Development, Histological, and Ultrastructural Evaluation of Sheep Hyperimmune Serum Therapy for COVID-19 / Desarrollo, Evaluación Histológica y Ultraestructural de la Terapia con Suero Hiperinmune de Oveja para COVID-19

SUMMARY: In response to the threat posed by new variants of SARS-CoV-2 and the urgent need for effective treatments in the absence of vaccines, the aim of this study was to develop a rapid and cost-effective hyperimmune serum (HS) derived from sheep and assess its efficacy. The utilization of a halal-certified, easily maintained in certain geographic regions, easy-to-handle animal such as sheep could provide a viable alternative to the expensive option of horses. Sheep were immunized with a whole inactivated SARS-CoV- 2 antigen to produce HS, which was evaluated for neutralizing potency using the PRNT50 assay. K18-hACE2 transgenic mice (n=35) were divided into three groups: control, SARS-CoV-2 exposure through inhalation, and SARS-CoV-2 exposed mice treated with HS. HS efficacy was assessed through serum proinflammatory cytokine levels, qRT-PCR analysis, histopathological examination of lungs and hearts, and transmission electron microscopy. Purified HS exhibited significant neutralizing activity (1/24,576). The SARS-CoV-2+HS group showed lower levels of TNF-α, IL-10, and IL-6 (P<0.01) and relatively lower levels of MCP-1 compared to the SARS-CoV-2 group. HS prevented death, reduced viral RNA levels in the lungs and hearts, protected against severe interstitial pneumonia, preserved lung tissue integrity, and prevented myocyte damage, while the SARS-CoV-2 group exhibited viral presence in the lungs. This study successfully developed a sheep-derived HS against the entire SARS-CoV-2 virus, resulting in a significant reduction in infection severity, inflammation, and systemic cytokine production. The findings hold promise for treating severe COVID-19 cases, including emerging viral variants, and immunocompromised patients.

En respuesta a la amenaza que suponen las nuevas variantes del SARS-CoV-2 y la urgente necesidad de tratamientos eficaces en ausencia de vacunas, el objetivo de este estudio fue desarrollar un suero hiperinmune (HS) rápido y rentable derivado de ovejas. y evaluar su eficacia. La utilización de un animal con certificación halal, de fácil mantenimiento en determinadas regiones geográficas y de fácil manejo, como las ovejas, podría proporcionar una alternativa viable a la costosa opción de los caballos. Las ovejas fueron inmunizadas con un antígeno de SARS-CoV-2 completamente inactivado para producir HS, cuya potencia neutralizante se evaluó mediante el ensayo PRNT50. Los ratones transgénicos K18-hACE2 (n = 35) se dividieron en tres grupos: control, exposición al SARS-CoV-2 mediante inhalación y ratones expuestos al SARS-CoV-2 tratados con HS. La eficacia de HS se evaluó mediante niveles de citoquinas proinflamatorias en suero, análisis qRT-PCR, examen histopatológico de pulmones y corazones y microscopía electrónica de transmisión. El HS purificado exhibió una actividad neutralizante significativa (1/24,576). El grupo SARS-CoV-2+HS mostró niveles más bajos de TNF-α, IL-10 e IL-6 (P<0,01) y niveles relativamente más bajos de MCP-1 en comparación con el grupo SARS-CoV-2. HS evitó la muerte, redujo los niveles de ARN viral en los pulmones y el corazón, protegió contra la neumonía intersticial grave, preservó la integridad del tejido pulmonar y evitó el daño de los miocitos, mientras que el grupo SARS-CoV-2 exhibió presencia viral en los pulmones. Este estudio desarrolló con éxito un HS derivado de ovejas contra todo el virus SARS-CoV-2, lo que resultó en una reducción significativa de la gravedad de la infección, la inflamación y la producción sistémica de citocinas. Los hallazgos son prometedores para el tratamiento de casos graves de COVID- 19, incluidas las variantes virales emergentes y los pacientes inmunocomprometidos.

[Safety profile of heterologous serum produced by the Butantan Institute, in São Paulo-SP, Brazil, from 2012 to 2015]. / Perfil de segurança dos soros heterólogos produzidos pelo Instituto Butantan, São Paulo-SP, no período de 2012 a 2015.

OBJECTIVE: to describe the safety profile of the heterologous serum produced by the Butantan Institute (BI) of São Paulo-SP, Brazil. METHODS: a descriptive study of adverse events (AEs) post-exposure to serum produced by the BI, encoded in the medical terminology of the Medical Dictionary for Regulatory Activities (MedDRA), and spontaneously reported to BI from 2012 to 2015. RESULTS: 52 individuals reported AEs, mainly related to Bothrops antivenom (n=11), diphtheria antitoxin (n=9) and unspecified snakebite serum (n=9); a mean of 3.2 AEs per individual was observed; among the total of 173 AEs, 63.0% were expected considering that they were described in the package insert; most of them were classified as skin and subcutaneous tissue disorders (30.6%); there were six deaths temporally related to the use of serum, but this association was discarded. CONCLUSION: in the studied period, the serum produced by the BI had no changes in their safety profiles, considering that the AEs were expected, according to the information previously described in the package insert.

Perfil de segurança dos soros heterólogos produzidos pelo Instituto Butantan, São Paulo-SP, no período de 2012 a 2015 / Safety profile of heterologous serum produced by the Butantan Institute, in São Paulo-SP, Brazil, from 2012 to 2015 / Perfil de seguridad de los sueros heterólogos producidos por el Instituto Butantan, São Paulo-SP, Brasil, en el período de 2012 al 2015

Resumo OBJETIVO: descrever o perfil de segurança dos soros heterólogos produzidos pelo Instituto Butantan (IB) de São Paulo-SP, Brasil. MÉTODOS: estudo descritivo dos relatos de eventos adversos (EA) pós-exposição aos soros produzidos pelo IB, codificados pela terminologia do Dicionário Médico para Atividades Regulatórias (MedDRA), notificados espontaneamente ao IB entre 2012 e 2015. RESULTADOS: foram notificados 52 usuários com algum evento adverso relacionado, principalmente, aos soros antibotrópico (n=11), antidiftérico (n=9) e antiofídico não especificado (n=9); observaram-se, em média, 3,2 EA por indivíduo; dos 173 EA notificados, 63,0% eram esperados por serem eventos descritos em bula; os EA mais notificados foram categorizados como afecções dos tecidos cutâneos e subcutâneos (30,6%); houve seis óbitos temporalmente relacionados ao uso de soros, porém essa associação foi descartada. CONCLUSÃO: no período estudado, os soros produzidos pelo IB não apresentaram alteração em seu perfil de segurança, já que os EA relatados eram esperados conforme informação descrita em bula.

Abstract OBJECTIVE: to describe the safety profile of the heterologous serum produced by the Butantan Institute (BI) of São Paulo-SP, Brazil. METHODS: a descriptive study of adverse events (AEs) post-exposure to serum produced by the BI, encoded in the medical terminology of the Medical Dictionary for Regulatory Activities (MedDRA), and spontaneously reported to BI from 2012 to 2015. RESULTS: 52 individuals reported AEs, mainly related to Bothrops antivenom (n=11), diphtheria antitoxin (n=9) and unspecified snakebite serum (n=9); a mean of 3.2 AEs per individual was observed; among the total of 173 AEs, 63.0% were expected considering that they were described in the package insert; most of them were classified as skin and subcutaneous tissue disorders (30.6%); there were six deaths temporally related to the use of serum, but this association was discarded. CONCLUSION: in the studied period, the serum produced by the BI had no changes in their safety profiles, considering that the AEs were expected, according to the information previously described in the package insert.

Resumen OBJETIVO: describir el perfil de seguridad de los sueros heterólogos producidos por el Instituto Butantan (IB) de São Paulo-SP, Brasil. MÉTODOS: estudio descriptivo de los informes de eventos adversos (EAs) post-exposición a los sueros del IB y codificados según el Diccionario Médico para Actividades Regulatorias (MedDRA). RESULTADOS: 52 usuarios presentaron EAs relacionados con los sueros antibotrópico (n=11), antidiftérico (n=9) y antiofídico no especificado (n=9); se observó, en los EAs, 3,2 de media por persona; de los 173 EAs reportados, 63,0% fueron "esperados", ya que figuran descritos en la bula farmacológica; los EAs más reportados fueron los trastornos de piel y tejido subcutáneo (30,6%); hubo seis muertes, pero se descartó la asociación con el uso de suero. CONCLUSIÓN: durante el período de estudio, los sueros del IB no mostraron ningún cambio en su perfil de seguridad, ya que los EAs reportados eran esperados conforme información descrita en la bula.

PROTECTIVE LEVELS OF VARICELLA-ZOSTER ANTIBODY DID NOT EFFECTIVELY PREVENT CHICKENPOX IN AN X-LINKED AGAMMAGLOBULINEMIA PATIENT.

We describe the case of an eight-year-old boy with X-linked agammaglobulinemia who developed mild varicella despite regular intravenous immunoglobulin (IVIG) therapy. He maintained protective antibody levels against varicella and the previous batches of IVIG that he received had adequate varicella-specific IgG levels. The case illustrates that IVIG may not prevent VZV infection.

PROTECTIVE LEVELS OF VARICELLA-ZOSTER ANTIBODY DID NOT EFFECTIVELY PREVENT CHICKENPOX IN AN X-LINKED AGAMMAGLOBULINEMIA PATIENT / Nível sérico adequado de anticorpo contra o vírus da varicela-zoster não foi suficiente para prevenir a infecção em criança com agamaglobulinemia ligada ao X

SUMMARY We describe the case of an eight-year-old boy with X-linked agammaglobulinemia who developed mild varicella despite regular intravenous immunoglobulin (IVIG) therapy. He maintained protective antibody levels against varicella and the previous batches of IVIG that he received had adequate varicella-specific IgG levels. The case illustrates that IVIG may not prevent VZV infection.

RESUMO Relatamos o caso de uma criança com agamaglobulinemia ligada ao X, sexo masculino, oito anos de idade, que desenvolveu quadro de varicela leve, apesar do tratamento regular com imunoglobulina intravenosa (IVIG). O paciente mantinha níveis adequados de imunoglobulina (IgG) contra varicela, assim como, os últimos lotes de IVIG por ele recebido também apresentavam níveis adequados do anticorpo específico. O caso ilustra que o tratamento regular com IVIG não é suficiente para prevenir a infecção pelo vírus da varicela-zoster.

Evaluación de la capacidad neutralizante de extractos de plantas de uso popular en Guatemala como antídotos para el envenenamiento por la mordedura de Bothrops asper / Neutralizing activities of ethanolic extracts of six plants traditionally used in Guatemala as antidotes for the envenomation caused by the snake Bothrops asper

Se determinó la capacidad de los extractos de seis plantas de uso etnomédico (Acacia hindsii, Aristolochia maxima, Cissampelos pareira, Hamelia patens, Piper peltatum y Sansevieria hyacinthoides) para neutralizar los efectos proteolítico y fosfolipasa A2 (PLA2) del veneno de Bothrops asper, la principal especie causante de envenenamiento en el país. Estos efectos, indicadores de la capacidad miotóxica, hemorrágica e inflamatoria del veneno, se evaluaron en ensayos controlados in vitro. Las plantas fueroncolectadas, secadas y extraídas por percolación con etanol. Los resultados demuestran que ninguno de los extractos posee actividad PLA2 o proteo-lítica intrínseca a las dosis estudiadas. Se determinó que tres de los extractos neutralizaron pobremente (< 50%) los efectos estudiados: S. hyacinthoides neutralizó 13.90 ± 6.41% del efecto PLA2 y P. peltatum y C. pareira el 32.98 ± 5.51% y 24.52 ± 7.45%, respectivamente, del efecto proteolítico. Por ello, ningún extracto se evaluó en pruebas de neutralización de la letalidad en ratones. Se concluye que no es recomendable el uso aislado de estas plantas en el tratamiento del envenenamiento por mordedura de B. asper, aunque posiblemente las que demostraron alguna actividad puedan resultar potenciadas al usarse en combinación con otras plantas, como se hace en las recetas tradicionales. Dada la complejidad de los componentes del veneno y sus efectos fisiopatológicos, falta investigar la capacidad de las plantas estudiadas para neutralizar las coagulopatías, edema y miotoxicidad producidas durante el envenenamiento.

Many plants are reported to be used in Guatemalan traditional medicine as antidotes against various effects of the snakebite; however, very few attempts have been made to evaluate their neutralizing capacity in controlled experiments. Six plants (Acacia hindsii, Cissampelos pareira; Hamelia patens, Piper peltatum, Sansevieria hyacinthoides and Aristolochia maxima) were evaluated in vitro for their ability to neutralize phospholipase A2 (PLA2) and proteolytic effects of the venom of Bothrops asper, the snake responsible for approximately half of the snakebite envenomations in Central America. These effects are indicatives of the ability of B. asper venom to produce myotoxicity, hemorrhage and inflammation. Plants were collected, dried and extracted by maceration with ethanol. After pre-incubation of several amounts of each extract with a challenge dose of venom, S. hyacinthoides demonstrated a low neutralizing capacity (< DE 50) of the PLA2 effect (13.90 ± 6.41%); C. pareira (32.98 ± 5.51%) and P. peltatum (24.52 ± 7.45%) neutralized less than 50% of the proteolytic effect. The results suggest that neither of the tested plants should be used individually to treat the main effects of B. asperenvenomation. However, the three low-active extracts might be potentiated when used in mixtures composed of several plants, as prepared by traditional healers. Given the complexity of the venom components and the multiple pathologic effects produced by B. asper envenomation, more tests are required to fully investigate the ability of this plants to neutralize the coagulant, fibrin(ogen)olytic, edematizing and myotoxic effects of the venom.

[Lactic acid and lactoserum in vulvovaginal infections]. / Acido láctico y lactoserum en infecciones vulvovaginales.

BACKGROUND: The discomfort of vaginal infection symptoms is a frequent cause of medical consultation. OBJECTIVE: To determine the degree of satisfaction reported by a group of Mexican patients used the combination lactoserum lactic acid and to the relief of vulvovaginal symptoms (odor, itching and burning) or not associated with infection and the safety of that combination when used for external genital hygiene daily. PATIENTS AND METHOD: Clinical observational, prospective, multicenter open and made with a group of Mexican women 18-60 years of age, residents in cities across the country with some or vulvovaginal symptoms: foul odor, itching, burning. All participants were told the application, once daily, 2.5 to 5 mL of shampoo, for 14-21 days. The patients rated the presence and severity of vulvar discomfort with visual analogue scale. The data were analyzed with Statistical Analysis System statistical package version 9.1, the quantitative variables by calculating the mean +/- standard deviation, categorical variables by calculating percentages. RESULTS: We included 559 patients who answered the questionnaires before and after treatment. The time-use shampoo study was variable, with less than seven days were 25 patients (4.5%), 8 to 14 days 123 patients (22.0%) of 15 to 21 days, 208 patients (37.3%) and 178 patients (31.9%) used it more than 21 days. The average time elapsed since the start of treatment until the disappearance of vulvar discomfort was 4.3 days, with a SD of 3.1. In symptomatic patients the average was 4.2 days (SD 3.3), while in asymptomatic patients was 4.4 days (SD 3.0), virtually the same between groups. At the time of the final visit, 92.2% of patients reported "feeling of cleanliness" in the genital area, 91.7% "fresh feeling" and 93.9% "general feeling of wellbeing". 97.2% felt that the product is "user friendly", 94.9% considered it "soft and gentle to the skin" and 95.2% described it with "pleasing aroma". CONCLUSIONS: Shampoo daily female external genital hygiene, the reason for this study is useful in the disappearance of the symptoms associated with infection or vulvovaginal.

Surto de tétano em búfalos (Buballus bubalis) no Estado do Pará / Outbreak of tetanus in bufalloes (Buballus bubalis) in Pará, Brazil

Descreve-se um surto de tétano em búfalos da raça Murrah em uma propriedade situada no município de São Caetano de Odivelas, localizado na região metropolitana de Belém, estado do Pará. Do rebanho de 250 bubalinos, 80 animais foram vacinados contra raiva por via intramuscular na região da garupa. Em um período de 15 a 19 dias após a vacinação quatro animais adoeceram, um morreu com dois dias de evolução, um foi eutanasiado in extremis no sétimo dia após o início dos sinais clínicos e os demais se recuperaram após tratamento. Nos bubalinos, o primeiro sinal clínico observado foi o prolapso da terceira pálpebra, em especial quando o animal era estimulado, seguido por andar rígido, manifestado por dificuldade de flexão dos membros e permanência em decúbito lateral com os membros estendidos, pálpebras muito abertas, sialorréia, hiperexcitabilidade, orelhas eretas, leve trismo e acúmulo de alimento na cavidade oral. À necropsia foi evidenciada uma área de coloração amarelada com presença de exsudação purulenta na musculatura da região da garupa, local de aplicação da vacina. Ao exame histopatológico não foram evidenciadas alterações significativas. Em dois animais foi realizado tratamento com penicilina por via intramuscular e soro antitetânico por via intramuscular e sub-aracnóide; após duas semanas esses animais se recuperaram. Diante do quadro clínico, dos dados epidemiológicos e da ausência de lesões histológicas foi feito o diagnóstico de tétano. Concluiu-se que o tétano é uma doença a ser considerada na bubalinocultura no Brasil. A infecção, provavelmente ocorreu durante o procedimento de vacinação, através injeções intramusculares utilizando agulhas contaminadas.

An outbreak of tetanus in Murrah buffaloes in Belém, state of Pará, Brazil, is described. The outbreak was observed in a flock of 250 buffaloes, 80 of which were vaccinated intra-muscularly in the croup against rabies. Four buffaloes fell ill 15 to 19 days after vaccination, one died after a clinical course of 2 days, one was submitted to euthanasia in extremis on the 7th day after the onset of symptoms, and two others recovered within 2 weeks after treatment with penicillin for 4 days and anti-tetanic serum. The first symptoms observed were prolapse of the third eyelid, especially when the animal was stimulated, followed by a rigid walk, as difficulty in inflexing the members and lateral recumbence with the members extended, besides very open eyelids, sialorrhea, hyperexcitability, erect ears and slight lockjaw, and food accumulation in the oral cavity. At post-mortem examination was found a focus of yellowish pus in the croup muscles, where the vaccine had been applied. At histopathology no significant alterations in internal organs were observed. Based on the clinical picture, the epidemic data and absence of histological lesions, the diagnosis of tetanus was established. The infection was considered to be due to the use of dirty needles used during vaccination.

Surto de tétano em búfalos (Buballus bubalis) no Estado do Pará / Outbreak of tetanus in bufalloes (Buballus bubalis) in Pará, Brazil

Descreve-se um surto de tétano em búfalos da raça Murrah em uma propriedade situada no município de São Caetano de Odivelas, localizado na região metropolitana de Belém, estado do Pará. Do rebanho de 250 bubalinos, 80 animais foram vacinados contra raiva por via intramuscular na região da garupa. Em um período de 15 a 19 dias após a vacinação quatro animais adoeceram, um morreu com dois dias de evolução, um foi eutanasiado in extremis no sétimo dia após o início dos sinais clínicos e os demais se recuperaram após tratamento. Nos bubalinos, o primeiro sinal clínico observado foi o prolapso da terceira pálpebra, em especial quando o animal era estimulado, seguido por andar rígido, manifestado por dificuldade de flexão dos membros e permanência em decúbito lateral com os membros estendidos, pálpebras muito abertas, sialorréia, hiperexcitabilidade, orelhas eretas, leve trismo e acúmulo de alimento na cavidade oral. À necropsia foi evidenciada uma área de coloração amarelada com presença de exsudação purulenta na musculatura da região da garupa, local de aplicação da vacina. Ao exame histopatológico não foram evidenciadas alterações significativas. Em dois animais foi realizado tratamento com penicilina por via intramuscular e soro antitetânico por via intramuscular e sub-aracnóide; após duas semanas esses animais se recuperaram. Diante do quadro clínico, dos dados epidemiológicos e da ausência de lesões histológicas foi feito o diagnóstico de tétano. Concluiu-se que o tétano é uma doença a ser considerada na bubalinocultura no Brasil. A infecção, provavelmente ocorreu durante o procedimento de vacinação, através injeções intramusculares utilizando agulhas contaminadas.(AU)

An outbreak of tetanus in Murrah buffaloes in Belém, state of Pará, Brazil, is described. The outbreak was observed in a flock of 250 buffaloes, 80 of which were vaccinated intra-muscularly in the croup against rabies. Four buffaloes fell ill 15 to 19 days after vaccination, one died after a clinical course of 2 days, one was submitted to euthanasia in extremis on the 7th day after the onset of symptoms, and two others recovered within 2 weeks after treatment with penicillin for 4 days and anti-tetanic serum. The first symptoms observed were prolapse of the third eyelid, especially when the animal was stimulated, followed by a rigid walk, as difficulty in inflexing the members and lateral recumbence with the members extended, besides very open eyelids, sialorrhea, hyperexcitability, erect ears and slight lockjaw, and food accumulation in the oral cavity. At post-mortem examination was found a focus of yellowish pus in the croup muscles, where the vaccine had been applied. At histopathology no significant alterations in internal organs were observed. Based on the clinical picture, the epidemic data and absence of histological lesions, the diagnosis of tetanus was established. The infection was considered to be due to the use of dirty needles used during vaccination.(AU)

Absence of experimental cross-protection induced by a Trypanosoma cruzi-like strain isolated from bats.

This study evaluated the possibility of inoculation and reinoculation with a trypanosomatid isolated from bats that is morphologically, biologically and molecularly similar to Trypanosoma cruzi, to protect against infection by virulent strains. Non-isogenic mice were divided into 24 groups that received from zero to three inoculations of Trypanosoma cruzi-like strain RM1, in the presence or absence of Freunds adjuvant, and were challenged with the VIC or JG strains of Trypanosoma cruzi. Parasitemia and survival were monitored and animals were sacrificed for histopathological analysis. Animals immunized with Trypanosoma cruzi-like strain RM1 presented decreased parasitemia, independently of the number of inoculations or the presence of adjuvant. In spite of this reduction, these animals did not present any protection against histopathological lesions. Severe eosinophilic infiltrate was observed and was correlated with the number of inoculations of Trypanosoma cruzi-like strain RM1. These findings suggest that prior inoculation with this strain did not protect against infection but, rather, aggravated the tissue inflammatory process.

Absence of experimental cross-protection induced by a Trypanosoma cruzi-like strain isolated from bats / Ausência de proteção cruzada experimental induzida por uma cepa Trypanosoma cruzi-like isolada de morcego

This study evaluated the possibility of inoculation and reinoculation with a trypanosomatid isolated from bats that is morphologically, biologically and molecularly similar to Trypanosoma cruzi, to protect against infection by virulent strains. Non-isogenic mice were divided into 24 groups that received from zero to three inoculations of Trypanosoma cruzi-like strain RM1, in the presence or absence of Freund’s adjuvant, and were challenged with the VIC or JG strains of Trypanosoma cruzi. Parasitemia and survival were monitored and animals were sacrificed for histopathological analysis. Animals immunized with Trypanosoma cruzi-like strain RM1 presented decreased parasitemia, independently of the number of inoculations or the presence of adjuvant. In spite of this reduction, these animals did not present any protection against histopathological lesions. Severe eosinophilic infiltrate was observed and was correlated with the number of inoculations of Trypanosoma cruzi-like strain RM1. These findings suggest that prior inoculation with this strain did not protect against infection but, rather, aggravated the tissue inflammatory process.

Este trabalho avaliou a possibilidade da inoculação e reinoculação de um tripanossomatídeo isolado de morcego, morfológica, biológica e molecularmente semelhante ao Trypanosoma cruzi, na proteção contra a infecção por cepas virulentas. Camundongos não-isogênicos foram divididos em 24 grupos, que receberam de zero a três inóculos da cepa RM1 de Trypanosoma cruzi-like, na presença ou ausência de adjuvante de Freund e desafiados com as cepas de Trypanosoma cruzi VIC ou JG. Acompanhou-se a parasitemia e a sobrevida e os camundongos foram sacrificados para análise histopatológica. Os animais imunizados com a cepa RM1 de Trypanosoma cruzi-like apresentaram redução da parasitemia, independente do número de inóculos ou presença de adjuvante. Apesar dessa redução, os animais não apresentaram proteção contra lesões histopatológicas e observaram-se intensos infiltrados eosinofílicos que foram correlacionados com o número de inóculos da cepa RM1 de Trypanosoma cruzi-like. Sugere-se que a inoculação prévia dessa cepa, ao invés de proteger contra a infecção, agravou o processo inflamatório tecidual.

Treatment of Argentine hemorrhagic fever.

Argentine hemorrhagic fever (AHF) is a rodent-borne illness caused by the arenavirus Junin that is endemic to the humid pampas of Argentina. AHF has had significant morbidity since its emergence in the 1950s, with a case-fatality rate of the illness without treatment between 15% and 30%. The use of a live attenuated vaccine has markedly reduced the incidence of AHF. Present specific therapy involves the transfusion of immune plasma in defined doses of neutralizing antibodies during the prodromal phase of illness. However, alternative forms of treatment are called for due to current difficulties in early detection of AHF, related to its decrease in incidence, troubles in maintaining adequate stocks of immune plasma, and the absence of effective therapies for severely ill patients that progress to a neurologic-hemorrhagic phase. Ribavirin might be a substitute for immune plasma, provided that the supply is guaranteed. Immune immunoglobulin or monoclonal antibodies should also be considered. New therapeutic options such as those being developed for systemic inflammatory syndromes should also be valuated in severe forms of AHF.

Lipoprotein NMB0928 from Neisseria meningitidis serogroup B as a novel vaccine candidate.

Polysaccharide-based vaccines for serogroup B Neisseria meningitidis have failed to induce protective immunity. As a result, efforts to develop vaccines for serogroup B meningococcal disease have mostly focused on outer membrane proteins (OMP). Vaccine candidates based on meningococcal OMP have emerged in the form of outer membrane vesicles (OMVs) or, more recently, purified recombinant proteins, as alternative strategies for serogroup B vaccine development. In our group, the protein composition of the Cuban OMVs-based vaccine VA-MENGOC-BC was elucidated using two-dimensional gel electrophoresis and mass spectrometry. The proteomic map of this product allowed the identification of new putative protective proteins not previously reported as components of an antimeningococcal vaccine. In the present study, we have determined the immunogenicity and protective capacity of NMB0928, one of those proteins present in the OMVs. The antigen was obtained as a recombinant protein in Escherichia coli, purified and used to immunize mice. The antiserum produced against the protein was capable to recognize the natural protein in different meningococcal strains by whole-cell ELISA and Western blotting. After immunization, recombinant NMB0928 induced bactericidal antibodies, and when the protein was administered inserted into liposomes, the elicited antibodies were protective in the infant rat model. These results suggest that NMB0928 is a novel antigen worth to be included in a broadly protective meningococcal vaccine.

Identification by genomic immunization of a pool of DNA vaccine candidates that confer protective immunity in mice against Neisseria meningitidis serogroup B.

We have shown previously that expression library immunization is viable alternative approach to induce protective immunity against Neisseria meningitidis serogroup B. In this study we report that few rounds of library screening allow identification of protective pools of defined antigens. A previously reported protective meningococcal library (L8, with 600 clones) was screened and two sub-libraries of 95 clones each were selected based on the induction of bactericidal and protective antibodies in BALB/c mice. After sequence analysis of each clone within these sub-libraries, we identified a pool of 20 individual antigens that induced protective immune responses in mice against N. meningitidis infection, and the observed protection was associated with the induction of bactericidal antibodies. Our studies demonstrate for the first time that ELI combined with sequence analysis is a powerful and efficient tool for identification of candidate antigens for use in a meningococcal vaccine.

Macrophage migration inhibitory factor is essential for allergic asthma but not for Th2 differentiation.

Macrophage migration inhibitory factor (MIF) is increased in asthmatic patients and plays a critical role in the pathogenesis of asthma. We show here that mice lacking MIF failed to develop airway hyper-responsiveness (AHR), tissue eosinophilia, and mucus metaplasia. Analysis of the bronchoalveolar fluids revealed a substantial reduction of IL-13, eotaxin and cysteinyl-leukotrienes. The lack of these cardinal features of asthma in MIF(-/-) mice occurs regardless of high concentrations of IL-4 in the lung and OVA-specific IgE in the serum. Antigen-specific lymphocyte proliferation and IL-13 production were similarly increased in the draining lymph nodes of OVA-immunized and challenged MIF(-/-) mice compared to WT, but were reduced in the spleen of MIF(-/-), thus indicating differential roles of MIF in these compartments. Stimulation of naive CD4(+) cells with anti-CD3 antibody demonstrated that MIF(-/-) cells produced increased amounts of IFN-gamma and IL-4 compared to WT CD4(+) cells. Finally, treatment of sensitized BALB/c mice with neutralizing anti-MIF antibody abrogated the development of ARH and airway inflammation without affecting the production of Th2 cytokines or IgE. The present study demonstrates that MIF is required for allergic inflammation, adding important elements to our knowledge of asthma pathogenesis and suggesting that neutralization of MIF might be of therapeutic value in asthma.

Varicella zoster virus antibody titers after intravenous zoster immune globulin in neonates, and the safety of this preparation.

AIMS: 1) To assess the safety of intravenous varicella zoster virus hyperimmune globulin G (IV-VZVIG) in neonates; 2) measure varicella zoster virus-specific IgG antibody (VZVIG) changes in newborn IV-VZVIG recipients. METHODS: Eighteen neonatal intensive care unit (NICU) neonates with varicella exposure were given 1 ml/kg (43 International Units (IU)/kg) Varitect IV-VZVIG. Serum VZVIG titers were assayed in neonatal recipients 0, 1, 7, 14, 21, 28, and 35 d after IV-VZVIG. Also, serum samples for VZV-IgM antibody determinations were obtained at 4 wk post-infusion. RESULTS: No varicella developed in the 18 infants. Infusion of 1 ml/kg (43 IU/kg) IV-VZVIG was generally safe. The IV-VZVIG dose resulted in seroconversion of a non-immune newborn. Five infants had low basal VZVIG titers, and two of them had a 0.6-log10 increase at 24 h post-infusion. Neonatal VZVIG titers (mean+/-SEM in log(10)) before IV-VZVIG and after by 1, 7, 14, 21, 28, and 35 d were 2.22+/-0.15, 2.17+/-0.17, 2.02+/-0.12, 0.87+/-0.2, 1.09+/-0.19, 2.33+/-0.07, and 2.16+/-0.1, respectively. CONCLUSIONS: One ml/kg (43 IU/kg) IV-VZVIG was generally safe. Our neonatal mean VZV-immune status did not significantly increase after the 1 ml/kg (43 IU/kg) IV-VZVIG dose, although no varicella developed and it caused a VZV-specific seroconversion.

Contribuição da FUNED para a produção de soros antivenenos e antitóxicos no Brasil

O artigo apresenta um breve relato da situação dos acidentes por animais

venenosos e produção de antivenenos.Destaca a produção de soros antiofídicos

e antitóxicos da FUNED, que foi iniciada juntamente com a sua fundação.

Enfatiza a relevante contribuição da FUNED para a auto-suficiência de

um produto estratégico e a manutenção de um programa bem sucedido

do Ministério da Saúde, o Programa Nacional de Imunizações, que garante,

gratuitamente, um produto de qualidade a pacientes em todo o território