São Paulo School of Advanced Sciences on Vaccines: an overview

Two years ago, we held an exciting event entitled the São Paulo School of Advanced Sciences on Vaccines (SPSASV). Sixty-eight Ph.D. students, postdoctoral fellows and independent researchers from 37 different countries met at the Mendes Plaza Hotel located in the city of Santos, SP - Brazil to discuss the challenges and the new frontiers of vaccinology. The SPSASV provided a critical and comprehensive view of vaccine research from basics to the current state-of-the-art techniques performed worldwide. For 10 days, we discussed all the aspects of vaccine development in 36 lectures, 53 oral presentations and 2 poster sessions. At the end of the course, participants were further encouraged to present a model of a grant proposal related to vaccine development against individual pathogens. Among the targeted pathogens were viruses (Chikungunya, HIV, RSV, and Influenza), bacteria (Mycobacterium tuberculosis and Streptococcus pyogenes), parasites (Plasmodium falciparum or Plasmodium vivax), and the worm Strongyloides stercoralis. This report highlights some of the knowledge shared at the SPSASV.(AU)

Biochemical and biological characterization of the Hypanus americanus mucus: A perspective on stingray immunity and toxins.

Stingrays skin secretions are largely studied due to the human envenoming medical relevance of the sting puncture that evolves to inflammatory events, including necrosis. Such toxic effects can be correlated to the biochemical composition of the sting mucus, according to the literature. Fish skin plays important biological roles, such as the control of the osmotic pressure gradient, protection against mechanical forces and microorganism infections. The mucus, on the other hand, is a rich and complex fluid, acting on swimming, nutrition and the innate immune system. The elasmobranch's epidermis is a tissue composed mainly by mucus secretory cells, and marine stingrays have already been described to present secretory glands spread throughout the body. Little is known about the biochemical composition of the stingray mucus, but recent studies have corroborated the importance of mucus in the envenomation process. Aiming to assess the mucus composition, a new non-invasive mucus collection method was developed that focused on peptides and proteins, and biological assays were performed to analyze the toxic and immune activities of the Hypanus americanus mucus. Pathophysiological characterization showed the presence of peptidases on the mucus, as well as the induction of edema and leukocyte recruitment in mice. The fractionated mucus improved phagocytosis on macrophages and showed antimicrobial activity against T. rubrumç. neoformans and C. albicans in vitro. The proteomic analyses showed the presence of immune-related proteins like actin, histones, hemoglobin, and ribosomal proteins. This protein pattern is similar to those reported for other fish mucus and stingray venoms. This is the first report depicting the Hypanus stingray mucus composition, highlighting its biochemical composition and importance for the stingray immune system and the possible role on the envenomation process.

The performance of serological tests for Leishmania infantum infection screening in dogs depends on the prevalence of the disease.

Dogs are considered the main reservoir of Leishmania infantum. This protozoan causes visceral leishmaniasis (VL), an uncontrolled urban zoonosis in Brazil. Serological tests and polymerase chain reaction (PCR) on peripheral blood were performed to identify infected dogs in scenarios of higher and lower prevalence of the disease (Teresina and Vitória). One-hundred infected and 57 non-infected animals from Teresina and 100 non-infected animals from Vitória were studied. Animal selection was not dependent on previous serology. The sensitivity (Teresina) and specificity (Teresina and Vitória) were as follows: indirect antibody fluorescence (IFAT) cut-off of 1:40 (IFAT 1:40): 96%, 18%, and 76%; IFAT 1:80: 90%, 33%, and 93%; direct agglutination test (DAT): 96%, 33%, and 98%; fast agglutination screening test (FAST): 93%, 68%, and 100%; immunochromatographic assay with a recombinant rK39 antigen (rK39): 88%, 74%, and 98%; enzyme linked immunosorbent assay (ELISA): 91%, 79%, and 98%; rapid dual-path platform test (TR DPP®): 98%, 60%, and 98%; and blood PCR: 29%, 93%, and 97%, respectively. In the high transmission area, none of the tests adequately discriminated L. infantum-infected from non-infected dogs. However, in the high transmission city, the area under the receiver operating characteristic (ROC) curve of FAST, DAT, ICrK39, ELISA and TR DPP® was high.

50 años dedicados al desarrollo de la Inmunología / 50 years dedicated to the development of Immunology

En el presente trabajo, dedicado a los 50 años de trabajo del Instituto de Hematología e Inmunología, se describe el desarrollo de la especialidad de Inmunología, sus principales alcances y logros científicos en las líneas de desarrollo fundamentales como: la introducción de técnicas de avanzada, el estudio inmunológico de hemopatías y el diagnóstico de las inmunodeficiencias primarias y secundarias. Dentro de estos se destacan la introducción de la citometría de flujo que permitió resultados relevantes en inmunología básica, en su aplicación al estudio de hemopatías como la anemia drepanocítica y el perfeccionamiento del diagnóstico inmunológico de las leucemias agudas del país. También, la caracterización de células madre procedentes de la médula ósea, utilizadas en la terapia celular regenerativa, y el estudio inmunológico del enfermo con insuficiencia renal crónica en espera de trasplante renal en Cuba(AU)

The present work, devoted to the 50 years of work of the Institute of Hematology and Immunology, describes the development of the Immunology specialty, its main achievements and scientific results on the fundamentals development lines, such as the introduction of advanced techniques, the immunological study of hemopathies and the diagnosis of primary and secondary immunodeficiencies. Among these, we highlight the introduction of flow cytometry that allowed relevant results in basic immunology, in its application to the study of hemopathies such as sickle cell anemia and the improvement of the immunological diagnosis of acute leukemias in the country. Also, the characterization of stem cells from the bone marrow used in regenerative cell therapy and the immunological study of the patient with chronic renal failure pending renal transplantation in Cuba(AU)

The performance of serological tests for Leishmania infantum infection screening in dogs depends on the prevalence of the disease

ABSTRACT Dogs are considered the main reservoir of Leishmania infantum. This protozoan causes visceral leishmaniasis (VL), an uncontrolled urban zoonosis in Brazil. Serological tests and polymerase chain reaction (PCR) on peripheral blood were performed to identify infected dogs in scenarios of higher and lower prevalence of the disease (Teresina and Vitória). One-hundred infected and 57 non-infected animals from Teresina and 100 non-infected animals from Vitória were studied. Animal selection was not dependent on previous serology. The sensitivity (Teresina) and specificity (Teresina and Vitória) were as follows: indirect antibody fluorescence (IFAT) cut-off of 1:40 (IFAT 1:40): 96%, 18%, and 76%; IFAT 1:80: 90%, 33%, and 93%; direct agglutination test (DAT): 96%, 33%, and 98%; fast agglutination screening test (FAST): 93%, 68%, and 100%; immunochromatographic assay with a recombinant rK39 antigen (rK39): 88%, 74%, and 98%; enzyme linked immunosorbent assay (ELISA): 91%, 79%, and 98%; rapid dual-path platform test (TR DPP®): 98%, 60%, and 98%; and blood PCR: 29%, 93%, and 97%, respectively. In the high transmission area, none of the tests adequately discriminated L. infantum-infected from non-infected dogs. However, in the high transmission city, the area under the receiver operating characteristic (ROC) curve of FAST, DAT, ICrK39, ELISA and TR DPP® was high.

Serological diagnosis of canine visceral leishmaniasis in Brazil: systematic review and meta-analysis.

OBJECTIVE: To evaluate the quality and accuracy of serological diagnosis of canine visceral leishmaniasis in the Americas. METHODS: A systematic review found original studies in the databases MEDLINE, EMBASE and LILACS up to November 2012 and in complementary sources up to February 2013. Studies were evaluated in accordance with QUADAS 2 and STARD parameters and recommended in accordance with GRADE parameters. Meta-analysis was carried out with Meta-DiSc software, using the random-effect model. RESULTS: Two hundred and eighty-four studies were identified, of which 25 met the inclusion criteria, comprising the final synthesis. All but one was conducted in Brazil, and only two were judged to be of good quality. 15 studies involving immuno-enzymatic tests with crude antigens (cELISA), 11 studies on indirect immunofluorescence tests (IFAT) and three on the immunochromatographic dual-path platform (DPP) test were meta-analysed. The combined results for sensitivity and specificity were cELISA: 0.89 (CI 95% 0.87-0.91) and 0.87 (CI 95% 0.86-0.88); IFAT: 0.88 (CI 95% 0.85-0.91) and 0.63 (CI 95% 0.61-0.65); and DPP: 0.83 (CI 95% 0.78-0.88) and 0.73 (CI 95% 0.70-0.75). CONCLUSION: Enzyme-linked immunosorbent assay with crude antigens and DPP tests have moderate accuracy for the diagnosis of canine visceral leishmaniasis, and the quality of the design, implementation and analysis of validation studies on diagnostic tests for this disease urgently require improvement. The recommendation for use of the evaluated tests is based on evidence that is scarce and restricted to Brazil.

Ferromagnetic resonance for the quantification of superparamagnetic iron oxide nanoparticles in biological materials

The aim of the present work is the presentation of a quantification methodology for the control of the amount of superparamagnetic iron oxide nanoparticles (SPIONs) administeredin biological materials by means of the ferromagnetic resonance technique (FMR) applied to studies both in vivo and in vitro. The in vivo study consisted in the analysis of the eliminationand biodistribution kinetics of SPIONs after intravenous administration in Wistar rats. The results were corroborated by X-ray fluorescence. For the in vitro study, a quantitative analysisof the concentration of SPIONs bound to the specific AC133 monoclonal antibodies was carriedout in order to detect the expression of the antigenic epitopes (CD133) in stem cells from human umbilical cord blood. In both studies FMR has proven to be an efficient technique forthe SPIONs quantification per volume unit (in vivo) or per labeled cell (in vitro).

Pruebas hormonales e inmunológicas para la evaluación de la función tiroidea / Hormonal and immunological tests for evaluating the thyroidal function

En la década de los años 80, con el desarrollo alcanzado en la tecnología de producción de anticuerpos monoclonales y en la concepción de un diseño de ensayo no competitivo tipo sandwich o de "2 sitios", comenzó el auge de los métodos inmunométricos (IMAs) en sus diferentes tipos en dependencia de la señal y reacción que la desencadena. Inicialmente fueron desarrollados los métodos inmunorradiométricos (IRMAs) de naturaleza isotópica y casi simultáneamente con estos los IMAs de naturaleza no isotópica. Entre estos últimos se encuentran los ensayos inmunoenzimométricos (IEMAs), los inmunofluorimétricos (IFMAs), los inmunoluminométricos (ILMAs) en sus subtipos de inmunoquimioluminiscentes (ICMAs), inmunobioluminiscentes (IBMAs) e inmuoelectroquimioluminiscentes (IECMAs). Los IMAs de naturaleza no isotópica han permitido el incremento de la automatización. Los IRMAs y IEMAs son considerados métodos de 2da. generación, y los IFMAs, ICMAs, IBMAs e IECMAs son de tercera generación(AU)

Desarrollo de una técnica serológica para el diagnostico temprano de lesión hepática / Development of a serological technique for the early diagnosis of hepatic lesion

Existen antecedentes en la literatura los cuales describen la obtención y purificación de antígenos hepato-específicos, así como su demostración en la circulación, sin embargo éstos antígenos no se han empleado como un método específico de diagnóstico en caso de lesiones hepáticas. En el presente trabajo obtuvimos antígenos hepáticos de los cuales demostramos su especialidad mediante el uso de técnicas inmunológicas, posteriormente animales de laboratório foram sometidos a técnicas de intoxicación para la obtención de daño hepático experimental el cual fue demostrado mediante estudios histopatológicos del hígado. Al mismo tiempo se demuestra la presencia de antígenos hepato-específicos en la circulación mediante diversas técnicas inmunológicas, demostrando que las técnicas de fijación de complemento e Inmunoelectroforesis bidimensional detectan la presencia de éstos antígenos en la circulación desde etapas tempranas del inicio de la lesión

Iridociclite näo granulomatosa bilateral em um caso de leptospirose / Non- granulumatous bilateral acute iridocidytis in a young patient with leptospirosis

Ilustra-se um caso de iridociclite aguda näo granulomatosa bilateral em um paciente jovem com suspeita clínica e laboratorial de Leptospirose. É uma doença em ascençäo, porém pouco comentada em nosso meio oftalmológico