulators of neutrophils influx in collagen-induced arthritis in the mouse

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Freqüência e variáveis associadas ao aleitamento materno em crianças com até 12 meses de idade no município de Araçatuba, São Paulo, Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Resistance to experimental autoimmune encephalomyelitis development in Lewis rats from a conventional animal facility

Experimental autoimmune encephalomyelitis (EAE) is an inflammatory disease of the brain and spinal cord that is mediated by CD4+ T lymphocytes specific to myelin components. In this study we compared development of EAE in Lewis rats from two colonies, one kept in pathogen-free conditions (CEMIB colony) and the other (Botucatu colony) kept in a conventional animal facility. Female Lewis rats were immunized with 100 µl of an emulsion containing 50 µg of myelin, associated with incomplete Freund's adjuvant plus Mycobacterium butyricum. Animals were daily evaluated for clinical score and weight. CEMIB colony presented high EAE incidence with clinical scores that varied from three to four along with significant weight losses. A variable disease incidence was observed in the Botucatu colony with clinical scores not higher than one and no weight loss. Immunological and histopathological characteristics were also compared after 20 days of immunization. Significant amounts of IFN-gamma, TNF-alpha and IL-10 were induced by myelin in cultures from CEMIB animals but not from the Botucatu colony. Significantly higher levels of anti-myelin IgG1 were detected in the CEMIB colony. Clear histopathological differences were also found. Cervical spinal cord sections from CEMIB animals showed typical perivascular inflammatory foci whereas samples from the Botucatu colony showed a scanty inflammatory infiltration. Helminths were found in animals from Botucatu colony but not, as expected, in the CEMIB pathogen-free animals. As the animals maintained in a conventional animal facility developed a very discrete clinical, and histopathological EAE in comparison to the rats kept in pathogen-free conditions, we believe that environmental factors such as intestinal parasites could underlie this resistance to EAE development, supporting the applicability of the hygiene hypothesis to EAE.

Taxa de ovulação e concentração plasmática de progesterona em fêmeas bovinas imunizadas com líquido folicular suíno

O presente estudo objetivou avaliar os efeitos da imunização ativa contra proteínas do líquido folicular suíno sobre a taxa de ovulação, duração do ciclo estral e concentração plasmática de progesterona em vacas e novilhas da raça Limousin. Realizaram-se duas imunizações com 15 dias de intervalo, aplicando-se um imunógeno composto do conteúdo protéico de 25ml de fluido folicular suíno adicionados de 1ml de gel de hidróxido de alumínio, via subcutânea. O número de ovulações foi mensurado por palpação retal e ultra-sonografia sete a nove dias após cada período de estro. Após a última imunização, os animais apresentaram maior incidência de ovulações duplas (41,7%). As médias de ovulações pré e pós-imunização foram 1,00+0,00 e 1,40+0,31 ovulações/ciclo, respectivamente (P<0,01;chi2). A imunização foi efetiva em aumentar o número de ovulações. Não foram observadas diferenças na duração do ciclo estral e na concentração de progesterona nos diferentes dias do ciclo. A manipulação das ações fisiológicas da inibina pode ser utilizada como alternativa para indução de ovulações múltiplas em bovinos.

Vacinação de idosos contra a influenza em um centro de saúde escola do interior do estado de São Paulo

Este estudo objetivou descrever o perfil dos idosos e avaliar informações que eles possuem sobre a vacinação contra a influenza. A coleta de dados foi realizada através de questionário, aplicado a 162 idosos de um Centro de Saúde Escola do interior de São Paulo, no período de abril a junho de 2006. Através da análise estatística descritiva, verificou-se que a maioria dos idosos tinha idade entre 70-79 anos e prevaleceu o sexo feminino (66%). em relação à vacinação, 83,8% dos entrevistados receberam a vacina e referiram não apresentar reação pós-vacinal. Os idosos obtiveram informação da vacinação por meio de rádio e televisão, e mencionaram que sua importância está relacionada com a prevenção. Concluiu-se que os idosos possuíam informações sobre a vacinação, porém ainda existe a necessidade de enfermeiros e outros profissionais da área, se responsabilizarem por uma orientação mais clara sobre a importância da vacinação contra influenza e suas complicações.