RESUMO
The aim of the present study was to investigate and compare muscle and tendon stiffness under active conditions in sprinters and untrained participants. In total, 14 sprinters and 24 untrained men participated in this study. Active muscle stiffness in the medial gastrocnemius muscle was calculated based on changes in estimated muscle force and fascicle length during fast stretching after submaximal isometric contractions. Tendon stiffness was determined during isometric plantar flexion using ultrasonography. No significant differences were observed in active muscle stiffness between sprinters and untrained men at any torque levels. Furthermore, no significant difference was noted in tendon stiffness between the two groups. These results suggest that muscle and tendon mechanical properties in the plantar flexors under active conditions are similar in sprinters and untrained participants.
Assuntos
Pé/fisiologia , Músculo Esquelético/fisiologia , Corrida/fisiologia , Tendões/fisiologia , Tendão do Calcâneo/fisiologia , Articulação do Tornozelo/fisiologia , Fenômenos Biomecânicos , Humanos , Contração Isométrica/fisiologia , Perna (Membro)/fisiologia , Masculino , Músculo Esquelético/anatomia & histologia , Aptidão Física , Tendões/anatomia & histologia , Tendões/diagnóstico por imagem , Torque , Ultrassonografia , Adulto JovemRESUMO
OBJECTIVE: To evaluate COBAS TaqMan MAI test misidentification of Mycobacterium lentiflavum as Mycobacterium intracellulare. MATERIALS AND METHODS: Preliminary comparative analysis identified 13 clinical isolates used in this study as COBAS Amplicor MAV and MIN-negative but COBAS TaqMan MAI-positive. The COBAS TaqMan MAI test limit of detection and reproducibility were evaluated by tenfold dilution series from 3 x 10(8) CFU/mL. Isolate 16S rDNA nucleotide sequences were compared with Mycobacterium avium and M. intracellulare. RESULTS: Discrepancies were observed between isolates identified as M. lentiflavum by 16S rDNA sequencing and as M. intracellulare by the COBAS TaqMan MAI test. The false-positive results were verified by sequence comparison of a randomly sampled clinical isolate and the M. intracellulare reference strain. Sequence analysis of M. lentiflavum and M. intracellulare 16S rDNA amplification products showed at least 3 mismatches between species. The high identity in the sequence was found for M. intracellulare by COBAS TaqMan MAI. CONCLUSION: In Japan, commercially available nucleic acid probe- and amplification-based tests cannot identify M. lentiflavum. Correct identification, though challenging, is possible using standard cultivation procedures for colony growth. Misleading results using the COBAS TaqMan MAI kit may lead to erroneous diagnoses.
Assuntos
Complexo Mycobacterium avium/genética , Complexo Mycobacterium avium/isolamento & purificação , Micobactérias não Tuberculosas/genética , Micobactérias não Tuberculosas/isolamento & purificação , Sequência de Bases , Kit de Reagentes para DiagnósticoRESUMO
BACKGROUND: Host effector mechanism against Mycobacterium tuberculosis (Mtb) infection is dependent on innate immune response by macrophages and neutrophils and the alterations in balanced adaptive immunity. Coordinated release of cytolytic effector molecules from NK cells and effector T cells and the subsequent granule-associated killing of infected cells have been documented; however, their role in clinical tuberculosis (TB) is still controversy. OBJECTIVE: To investigate whether circulating granulysin and other effector molecules are associated with the number of NK cells, iNKT cells, Vγ9(+)Vδ2(+) T cells, CD4(+) T cells and CD8(+) T cells, and such association influences the clinical outcome of the disease in patients with pulmonary TB and HIV/TB coinfection. METHODS: Circulating granulysin, perforin, granzyme-B and IFN-γ levels were determined by ELISA. The isoforms of granulysin were analyzed by Western blot analysis. The effector cells were analyzed by flow cytometry. RESULTS: Circulating granulysin and perforin levels in TB patients were lower than healthy controls, whereas the granulysin levels in HIV/TB coinfection were much higher than in any other groups, TB and HIV with or without receiving HAART, which corresponded to the number of CD8(+) T cells which kept high, but not with NK cells and other possible cellular sources of granulysin. In addition, the 17kDa, 15kDa and 9kDa isoforms of granulysin were recognized in plasma of HIV/TB coinfection. Increased granulysin and decreased IFN-γ levels in HIV/TB coinfection and TB after completion of anti-TB therapy were observed. CONCLUSION: The results suggested that the alteration of circulating granulysin has potential function in host immune response against TB and HIV/TB coinfection. This is the first demonstration so far of granulysin in HIV/TB coinfection.
Assuntos
Antígenos de Diferenciação de Linfócitos T/fisiologia , Infecções por HIV/complicações , Subpopulações de Linfócitos , Tuberculose/complicações , Adulto , Western Blotting , Feminino , Citometria de Fluxo , Infecções por HIV/fisiopatologia , Humanos , Masculino , Pessoa de Meia-Idade , Tuberculose/fisiopatologiaRESUMO
OBJECTIVE: Fluorescent staining is of paramount importance, not only for confirming the presence of mycobacteria in a given specimen but also for providing an estimated growth quantification. In this study, for rapidly growing Mycobacterium fortuitum, we evaluated the effectiveness of a rapid fluorescent staining method employing auramine-rhodamine (AR) fluorescent stain and acridine-orange (AO) fluorescent stain compared to that of the standard Ziehl-Neelsen (ZN) stain currently in use in our laboratory. METHOD: We evaluated the acid-fast nature of M. fortuitum strain ATCC6841 and 42 clinical isolates from each patient diagnosed at NHO Kinki-chuo Chest Medical Center. These isolates were preliminarily identified as M. fortuitum using DNA-DNA hybridization (DDH Mycobacteria; Kyokuto Pharmaceutical, Tokyo, Japan). These isolates were further identified by comparative sequence analysis of the ITS regions and the partial 16S rRNA gene. RESULTS: A total of 26 M. fortuitum strains (61.9%) demonstrated the lack of an acid-fast nature by AR staining, and slightly fewer demonstrated the same by AO staining. Sequence analysis of these 42 clinical isolates led to the identification of 35 M. fortuitum subsp. acetamidolyticum isolates (83.3%) and 7 closely M. fortuitum isolates. DISCUSSION: This work reported the loss of the acid-fast nature of specific M. fortuitum strains. It is likely that both the specific cell envelope of M. fortuitum and the staining mechanics could have been responsible for the loss of the acid-fast nature since the 2 different fluorescent stains yielded the same results. M. fortuitum is a mycobacterium species that does not stain with the commonly used fluorescence microscopy technique. Therefore, we suggested the use of an identification scheme for these organisms that employs ZN staining and the study of cultural characteristics (growth rate, temperature, and pigment production).
Assuntos
Corantes , Fluorescência , Mycobacterium fortuitum/isolamento & purificação , Humanos , Mycobacterium fortuitum/citologia , Análise de Sequência de RNA , Fatores de TempoRESUMO
We elucidated the relationship between prognosis of non-small-cell lung cancer (NSCLC) and Wilms' tumor gene (WT1) mRNA expression in tumor tissue. The WT1 mRNA expression levels of the fatal cases were lower as compared with those of the survival cases. Overall survival (OS) and disease-free survival (DFS) of the high WT1 expression group were longer than of the low expression group. As for squamous cell lung cancer (SQLC), low WT1 expression was significantly associated with lymph node metastasis. Cox analysis revealed that the gene level was a significant prognostic factor in OS and DFS. Low WT1 expression predicted poor prognosis in patients with NSCLC.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/genética , Genes do Tumor de Wilms , Neoplasias Pulmonares/genética , Proteínas WT1/biossíntese , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Intervalo Livre de Doença , Feminino , Expressão Gênica , Humanos , Neoplasias Pulmonares/metabolismo , Masculino , Pessoa de Meia-Idade , Prognóstico , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Proteínas WT1/genéticaRESUMO
SUBJECTS & METHODS: We reviewed the patient characteristics, radiological findings, treatments, and clinical outcomes in 12 cases of pulmonary Mycobacterium szulgai disease diagnosed at our hospital from April 1998 to March 2008. In addition, drug susceptibility testing of the causative isolates was performed with several antibiotics, including clarithromycin (CAM) and rifampicin (RFP), using BrothMIC NTM. RESULTS: The patients included 10 men and 2 women, with a mean age of 57.2 years. Among them, 10 were smokers, 5 previously had pulmonary tuberculosis, 3 had a history of gastric ulcers, and 1 had a history of esophageal cancer surgery. All patients had been previously treated with various chemotherapeutic regimens. Six of them were treated with chemotherapy, including CAM, and they improved both radiologically and bacteriologically. The minimal inhibitory concentration of CAM for all the strains tested was less than 0.25 microg/mL, which is the likely critical concentration for clinical efficacy of CAM. The present study suggested that, in addition to smoking and a history of pulmonary tuberculosis, gastroesophageal disorders were relevant underlying conditions in patients with pulmonary M. szulgai disease. CONCLUSION: Chemotherapeutic drugs, including CAM, are clinically and bacteriologically effective for pulmonary M. szulgai disease.
Assuntos
Pneumopatias , Infecções por Mycobacterium não Tuberculosas , Adulto , Idoso , Feminino , Humanos , Pneumopatias/tratamento farmacológico , Pneumopatias/microbiologia , Masculino , Pessoa de Meia-Idade , Infecções por Mycobacterium não Tuberculosas/tratamento farmacológico , Infecções por Mycobacterium não Tuberculosas/microbiologia , Estudos RetrospectivosRESUMO
PURPOSE: We aimed to investigate the prevalence and possible transmission routes of rifampicin (RFP) mono-resistant Mycobacterium tuberculosis strains. METHODS: Drug susceptibility testing was used to identify 15 RFP-resistant strains out of 4633 M. tuberculosis isolates. Sequencing of the rpoB gene and VNTR analysis were performed to further confirm the genetic classification. RESULTS: Resistance-conferring mutations in the RFP resistance-determining region (RRDR) of the rpoB gene were found in 14 of the 15 strains with phenotypic RFP mono-resistance. VNTR analysis revealed 2 clusters of 5 identical strains each. CONCLUSIONS: Although the community prevalence of RFP mono-resistant M. tuberculosis is low, the results of VNTR analysis suggested that rather than being recently transmitted, these strains may have been widely transmitted as latent infections in the population.
Assuntos
Antibióticos Antituberculose/farmacologia , Mycobacterium tuberculosis/genética , Rifampina/farmacologia , Proteínas de Bactérias/genética , RNA Polimerases Dirigidas por DNA , Farmacorresistência Bacteriana/genética , Mycobacterium tuberculosis/efeitos dos fármacosRESUMO
Granulysin and interferon-gamma (IFN-γ) have broad antimicrobial activity which controls Mycobacterium tuberculosis (M. tuberculosis) infection. Circulating granulysin and IFN-γ concentrations were measured and correlated with clinical disease in Thai patients with newly diagnosed, relapsed and chronic tuberculosis (TB). Compared to controls, patients with newly diagnosed, relapsed and chronic TB had lower circulating granulysin concentrations, these differences being significant only in newly diagnosed and relapsed TB (P < 0.001 and 0.004, respectively). Granulysin concentrations in patients with newly diagnosed and relapsed TB were significantly lower than in those with chronic TB (P= 0.003 and P= 0.022, respectively). In contrast, significantly higher circulating IFN-γ concentrations were found in patients with newly diagnosed and relapsed TB compared to controls (P < 0.001). The IFN-γ concentrations in newly diagnosed and relapsed patients were not significantly different from those of patients with chronic TB. However, in vitro stimulation of peripheral blood mononuclear cells (PBMCs) from patients with newly diagnosed, relapsed and chronic TB with purified protein derivative (PPD) or heat killed M. tuberculosis (H37Ra) enhanced production of granulysin by PBMCs. In vitro, stimulation of PBMCs of newly diagnosed TB patients with PPD produced greater amounts of IFN-γ than did controls, while those stimulated with H37Ra did not. The results demonstrate that patients with active pulmonary TB have low circulating granulysin but high IFN-γ concentrations, suggesting possible roles in host defense against M. tuberculosis for these agents.
Assuntos
Antígenos de Diferenciação de Linfócitos T/sangue , Interferon gama/sangue , Plasma/química , Tuberculose/diagnóstico , Tuberculose/imunologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Doença Crônica , Feminino , Humanos , Masculino , Mycobacterium tuberculosis , Recidiva , Tailândia , Tuberculose/microbiologia , Adulto JovemRESUMO
Objective. Our aim was to investigate the effects of IL-6 blockade on the progression of Mycobacterium tuberculosis (TB) and compare them with those of TNF-α blockade in mice. Methods. Mice were intravenously infected with TB and injected with antibodies. Survival was monitored and histological and immunological studies were carried out. Results. All anti-IL-6R Ab-treated mice and 8 of 10 control mice survived until sacrificed 224 days after TB challenge, whereas anti-TNF-α Ab-treated mice all died between 120 and 181 days. Anti-IL-6R Ab-treated mice exhibited no significant differences in TB CFU in organs, including the lungs, and no deterioration in histopathology compared to control mice at 4 weeks. In contrast, anti-TNF-α Ab-treated mice exhibited increased TB CFU and greater progression of histopathological findings in organs than control mice. Spleen cells from anti-TNF-α Ab-treated mice had decreased antigen-specific response in IFN-γ release and proliferation assays. The results in anti-IL-6R Ab-treated mice suggest that spleen cell responses were decreased to a lesser degree. Similar results were obtained in IL-6 knockout (KO) mice, compared with TNF receptor 1 (TNFR1) KO and TNFR1/IL-6 double KO (DKO) mice. Conclusion. IL-6R blockade promotes the progression of TB infection in mice far less than TNF-α blockade.
Assuntos
Anticorpos/farmacologia , Mycobacterium tuberculosis/efeitos dos fármacos , Receptores de Interleucina-6/imunologia , Tuberculose/patologia , Fator de Necrose Tumoral alfa/imunologia , Animais , Citocinas/metabolismo , Feminino , Interleucina-6/antagonistas & inibidores , Interleucina-6/genética , Fígado/microbiologia , Fígado/patologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos DBA , Camundongos Knockout , Mycobacterium tuberculosis/crescimento & desenvolvimento , Proteína Amiloide A Sérica/metabolismo , Baço/metabolismo , Baço/microbiologia , Análise de Sobrevida , Linfócitos T/imunologia , Tuberculose/mortalidadeRESUMO
OBJECTIVE: Mycobacterium tuberculosis infection is a major global threat to human health. The only tuberculosis (TB) vaccine currently available is bacillus Calmette-Guérin (BCG), although it has no efficacy in adults. Therefore, the development of a novel vaccine against TB for adults is desired. METHOD: A novel TB vaccine expressing mycobacterial heat shock protein 65 (HSP65) and interleukin-12 (IL-12) delivered by the hemagglutinating virus of Japan- (HVJ)- envelope was evaluated against TB infection in mice. Bacterial load reductions and histopathological assessments were used to determine efficacy. RESULTS: Vaccination by BCG prime with IgHSP65+murine IL-12/HVJ-envelope boost resulted in significant protective efficacy (>10, 000-fold versus BCG alone) against TB infection in the lungs of mice. In addition to bacterial loads, significant protective efficacy was demonstrated by histopathological analysis of the lungs. Furthermore, the vaccine increased the number of T cells secreting IFN-γ. CONCLUSION: This vaccine showed extremely significant protection against TB in a mouse model, consistent with results from a similar paper on cynomolgus monkeys. The results suggest that further development of the vaccine for eventual testing in clinical trials may be warranted.
Assuntos
Mycobacterium tuberculosis/imunologia , Vírus Sendai/genética , Vacinas contra a Tuberculose , Tuberculose Pulmonar/imunologia , Adulto , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Proteínas de Bactérias/metabolismo , Células Cultivadas , Chaperonina 60/genética , Chaperonina 60/imunologia , Chaperonina 60/metabolismo , Modelos Animais de Doenças , Vetores Genéticos , Humanos , Imunização Secundária , Interferon gama/metabolismo , Interleucina-12/genética , Interleucina-12/imunologia , Interleucina-12/metabolismo , Pulmão/imunologia , Pulmão/metabolismo , Pulmão/microbiologia , Pulmão/patologia , Camundongos , Camundongos Endogâmicos BALB C , Mycobacterium tuberculosis/patogenicidade , Linfócitos T/imunologia , Linfócitos T/metabolismo , Linfócitos T/patologia , Tuberculose Pulmonar/prevenção & controleRESUMO
PURPOSE: BCG is not efficacious against M. tuberculosis (TB) in adult. Therefore, novel TB vaccines were established by using three kinds of animal models (cynomolgus monkey model which is the best animal model of human TB, IL-2R knock out SCID mice as a human immune model, and granulysin transgenic mouse). METHODS AND RESULTS: DNA vaccine expressing TB Hsp65 and IL-12 was delivered by the hemagglutinating virus of Japan (HVJ)-envelope. The BCG prime followed by Hsp65+IL-12/HVJ vaccine boost showed a synergistic effect in the TB-infected cynomolgus monkey (100% survival). In contrast, 33% of monkeys were alive in BCG alone group. Furthermore, the prolongation of survival period of the monkey was observed by the combination of BCG and DNA vaccine even when the boost was performed after long-term period (4month) from prime. This combination also improved the erythrocyte sedimentation rate (ESR), increased the body weight, and augmented the proliferation of PBL and IL-12 production at higher levels than BCG alone or saline. Furthermore, this vaccine exerted therapeutic efficacy in IL-2R knock out SCID-PBL/hu mice, which were transplanted with human T cells. Granulysin is an important defensive molecule expressed by human T cells and NK cells and has a cytolytic activity against microbes including Mycobacterium tuberculosis (TB) and tumors. Expression of 15kD (15K) granulysin protein and mRNA in CD8 positive T cells in the patients infected with drug sensitive (TB) or multi-drug resistant (MDR-TB) M. tuberculosis were lower than that in the healthy volunteers, suggesting that granulysin treatment might improve the tuberculous disease in human. Therefore, we established two kinds of granulysin transgenic mice (15K granulysin transgenic mice and 9K granulysin transgenic mice). It was demonstrated that 15K granulysin transgenic mice as well as 9K granulysin transgenic mice exerted in vivo anti-TB effect, including the decrease of the number of TB and augmentation of the CTL activity. These are the first findings which demonstrate in vivo effects of 15K granulysin and 9K granulysin against TB infection. Moreover, DNA vaccine expressing 15K granulysin showed a therapeutic activity against TB in mice. CONCLUSION: These data indicate that monkey, IL-2R gene-knock out SCID-PBL/hu and granulysin transgenic mice models provide useful tools for the development of novel vaccines (HVJ-Envelope/Hsp65 DNA + IL-12 DNA vaccine and granulysin vaccine) against TB.
Assuntos
Proteínas de Bactérias/imunologia , Chaperonina 60/imunologia , Mycobacterium tuberculosis/imunologia , Vacinas contra a Tuberculose/administração & dosagem , Vacinas contra a Tuberculose/imunologia , Animais , Proteínas de Bactérias/genética , Proliferação de Células , Chaperonina 60/genética , Modelos Animais de Doenças , Imunização Secundária/métodos , Interleucina-12/genética , Interleucina-12/imunologia , Leucócitos Mononucleares/imunologia , Macaca fascicularis , Camundongos , Camundongos SCID , Camundongos Transgênicos , Mycobacterium tuberculosis/genética , Doenças dos Primatas/imunologia , Doenças dos Primatas/prevenção & controle , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Doenças dos Roedores/imunologia , Doenças dos Roedores/prevenção & controle , Vacinação/métodosRESUMO
PURPOSE: Multi-drug resistant (MDR) Mycobacterium Tuberculosis (M.TB) is a big problem in the world. We have developed novel TB therapeutic vaccines. METHODS AND RESULTS: DNA vaccine expressing mycobacterial heat shock protein 65 and IL-12 was delivered by the hemagglutinating virus of Japan (HVJ)-envelope. M. TB, MDR-TB or extremenly drug resistant (XDR-TB) was injected i.v. into DBA/1 mice, and treated with the vaccine three times. This HVJ-E/Hsp65DNA+IL-12DNA vaccine provided strong therapeutic efficacy against MDR-TB and XDR-TB (prolongation of survival time and the decrease in the number of TB) in mice. Therapeutic effect of this vaccine on TB infection was also demonstrated in chronic TB infection murine model using aerosol infection intratracheally. On the other hand, granulysin protein produced from CTL has lethal activity against TB. Granulysin protein vaccine also exerted strong therapeutic effect. Furthermore, we extended our studies to monkey model, which is currently the best animal model of human TB. Hsp65DNA+IL-12 DNA vaccine exerted strong therapeutic efficacy (100% survival and augmentation of immune responses) in the TB-infected monkeys. In contrast, the survival of the saline control group was 60% at 16 week post-challenge. HVJ-Envelope/HSP65 DNA+IL-12 DNA vaccine increased the body weight of TB-infected monkeys, improved the erythrocyte sedimentation rate, and augmentated the immune responses (proliferation of PBL and IL-2 production). The enhancement of IL-2 production from monkeys treated with this vaccine was correlated with the therapeutic efficacy of the vaccine. CONCLUSION: These data indicate that novel vaccines might be useful against TB including XDR-TB and MDR-TB for human therapeutic clinical trials.
Assuntos
Adjuvantes Imunológicos/administração & dosagem , Antígenos de Diferenciação de Linfócitos T/administração & dosagem , Imunoterapia/métodos , Vacinas contra a Tuberculose/imunologia , Tuberculose Resistente a Múltiplos Medicamentos/terapia , Vacinas de DNA/imunologia , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Chaperonina 60/genética , Chaperonina 60/imunologia , Modelos Animais de Doenças , Humanos , Interleucina-12/genética , Macaca fascicularis , Doenças dos Primatas/microbiologia , Doenças dos Primatas/terapia , Doenças dos Roedores/microbiologia , Doenças dos Roedores/terapia , Análise de Sobrevida , Resultado do Tratamento , Vacinas contra a Tuberculose/genética , Tuberculose Resistente a Múltiplos Medicamentos/imunologia , Vacinas de DNA/genéticaRESUMO
The purpose of this study was to investigate muscle and tendon properties in highly trained sprinters and their relations to running performance. Fifteen sprinters and 15 untrained subjects participated in this study. Muscle thickness and tendon stiffness of knee extensors and plantar flexors were measured. Sprinter muscle thickness was significantly greater than that of the untrained subjects for plantar flexors, but not for knee extensors (except for the medial side). Sprinter tendon stiffness was significantly lower than that of the untrained subjects for knee extensors, but not for plantar flexors. The best official record of a 100-m race was significantly correlated to the muscle thickness of the medial side for knee extensors. In conclusion, the tendon structures of highly trained sprinters are more compliant than those of untrained subjects for knee extensors, but not for plantar flexors. Furthermore, a thicker medial side of knee extensors was associated with greater sprinting performance.
Assuntos
Articulação do Tornozelo/fisiologia , Articulação do Joelho/fisiologia , Músculo Esquelético/diagnóstico por imagem , Músculo Esquelético/fisiologia , Corrida/fisiologia , Tendões/diagnóstico por imagem , Tendões/fisiologia , Articulação do Tornozelo/diagnóstico por imagem , Módulo de Elasticidade/fisiologia , Humanos , Articulação do Joelho/diagnóstico por imagem , Masculino , Aptidão Física/fisiologia , Ultrassonografia , Adulto JovemRESUMO
We compared rifabutin susceptibility and rpoB mutations in 98 multi-drug-resistant strains of Mycobacterium tuberculosis (MDR-TB) by DNA sequencing and with a line probe assay using the commercially available INNO-LiPA Rif. TB kit (the LiPA). Our results indicated that rifabutin continues to remain active against MDR-TB strains harboring certain genetic alterations and also that the LiPA might be useful in identifying MDR-TB strains susceptible to rifabutin.
Assuntos
Proteínas de Bactérias/genética , Mycobacterium tuberculosis/efeitos dos fármacos , Mycobacterium tuberculosis/genética , Rifabutina/farmacologia , Tuberculose Resistente a Múltiplos Medicamentos/genética , Tuberculose Resistente a Múltiplos Medicamentos/microbiologia , Antibióticos Antituberculose/farmacologia , Análise Mutacional de DNA/métodos , RNA Polimerases Dirigidas por DNA , Farmacorresistência Bacteriana Múltipla , Humanos , Testes de Sensibilidade Microbiana/métodos , Mutação/genética , Hibridização de Ácido Nucleico/métodosRESUMO
A third of the world's population is infected with Mycobacterium tuberculosis, and 2 million people die from tuberculosis every year. The only tuberculosis vaccine currently available is an attenuated strain of Mycobacterium bovis BCG, although its efficacy against adult tuberculosis disease remains controversial. Furthermore multi-drug resistant tuberculosis is becoming big problems in the world. Therefore, the development of novel therapeutic vaccine as well as novel prophylactic vaccine against tuberculosis is required. This review provides a summary of novel vaccines (especially DNA vaccines) in preclinical stage using mouse, guinea pig and monkey models. In several promising novel vaccines, the studies were extended to a cynomolgus monkey model, which is currently the best animal model of human tuberculosis. The review also provides recent advances of the precise studies of induction of immunity including CD8 positive cytotoxic T cells and effector molecules such as granulysin by these vaccines, against multi-drug resistant tuberculosis and extremely drug resistant tuberculosis.
Assuntos
Vacinas contra a Tuberculose/imunologia , Vacinas contra a Tuberculose/uso terapêutico , Tuberculose/tratamento farmacológico , Tuberculose/prevenção & controle , Vacinas de DNA/imunologia , Animais , Modelos Animais de Doenças , Cobaias , Humanos , Macaca fascicularis , Camundongos , Mycobacterium tuberculosis/imunologia , Linfócitos T Citotóxicos/imunologia , Vacinas contra a Tuberculose/genética , Vacinas de DNA/genéticaRESUMO
A third of world's population infected with Mycobacterium tuberculosis, and 2 million people die from tuberculosis every year. It is well established that protective to M. tuberculosis depends on both CD4+ and CD8+ T cells. In particular, acquired immunity (cytotoxic T cell, Th1 helper T cell and M phi) play an important role for TB infection. Recently, natural immunity also play a very attractive role for the development of TB immunity. We found that memory CTL is most important for the protection against TB using several kinds of mice. It was demonstrated that DBA/1 mice are more sensitive to TB infection than BALB/c mice (Th2 prone mice). Induction of memory CTL in DBA/1 mice was lower than BALB/c. In contrast, IFN-gamma production of DBA/1 mice was higher than BALB/c. Therefore, famous researchers in the fields of TB immunity reviewed the recent advances of TB immunity, such as (1) T cell immunity and recognition against TB antigen, (2) TLR9 and CpG motif, (3) lipocalin2 and SLPI in natural TB immunity, (4) acquired immunity (CTL) and granulysin. The development novel vaccine (HSP65+IL-12 DNA vaccine), (5) The mechanism of protection against TB, in this mini-review series.
Assuntos
Mycobacterium tuberculosis/imunologia , Animais , Humanos , Imunidade Inata , Tuberculose/imunologiaRESUMO
CDC and ACET in U.S.A. reported that novel vaccines instead of BCG are required for the protection against infection of Mycobacterium tuberculosis worldwide. However, no novel vaccine for clinical use has not yet been developed in the world including U.S.A. and Europe. We have developed a novel tuberculosis (TB) vaccine; a combination of the DNA vaccines expressing mycobacterial heat shock protein 65 (HSP65) and interleukin 12 (IL-12) delivered by the hemagglutinating virus of Japan (HVJ)-envelope and -liposome (HSP65+IL-12/HVJ). This vaccine provided remarkable protective efficacy in mouse compared to the BCG vaccine on the basis of C.F.U of number of TB, survival, an induction of the CD8 positive CTL activity and improvement of the histopathological tuberculosis lesions. This vaccine also provided therapeutic efficacy against multidrug resistant TB (MDR-TB) and extremely drug resistant TB (XDR-TB) in murine models. Furthermore, we extended our studies to a cynomolgus monkey model, which is currently the best animal model of human tuberculosis. This novel vaccine provided a higher level of the protective efficacy than BCG based upon the assessment of mortality, the ESR, body weight, chest X-ray findings and immune responses. Furthermore, the BCG priming and HSP65+IL-12/HVJ vaccine (booster) by the priming-booster method showed a synergistic effect in the TB-infected cynomolgus monkey (100% survival). Furthermore, this vaccine exerted therapeutic efficacy (100% survival) and augmentation of immune responses in the TB-infected monkeys. These data indicate that our novel DNA vaccine might be useful against Mycobacterium tuberculosis including XDR-TB and MDR-TB for human therapeutic clinical trials. The review also provides recent advances of the precise studies of induction of immunity including CD8 positive cytotoxic T cells and effector molecules such as granulysin by these vaccines, against multi-drug resistant tuberculosis and extremely drug resistant tuberculosis.
Assuntos
Antígenos de Diferenciação de Linfócitos T/imunologia , Proteínas de Choque Térmico/imunologia , Interleucina-12/imunologia , Células T Matadoras Naturais/imunologia , Vacinas contra a Tuberculose/imunologia , Tuberculose/imunologia , Animais , DNA , Proteínas de Choque Térmico/genética , Humanos , Interleucina-12/genética , CamundongosRESUMO
PURPOSE: TB among foreigners is presently a serious issue in some developed countries and could become so in Japan. The purpose of this report is to assess the epidemiological situation of TB among foreigners in Japan. MATERIALS AND METHODS: The trend of TB reporting among foreigners in Japan was examined with regard to work status. RESULTS: The number of reported TB cases among employees and students in Japan increased between 1998 and 2008, but that among housekeepers was level throughout the same period. The increase among employees and students might be due to the increased numbers of foreign employees and students. In the case of housekeepers, the increase in the number of permanent residents did not lead to an increase in TB among these housekeepers. Estimates of TB reporting rates decreased during the study period, so the changes in reporting rates would not have caused the increase in TB cases. This downward trend may have been caused by an increase in longer-term residents and a decrease in TB incidence in home countries. Even though the TB reporting rate is decreasing, the rates in those countries are much higher than in Japan in the same work categories. DISCUSSION: To control the spread of TB, it is important to identify high-risk individuals. The Japanese TB control program should further strengthen mass health examination programs for foreign housekeepers and employees (especially temporary and daily employees), case-finding based on individuals' access to hospitals or clinics when suffering from TB symptoms, and flexible and periodic adjustment of TB control activities for foreigners according to future changes in the number and distribution of foreigners in Japan. Furthermore, improving the TB epidemiological situation in home countries might contribute to the downward trend of TB reporting rates among foreigners in Japan. Therefore, Japanese assistance in TB control activities in surrounding countries such as China, South Korea, and the Philippines might contribute to TB control activities for foreigners in Japan.
Assuntos
Emigrantes e Imigrantes/estatística & dados numéricos , Emprego , Tuberculose/epidemiologia , Feminino , Humanos , Japão/epidemiologia , Masculino , EstudantesRESUMO
OBJECTS: To analyze the molecular epidemiology of Mycobacterium gordonae strains from patients and environments in the hospital. SUBJECTS: A total of 46 clinical strains were obtained from patients registered at the NHO Kinki-chuo Chest Medical Center and 3 strains from hospital environments. METHODS: By using genetic data from the 16S rRNA gene and hsp65PRA, pulsed-field gel electrophoresis (PFGE) assessment of their intraspecies variability and epidemiology was carried out. RESULTS: Strains from six patients and environmental cultures exhibited the different genotypes of 16S rRNA gene sequencing and the hsp65PRA type. The PFGE analysis suggested no pseudo-outbreak and showed a polyclonal infection in one patient. CONCLUSION: These findings suggest that we should maintain effective surveillance of environments in the hospital and continuously perform molecular epidemiological investigations for infection control of M. gordonae.
Assuntos
Infecções por Mycobacterium não Tuberculosas/epidemiologia , Micobactérias não Tuberculosas/genética , Micobactérias não Tuberculosas/isolamento & purificação , Adulto , Idoso , Idoso de 80 Anos ou mais , Proteínas de Bactérias/análise , Chaperonina 60/análise , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , RNA Ribossômico 16S/análiseRESUMO
PURPOSE: Evaluation of the INNO-LiPA MYCOBACTERIA v2 (the INNO-LiPA assay) for mycobacterial identification. MATERIALS AND METHODS: The laboratory identifications consisting of Cobas Amplicor systems, AccuProbe, and DDH, are commonly used to identify mycobacterial isolates in Japan. We compared the results between the INNO-LiPA assay and the common methods. A total of 122 clinical isolates from NHO Kinki-chuo Chest Medical Center from 1 February to 30 June 2006 were tested. RESULTS: There was agreement between the INNO-LiPA assay and the common methods for 112 mycobacterium isolates. The six discordant isolates have showed same results between sequencings and the INNO-LiPA assay. The one M. fortuitum isolates was indicated correctness by DDH and the one M. intracellulare isolates was recognized by Cobas Amplicor systems and as MAC by AccuProbe MAC. Moreover, discrepant results between sequencings and mycobacterial identifications including the INNO-LiPA assay were 2 isolates (M. paraffinicum, M. mucogenicum variant type). CONCLUSION: The INNO-LiPA assay could provide rapid and correct identification results with clear-cut and easy interpretation.