Patterns of Gene Expression in Peripheral Blood Mononuclear Cells and Outcomes from Patients with Sepsis Secondary to Community Acquired Pneumonia

Mechanisms governing the inflammatory response during sepsis have been shown to be complex, involving cross-talk between diverse signaling pathways. Current knowledge regarding the mechanisms underlying sepsis provides an incomplete picture of the syndrome, justifying additional efforts to understand this condition. Microarray-based expression profiling is a powerful approach for the investigation of complex clinical conditions such as sepsis. in this study, we investigate whole-genome expression profiles in mononuclear cells from survivors (n = 5) and non-survivors (n = 5) of sepsis. To circumvent the heterogeneity of septic patients, only patients admitted with sepsis caused by community-acquired pneumonia were included. Blood samples were collected at the time of sepsis diagnosis and seven days later to evaluate the role of biological processes or genes possibly involved in patient recovery. Principal Components Analysis (PCA) profiling discriminated between patients with early sepsis and healthy individuals. Genes with differential expression were grouped according to Gene Ontology, and most genes related to immune defense were up-regulated in septic patients. Additionally, PCA in the early stage was able to distinguish survivors from non-survivors. Differences in oxidative phosphorylation seem to be associated with clinical outcome because significant differences in the expression profile of genes related to mitochondrial electron transport chain (ETC) I-V were observed between survivors and non-survivors at the time of patient enrollment. Global gene expression profiles after seven days of sepsis progression seem to reproduce, to a certain extent, patterns collected at the time of diagnosis. Gene expression profiles comparing admission and follow-up samples differed between survivors and non-survivors, with decreased expression of genes related to immune functions in non-survivors. in conclusion, genes related to host defense and inflammatory response ontology were up-regulated during sepsis, consistent with the need for a host response to infection, and the sustainability of their expression in follow-up samples was associated with outcomes.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Albert Einstein Research and Education Institute - Hospital Israelita Albert EinsteinHosp Israelita Albert Einstein, Inst Israelita Ensino & Pesquisa, Ctr Expt Res, São Paulo, BrazilHosp Israelita Albert Einstein, Intens Care Unit, São Paulo, BrazilUniversidade Federal de São Paulo Unifesp, EPM, Hosp São Paulo, Div Infect Dis, São Paulo, BrazilHosp Sirio Libanes, Intens Care Unit, São Paulo, BrazilUniversidade Federal de São Paulo Unifesp, EPM, Dept Gynecol, São Paulo, BrazilUniversidade Federal de São Paulo Unifesp, Intens Care Unit, Hosp São Paulo, São Paulo, BrazilUniversidade Federal de São Paulo Unifesp, EPM, Hosp São Paulo, Div Infect Dis, São Paulo, BrazilUniversidade Federal de São Paulo Unifesp, EPM, Dept Gynecol, São Paulo, BrazilUniversidade Federal de São Paulo Unifesp, Intens Care Unit, Hosp São Paulo, São Paulo, BrazilFAPESP: FAPESP 2006/58744-1Web of Scienc

Influence of GB virus C on IFN-γ and IL-2 production and CD38 expression in T lymphocytes from chronically HIV-infected and HIV-HCV-co-infected patients

This study was designed to assess the effect of GB virus (GBV)-C on the immune response to human immunodeficiency virus (HIV) in chronically HIV-infected and HIV- hepatitis C virus (HCV)-co-infected patients undergoing antiretroviral therapy. A cohort of 159 HIV-seropositive patients, of whom 52 were HCV-co-infected, was included. Epidemiological data were collected and virological and immunological markers, including the production of interferon gamma (IFN-γ) and interleukin (IL)-2 by CD4, CD8 and Tγδ cells and the expression of the activation marker, CD38, were assessed. A total of 65 patients (40.8%) presented markers of GBV-C infection. The presence of GBV-C did not influence HIV and HCV replication or TCD4 and TCD8 cell counts. Immune responses, defined by IFN-γ and IL-2 production and CD38 expression did not differ among the groups. Our results suggest that neither GBV-C viremia nor the presence of E2 antibodies influence HIV and HCV viral replication or CD4 T cell counts in chronically infected patients. Furthermore, GBV-C did not influence cytokine production or CD38-driven immune activation among these patients. Although our results do not exclude a protective effect of GBV-C in early HIV disease, they demonstrate that this effect may not be present in chronically infected patients, who represent the majority of patients in outpatient clinics.Universidade Federal de São Paulo (UNIFESP) Laboratório de Virologia e Imunologia Disciplina de InfectologiaFleury Medicina DiagnósticaUNIFESP, Laboratório de Virologia e Imunologia Disciplina de InfectologiaSciEL

Evaluation of parameters of immune response in co-infection with HIV-1 virus and hepatitis C and G (HGV / GBV-C)

GBV-C co-infection is frequent in humans and could last for years without clinical symptoms. GBV-C is a flavivirus composed by a single positive RNA strain, closely related to HCV. Initially in its discovery GBV-C was associated with non-A-B hepatitis. However, subsequent studies failed to associate GBV-C with any known human disease and the virus became neglected until a series of studies associated the virus with prolonged survival in HIV infected recipients. Studies evaluating the co-infection presents conflicting results whereas triple HIV-HCV-GBV-C infection remains to be clarified. With the aim to evaluate the effect of GBV-C upon HIV and HIV-HCV co-infected patients, we included a cohort of 159 HIV-seropositive patients from CCDI-UNIFESP. The patients were tested for the presence of anti-E2 antibodies and GBV-C RNA. Of the 107 HIV seropositive patients negative to HCV infection, 41 (38,3%) were positive to GBV-C infection markers, of whom 17 (15,8%) were GBV-C viremic and 24 (22,4%) were positive to anti-E2 antibodies. Of the 52 HIV-HCV co-infected patients, 24 (46,1%) were positive to GBV-C infection markers, of whom 14 (26,9%) were viremic and 10 (19,2%) presented anti-E2 antibodies. Epidemiological data were collected; virological and immunological markers and also hepatic function were evaluated. Besides, IFN- and IL-2 production on CD4, T CD8 and T cells and CD38 activation marker on T CD4 and T CD8 cells were also evaluated. No significant differences on HIV viral load nor on T CD4 and T CD8 cell counts were observed, according to the infection profile. Immune response, evaluated by the IFN- and IL-2 production and by the CD38 expression did not differ among the groups studied. Univariate analysis demonstrated higher ALT, AST and GGT levels in the HIV-HCV co-infetced and HIVHCV- GBV-C triple infected patients. The multivariate analysis demonstrated that the GBV-C influences ALT levels on triple infected patients. Our results demonstrate that GBV-C does not exerts beneficial effects upon chronically HIV infected patients, unlike, it can cause hepatic overload, as demonstrated by the higher ALT levels in the HIV-HCV-GBV-C triple infected patients. In the light of these results, is reasonable to prudently consider this interaction until the possible pathological role of GBV-C on this situation is completely excluded.A infecção pelo GBV-C é freqüente em indivíduos saudáveis e pode permanecer por longos períodos sem a manifestação de sinais e sintomas clínicos. O GBV-C é um flavivírus composto por uma única fita de RNA de polaridade positiva, intimamente relacionado ao HCV. Inicialmente, o GBV-C foi associado à casos de hepatite fulminante de etiologia desconhecida. Estudos posteriores, no entanto, falharam em associar o GBV-C a qualquer doença humana conhecida e o vírus foi negligenciado por um longo período até que estudos sugeriram um efeito benéfico da co-infecção em pacientes HIV soropositivos. Os estudos envolvendo a co-infecção HIV-GBV-C apresentam resultados controversos enquanto trabalhos avaliando a tripla infecção HIV-HCV-GBV-C ainda são raros. Com o intuito de avaliar o efeito do GBV-C sobre pacientes HIV e HIV-HCV co-infectados crônicos, incluímos uma coorte de 159 pacientes HIV soropositivos triados a partir do CCDI-UNIFESP. Os pacientes foram testados para a presença de anticorpos anti-E2 e RNA do GBV-C. Dos 107 pacientes HIV, negativos para o HCV, 41 (38,3 por cento) apresentaram marcadores de infecção pelo GBV-C, dos quais 17 (15,8 por cento) eram virêmicos e 24 (22,4 por cento) positivos para anticorpos anti-E2. Dos 52 pacientes HIV-HCV co-infectados, 24 (46,1 por cento) apresentaram marcadores de infecção pelo GBV-C, dos quais 14 (26,9 por cento) apresentaram viremia e 10 (19,2 por cento) foram positivos para anticorpos anti-E2 do GBV-C. Foram coletados dados epidemiológicos e avaliados marcadores virológicos, imunológicos e de função hepática, além da produção de IFN-γ e IL-2 em células T CD4, T CD8 e Tγδ e da avaliação do marcador de ativação celular CD38 em células T CD4 e T CD8. Não foram observadas diferenças estatísticas nos níveis de CV do HIV e nem na contagem de linfócitos T CD4 e T CD8, de acordo com o perfil de infecção. A resposta imune, avaliada pela produção de citocinas IFN-γ e IL-2 e da expressão do marcador de ativação celular CD38 não diferiu entre os grupos avaliados. A análise univariada demonstrou aumento dos níveis de ALT, AST e GGT no grupo de pacientes HIV-HCV co-infectados e no grupo triplo infectado HIV-HCV- GBV-C. A análise multivariada revelou a influência do GBV-C sobre o aumento da ALT nos pacientes com tripla infecção. Os nossos dados demonstram que o GBV-C não exerce influência positiva sobre a infecção pelo HIV e pode causar sobrecarga hepática, como demonstrado pela elevação da ALT, em pacientes com tripla infecção. Dessa forma, esta interação deve ser vista com cautela até que se exclua completamente a possibilidade de patogenicidade do GBV-C nessa situação.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAPESP: 05/57611-5TEDEBV UNIFESP: Teses e dissertaçõe

HIV-GB virus C co-infection: an overview

GB virus C (GBV-C) and hepatitis G virus (HGV) are two isolates of the same virus, independently identified in humans in the 1990s by two research laboratories, and were initially considered a potential cause of liver disease. Studies failed to associate the virus with hepatitis or any known human disease. GBV-C reappeared in the scientific scene when some research groups, in an attempt to find the interference of the virus among HIV seropositive patients, reported a lower mortality rate and slower disease progression among co-infected patients. From then on, several mechanisms have been proposed to clarify this putative benefit; however, the question whether GBV-C exerts a protective effect in HIV-infected patients remains to be resolved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo, Lab Virol & Immunol 1, Virol Lab, Infect Dis Discipline, BR-04038032 São Paulo, BrazilFleury Med & Hlth, São Paulo, BrazilUniversidade Federal de São Paulo, Lab Virol & Immunol 1, Virol Lab, Infect Dis Discipline, BR-04038032 São Paulo, BrazilFAPESP: 05/576115-5FAPESP: 05/58901-7Web of Scienc

BACTERIAL SENSING, CELL SIGNALING, and MODULATION of the IMMUNE RESPONSE DURING SEPSIS

Since the definition of systemic inflammatory response syndrome/sepsis was originally proposed, a large amount of new information has been generated showing a much more complex scenario of inflammatory and counterinflammatory responses during sepsis. Moreover, some fundamental mechanisms of sensing and destroying invading microorganisms have been uncovered, which include the discovery of TLR4 as the lipopolysaccharide (LPS) gene, implications of innate immune cells as drivers of the adaptive response to infection, and the modulation of multiple accessory molecules that stimulate or inhibit monocyte/macrophage and lymphocyte interactions. the complexity of the infection/injury-induced immune response could be better appreciated with the application of genomics and proteomics studies, and LPS was a useful tool in many of these studies. in this review, we discuss aspects of bacterial recognition and induced cellular activation during sepsis. Because of the relevance of endotoxin (LPS) research in the field, we focus on LPS and host interactions as a clue to understand microorganisms sensing and cell signaling, then we discuss how this response is modulated in septic patients.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo, Escola Paulista Med, Hosp São Paulo, Div Infect Dis,Dept Med, São Paulo, BrazilMax Planck Inst Immunobiol, Freiburg, GermanyUniversidade Federal de São Paulo, Escola Paulista Med, Hosp São Paulo, Div Infect Dis,Dept Med, São Paulo, BrazilFAPESP: 2006/58744-1CNPq: 303635/2008-8FAPESP: 2008/07511-2Web of Scienc

Differential expression of toll-like receptor signaling cascades in LPS-tolerant human peripheral blood mononuclear cells

Pre-exposure to low doses of LPS induces resistance to a lethal challenge, a phenomenon known as endotoxin tolerance. in this study, tolerance was induced in human PBMC by culturing cells with 1 ng/mL LPS for 48 h. Cells were subsequently challenged with 100 ng/mL LPS for 2, 6 and 24 h, and the expression of 84 genes encoding proteins involved in the TLR signaling pathway was evaluated at each time point by PCR array. LPS pretreatment did not modulate the expression of TLR4 and CD14 on the surface of monocytes. A gene was defined as tolerized when LPS pretreatment reversed the effect of LPS challenge on the expression of the gene or as non-tolerized when LPS pretreatment did not reverse the effects of LPS challenge. We observed impaired signal transduction through the NF-kappa B, JNK, ERK and TRIF pathways, whereas expression of p38 pathway-related genes was preserved in LPS-tolerant cells. These results show a distinct regulation of the TLR pathway cascades during tolerance; this may account for the differential gene expression of some inflammatory mediators, such as up-regulation of IL-10 and COX2 as well as down-regulation of INF-alpha and IL-12. Depending on the effect of LPS-induced gene up-regulation or down-regulation, tolerance, as a reversion of such LPS effects, may result in repression or induction of gene expression. (C) 2010 Elsevier GmbH. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo, Div Infect Dis, Escola Paulista Med, Dept Med, BR-04039032 São Paulo, BrazilUniversidade Federal de São Paulo, Div Infect Dis, Escola Paulista Med, Dept Med, BR-04039032 São Paulo, BrazilFAPESP: 2006/58744-1FAPESP: 2007/56960-7Web of Scienc

Gene expression profiling of mononuclear cells from patients with sepsis secondary to community-acquired pneumonia

Mechanisms governing the inflammatory response during sepsis involve crosstalk between diverse signaling pathways, but current knowledge provides an incomplete picture of the syndrome. Microarray-based expression profiling is a powerful approach for the investigation of complex clinical conditions such as sepsis. In this study, we investigated whole-genome expression profiles in mononuclear cells from septic patients admitted in intensive care units with community-acquired pneumonia. Blood samples were collected at the time of sepsis diagnosis and seven days later since we aimed to evaluate the role of biological processes or genes possibly involved in patient recovery. Here we provide a detailed description of the study design, including clinical information, experimental methods and procedures regarding data analysis. Metadata corresponding to microarray results deposited in the database Gene Expression Omnibus (GEO) under the accession number GSE48080 are also described in this report. Our dataset allows the identification of genes possibly associated with host defense to infection as well as gene expression patterns associated with patient outcome

Global gene expression and patient outcome.

<p>Unsupervised classification by principal components analysis (PCA) of septic patients considering the outcome. Two separate clusters depict survivors and non-survivors, and this result is independent of the date of sample collection. Numbers refers to patients' identification and the day of sample collection (0 or 7).</p