Detecção de proteínas imunorreativas de Rickettsia sp. cepa Mata Atlântica / Detection of immunoreactive proteins of Rickettsia sp. Atlantic Forest strain

A Febre Maculosa Brasileira (FMB) é uma doença infecciosa, transmitida por carrapatos ao homem. Uma nova riquetsiose humana foi descrita como causadora de Febre Maculosa no Estado de São Paulo, sendo denominada de Rickettsia sp. cepa Mata Atlântica. O presente trabalho teve como objetivo detectar e identificar proteínas com potencial de estimular o sistema imune de hospedeiro mamífero, desta nova cepa descrita. Para tanto, foi realizado a extração proteica total de Rickettsia sp. cepa Mata Atlântica. As proteínas extraídas foram fracionadas por eletroforese. As bandas proteicas foram transferidas para membranas de nitrocelulose por migração elétrica e submetidas à técnica de Western-blot, para detecção proteica. Ao todo sete proteínas imunorreativas foram detectadas. Duas proteínas apresentaram maior abundancia, com peso molecular, de 200 e 130 kDa respectivamente. Através da comparação de mapas proteômicos existentes e pelo peso molecular que estas proteínas apresentaram, sugere-se que as duas proteínas detectadas representem rOmpA (200 kDa) e rOmpB (130 kDa). As demais proteínas detectadas apresentaram menor ocorrência e peso molecular inferior a 78 kDa, podendo representar membros da família de antígenos de superfície celular (Sca - Surface cell antigen). As proteínas detectadas poderão servir como base de estudo na elaboração de métodos diagnósticos sensíveis e específicos, no desenvolvimento de vacinas, além de possibilitarem novos estudos para terapias mais eficazes.(AU)

Brazilian Spotted Fever (BSF) is an infectious disease transmitted by ticks to humans. A new human rickettsial infection was reported to cause spotted fever in the State of São Paulo and was named Rickettsia sp. Strain Atlantic Forest. This study aimed to detect and identify proteins with potential to stimulate the immune system of mammalian host of this new strain described. Therefore, we performed total protein extraction Rickettsia sp. Strain Atlantic Forest. The extracted proteins were fractionated by electrophoresis. The protein bands were transferred to nitrocellulose membrane by electrical migration and subjected to Western blot for protein detection. In all, seven immunoreactive proteins were detected. Two proteins showed higher abundance, with molecular weight of 200 and 130 kDa respectively. By comparing existing proteomic maps and the molecular weight of these proteins showed that, it is suggested that the two proteins detected representing rOmpA (200 kDa) and rOmpB (130 kDa). The other detected proteins had lower occurrence and molecular weight less than 78 kDa, which may represent members of the cell surface antigens Family (Sca - Surface cell antigen). The detected proteins may serve as a study based on the development of sensitive and specific diagnostic methods in the development of vaccines and they enable further studies to more effective therapies.(AU)

Research Progress in Targeted Drug Delivery System for Anti-intracellular Bacteria / 中国药师

Infection of intracellular pathogens remains a serious challenge for clinical anti-infective treatment. The most commonly used strategy for the treatment of intracellular bacteria is long-term use of antimicrobials at high concentration,which not only brings se-rious side effects to patients,but also causes great economic pressure to the whole society. Thus,the development of new strategies to better kill intracellular bacteria by the antimicrobials which showed good bactericidal effect in in vitro experiments is extremely impor-tant. With appropriate carriers to embed or encapsulate drugs can better control drugs' pharmacokinetic behavior resulting in effective bioavailability increase of antimicrobials. The current studies on targeted drug delivery systems for anti-intracellular bacteria were re-viewed in the paper.

Detection of macrophages infected with Mycobacterium avium subspecies paratuberculosis in a cow with clinical stage IV of Johne's disease. A case report / Detección de macrófagos infectados con Mycobacterium avium subspecies paratuberculosis en una vaca en fase clínica IV de la enfermedad de Johne. Reporte de un caso / Detecção de macrófagos infectados com Mycobacterium avium paratuberculosis subespécie em uma vaca com uma fase clínica IV da doença de Johne. Um relato de caso

Anamnesis: a six-year-old milking Holstein cow (Bos taurus taurus) was diagnosed with stage IV Johne's Disease (JD). Clinical and Laboratory findings: the cow suffered intermittent diarrhea during 6 months with no response to antibiotic treatment. Consequently, the cow was subjected to euthanasia. Treatment approach: antemortem milk and peripheral blood samples and postmortem colon, mediastinic, mesenteric lymph nodes, and spleen samples were processed for macrophages isolation. Total DNA was extracted from macrophages and used to diagnose IS900 of Mycobacterium avium paratuberculosis (MAP) through real time PCR. The MAP IS900 segment was successfully amplified from cells of all samples, indicating that these cells were MAP-infected macrophages. Conclusion: macrophages of cows suffering from JD can be used for amplification of the MAP IS900 segment.

Anamnesis: una vaca Holstein de 6 años de edad (Bos taurus taurus) presentó sintomatología de la fase IV de la enfermedad de Johne. Hallazgos clínicos y de laboratorio: la vaca presentó diarrea intermitente durante 6 meses sin respuesta al tratamiento con antibióticos. En consecuencia, la vaca fue sometida a eutanasia. Esquema de tratamiento: muestras de leche y de sangre periférica se tomaron ante-mortem; muestras de la mucosa del intestino, bazo y linfonodos mediastínico y mesentérico, se tomaron post-mortem, todas para aislamiento de macrófagos. El ADN total de los macrófagos fue usado para la amplificación del segmento IS900 de Mycobacterium avium paratuberculosis (MAP) por PCR en tiempo real. Conclusión: los macrófagos de vacas con la enfermedad de Johne pueden ser usados para la amplificación del segmento IS900 de MAP.

Anamnese: uma vaca Holstein com 6 anos de idade (Bos taurus taurus) apresentou os sintomas da fase IV da doença de Johne. Achados clínicos e laboratoriais: a vaca teve diarreia intermitente por seis meses sem resposta ao tratamento com antibióticos. Por conseguinte, a vaca foi submetida à eutanásia. Abordagem de tratamento: amostras de leite e de sangue periférico foram retiradas ante-mortem, enquanto as amostras da mucosa intestinal, baço e linfonodos mesentéricos e mediastinais foram tomadas todas post-mortem para o isolamento de macrófagos. O ADN total de macrófagos foi utilizado para amplificação do segmento IS900 do Mycobacterium avium paratuberculosis (MAP) por PCR em tempo real. Conclusão: macrófagos isolados a partir de vacas com doença de Johne podem ser utilizados para a amplificação da segmento IS900 do MAP.

Intracellular Bacterial Infection and Invariant NKT Cells

The invariant (i) natural killer (NK)T cells represent a unique subset of T lymphocytes which express the V alpha14 chain of the T cell receptor (TCR), that recognizes glycolipid antigens presented by the nonpolymorphic major histocompatibility complex (MHC) class I-like antigen presentation molecule CD1d, and they participate in protection against some microbial pathogens. Although iNKT cells have originally been regarded as T cells co-expressing NKR-P1B/C (NK1.1: CD 161), they do not seem to consistently express this marker, since NK1.1 surface expression on iNKT cells undergoes dramatic changes following facultative intracellular bacterial infection, which is correlated with functional changes of this cell population. Accumulating evidence suggests that NK1.1 allows recognition of "missing-self", thus controling activation/inhibition of NK1.1-expressing cells. Therefore, it is tempting to suggest that iNKT cells participate in the regulation of host immune responses during facultative intracellular bacterial infection by controlling NK1.1 surface expression. These findings shed light not only on the unique role of iNKT cells in microbial infection, but also provide evidence for new aspects of the NK1.1 as a regulatory molecule on these cells.