Antibody and cytokine serum levels in patients subjected to anti-rabies prophylaxis with serum-vaccination

Rabies is considered a fatal disease once clinical symptoms have developed. The aim of this study was to evaluate epidemiological aspects and immune response in patients attacked by domestic and wild animals and subjected to post-exposure rabies treatment with equine serum and associated vaccine. Thirty-three patients were evaluated; they were between 13 and 65 years old, 75.8% were male and 24.2% female, and from the Botucatu neighborhood. Twenty healthy control individuals with the same age range were also studied. Specific antibodies to equine immunoglobulins and IFN-γ, IL-2, IL-4, and IL-10 production were evaluated by ELISA. IgM, IgE, IgG and subclasses, and rabies virus antibodies serum levels were determined by nephelometry and seroneutralization methods, respectively. No anaphylactic or serum sickness allergic reactions were observed in patients after treatment. Anti-equine IgG levels were significantly higher than those of IgM after 14 and 28 days of treatment. Protective antibodies to rabies virus > 0.5 UI/ml were detected in 84.6% and 75% of patients at days 14 and 28, respectively. IFN-γ, IL-2 and IL-10 levels in patients before and 48h after treatment were significantly higher than in controls suggesting that both Th1 and Th2 cells were activated in the patients. Serum IgM levels were higher at day 14, and IgG 2 and IgE levels were higher at day 28 of treatment. These results suggest that post-exposure rabies treatment in humans induces significant alterations in patient immune response characterized by increased levels of cytokines, serum levels of specific rabies virus antibodies, and the equine serum components employed in the treatment.

Retrospective analysis of post-exposure to human anti-rabies treatment in Botucatu, São Paulo State, Brazil

This analysis aimed to identify characteristics of accidents that would, probably, provoke rabies infection. A total of 14,409 survey questionnaires for surveillance of human rabies from the Brazilian Information System for Disease Notification (SINAN), from 2000 to 2005, were analyzed. Regarding demographics, it was observed that 7,377 (51.5%) of the victims were white, 4,458 (30.93%) were children and 8,008 (55.58%) were males. Urban cases were prevalent (88.10%) while dogs were the animals most frequently involved in accidents, in 11,700 cases (81.19%). Bites (84.35%) and scratches (19.15%) were the most prevalent exposure types, and occurred predominantly on victims' extremities (38.79%). The prophylactic measure taken in 6,179 cases comprised anti-rabies vaccine; of these victims, 421 (2.92%) showed systemic reactions while 693 (4.80%) reported no response. The importance of developing awareness in professionals that should correctly report post-exposure immunoprophylaxis cases is emphasized given the high number of individuals who receive this type of treatment annually.

Subconjunctival Delivery of Antibiotics in a Controlled-Release System A Novel Anti-infective Prophylaxis Approach for Cataract Surgery

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

Evaluation of cytokines concentration and percentage of survival of rabies virus-infected mice submitted to anti-rabies Vero-cell propagated vaccine and P-acnes

Previously, survival of rabies infection was shown to correlate with low IL-6 serum concentration in mice subjected to post-exposure treatment with the Fuenzalida Palacios rabies vaccine in conjunction with the immunomodulator Propionibacterium acnes, previously Corynebacterium parvum. Considering the substitution of the Fuenzalida Palacios rabies vaccine by the Vero cell raised anti-rabies vaccine in almost all countries, the objective of this work was to evaluate the survival and cytokine serum concentration of rabies virus-infected mice treated with P. acnes in conjunction with or the anti-rabies-VERO vaccine. For this, Swiss mice were experimentally infected with street rabies virus and subjected to vaccine and/or P. acnes following infection. Animals were killed at different times and serum was collected to evaluate cytokines. The greatest survival was observed in animals given one or two does of P. acnes in the absence of vaccination. Animals given anti-rabies VERO vaccine alone or with three doses of P. acnes had the second highest survival rate. The group that had the highest percentage of mortality also had the highest IL-6 concentration on the 10th day, a time correlating with clinical symptoms of the animals. The results reinforce the inefficacy of anti-rabies vaccine in only one dose as a post-exposure treatment irrespective of the type of vaccine used, the immunomodulation activity of P. acnes in rabies post-exposure treatment and suggest a role for IL-6 in rabies virus pathogenesis. (c) 2006 Elsevier B.V. All rights reserved.

EFFECTS of SELENIUM SUPPLEMENTATION on CATTLE ANTI-RABIES HUMORAL IMMUNE RESPONSE and LEVELS of SERUM SELENIUM and CORTISOL

This study evaluated the effect of different concentrations of selenium (Se) supplementation on cattle anti-rabies humoral immune response, serum Se concentrations and cortisol levels. Sixty uncastrated male Nelore calves from 10 to 12 months grazing on Brachiaria decumbens forage were studied. The animals were assigned to one of four groups (n = 15 each), which received non-supplemented diets (Gc) or supplemented with daily and individual Selenium ( Se) concentrations of 3.6 mg (G(3.6)), 5.4 mg (G(5.4)) or 6.4 mg (G(6.4)). The calves were immunized on day 0 with one dose of commercial liquid inactivated rabies vaccination. on days 15, 30, 60, 90 and 120, the cattle underwent the same stressing procedures used for vaccination in the corral. Cattle blood samples were collected after vaccination and stressing procedures to determine serum Se levels, rabies antibody titers and serum cortisol. Se levels were also determined in forage samples collected from the paddocks in which the cattle were held. Se concentration in B. decumbens was 0.04 mg of Se/kg dry matter. Baseline Se levels obtained on day 0 were higher in Gc than in G(5.4) and G(6.4) (P = 0.005). Serum Se levels decreased in Gc throughout the experiment (P < 0.004), increased in G(3.6) (P < 0.000) and G(5.4) (P < 0.000) and were kept high from day 60 on in group G(6.4) (P < 0.002). Rabies antibody titers did not differ among control and supplemented groups. However, 120 days after vaccination rabies antibody titers were kept above protective levels (>= 0.5 UI/mL) only in group G(3.6) (P < 0.00002), whereas they dropped in the other groups (P < 0.05). Serum cortisol levels did not differ among the experimental groups (P = 0.79), reached peak levels on day 90 and returned close to baseline levels on day 120. Se and cortisol levels were not markedly correlated. Serum cortisol and rabies antibody titers were correlated only in group G(6.4), on day 60 (R = 0.513; P = 0.05) and 120 (R = 0.644; P = 0.009). Serum Se and rabies antibody titers were correlated only in group G(6.4), on day 60 (R = -0.580; P = 0.023). In conclusion: a) the profile of Se variation is different among groups receiving different concentrations of this element; b) the supplementation dosage of 3.6 mg Se/animal/day is efficient to treat/prevent marginal Se deficiency; c) individual supplementation with daily concentrations of 3.6 mg Se enhances the maintenance of rabies antibody titers in cattle; d) individual supplementation with daily concentrations of 3.6; 5.4 and 6.4 mg Se are ineffective in reducing serum cortisol; e) repeated cattle handling in corrals stress animals that adapt to these procedures, although serum cortisol does not return to baseline levels by 120 days; and f) the stress generated by repeated management in cattle in the corral does not diminish antibody titers after vaccination against rabies.

Experimental antiviral therapy against different rabies virus lineages using transfection with anti-rabies antibodies

Financial Support: FAPESP

Sorovacinação antirrabica: aspectos epidemiologicos e implicações para assistência de enfermagem

This study was developed with the purpose to search for relevant knowledge concerning nursing care for individuals exposed to the rabies virus and submitted to post-exposure anti-rabies serovaccination. The authors aimed at evaluating the epidemiological aspects of accidents involving household and wild animals occurring in 2007 and patients assisted at a reference hospital located in the mid-southern region of São Paulo State. They also aimed at identifying the relevance of nurses' actions by describing aspects of care provision. The method adopted was exploratory, retrospective and descriptive of the epidemiological aspects of the accidents and of the care provided to these patients by referring to information in their medical charts. Fifty-one charts of patients aged 17 to 81 years, considered to be at risk and with indication for post-exposure prophylaxis against human rabies were evaluated. It was found that nursing care provision to these patients presents low complexity although it requires properly trained professionals. It was possible to identify the relevance of nurses' actions in care provision and procedures related to anti-rabies prophylaxis; however, consistent information concerning nursing care was not observed.

Avaliação dos registros de profilaxia anti-rábica humana pós-exposição no Município de Jaboticabal, são Paulo, no período de 2000 a 2006

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Pre-clinical toxicity & immunobiological evaluation of DNA rabies vaccine & combination rabies vaccine in rhesus monkeys (Macaca mulatta)

Background & objectives: Pre-clinical toxicology evaluation of biotechnology products is a challenge to the toxicologist. The present investigation is an attempt to evaluate the safety profile of the first indigenously developed recombinant DNA anti-rabies vaccine DRV (100 mu g)] and combination rabies vaccine CRV (100 mu g DRV and 1.25 IU of cell culture-derived inactivated rabies virus vaccine)], which are intended for clinical use by intramuscular route in Rhesus monkeys. Methods: As per the regulatory requirements, the study was designed for acute (single dose - 14 days), sub-chronic (repeat dose - 28 days) and chronic (intended clinical dose - 120 days) toxicity tests using three dose levels, viz. therapeutic, average (2x therapeutic dose) and highest dose (10 x therapeutic dose) exposure in monkeys. The selection of the model i.e. monkey was based on affinity and rapid higher antibody response during the efficacy studies. An attempt was made to evaluate all parameters which included physical, physiological, clinical, haematological and histopathological profiles of all target organs, as well as Tiers I, II, III immunotoxicity parameters. Results: In acute toxicity there was no mortality in spite of exposing the monkeys to 10XDRV. In sub chronic and chronic toxicity studies there were no abnormalities in physical, physiological, neurological, clinical parameters, after administration of test compound in intended and 10 times of clinical dosage schedule of DRV and CRV under the experimental conditions. Clinical chemistry, haematology, organ weights and histopathology studies were essentially unremarkable except the presence of residual DNA in femtogram level at site of injection in animal which received 10X DRV in chronic toxicity study. No Observational Adverse Effects Level (NOAEL) of DRV is 1000 ug/dose (10 times of therapeutic dose) if administered on 0, 4, 7, 14, 28th day. Interpretation & conclusions: The information generated by this study not only draws attention to the need for national and international regulatory agencies in formulating guidelines for pre-clinical safety evaluation of biotech products but also facilitates the development of biopharmaceuticals as safe potential therapeutic agents.

Preparation and mass spectrometric study of egg yolk antibody (IgY) against rabies virus

Rabies virus was used as the antigen to immunize laying chickens. Anti-rabies virus immunoglobulin Y(IgY) was isolated from yolks of the eggs laid by these chickens using a two-step salt precipitation and one-step gel filtration protocol. The purified IgY was reduced with dithiothreitol, and heavy chains (HC) and light chains (LC) were obtained. In addition, the purified IgY was digested with pepsin and the fragment with specific antigen binding properties (Fab) was produced. Using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOFMS), the average molecular weights of IgY, HC, LC, and Fab were determined as 167 250, 65 105, 18 660, and 45,359 Da, respectively. IgY has two structural differences compared with mammalian IgGs. First, the molecular weight of the heavy chain of IgY is larger than that of its mammalian counterpart, while the molecular weight of the light chain of IgY is smaller. Second, upon pepsin digestion, anti-rabies virus IgY is degraded into Feb, in contrast to mammalian IgG, which has been reported to be degraded into F(ab')(2) under the same conditions. Copyright (C) 2001 John Wiley & Sons, Ltd.

Ocorrência de anticorpos anti-Neospora caninum em cães da microrregião da Serra de Botucatu, Estado de São Paulo, Brasil

Neosporose é uma enfermidade parasitária causada pelo protozoário Neospora caninum reconhecido como importante causa de abortamento bovino e neuropatia canina. Considerando o isolamento de N. caninum e a sorologia freqüente em bovinos em nossa região, os objetivos do presente trabalho foram avaliar a ocorrência de anticorpos anti-N. caninum em cães da Microrregião da Serra de Botucatu, Estado de SãoPaulo, e sua associação ao sexo, idade e procedência quanto à zona urbana (exclusivamente cidade), rural (somente chácaras e sítios) e peri-urbana (acesso à zona urbana e rural) dos cães estudados. Foram analisados 963 cães, com ou sem raça definida, de ambos os sexos e diferentes idades, sem apresentação de qualquer sintomatologia clínica. Os animais foram selecionados aleatoriamente durante a campanha de vacinação anti-rábica da microrregião da Serra de Botucatu, no período de maio a setembro de 1998. O soro obtido dos animais foi avaliado por meio da Reação de Imunofluorescência Indireta (RIFI) utilizando como antígeno a cepa padrão NC-1 deN. caninum. Observaram-se 245 animais reagentes (25,4% de positividade), sendo 161 (27,5%) machos e, 84 (22,3%) fêmeas. Dos animais de zona urbana, rural e mista 223 (25,8%),11(16,9%) e 11(33,3%), respectivamente, foram reagentes à prova de RIFI. Todos os 11 municípios apresentaram cães soropositivos com valores de ocorrência que variaram de 8,9% a 53,5%. Observou-se percentual de positividade menor em cães até um ano (16,2%) quando comparados àqueles entre 1 a 4 anos e superior a 4 anos (28,4 % e 28,0%, respectivamente) que não apresentaram diferença entre si. Os resultados obtidos caracterizaram soropositividade para N. caninum em cães pertencentes a todos os municípios da Microrregião da Serra de Botucatu evidenciando a ampla distribuição do agente na região.

Avaliação da vacinação anti-rábica e da suplementação com probiótico na resposta imune humoral em bovinos

This study evaluated the humoral immune response of a new rabies vaccine developed by the Instituto Butantan (potency of 3.27 UI/ml) in primovaccinated cattle and the effect of probiotic on this response. Thirty-four 15-month old Nelore cattle were randomly divided into 2 groups (17 animals/group). All the animals were vaccinated on day 0 (zero) and then animals in one group received probiotic added to a mineral mixture (GP) while the others were given only the mineral mixture (GC). Blood samples were collected on days 0, 75 and 150 for rabies neutralizing antibodies titers by seroneutralization assay on BHK21 cells (RFFIT). Protective antibody titers (>= 0.5 UI/mL) were found in 82.4% of the animals from GP and in 76.5% of the animals from GC and no statistical difference (p>0.05) between antibody titers in GP and GC was detected on days 75 and 150. It was also observed that in both groups antibody titers was decreased on day 150 (p<0.01). In conclusion, the tested rabies vaccine promotes efficient soroconversion and keeps antibody levels in primovaccinated cattle, but probiotic does not affect the humoral anti-rabies immune response.

Serological profile of anti-Toxoplasma gondii antibodies in apparently healthy dogs of the city of Botucatu, São Paulo state, Brazil

Toxoplasmosis is a zoonosis caused by Toxoplasma gondii, a coccidian protozoan of worldwide distribution. The seroprevalence in canine population can be an alternative for measuring T. gondii urban spreading. A total of 780 blood samples from dogs were collected, during the yearly anti-rabies campaign, carried out by the Department of Veterinary Hygiene and Public Health, School of Veterinary Medicine and Animal Husbandry (FMVZ), São Paulo State University, UNESP, together with the county health authorities, in August 1999. Using Indirect Fluorescent Antibody Test (IFAT) for detecting antibodies anti-T. gondii in the sera samples, we observed that 258 dogs (33.1%) were positive. The associations between the serological results and the epidemiological variables were studied. Statistically significant differences were not found regarding sex (32.2% male and 34.3% female reactors). Dogs without a defined breed showed seropositivity statistically higher than the pedigreed group. The occurrence of infection was considered higher with age.

A Bayesian approach to estimate the accuracy of in-house ELISA assay to measure rabies antibodies from compulsory vaccinated dogs and cattle

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Eficácia terapêutica de RNAs de interferência (siRNAS) e avaliação da resposta imune em camundongos infectados com vírus da raiva de origem de cão e de morcego

Pós-graduação em Cirurgia Veterinária - FCAV