RESUMO
Scrub typhus is a potentially lethal infection caused by the obligate intracellular bacterium Orientia tsutsugamushi Reports on the emergence of doxycycline-resistant strains highlight the urgent need to develop novel antiinfectives against scrub typhus. Corallopyronin A (CorA) is a novel α-pyrone compound synthesized by the myxobacterium Corallococcus coralloides that was characterized as a noncompetitive inhibitor of the switch region of the bacterial RNA polymerase (RNAP). We investigated the antimicrobial action of CorA against the human-pathogenic Karp strain of O. tsutsugamushiin vitro and in vivo The MIC of CorA against O. tsutsugamushi was remarkably low (0.0078 µg/ml), 16-fold lower than that against Rickettsia typhi In the lethal intraperitoneal O. tsutsugamushi mouse infection model, a minimum daily dose of 100 µg CorA protected 100% of infected mice. Two days of treatment were sufficient to confer protection. In contrast to BALB/c mice, SCID mice succumbed to the infection despite treatment with CorA or tetracycline, suggesting that antimicrobial treatment required synergistic action of the adaptive immune response. Similar to tetracycline, CorA did not prevent latent infection of O. tsutsugamushiin vivo However, latency was not caused by acquisition of antimicrobial resistance, since O. tsutsugamushi reisolated from latently infected BALB/c mice remained fully susceptible to CorA. No mutations were found in the CorA-binding regions of the ß and ß' RNAP subunit genes rpoB and rpoC Inhibition of the RNAP switch region of O. tsutsugamushi by CorA is therefore a novel and highly potent target for antimicrobial therapy for scrub typhus.
Assuntos
Antibacterianos/uso terapêutico , RNA Polimerases Dirigidas por DNA/antagonistas & inibidores , Lactonas/uso terapêutico , Orientia tsutsugamushi/patogenicidade , Tifo por Ácaros/tratamento farmacológico , Animais , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos SCID , Myxococcales/efeitos dos fármacos , Myxococcales/patogenicidade , Orientia tsutsugamushi/efeitos dos fármacos , Rickettsia typhi/efeitos dos fármacos , Rickettsia typhi/patogenicidade , Tifo por Ácaros/microbiologiaRESUMO
Serology is the preferred method to confirm a Chagas disease diagnosis and to screen blood donors. A battery of assays is often required due to the limited accuracy of single assays. The Elecsys Chagas assay is a newly developed, double-antigen sandwich assay for use on the Elecsys and cobas e immunoassay analyzers, intended to identify individuals infected with Trypanosoma cruzi, for diagnosis and screening. The performance of the Elecsys Chagas assay was evaluated in comparison with those of other widely used T. cruzi antibody assays, at multiple sites (Europe/Latin America). Relative sensitivity and specificity were assessed by using samples from blood donors, pregnant women, and hospitalized patients from regions where Chagas disease is endemic and from regions of nonendemicity. The Elecsys Chagas assay had an overall relative sensitivity of 100% (n = 674). Overall relative specificities were 99.90% (n = 14,681), 100% (n = 313), and 100% (n = 517) for samples from blood donors, pregnant women, and hospitalized patients, respectively. The analytical specificity was 99.83% (n = 594). The Elecsys Chagas assay detected T. cruzi antibodies in two World Health Organization (WHO) standard T. cruzi reference panels (panels 09/188 and 09/186) at a 1:512 dilution, corresponding to a cutoff sensitivity of approximately 1 mIU/ml. The Elecsys Chagas assay demonstrated robust performance under routine conditions at multiple sites in Europe and Latin America. In contrast to other available Chagas assays, the Elecsys assay uses a reduced number of recombinant T. cruzi antigens, resulting in a significantly smaller number of cross-reactions and improved analytical specificity while being highly sensitive.
Assuntos
Anticorpos Antiprotozoários/sangue , Doença de Chagas/diagnóstico , Imunoensaio/métodos , Testes Sorológicos/métodos , Trypanosoma cruzi/imunologia , Europa (Continente) , Feminino , Humanos , América Latina , Masculino , Gravidez , Sensibilidade e EspecificidadeRESUMO
Rickettsia typhi is the causative agent of endemic typhus, a disease with increasing incidence worldwide that can be fatal. Because of its obligate intracellular life style, genetic manipulation of the pathogen is difficult. Nonetheless, in recent years, genetic manipulation tools have been successfully applied to rickettsiae. We describe here for the first time the transformation of R. typhi with the pRAM18dRGA plasmid that originally derives from Rickettsia amblyommatis and encodes the expression of GFPuv (green fluorescent protein with maximal fluorescence when excited by UV light). Transformed R. typhi (R. typhiGFPuv) bacteria are viable, replicate with kinetics similar to those of wild-type R. typhi in cell culture, and stably maintain the plasmid and GFPuv expression under antibiotic treatment in vitro and in vivo during infection of mice. CB17 SCID mice infected with R. typhiGFPuv succumb to the infection with kinetics similar to those for animals infected with wild-type R. typhi and develop comparable pathology and bacterial loads in the organs, demonstrating that the plasmid does not influence pathogenicity. In the spleen and liver of infected CB17 SCID mice, the bacteria are detectable by immunofluorescence microscopy in neutrophils and macrophages by histological staining. Finally, we show for the first time that transformed rickettsiae can be used for the detection of CD8+ T cell responses. GFP-specific restimulation of spleen cells from R. typhiGFPuv-infected BALB/c mice elicits gamma interferon (IFN-γ), tumor necrosis factor alpha (TNF-α), and interleukin 2 (IL-2) secretion by CD8+ T cells. Thus, R. typhiGFPuv bacteria are a novel, potent tool to study infection with the pathogen in vitro and in vivo and the immune response to these bacteria.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Citocinas/imunologia , Rickettsia typhi/patogenicidade , Tifo Endêmico Transmitido por Pulgas/imunologia , Animais , Proteínas de Fluorescência Verde/genética , Fígado/microbiologia , Macrófagos/microbiologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos SCID , Neutrófilos/microbiologia , Plasmídeos , Rickettsia typhi/genética , Baço/microbiologia , Transformação Bacteriana , Tifo Endêmico Transmitido por Pulgas/microbiologiaRESUMO
Rickettsioses are emerging febrile diseases caused by obligate intracellular bacteria belonging to the family Rickettsiaceae. Rickettsia typhi belongs to the typhus group (TG) of this family and is the causative agent of endemic typhus, a disease that can be fatal. In the present study, we analyzed the course of R. typhi infection in C57BL/6 RAG1(-/-) mice. Although these mice lack adaptive immunity, they developed only mild and temporary symptoms of disease and survived R. typhi infection for a long period of time. To our surprise, 3 to 4 months after infection, C57BL/6 RAG1(-/-) mice suddenly developed lethal neurological disorders. Analysis of these mice at the time of death revealed high bacterial loads, predominantly in the brain. This was accompanied by a massive expansion of microglia and by neuronal cell death. Furthermore, high numbers of infiltrating CD11b(+) macrophages were detectable in the brain. In contrast to the microglia, these cells harbored R. typhi and showed an inflammatory phenotype, as indicated by inducible nitric oxide synthase (iNOS) expression, which was not observed in the periphery. Having shown that R. typhi persists in immunocompromised mice, we finally asked whether the bacteria are also able to persist in resistant C57BL/6 and BALB/c wild-type mice. Indeed, R. typhi could be recultivated from lung, spleen, and brain tissues from both strains even up to 1 year after infection. This is the first report demonstrating persistence and reappearance of R. typhi, mainly restricted to the central nervous system in immunocompromised mice.
Assuntos
Infecções do Sistema Nervoso Central/microbiologia , Infecções do Sistema Nervoso Central/patologia , Inflamação/patologia , Rickettsia typhi/isolamento & purificação , Tifo Endêmico Transmitido por Pulgas/microbiologia , Tifo Endêmico Transmitido por Pulgas/patologia , Animais , Encéfalo/parasitologia , Encéfalo/patologia , Doença Crônica , Proteínas de Homeodomínio/genética , Pulmão/parasitologia , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , Óxido Nítrico Sintase Tipo II/análise , Baço/parasitologiaRESUMO
Scrub typhus is a potentially lethal infection that is caused by the obligate intracellular bacterium Orientia tsutsugamushi The roles of Toll-like receptor 2 (TLR2) and TLR4 in innate recognition of O. tsutsugamushi have not been elucidated. By overexpression of TLR2 or TLR4 in HEK293 cells, we demonstrated that TLR2, but not TLR4, recognizes heat-stable compounds of O. tsutsugamushi that were sensitive to treatment with sodium hydroxide, hydrogen peroxide, and proteinase K. TLR2 was required for the secretion of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) by dendritic cells. In an intradermal mouse infection model, TLR2-deficient mice did not show impaired control of bacterial growth or reduced survival. Moreover, after intraperitoneal infection, TLR2-deficient mice were even more resistant to lethal infection than C57BL/6 wild-type mice, which showed stronger symptoms and lower survival rates during the convalescent phase. Compared to the time of reduction of bacterial loads in TLR2-deficient mice, the reduction of bacterial loads in infected organs was accelerated in wild-type mice. The higher mortality of wild-type mice was associated with increased concentrations of serum alkaline phosphatase but not aspartate aminotransferase. The transcription of mRNA for TNF-α and IL-6 decreased more rapidly in peritoneum samples from wild-type mice than in those from TLR2-deficient mice and was therefore not a correlate of increased susceptibility. Thus, although TLR2 is an important mediator of the early inflammatory response, it is dispensable for protective immunity against O. tsutsugamushi Increased susceptibility to O. tsutsugamushi infection in TLR2-competent mice rather suggests a TLR2-related immunopathologic effect.
Assuntos
Predisposição Genética para Doença , Orientia tsutsugamushi , Tifo por Ácaros/genética , Tifo por Ácaros/metabolismo , Receptor 2 Toll-Like/metabolismo , Animais , Células da Medula Óssea , Células Dendríticas , Feminino , Regulação da Expressão Gênica , Células HEK293 , Humanos , Interleucina-6/genética , Interleucina-6/metabolismo , Ligantes , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptor 2 Toll-Like/genética , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/metabolismo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismoRESUMO
We report 2 cases of patients who were successfully treated surgically for remaining or recurrent mitral regurgitation after MitraClip (Abbott Vascular, Santa Clara, CA, USA) implantation. In one patient, intervention with MitraClip was indicated because of the extremely poor heart function and poor general status of the patient. However, the severe mitral insufficiency remained after the MitraClip treatment and valve replacement was required. In the other high-risk patient, severe mitral regurgitation recurred 3 years after a successful MitraClip treatment due to infective endocarditis. Our experience suggests that some patients who are considered "high-risk" before MitraClip treatment might be reasonable candidates for a straight forward mitral valve surgery, even in the re-intervention setting. We conclude that patients considered for MitraClip implantation should undergo detailed risk stratification, because we have to keep in mind that after failed clip implantation the perioperative risk increases and the chance of mitral repair decreases.
Assuntos
Implante de Prótese de Valva Cardíaca , Insuficiência da Valva Mitral/cirurgia , Idoso , Humanos , Masculino , Pessoa de Meia-Idade , Insuficiência da Valva Mitral/diagnóstico , Insuficiência da Valva Mitral/etiologia , Reoperação , Falha de TratamentoRESUMO
The implantation of cardiac assist devices is associated with poor outcome in patients with multiple organ failure and unknown neurologic status. Therefore, temporary left ventricular assist devices (LVAD) using, for example, extracorporeal centrifugal pumps may provide the chance to further evaluate the patient's clinical course and a potential qualification for implantable LVAD therapy. On the other hand, a main disadvantage of the temporary LVAD implantation is the need for redo surgery, increasing the risk of the final LVAD Implantation. To minimize this drawback of the temporary LVAD implantation, we implanted the temporary LVAD using a minimally invasive technique. The operation was done without cardiopulmonary bypass support, and the temporary LVAD was implanted through upper hemisternotomy and left anterior mini-thoracotomy. The patient recovered from multiple organ failure and was successfully bridged to a permanent LVAD therapy.
Assuntos
Oxigenação por Membrana Extracorpórea/métodos , Insuficiência Cardíaca/cirurgia , Coração Auxiliar , Procedimentos Cirúrgicos Minimamente Invasivos , Implantação de Prótese , Coração/fisiopatologia , Insuficiência Cardíaca/fisiopatologia , Humanos , Masculino , Pessoa de Meia-Idade , Resultado do TratamentoRESUMO
We analyzed the long-term results of two surgical techniques (beating versus non-beating) for isolated tricuspid valve (TV) surgery.The long-term results of 92 consecutive patients who underwent isolated TV surgery were analyzed. We compared patients with beating heart (BH) surgery (n = 48) with patients undergoing arrested heart (AH) surgery (n = 44).BH surgery was more frequently chosen in urgent/emergent operations (P = 0.029) and in redo-operations (P < 0.001). Preoperatively, the rates of renal insufficiency (P = 0.002) and EuroSCORE (P = 0.019) were higher in the BH group than in the AH group. There were no differences in perioperative outcomes and 30-day mortality between the groups. However, freedom from reoperation was significantly lower in the BH group compared to the AH group (P = 0.039). We observed a trend towards lower survival rates at 1, 5, and 10 years in the BH group (77%, 54%, and 41%) compared to those of the AH group (86%, 75%, and 72%, P = 0.062). Multivariate Cox hazard model analysis revealed preoperative heart rhythm (P = 0.014, odds ratio [OR] = 2.296) and EuroSCORE (P = 0.022, OR = 1.049) as independent risk factors for mortality after isolated TV surgery.The superiority of BH surgery over AG surgery was not proven. Surgical intervention should be considered early, since patients with elevated EuroSCORES and arrhythmia have significantly higher mortality rates.
Assuntos
Parada Cardíaca Induzida/métodos , Frequência Cardíaca , Doenças das Valvas Cardíacas , Implante de Prótese de Valva Cardíaca , Valva Tricúspide , Adulto , Idoso , Pesquisa Comparativa da Efetividade , Feminino , Seguimentos , Alemanha , Doenças das Valvas Cardíacas/diagnóstico , Doenças das Valvas Cardíacas/fisiopatologia , Doenças das Valvas Cardíacas/cirurgia , Implante de Prótese de Valva Cardíaca/efeitos adversos , Implante de Prótese de Valva Cardíaca/métodos , Implante de Prótese de Valva Cardíaca/mortalidade , Humanos , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Modelos de Riscos Proporcionais , Reoperação/métodos , Reoperação/estatística & dados numéricos , Fatores de Risco , Tempo , Resultado do Tratamento , Valva Tricúspide/patologia , Valva Tricúspide/cirurgiaRESUMO
During August 2010-December 2012, we conducted a study of patients in Ghana who had Buruli ulcer, caused by Mycobacterium ulcerans, and found that 23% were co-infected with Mansonella perstans nematodes; 13% of controls also had M. perstans infection. M. perstans co-infection should be considered in the diagnosis and treatment of Buruli ulcer.
Assuntos
Úlcera de Buruli/epidemiologia , Mansonella/isolamento & purificação , Mansonelose/epidemiologia , Mycobacterium ulcerans/isolamento & purificação , Adolescente , Adulto , Animais , Úlcera de Buruli/diagnóstico , Úlcera de Buruli/microbiologia , Criança , Coinfecção , Feminino , Gana/epidemiologia , Humanos , Incidência , Masculino , Mansonelose/diagnóstico , Mansonelose/parasitologia , Pessoa de Meia-Idade , Estudos RetrospectivosRESUMO
Protection against malaria can be achieved by induction of a strong CD8(+) T-cell response against the Plasmodium circumsporozoite protein (CSP), but most subunit vaccines suffer from insufficient memory responses. In the present study, we analyzed the impact of postimmunization sporozoite challenge on the development of long-lasting immunity. BALB/c mice were immunized by a heterologous prime/boost regimen against Plasmodium berghei CSP that induces a strong CD8(+) T-cell response and sterile protection, which is short-lived. Here, we show that protective immunity is prolonged by a sporozoite challenge after immunization. Repeated challenges induced sporozoite-specific antibodies that showed protective capacity. The numbers of CSP-specific CD8(+) T cells were not substantially enhanced by sporozoite infections; however, CSP-specific memory CD8(+) T cells of challenged mice displayed a higher cytotoxic activity than memory T cells of immunized-only mice. CD4(+) T cells contributed to protection as well; but CD8(+) memory T cells were found to be the central mediator of sterile protection. Based on these data, we suggest that prolonged protective immunity observed after immunization and infection is composed of different antiparasitic mechanisms including CD8(+) effector-memory T cells with increased cytotoxic activity as well as CD4(+) memory T cells and neutralizing antibodies.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Imunidade Humoral , Memória Imunológica , Plasmodium berghei/imunologia , Esporozoítos/imunologia , Animais , Anticorpos Antiprotozoários/imunologia , Especificidade de Anticorpos/imunologia , Linfócitos T CD4-Positivos/imunologia , Modelos Animais de Doenças , Feminino , Imunização , Malária/imunologia , Malária/parasitologia , Malária/prevenção & controle , Vacinas Antimaláricas/administração & dosagem , Vacinas Antimaláricas/imunologia , Camundongos , Camundongos Endogâmicos BALB C , FenótipoRESUMO
SOCS3 is a feedback regulator of cytokine signaling that affects T-cell polarization. Human tuberculosis is accompanied by increased SOCS3 expression in T cells, and this may influence susceptibility against Mycobacterium tuberculosis. Because the role of SOCS3 in human T-cell function is not well defined, we characterized cytokine expression and proliferation of human T cells with differential SOCS3 expression in the present study. We established a flow cytometry-based method for SOCS3 protein quantification and detected higher SOCS3 levels induced by M tuberculosis specific T-cell activation and a transient decrease of SOCS3 expression in the presence of mycobacteria-infected macrophages. Notably increased SOCS3 expression was detected in IL-17-expressing T-cell clones and in CD161(+) T helper type 17 cells ex vivo. Ectopic SOCS3 expression in primary CD4(+) T cells by lentiviral transduction induced increased IL-17 production but diminished proliferation and viability. Recombinant IL-7 inhibited SOCS3 expression and reduced IL-17-expressing T-cell proportions. We concluded that higher SOCS3 expression in human T cells favors T helper type 17 cells. Therefore, increased SOCS3 expression in human tuberculosis may reflect polarization toward IL-17-expressing T cells as well as T-cell exhaustion marked by reduced proliferation.
Assuntos
Interleucina-17/genética , Proteínas Supressoras da Sinalização de Citocina/fisiologia , Linfócitos T/metabolismo , Linfócitos T CD4-Positivos/efeitos dos fármacos , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD4-Positivos/fisiologia , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Citometria de Fluxo , Regulação da Expressão Gênica , Humanos , Interleucina-17/metabolismo , Interleucina-17/farmacologia , Ativação Linfocitária/genética , Mycobacterium tuberculosis/imunologia , Proteína 3 Supressora da Sinalização de Citocinas , Proteínas Supressoras da Sinalização de Citocina/genética , Proteínas Supressoras da Sinalização de Citocina/metabolismo , Especificidade do Receptor de Antígeno de Linfócitos T/efeitos dos fármacos , Especificidade do Receptor de Antígeno de Linfócitos T/genética , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia , Linfócitos T/fisiologia , Transfecção , Tuberculose/imunologia , Regulação para Cima/genéticaRESUMO
Transgenic over-expression of CD83 on B cells leads to a reduced response to BCR engagement but to an enhanced secretion of IL-10 upon LPS stimulation. In this study, we analyzed the differential influence of CD83 on the stimulation of different B cell subsets via the BCR or TLR4. Neither wild type nor CD83 transgenic (CD83tg) B cells produced any IL-10 in response to BCR stimulation. BCR engagement led to reduced activation of LYN, SYK and ERK1/2 resulting in reduced numbers of proliferating cells in all CD83tg B cell subsets. Moreover, CD83tg follicular (FO) but not marginal zone (MZ) or transitional (TN) B cells showed significantly enhanced cell death. In contrast, LPS stimulation led to normal frequencies of proliferating CD83tg FO, MZ and TN B cells although TLR4 engagement did not rescue FO B cells from apoptosis. Furthermore, LPS stimulation led to high IL-10 production derived from CD83tg MZ B cells that reacted to LPS stimulation with enhanced ERK1/2 activation. Finally, we show that CD83 co-localizes with the BCR complex as well as with the LPS receptor complex suggesting that CD83 interacts with components of both signaling complexes. Taken together, the results of this study show that CD83 already inhibits the initiation of BCR signaling leading to insufficient activation signals in all B cells and reduced survival especially of FO B cells. On the other hand, CD83 supports TLR4-mediated IL-10 release exclusively in MZ B cells. Thus, CD83 differentially modulates FO and MZ B cell responses.
Assuntos
Antígenos CD/metabolismo , Linfócitos B/imunologia , Linfócitos B/metabolismo , Imunoglobulinas/metabolismo , Glicoproteínas de Membrana/metabolismo , Baço/citologia , Baço/imunologia , Animais , Antígenos CD/imunologia , Imunoglobulinas/imunologia , Glicoproteínas de Membrana/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Antígeno CD83RESUMO
Malaria is still responsible for up to 1 million deaths per year worldwide, highlighting the need for protective malaria vaccines. Helminth infections that are prevalent in malaria endemic areas can modulate immune responses of the host. Here we show that Strongy-Ioides ratti, a gut-dwelling nematode that causes transient infections, did not change the efficacy of vaccination against Plasmodium berghei. An ongoing infection with Litomosoides sigmodontis, a tissue-dwelling filaria that induces chronic infections in BALB/c mice, significantly interfered with vaccination efficacy. The induction of P. berghei circumspor-ozoite protein (CSP)-specific CD8(+) T cells, achieved by a single immunization with a CSP fusion protein, was diminished in L. sigmodontis-infected mice. This modulation was reflected by reduced frequencies of CSP-specific CD8(+) T cells, reduced CSP-specific IFN-y and TNF-a production, reduced CSP-specific cytotoxicity, and reduced protection against P. berghei challenge infection. Implementation of a more potent vaccine regime, by first priming with CSP-expressing recombinant live Salmonella prior to CSP fusion protein immunization, restored induction of CSP-specific CD8(+) T cells and conferred almost sterile immunity to P. berghei challenge infection also in L. sigmodontis-infected mice. In summary, we show that appropriate vaccination regimes can overcome helminth-induced interference with vaccination efficacy.
Assuntos
Antígenos de Protozoários/farmacologia , Linfócitos T CD8-Positivos/imunologia , Filariose/imunologia , Filarioidea/imunologia , Malária/imunologia , Plasmodium berghei/imunologia , Proteínas de Protozoários/farmacologia , Strongyloides ratti/imunologia , Estrongiloidíase/imunologia , Animais , Antígenos de Protozoários/imunologia , Imunização , Interferon gama/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Proteínas de Protozoários/imunologia , Ratos , Ratos Wistar , Salmonella/imunologia , Sigmodontinae , Fator de Necrose Tumoral alfa/imunologiaRESUMO
BACKGROUND: Systematic long-term data on tricuspid valve (TV) endocarditis are limited. The aim of this study was to investigate the outcome of surgery for isolated TV endocarditis. METHODS AND RESULTS: A total of 637 patients who underwent TV surgery between June 1996 and September 2012 at Hannover Medical School were retrospectively investigated. Of the 637 patients, 33 (14 female, mean age, 49 ± 21 years) underwent isolated TV surgery for endocarditis: biological TV replacement, n=14; mechanical TV replacement, n=4; TV reconstruction, n=15. A total of 28 cases were associated with i.v. drug abuse (n=14) or pacemaker infection (n=14). Staphylococcus (S.) aureus was the most common microorganism detected on preoperative blood culture. Mean follow-up was 6.0 ± 4.1 years (83% completed). Three patients (9%) died during the first 30 postoperative days. Survival at 1, 5 and 10 years was 88%, 73%, and 73%, respectively. Freedom from reoperation was 100%, 95%, and 88%, respectively. During follow-up New York Heart Association class improved significantly, and echocardiography identified remaining TV insufficiency grade ≥ II° only in 2 patients. Statistical analysis identified advanced age, logistic EuroSCORE and positive blood culture for S. aureus as significant risk factors for long-term mortality. CONCLUSIONS: Isolated TV endocarditis is strongly associated with i.v. drug abuse or pacemaker infection. Long-term outcome is acceptable, independent of the surgical procedure.
Assuntos
Endocardite Bacteriana , Doenças das Valvas Cardíacas , Infecções Estafilocócicas , Staphylococcus aureus , Valva Tricúspide , Adulto , Idoso , Intervalo Livre de Doença , Endocardite Bacteriana/etiologia , Endocardite Bacteriana/microbiologia , Endocardite Bacteriana/mortalidade , Endocardite Bacteriana/cirurgia , Feminino , Seguimentos , Doenças das Valvas Cardíacas/microbiologia , Doenças das Valvas Cardíacas/mortalidade , Doenças das Valvas Cardíacas/cirurgia , Próteses Valvulares Cardíacas/microbiologia , Humanos , Masculino , Pessoa de Meia-Idade , Marca-Passo Artificial/microbiologia , Estudos Retrospectivos , Infecções Estafilocócicas/etiologia , Infecções Estafilocócicas/patologia , Infecções Estafilocócicas/cirurgia , Transtornos Relacionados ao Uso de Substâncias/complicações , Transtornos Relacionados ao Uso de Substâncias/mortalidade , Transtornos Relacionados ao Uso de Substâncias/cirurgia , Taxa de Sobrevida , Valva Tricúspide/microbiologia , Valva Tricúspide/cirurgiaRESUMO
One third of the human population is infected with helminth parasites. To promote their longevity and to limit pathology, helminths have developed several strategies to suppress the immune response of their host. As this immune suppression also acts on unrelated third-party Ags, a preexisting helminth infection may interfere with vaccination efficacy. In this study, we show that natural infection with Litomosoides sigmodontis suppressed the humoral response to thymus-dependent but not to thymus-independent model Ags in C57BL/6 mice. Thereby, we provide evidence that reduced humoral responses were mediated by interference with Th cell function rather than by direct suppression of B cells in L. sigmodontis-infected mice. We directly demonstrate suppression of Ag-specific proliferation in OVA-specific Th cells after adoptive transfer into L. sigmodontis-infected mice that led to equally reduced production of OVA-specific IgG. Transferred Th cells displayed increased frequencies of Foxp3(+) after in vivo stimulation within infected but not within naive mice. Helminth-mediated suppression was induced by established L. sigmodontis infections but was completely independent of the individual worm burden. Using DEREG mice, we rule out a central role for host-derived regulatory T cells in the suppression of transferred Th cell proliferation. In contrast, we show that L. sigmodontis-induced, host-derived IL-10 mediated Foxp3 induction in transferred Th cells and significantly contributed to the observed Th cell hypoproliferation within infected mice.
Assuntos
Antígenos/imunologia , Filariose/imunologia , Imunidade Humoral/imunologia , Interleucina-10/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Transferência Adotiva , Animais , Linfócitos B/imunologia , Separação Celular , Ensaio de Imunoadsorção Enzimática , Filarioidea/imunologia , Citometria de Fluxo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos KnockoutRESUMO
Granzyme (gzm) A and B, proteases of NK cells and T killer cells, mediate cell death, but also cleave extracellular matrices, inactivate intracellular pathogens, and induce cytokines. Moreover, macrophages, Th2 cells, regulatory T cells, mast cells, and B cells can express gzms. We recently reported gzm induction in human filarial infection. In this study, we show that in rodent filarial infection with Litomosoides sigmodontis, worm loads were significantly reduced in gzmA × B and gzmB knockout mice during the whole course of infection, but enhanced only early in gzmA knockout compared with wild-type mice. GzmA/B deficiency was associated with a defense-promoting Th2 cytokine and Ab shift, enhanced early inflammatory gene expression, and a trend of reduced alternatively activated macrophage induction, whereas gzmA deficiency was linked with reduced inflammation and a trend toward increased alternatively activated macrophages. This suggests a novel and divergent role for gzms in helminth infection, with gzmA contributing to resistance and gzmB promoting susceptibility.
Assuntos
Filariose/enzimologia , Filariose/imunologia , Filarioidea/imunologia , Granzimas/fisiologia , Animais , Anticorpos Anti-Helmínticos/biossíntese , Feminino , Filariose/patologia , Granzimas/deficiência , Granzimas/genética , Imunidade Inata , Inflamação/enzimologia , Inflamação/imunologia , Inflamação/prevenção & controle , Ativação de Macrófagos/genética , Ativação de Macrófagos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Sigmodontinae , Células Th2/enzimologia , Células Th2/imunologia , Células Th2/patologiaRESUMO
Local inflammation during cutaneous leishmaniasis is accompanied by accumulation of CD11b(+) cells at the site of the infection. A functional role for these monocytic cells in local angiogenesis in leishmaniasis has not been described so far. Here, we show that CD11b(+) cells express high levels of the myeloid differentiation antigen carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1). In experimental cutaneous leishmaniasis in C57BL/6 wild-type (B6.WT) and B6.Ceacam1(-/-) mice, we found that only B6.Ceacam1(-/-) mice develop edemas and exhibit impairment of both hemangiogenesis and lymphangiogenesis. Because CEACAM1 expression correlates with functional angiogenesis, we further analyzed the role of the CD11b(+) population. In B6.Ceacam1(-/-) mice, we found systemic reduction of Ly-6C(high)/CD11b(high) monocyte precursors. To investigate whether CEACAM1(+) myeloid cells are causally related to efficient angiogenesis, we used reverse bone marrow transplants (BMTs) to restore CEACAM1(+) or CEACAM1(-) bone marrow in B6.Ceacam1(-/-) or B6.WT recipients, respectively. We found that angiogenesis was restored by CEACAM1(+) BMT only. In addition, we observed reduced morphogenic potential of inflammatory cells in Matrigel implants in CEACAM1(-) backgrounds or after systemic depletion of CD11b(high) macrophages. Taken together, we show for the first time that CEACAM1(+) myeloid cells are crucial for angiogenesis in inflammation.
Assuntos
Antígeno Carcinoembrionário/análise , Inflamação/fisiopatologia , Leishmaniose Cutânea/fisiopatologia , Células Mieloides/fisiologia , Neovascularização Patológica/fisiopatologia , Animais , Anticorpos Antiprotozoários/biossíntese , Transplante de Medula Óssea , Antígeno CD11b/análise , Antígeno Carcinoembrionário/biossíntese , Antígeno Carcinoembrionário/genética , Colágeno , Combinação de Medicamentos , Edema/etiologia , Edema/patologia , Glicoproteínas/biossíntese , Imunidade Celular , Implantes Experimentais , Inflamação/etiologia , Inflamação/imunologia , Interferon gama/biossíntese , Laminina , Leishmania major/imunologia , Leishmaniose Cutânea/complicações , Leishmaniose Cutânea/imunologia , Leishmaniose Cutânea/patologia , Vasos Linfáticos/metabolismo , Macrófagos/parasitologia , Macrófagos/fisiologia , Proteínas de Membrana Transportadoras , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Células Mieloides/química , Células Mieloides/classificação , Neovascularização Patológica/patologia , Proteoglicanas , Quimera por Radiação , Células Th1/imunologiaRESUMO
The transmembrane glycoprotein CD83 is an important regulator of both thymic T cell maturation and peripheral T cell response. Recent studies suggested that CD83 is also involved in the regulation of B cell maturation, activation, and homeostasis. In this study, we show that in vivo overexpression of CD83 dose dependently interfered with the Ig response to thymus-dependent and thymus-independent model Ag immunization. CD83 deficiency, in contrast, which was restricted to B cells in mixed bone marrow chimeras, led to unchanged or even slightly increased Ig responses. Strikingly, the engagement of CD83 that is naturally up-regulated on wild-type B cells by injection of anti-CD83 mAb in vivo induced a 100-fold increase in the IgG1 response to immunization. Kinetic analysis revealed that CD83 had to be engaged simultaneously or shortly after the B cell activation through injection of Ag, to modulate the IgG1 secretion. Furthermore, using mixed bone marrow chimeras in which either selectively the B cells or the dendritic cells were CD83 deficient, we demonstrate that anti-CD83 mAb mediated its biologic effect by engaging CD83 on B cells and not on CD11c(+) dendritic cells. Taken together, we provide strong evidence that CD83 transduces regulatory signals into the very B cell on which it is expressed.
Assuntos
Antígenos CD/imunologia , Antígenos CD/metabolismo , Linfócitos B/imunologia , Linfócitos B/metabolismo , Imunoglobulinas/imunologia , Imunoglobulinas/metabolismo , Glicoproteínas de Membrana/imunologia , Glicoproteínas de Membrana/metabolismo , Animais , Formação de Anticorpos/genética , Antígenos CD/biossíntese , Relação Dose-Resposta Imunológica , Ficoll/administração & dosagem , Ficoll/imunologia , Haptenos/administração & dosagem , Haptenos/imunologia , Hemocianinas/administração & dosagem , Hemocianinas/imunologia , Imunoglobulina G/biossíntese , Imunoglobulina G/sangue , Imunoglobulina M/biossíntese , Imunoglobulina M/sangue , Imunoglobulinas/biossíntese , Imunoglobulinas/deficiência , Terapia de Imunossupressão , Ativação Linfocitária/genética , Ativação Linfocitária/imunologia , Glicoproteínas de Membrana/biossíntese , Glicoproteínas de Membrana/deficiência , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Nitrofenóis/administração & dosagem , Nitrofenóis/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Linfócitos T Auxiliares-Indutores/metabolismo , Antígeno CD83RESUMO
Cerebral malaria (CM) associated with Plasmodium berghei ANKA (PbA) infection is an accepted model of human CM. CM during PbA infection critically depends on sequestration of T cells into the brain. Several studies aimed to address the role of regulatory T cells (T(reg)) in modulating this pathogenic T cell response. However, these studies are principally hampered due to the fact that until recently no reagents were available to deplete Foxp3(+) T(reg) specifically. To study the function of T(reg) in the genesis of CM, we used depletion of T(reg) mice that are transgenic for a bacterial artificial chromosome expressing a diphtheria toxin receptor-enhanced GFP fusion protein under the control of the foxp3 gene locus. These mice allow for a selective depletion of Foxp3(+) T(reg) by diphtheria toxin injection, and also their specific detection and purification during an ongoing infection. Using depletion of T(reg) mice, we found only a small increase in the absolute numbers of Foxp3(+) T(reg) during PbA infection and, consequently, the ratio of T(reg) to T effector cells (T(eff)) decreased due to the rapid expansion of T(eff). Although the latter sequester in the brains of infected mice, almost no T(reg) were found in the brains of infected mice. Furthermore, we demonstrate that depletion of T(reg) has no influence on sequestration of T(eff) and on the clinical outcome, and only minor influence on T cell activation. Using ex vivo analysis of purified T(reg) from either naive mice or PbA-infected mice, we found that both exhibit similar inhibitory capacity on T(eff).