Identification of the Human Papillomavirus Genotypes, According to the Human Immunodeficiency Virus Status in a Cohort of Women from Maputo, Mozambique

Background: Human papillomavirus (HPV) infection is now a well-established cause of cervical cancer and other anogenital cancers. An association between human immunodeficiency virus (HIV) infection and higher HPV incidence and prevalence are commonly reported. This study was conducted to demonstrate HPV prevalence, genotypes and its characteristics, according to the HIV status in women from Maputo in Mozambique. Methods: A total of 233 participants with ages ranging from fourteen to forty-five were included. Cervical samples were collected, DNA extracted, and HPV genotyping was performed using the HPV Direct Flow CHIP Kit. Results: In total, 177 HIV-negative and 56 HIV-positive women were included in the analysis. The overall HPV prevalence was 63% and was significantly higher among HIV-positive women (79% versus 58% among HIV-negative women; p = 0.005). The prevalence of multiple HPV type infections was 32%. High-risk HPV types 52, 68, 35, 18 and 16 were the most frequent. A higher proportion of HIV-positive women had multiple HPV types compared with HIV-negative women. Conclusions: This study demonstrated a high prevalence of HPV in the study cohort. HIV-positive women were identified as having the highest HPV prevalence and infection with multiple HPV types across all ages. High-risk genotypes were the most commonly found

Differential interactions of virulent and non-virulent H. parasuis strains with naïve or swine influenza virus pre-infected dendritic cells

Pigs possess a microbiota in the upper respiratory tract that includes Haemophilus parasuis. Pigs are also considered the reservoir of influenza viruses and infection with this virus commonly results in increased impact of bacterial infections, including those by H. parasuis. However, the mechanisms involved in host innate responses towards H. parasuis and their implications in a co-infection with influenza virus are unknown. Therefore, the ability of a non-virulent H. parasuis serovar 3 (SW114) and a virulent serovar 5 (Nagasaki) strains to interact with porcine bone marrow dendritic cells (poBMDC) and their modulation in a co-infection with swine influenza virus (SwIV) H3N2 was examined. At 1 hour post infection (hpi), SW114 interaction with poBMDC was higher than that of Nagasaki, while at 8 hpi both strains showed similar levels of interaction. The co-infection with H3N2 SwIV and either SW114 or Nagasaki induced higher levels of IL-1β, TNF-α, IL-6, IL-12 and IL-10 compared to mock or H3N2 SwIV infection alone. Moreover, IL-12 and IFN-α secretion differentially increased in cells co-infected with H3N2 SwIV and Nagasaki. These results pave the way for understanding the differences in the interaction of non-virulent and virulent strains of H. parasuis with the swine immune system and their modulation in a viral co-infection

Estudo das frequências alélicas de Short Tandem Repeats (STRs) autossómicos na população de Moçambique

Poster apresentado na III Conferência do Instituto Nacional de Medicina Legal e Ciências Forense, Coimbra, 2016Os marcadores genéticos Short Tandem Repeats (STRs) são sequências nucleótidicas altamente polimórficas e repetitivas, encontradas no ácido desoxirribonucleico (ADN), que devido à sua elevada variabilidade e ao facto de serem facilmente amplificados por Reação em Cadeia da Polimerase (PCR), têm sido utilizadas com diferentes fins, nomeadamente, em estudos de genética populacional e de identificação humana. A identificação humana utilizando o ADN é um grande desafio para Moçambique, uma vez que não existem bases de dados genéticos populacionais. Assim, o objetivo deste estudo consiste na contribuição genética para estas bases de dados através da tipagem de 21 loci de STRs autossómicos em dadores dos bancos de sangue de Moçambique.N/

Swine, human or avian influenza viruses differentially activates porcine dendritic cells cytokine profile

Swine influenza virus (SwIV) is considered a zoonosis and the fact that swine may act as an intermediate reservoir for avian influenza virus, potentially infectious for humans, highlights its relevance and the need to understand the interaction of different influenza viruses with the porcine immune system. Thus, in vitro porcine bone marrow-derived dendritic cell (poBMDCs) were infected with a circulating SwIV A/Swine/Spain/SF32071/2007(H3N2), 2009 human pandemic influenza virus A/Catalonia/63/2009(H1N1), low pathogenic avian influenza virus (LPAIV) A/Anas plathyrhynchos/Spain/1877/2009(aH7N2) or high pathogenic avian influenza virus (HPAIV) A/Chicken/Italy/5093/1999(aH7N1). Swine influenza virus H3N2 infection induced an increase of SLA-I and CD80/86 at 16 and 24 h post infection (hpi), whereas the other viruses did not. All viruses induced gene expression of NF-κB, TGF-β, IFN-β and IL-10 at the mRNA level in swine poBMDCs to different extents and in a time-dependent manner. All viruses induced the secretion of IL-12 mostly at 24 hpi whereas IL-18 was detected at all tested times. Only swH3N2 induced IFN-α in a time-dependent manner. Swine H3N2, aH7N2 and aH7N1 induced secretion of TNF-α also in a time-dependent manner. Inhibition of NF-κB resulted in a decrease of IFN-α and IL-12 secretion by swH3N2-infected poBMDC at 24 hpi, suggesting a role of this transcription factor in the synthesis of these cytokines. Altogether, these data might help in understanding the relationship between influenza viruses and porcine dendritic cells in the innate immune response in swine controlled through soluble mediators and transcription factors.This work was partly funded by the Project No. CSD 2006-00007, AGL2006-13809-C03-01, AGL2009-12945-C02-01 and AGL2010-22200-C02-01 by the Spanish Government. PhD studies of Mrs. Tufária Mussá and Masssimiliano Baratelli are funded by doctoral grants from the AECID and MICIN respectively.Peer reviewe

Immunogenicity of PE18, PE31, and PPE26 proteins from Mycobacterium tuberculosis in humans and mice

IntroductionThe large family of PE and PPE proteins accounts for as much as 10% of the genome of Mycobacterium tuberculosis. In this study, we explored the immunogenicity of three proteins from this family, PE18, PE31, and PPE26, in humans and mice.MethodsThe investigation involved analyzing the immunoreactivity of the selected proteins using sera from TB patients, IGRA-positive household contacts, and IGRA-negative BCG vaccinated healthy donors from the TB endemic country Mozambique. Antigen-recall responses were examined in PBMC from these groups, including the evaluation of cellular responses in healthy unexposed individuals. Moreover, systemic priming and intranasal boosting with each protein, combined with the Quil-A adjuvant, were conducted in mice.ResultsWe found that all three proteins are immunoreactive with sera from TB patients, IGRA-positive household contacts, and IGRA-negative BCG vaccinated healthy controls. Likewise, antigen-recall responses were induced in PBMC from all groups, and the proteins stimulated proliferation of peripheral blood mononuclear cells from healthy unexposed individuals. In mice, all three antigens induced IgG antibody responses in sera and predominantly IgG, rather than IgA, responses in bronchoalveolar lavage. Additionally, CD4+ and CD8+ effector memory T cell responses were observed in the spleen, with PE18 demonstrating the ability to induce tissue-resident memory T cells in the lungs.DiscussionHaving demonstrated immunogenicity in both humans and mice, the protective capacity of these antigens was evaluated by challenging immunized mice with low-dose aerosol of Mycobacterium tuberculosis H37Rv. The in vitro Mycobacterial Growth Inhibition Assay (MGIA) and assessment of viable bacteria in the lung did not demonstrate any ability of the vaccination protocol to reduce bacterial growth. We therefore concluded that these three specific PE/PPE proteins, while immunogenic in both humans and mice, were unable to confer protective immunity under these conditions

Assessment of measles immunity among infants in Maputo City, Mozambique

<p>Abstract</p> <p>Background</p> <p>The optimum age for measles vaccination varies from country to country and thus a standardized vaccination schedule is controversial. While the increase in measles vaccination coverage has produced significant changes in the epidemiology of infection, vaccination schedules have not been adjusted. Instead, measures to cut wild-type virus transmission through mass vaccination campaigns have been instituted. This study estimates the presence of measles antibodies among six- and nine-month-old children and assesses the current vaccination seroconversion by using a non invasive method in Maputo City, Mozambique.</p> <p>Methods</p> <p>Six- and nine-month old children and their mothers were screened in a cross-sectional study for measles-specific antibodies in oral fluid. All vaccinated children were invited for a follow-up visit 15 days after immunization to assess seroconversion. </p> <p>Results</p> <p>82.4% of the children lost maternal antibodies by six months. Most children were antibody-positive post-vaccination at nine months, although 30.5 % of nine month old children had antibodies in oral fluid before vaccination. We suggest that these pre-vaccination antibodies are due to contact with wild-type of measles virus. The observed seroconversion rate after vaccination was 84.2%. </p> <p>Conclusion</p> <p>These data indicate a need to re-evaluate the effectiveness of the measles immunization policy in the current epidemiological scenario.</p

Global respiratory syncytial virus–related infant community deaths

Background Respiratory syncytial virus (RSV) is a leading cause of pediatric death, with >99% of mortality occurring in low- and lower middle-income countries. At least half of RSV-related deaths are estimated to occur in the community, but clinical characteristics of this group of children remain poorly characterized. Methods The RSV Global Online Mortality Database (RSV GOLD), a global registry of under-5 children who have died with RSV-related illness, describes clinical characteristics of children dying of RSV through global data sharing. RSV GOLD acts as a collaborative platform for global deaths, including community mortality studies described in this supplement. We aimed to compare the age distribution of infant deaths <6 months occurring in the community with in-hospital. Results We studied 829 RSV-related deaths <1 year of age from 38 developing countries, including 166 community deaths from 12 countries. There were 629 deaths that occurred <6 months, of which 156 (25%) occurred in the community. Among infants who died before 6 months of age, median age at death in the community (1.5 months; IQR: 0.8−3.3) was lower than in-hospital (2.4 months; IQR: 1.5−4.0; P < .0001). The proportion of neonatal deaths was higher in the community (29%, 46/156) than in-hospital (12%, 57/473, P < 0.0001). Conclusions We observed that children in the community die at a younger age. We expect that maternal vaccination or immunoprophylaxis against RSV will have a larger impact on RSV-related mortality in the community than in-hospital. This case series of RSV-related community deaths, made possible through global data sharing, allowed us to assess the potential impact of future RSV vaccines

Immunoregulation of porcine dendritic cells by influenza viruses and Haemophilus parasuis: lessons from in vitro studies

El virus de la gripe porcina (SwIV) es una zoonosis. El hecho de que los cerdos pueden actuar como "cocteleras virales" del virus de la gripe potencialmente infecciosos para el ser humano pone de manifiesto su relevancia y la necesidad de comprender la interacción de los diferentes virus de la gripe y otros patógenos respiratorios con el sistema inmune porcino. Además, el estado clínico de los cerdos infectados con gripe puede agravarse debido a una infección bacteriana secundaria. Algunas de estas bacterias como el Haemophilus parasuis son comensales del tracto respiratorio y producen la enfermedad de Glässer, una enfermedad de las granjas de cerdos con alto impacto económico. Por otro lado, las células dendríticas (DCs) son importantes en la inducción de una respuesta inmune efectiva contra los patógenos. Su localización en las vías respiratorias como células centinelas, les convierte en dianas de los patógenos. Sin embargo, su interacción con los virus de la gripe o su papel en las co-infecciones no ha sido totalmente caracterizado. Con el objetivo de entender al nivel celular la interacción de los virus de la gripe y el H. parasuis como patógenos comunes del trato respiratorio de los cerdos, tres estudios fueron desarrollados y presentados en esta tesis. En el primer estudio, las células dendríticas derivadas de la médula ósea de cerdos (poBMDCs) fueron expuestas a una cepa circulante de la gripe porcina H3N2. En el segundo estudio, las poBMDCs fueron infectados con diferentes virus de la gripe que el sistema inmune porcino puede encontrar en la naturaleza como son: el A/Swine/Spain/SF32071/2007(H3N2), el virus de la gripe pandémica de 2009 A/Catalonia/63/2009(H1N1), el virus de la gripe aviar de baja patogenicidad Anas/plathyrhynchos/Spain/1877/2009(H7N2) y el virus de la gripe aviar de alta patogenicidad A/Chicken/Italy/5093/1999(H7N1); y finalmente en el tercer estudio, las poBMDCs fueron infectadas o co-infectadas con el H3N2-SwIV y H. parasuis no virulenta (SW114) o virulenta (Nagasaki). De estos estudios, tres conclusiones generales se pueden extraer: (i) las poBMDCs se infectan con el virus de la gripe porcina H3N2 y transmiten la infección a las células permisivas sólo cuando se favorece la interacción célula-célula, (ii) el perfil de secreción de citoquinas, la expresión de los mRNA de citoquinas y factores de transcripción después de la infección con los virus H3N2 SwIV, hH1N1 y aH7N2 o aH7N1 pueden indicar una activación de la respuesta inmune específica contra estos virus, (iii) la previa infección de las poBMDCs con el H3N2-SwIV seguida por la infección con H. parasuis resultó en un perfil de citoquinas sesgada mayoritariamente hacia una respuesta inflamatoria. En conjunto, estos datos nos ayudan a comprender la interacción entre los patógenos respiratorios como los virus de la gripe porcina y/o H. paraseis, con las células dendríticas así como conocer posibles mecanismos involucrados en el desarrollo de la respuesta inmune protectora.Swine influenza virus (SwIV) is considered a zoonosis and the fact that swine may act as “mixing vessels” of influenza viruses potentially infectious for humans illustrates its relevance and the need to understand the interaction of different influenza viruses and other respiratory pathogens with the porcine immune system. Moreover, the clinical status of influenza infected animals may deteriorate due to secondary bacterial infection. Some of these bacteria such as Haemophilus parasuis are commensal of the respiratory tract of pigs and the causal agent of Glässer’s disease, an important disease in swine herds that results in high economical loses. In other hand, dendritic cells (DCs) mediate the induction of immunity against pathogens and the location of DCs beneath the epithelium of respiratory organs makes them suitable targets for pathogens. However, their interaction with different influenza viruses or the role of DCs in co-infections has not been fully characterized. With the aim to understand at cellular level the interaction of influenza virus and H. parasuis as common pathogens of pig’s respiratory tract, three studies were undertaken and are presented in this thesis. In the first study, porcine bone marrow derived DCs (poBMDCs) were exposed to a circulating strain of H3N2-SwIV. In the second study, poBMDCs were infected with different influenza viruses that would enter in contact with the porcine immune system, such as: SwIV A/Swine/Spain/SF32071/2007(H3N2), the 2009 pandemic influenza virus A/Catalonia/63/2009(H1N1), the low pathogenic avian influenza virus A/Anas plathyrhynchos/Spain/1877/2009(H7N2) and the high pathogenic avian influenza virus A/Chicken/Italy/5093/1999(H7N1); and finally in the third study, poBMDCs were infected or co-infected with the H3N2-SwIV and H. parasuis non-virulent (SW114) or virulent (Nagasaki). From these studies, three general conclusions can be drawn: (i) poBMDCs are infected by H3N2-SwIV and they transmit the infection to permissive cells only when cell-to-cell contact was favoured; (ii) the cytokine profile and the mRNA expression of transcription factors after H3N2-SwIV, hH1N1 and aH7N2 or aH7N1 may indicate a specific immune response activation against these viruses; (iii) previous infection of poBMDCs with H3N2-SwIV followed by H. parasuis infection resulted in a profile of cytokines biased mainly towards inflammatory response. Overall, these data pave the way for understanding the relation between respiratory pathogens such as influenza viruses and/or H. parasuis and porcine dendritic cells for triggering possible mechanisms driving to protective immune response

Immunoregulation of porcine dendritic cells by influenza viruses and Haemophilus parasuis: lessons from in vitro studies

El virus de la gripe porcina (SwIV) es una zoonosis. El hecho de que los cerdos pueden actuar como "cocteleras virales" del virus de la gripe potencialmente infecciosos para el ser humano pone de manifiesto su relevancia y la necesidad de comprender la interacción de los diferentes virus de la gripe y otros patógenos respiratorios con el sistema inmune porcino. Además, el estado clínico de los cerdos infectados con gripe puede agravarse debido a una infección bacteriana secundaria. Algunas de estas bacterias como el Haemophilus parasuis son comensales del tracto respiratorio y producen la enfermedad de Glässer, una enfermedad de las granjas de cerdos con alto impacto económico. Por otro lado, las células dendríticas (DCs) son importantes en la inducción de una respuesta inmune efectiva contra los patógenos. Su localización en las vías respiratorias como células centinelas, les convierte en dianas de los patógenos. Sin embargo, su interacción con los virus de la gripe o su papel en las co-infecciones no ha sido totalmente caracterizado. Con el objetivo de entender al nivel celular la interacción de los virus de la gripe y el H. parasuis como patógenos comunes del trato respiratorio de los cerdos, tres estudios fueron desarrollados y presentados en esta tesis. En el primer estudio, las células dendríticas derivadas de la médula ósea de cerdos (poBMDCs) fueron expuestas a una cepa circulante de la gripe porcina H3N2. En el segundo estudio, las poBMDCs fueron infectados con diferentes virus de la gripe que el sistema inmune porcino puede encontrar en la naturaleza como son: el A/Swine/Spain/SF32071/2007(H3N2), el virus de la gripe pandémica de 2009 A/Catalonia/63/2009(H1N1), el virus de la gripe aviar de baja patogenicidad Anas/plathyrhynchos/Spain/1877/2009(H7N2) y el virus de la gripe aviar de alta patogenicidad A/Chicken/Italy/5093/1999(H7N1); y finalmente en el tercer estudio, las poBMDCs fueron infectadas o co-infectadas con el H3N2-SwIV y H. parasuis no virulenta (SW114) o virulenta (Nagasaki). De estos estudios, tres conclusiones generales se pueden extraer: (i) las poBMDCs se infectan con el virus de la gripe porcina H3N2 y transmiten la infección a las células permisivas sólo cuando se favorece la interacción célula-célula, (ii) el perfil de secreción de citoquinas, la expresión de los mRNA de citoquinas y factores de transcripción después de la infección con los virus H3N2 SwIV, hH1N1 y aH7N2 o aH7N1 pueden indicar una activación de la respuesta inmune específica contra estos virus, (iii) la previa infección de las poBMDCs con el H3N2-SwIV seguida por la infección con H. parasuis resultó en un perfil de citoquinas sesgada mayoritariamente hacia una respuesta inflamatoria. En conjunto, estos datos nos ayudan a comprender la interacción entre los patógenos respiratorios como los virus de la gripe porcina y/o H. paraseis, con las células dendríticas así como conocer posibles mecanismos involucrados en el desarrollo de la respuesta inmune protectora.Swine influenza virus (SwIV) is considered a zoonosis and the fact that swine may act as "mixing vessels" of influenza viruses potentially infectious for humans illustrates its relevance and the need to understand the interaction of different influenza viruses and other respiratory pathogens with the porcine immune system. Moreover, the clinical status of influenza infected animals may deteriorate due to secondary bacterial infection. Some of these bacteria such as Haemophilus parasuis are commensal of the respiratory tract of pigs and the causal agent of Glässer's disease, an important disease in swine herds that results in high economical loses. In other hand, dendritic cells (DCs) mediate the induction of immunity against pathogens and the location of DCs beneath the epithelium of respiratory organs makes them suitable targets for pathogens. However, their interaction with different influenza viruses or the role of DCs in co-infections has not been fully characterized. With the aim to understand at cellular level the interaction of influenza virus and H. parasuis as common pathogens of pig's respiratory tract, three studies were undertaken and are presented in this thesis. In the first study, porcine bone marrow derived DCs (poBMDCs) were exposed to a circulating strain of H3N2-SwIV. In the second study, poBMDCs were infected with different influenza viruses that would enter in contact with the porcine immune system, such as: SwIV A/Swine/Spain/SF32071/2007(H3N2), the 2009 pandemic influenza virus A/Catalonia/63/2009(H1N1), the low pathogenic avian influenza virus A/Anas plathyrhynchos/Spain/1877/2009(H7N2) and the high pathogenic avian influenza virus A/Chicken/Italy/5093/1999(H7N1); and finally in the third study, poBMDCs were infected or co-infected with the H3N2-SwIV and H. parasuis non-virulent (SW114) or virulent (Nagasaki). From these studies, three general conclusions can be drawn: (i) poBMDCs are infected by H3N2-SwIV and they transmit the infection to permissive cells only when cell-to-cell contact was favoured; (ii) the cytokine profile and the mRNA expression of transcription factors after H3N2-SwIV, hH1N1 and aH7N2 or aH7N1 may indicate a specific immune response activation against these viruses; (iii) previous infection of poBMDCs with H3N2-SwIV followed by H. parasuis infection resulted in a profile of cytokines biased mainly towards inflammatory response. Overall, these data pave the way for understanding the relation between respiratory pathogens such as influenza viruses and/or H. parasuis and porcine dendritic cells for triggering possible mechanisms driving to protective immune response