In situ Immune Signatures and Microbial Load at the Nasopharyngeal Interface in Children With Acute Respiratory Infection

Acute respiratory infection (ARI) is the most frequent cause for hospitalization in infants and young children. Using multiplexed nCounter technology to digitally quantify 600 human mRNAs in parallel with 14 virus- and 5 bacterium-specific RNAs, we characterized viral and bacterial presence in nasopharyngeal aspirates (NPA) of 58 children with ARI and determined the corresponding in situ immune profiles. NPA contained different groups of organisms and these were classified into bacterial (n = 27), viral (n = 5), codetection [containing both viral and bacterial transcripts (n = 21), or indeterminate intermediate where microbial load is below threshold (n = 5)]. We then identified differentially expressed immune transcripts (DEITs) comparing NPAs from symptomatic children vs. healthy controls, and comparing children presenting NPAs with detectable microbial load vs. indeterminate. We observed a strong innate immune response in NPAs, due to the presence of evolutionarily conserved type I Interferon (IFN)-stimulated genes (ISG), which was correlated with total bacterial and/or viral load. In comparison with indeterminate NPAs, adaptive immunity transcripts discriminated among viral, bacterial, and codetected microbial profiles. In viral NPAs, B cell transcripts were significantly enriched among DEITs, while only type III IFN was correlated with viral load. In bacterial NPAs, myeloid cells and coinhibitory transcripts were enriched and significantly correlated with bacterial load. In conclusion, digital nCounter transcriptomics provide a microbial and immunological in situ “snapshot” of the nasopharyngeal interface in children with ARI. This enabled discrimination among viral, bacterial, codetection, and indeterminate transcripts in the samples using non-invasive sampling

DETC Induces Leishmania Parasite Killing in Human In Vitro and Murine In Vivo Models: A Promising Therapeutic Alternative in Leishmaniasis

Background: Chemotherapy remains the primary tool for treatment and control of human leishmaniasis. However, currently available drugs present serious problems regarding side-effects, variable efficacy, and cost. Affordable and less toxic drugs are urgently needed for leishmaniasis. Methodology/Principal Findings: We demonstrate, by microscopy and viability assays, that superoxide dismutase inhibitor diethyldithiocarbamate (DETC) dose-dependently induces parasite killing (p,0.001) and is able to ??????sterilize?????? Leishmania amazonensis infection at 2 mM in human macrophages in vitro. We also show that DETC-induced superoxide production (p,0.001) and parasite destruction (p,0.05) were reverted by the addition of the antioxidant N-acetylcysteine, indicating that DETC-induced killing occurs through oxidative damage. Furthermore, ultrastructural analysis by electron microscopy demonstrates a rapid and highly selective destruction of amastigotes in the phagosome upon DETC treatment, without any apparent damage to the host cell, including its mitochondria. In addition, DETC significantly induced parasite killing in Leishmania promastigotes in axenic culture. In murine macrophages infected with Leishmania braziliensis, DETC significantly induced in vitro superoxide production (p = 0.0049) and parasite killing (p = 0.0043). In vivo treatment with DETC in BALB/C mice infected with Leishmania braziliensis caused a significant decrease in lesion size (p,0.0001), paralleled by a 100-fold decrease (p = 0.0087) in parasite burden. Conclusions/Significance: Due to its strong leishmanicidal effect in human macrophages in vitro, its in vivo effectiveness in a murine model, and its previously demonstrated in vivo safety profile in HIV treatment, DETC treatment might be considered as a valuable therapeutic option in human leishmaniasis, including HIV/Leishmania co-infection

Trends and predictors of transmitted drug resistance (TDR) and clusters with TDR in a local Belgian HIV-1 epidemic

We aimed to study epidemic trends and predictors for transmitted drug resistance (TDR) in our region, its clinical impact and its association with transmission clusters. We included 778 patients from the AIDS Reference Center in Leuven (Belgium) diagnosed from 1998 to 2012. Resistance testing was performed using population-based sequencing and TDR was estimated using the WHO-2009 surveillance list. Phylogenetic analysis was performed using maximum likelihood and Bayesian techniques. The cohort was predominantly Belgian (58.4%), men who have sex with men (MSM) (42.8%), and chronically infected (86.5%). The overall TDR prevalence was 9.6% (95% confidence interval (CI): 7.7-11.9), 6.5% (CI: 5.0-8.5) for nucleoside reverse transcriptase inhibitors (NRTI), 2.2% (CI: 1.4-3.5) for non-NRTI (NNRTI), and 2.2% (CI: 1.4-3.5) for protease inhibitors. A significant parabolic trend of NNRTI-TDR was found (p = 0.019). Factors significantly associated with TDR in univariate analysis were male gender, Belgian origin, MSM, recent infection, transmission clusters and subtype B, while multivariate and Bayesian network analysis singled out subtype B as the most predictive factor of TDR. Subtype B was related with transmission clusters with TDR that included 42.6% of the TDR patients. Thanks to resistance testing, 83% of the patients with TDR who started therapy had undetectable viral load whereas half of the patients would likely have received a suboptimal therapy without this test. In conclusion, TDR remained stable and a NNRTI up-and-down trend was observed. While the presence of clusters with TDR is worrying, we could not identify an independent, non-sequence based predictor for TDR or transmission clusters with TDR that could help with guidelines or public health measures

In situ Immune Signatures and Microbial Load at the Nasopharyngeal Interface in Children With Acute Respiratory Infection

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2018-12-18T13:36:14Z No. of bitstreams: 1 Fukutani K. In situ immune signatures...2018.pdf: 4134573 bytes, checksum: 5a4970b64bcb3bbe5749570022e8cdaa (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2018-12-18T14:08:09Z (GMT) No. of bitstreams: 1 Fukutani K. In situ immune signatures...2018.pdf: 4134573 bytes, checksum: 5a4970b64bcb3bbe5749570022e8cdaa (MD5)Made available in DSpace on 2018-12-18T14:08:09Z (GMT). No. of bitstreams: 1 Fukutani K. In situ immune signatures...2018.pdf: 4134573 bytes, checksum: 5a4970b64bcb3bbe5749570022e8cdaa (MD5) Previous issue date: 2018Fundação de Amparo à Pesquisa do Estado da Bahia (FAPESB; Grant PNX0019/2009 to AB) and Belgian FWO (Grant G0D6817N). KF was supported by a fellowship from Science without Borders (PVE Anne- Mieke Vandamme). TD was supported by the Institute for the Promotion of Innovation through Science and Technology Flanders (IWT-Vlaanderen), project 141614. CN-C, AB, and CO were senior investigators from Conselho Nacional de Pesquisa Científica (CNPq). HN was supported by FAPESP (Grant Nos. 2012/19278-6 and 2017/50137-3). BA was supported by the National Institutes of Health (U01AI115940).Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.Federal University of Bahia. School of Medicine. Salvador, BA, Brazil / Federal University of Bahia. School of Medicine. Department of Pediatrics. Salvador, BA, Brazil.Federal University of Bahia. School of Medicine. Department of Pediatrics. Salvador, BA, Brazil.Federal University of Bahia. School of Medicine. Department of Pediatrics. Salvador, BA, Brazil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Federal University of Bahia. School of Medicine. Department of Pediatrics. Salvador, BA, Brazil.Rega Institute for Medical Research. Department of Microbiology and Immunology. KU Leuven, Leuven, Belgium.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Federal University of Bahia. School of Medicine. Department of Pediatrics. Salvador, BA, Brazil.University of São Paulo. School of Pharmaceutical Sciences. Department of Clinical and Toxicological Analyses. São Paulo, SP, Brazil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Fundação José Silveira. Multinational Organization Network Sponsoring Translational and Epidemiological Research Initiative. Salvador, BA, Brasil.Rega Institute for Medical Research. Department of Microbiology and Immunology. KU Leuven, Leuven, Belgium.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Federal University of Bahia. School of Medicine. Department of Pediatrics. Salvador, BA, Brazil.Acute respiratory infection (ARI) is the most frequent cause for hospitalization in infants and young children. Using multiplexed nCounter technology to digitally quantify 600 human mRNAs in parallel with 14 virus- and 5 bacterium-specific RNAs, we characterized viral and bacterial presence in nasopharyngeal aspirates (NPA) of 58 children with ARI and determined the corresponding in situ immune profiles. NPA contained different groups of organisms and these were classified into bacterial (n = 27), viral (n = 5), codetection [containing both viral and bacterial transcripts (n = 21), or indeterminate intermediate where microbial load is below threshold (n = 5)]. We then identified differentially expressed immune transcripts (DEITs) comparing NPAs from symptomatic children vs. healthy controls, and comparing children presenting NPAs with detectable microbial load vs. indeterminate. We observed a strong innate immune response in NPAs, due to the presence of evolutionarily conserved type I Interferon (IFN)-stimulated genes (ISG), which was correlated with total bacterial and/or viral load. In comparison with indeterminate NPAs, adaptive immunity transcripts discriminated among viral, bacterial, and codetected microbial profiles. In viral NPAs, B cell transcripts were significantly enriched among DEITs, while only type III IFN was correlated with viral load. In bacterial NPAs, myeloid cells and coinhibitory transcripts were enriched and significantly correlated with bacterial load. In conclusion, digital nCounter transcriptomics provide a microbial and immunological in situ "snapshot" of the nasopharyngeal interface in children with ARI. This enabled discrimination among viral, bacterial, codetection, and indeterminate transcripts in the samples using non-invasive sampling

In situ Immune Signatures and Microbial Load at the Nasopharyngeal Interface in Children With Acute Respiratory Infection

Acute respiratory infection (ARI) is the most frequent cause for hospitalization in infants and young children. Using multiplexed nCounter technology to digitally quantify 600 human mRNAs in parallel with 14 virus- and 5 bacterium-specific RNAs, we characterized viral and bacterial presence in nasopharyngeal aspirates (NPA) of 58 children with ARI and determined the corresponding in situ immune profiles. NPA contained different groups of organisms and these were classified into bacterial (n = 27), viral (n = 5), codetection [containing both viral and bacterial transcripts (n = 21), or indeterminate intermediate where microbial load is below threshold (n = 5)]. We then identified differentially expressed immune transcripts (DEITs) comparing NPAs from symptomatic children vs. healthy controls, and comparing children presenting NPAs with detectable microbial load vs. indeterminate. We observed a strong innate immune response in NPAs, due to the presence of evolutionarily conserved type I Interferon (IFN)-stimulated genes (ISG), which was correlated with total bacterial and/or viral load. In comparison with indeterminate NPAs, adaptive immunity transcripts discriminated among viral, bacterial, and codetected microbial profiles. In viral NPAs, B cell transcripts were significantly enriched among DEITs, while only type III IFN was correlated with viral load. In bacterial NPAs, myeloid cells and coinhibitory transcripts were enriched and significantly correlated with bacterial load. In conclusion, digital nCounter transcriptomics provide a microbial and immunological in situ "snapshot" of the nasopharyngeal interface in children with ARI. This enabled discrimination among viral, bacterial, codetection, and indeterminate transcripts in the samples using non-invasive sampling.status: publishe

Zika Virus Infection and Stillbirths: A Case of Hydrops Fetalis, Hydranencephaly and Fetal demise

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2016-04-08T19:02:18Z No. of bitstreams: 1 Sarno M Zika Virus Infection and Stillbirths....pdf: 309667 bytes, checksum: bfc7d6ecd8b93b6ac4cee8fcc7cbc5d5 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2016-04-08T19:37:02Z (GMT) No. of bitstreams: 1 Sarno M Zika Virus Infection and Stillbirths....pdf: 309667 bytes, checksum: bfc7d6ecd8b93b6ac4cee8fcc7cbc5d5 (MD5)Made available in DSpace on 2016-04-08T19:37:02Z (GMT). No. of bitstreams: 1 Sarno M Zika Virus Infection and Stillbirths....pdf: 309667 bytes, checksum: bfc7d6ecd8b93b6ac4cee8fcc7cbc5d5 (MD5) Previous issue date: 2016Hospital Geral Roberto Santos, Secretaria Estadual da Saúde da Bahia., Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Medicina. Instituto da Saúde Coletiva. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilHospital Geral Roberto Santos, Secretaria Estadual da Saúde da Bahia., Salvador, BA, BrasilUniversidade Federal da Bahia. Faculdade de Medicina. Instituto da Saúde Coletiva. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Yale School of Public Health. Department of Epidemiology of Microbial Diseases. New Haven, Connecticut, USAHospital Geral Roberto Santos, Secretaria Estadual da Saúde da Bahia., Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilThe University of Texas Medical Branch. Institute for Human Infections and Immunity. Department of Pathology and Center of Biodefense and Emerging Infectious Diseases. Galveston, Texas, USAFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Yale School of Public Health. Department of Epidemiology of Microbial Diseases. New Haven, Connecticut, USAHospital Geral Roberto Santos, Secretaria Estadual da Saúde da Bahia., Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Medicina. Instituto da Saúde Coletiva. Salvador, BA, BrasilThe rapid spread of Zika virus in the Americas and outbreak of microcephaly in Brazil has raised attention to the possible deleterious effects that the virus may have on fetuses. We report a case of a 20-year-old pregnant woman from Salvador, Brazil whose fetus had developed hydrops fetalis, a condition where there is abnormal accumulation of fluid in the fetus, as well as severe central nervous system defects such as microcephaly and hydranencephaly. After fetal demise, ZIKV RNA was detected in central nervous system tissues and amniotic fluid. The case report provides evidence that in addition to microcephaly, there may be a link between Zika virus infection and hydrops fetalis and fetal demise. Given the recent spread of the virus, systematic investigation of spontaneous abortions and stillbirths may be warranted to evaluate the risk that ZIKV infection imparts on these outcomes

Maternal Immune Response to ZIKV Triggers High-Inflammatory Profile in Congenital Zika Syndrome

The immunological mechanisms involved in the development of congenital Zika syndrome (CZS) have yet to be fully clarified. This study aims to assess the immuno-inflammatory profile of mothers and their children who have been diagnosed with CZS. Blood samples, which were confirmed clinically using the plaque reduction neutralization test (PRNT), were collected from children with CZS and their mothers (CZS+ group). Samples were also collected from children who did not develop CZS and had a negative PRNT result and from their mothers (CZS- group). The data demonstrated a correlation between the leukocyte profile of CZS+ children and their mothers, more evident in monocytes. Monocytes from mothers of CZS+ children showed low expression of HLA and elevated hydrogen peroxide production. CZS+ children presented standard HLA expression and a higher hydrogen peroxide concentration than CZS- children. Monocyte superoxide dismutase activity remained functional. Moreover, when assessing the monocyte polarization, it was observed that there was no difference in nitrite concentrations; however, there was a decrease in arginase activity in CZS+ children. These data suggest that ZIKV infection induces a maternal immuno-inflammatory background related to the child’s inflammatory response after birth, possibly affecting the development and progression of congenital Zika syndrome

Maternal Th17 Profile after Zika Virus Infection Is Involved in Congenital Zika Syndrome Development in Children

Brazil is one of the countries that experienced an epidemic of microcephaly and other congenital manifestations related to maternal Zika virus infection which can result in Congenital Zika Syndrome (CZS). Since the Zika virus can modulate the immune system, studying mothers’ and children’s immune profiles become essential to better understanding CZS development. Therefore, we investigated the lymphocyte population profile of children who developed CZS and their mothers’ immune response in this study. The study groups were formed from the Plaque Reduction Neutralization Test (PRNT) (CZS+ group) result. To evaluate the lymphocyte population profile, we performed phenotyping of peripheral lymphocytes and quantification of serum cytokine levels. The immunophenotyping and cytokine profile was correlated between CSZ+ children and their mothers. Both groups exhibited increased interleukin-17 levels and a reduction in the subpopulation of CD4+ T lymphocytes. In contrast, the maternal group showed a reduction in the population of B lymphocytes. Thus, the development of CZS is related to the presence of an inflammatory immune profile in children and their mothers characterized by Th17 activation

Transient hearing loss in adults associated with Zika virus infection

This work was supported by the National Institutes of Health (grant numbers 1 R01 AI121207 and R24 AI120942). R. K. and L. A. S. were supported by “Programa Ciências sem fronteiras,” Conselho Nacional de Desenvolvimento Cientifico e Tecnológico of Brazil.Hospital Santa Izabel. Salvador, BA, Brazil.Ministério da Saúde. Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brazil.Hospital Santa Izabel. Salvador, BA, Brazil.Hospital Santa Izabel. Salvador, BA, Brazil.Hospital Santa Izabel. Salvador, BA, Brazil.Hospital Santa Izabel. Salvador, BA, Brazil.Ministério da Saúde. Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. World Health Organization Collaborating Center for Reference and Research on Arbovirus. Ananindeua, PA, Brasil.University of Texas Medical Branch. Center for Biodefense and Emerging Infectious Diseases. Department of Pathology. Galveston, TX, USA.University of Texas Medical Branch. Center for Biodefense and Emerging Infectious Diseases. Department of Pathology. Galveston, TX, USA.Ministério da Saúde. Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brazil / Yale School of Public Health. Department of Epidemiology of Microbial Diseases. New Haven, CT, USA.Ministério da Saúde. Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brazil / José Silveira Foundation. Research Initiative. Multinational Organization Network Sponsoring Translational and Epidemiological. Salvador, BA, BrazilMinistério da Saúde. Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brazil.Ministério da Saúde. Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brazil / KU Leuven. Rega Institute for Medical Research. Department of Microbiology and Immunology. Laboratory of Clinical and Epidemiological Virology. Leuven, Belgium.Hospital Santa Izabel. Salvador, BA, Brazil / Ministério da Saúde. Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brazil / Universidade Federal da Bahia. Faculdade de Medicina da Bahia. Salvador, BA, Brazil.In 2015, during the outbreak of Zika virus (ZIKV) in Brazil, we identified 3 cases of acute hearing loss after exanthematous illness. Serology yielded finding compatible with ZIKV as the cause of a confirmed (n = 1) and a probable (n = 2) flavivirus infection, indicating an association between ZIKV infection and transient hearing loss