Permicibility of cellular lines of bats (rousettus aegyptiacus and eidolon helvum) to different human viruses

Orientador: Clarice Weis ArnsDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências MédicasResumo: Frente às epidemias do ultimo século é possível observar que o aumento da interação entre humanos e animais silvestres tem facilitado o surgimento de cepas virais de importância para a saúde pública. Dentre os animais silvestres, os morcegos (ordem Chiroptera) se destacam pelo seu modo de via antropofílico, e são descritos corriqueiramente como reservatórios e potencias transmissores para diversos patógenos virais humanos. Desse modo, a presente dissertação pretende caracterizar a permissibilidade e a replicação de diversos vírus humanos em linhagens celulares de morcego Rousettus aegyptiacus e Eidolon helvum. Foram realizadas sucessivas passagens de amostras virais nas células em busca da observação de efeito citopático. Em seguida foi avaliado a cinética de multiplicação viral das células de morcego em comparação com as células padrões de crescimento de cada vírus sendo esse quantificado por meio de Real Time qPCR. As linhagens celulares foram testadas para os vírus da Dengue, Zika vírus, Vírus Sincicial Respiratório e Herpes vírus tipo 1, em parte devido a sua prevalência e importância na saúde pública nos últimos anos e pelo fato de alguns desses agentes possuírem padrões sazonais de circulação em humanos, o que indica a possibilidade da presença de reservatórios animais assintomáticos. Com os resultados obtidos podemos dizer que as linhagens RoNi/7 e EidNi/41 foram permissivas e promoveram a replicação dos vírus da Dengue e Herpes vírus Tipo 1, oque demonstra um potencial envolvimento dos morcegos na epidemiologia dessas doenças reforçando a necessidade de mais estudos. Para o Zika vírus a linhagem celular EidNi/41 foi capaz de promover replicação viral já a linhagem RoNi/7 não obteve mesmo sucesso. Para o Vírus Sincicial Respiratório, ambas as linhagens não foram permissivas e não foram capazes de promover a replicação viralAbstract: Analyzing the epidemics of the last century, it is possible to the increase of the interaction between humans and a wild animal has facilitated the emergence of viral strains of importance for public health. Among the wild animals, bats (order Chiroptera) stand out by their anthropophilic way of way, and are described as reservoirs and transmitting powers for several human viral pathogens. This present dissertation aims to characterize the permissibility and replication of human viruses in Rousettus aegyptiacus and Eidolon helvum bat cell lines. Successive passages of viral samples were performed on the cells in search of cytopathic effect. Then, the kinetics of viral multiplication of the bat cells were evaluated in comparison with the standard growth cells of each virus and this was quantified by means of Real Time qPCR. Cell lines were tested for Dengue virus, Zika virus, Respiratory Syncytial Virus and Herpes virus type 1, in part because of their prevalence and importance in public health in recent years and because some of these agents have seasonal circulation patterns in humans, which indicates the possibility of the presence of asymptomatic animal reservoirs. With the results obtained we can say that the lines RoNi/7 and EidNi/41 were permissive and promoted the replication of the Dengue virus and Herpes virus type 1, which demonstrates a potential involvement of the bats in the epidemiology of these diseases reinforcing the need for further studies. For the Zika virus the EidNi/41 cell lineage was able to promote viral replication but RoNi/7 lineage was not even successful. For Respiratory Syncytial Virus, both strains were not permissive and were not able to promote viral replicationMestradoClinica MedicaMestra em Ciências3370/2017CAPE

A lipidomics approach in the characterization of zika-infected mosquito cells: potential targets for breaking the transmission cycle

Recent outbreaks of Zika virus in Oceania and Latin America, accompanied by unexpected clinical complications, made this infection a global public health concern. This virus has tropism to neural tissue, leading to microcephaly in newborns in a significant proportion of infected mothers. The clinical relevance of this infection, the difficulty to perform accurate diagnosis and the small amount of data in literature indicate the necessity of studies on Zika infection in order to characterize new biomarkers of this infection and to establish new targets for viral control in vertebrates and invertebrate vectors. Thus, this study aims at establishing a lipidomics profile of infected mosquito cells compared to a control group to define potential targets for viral control in mosquitoes. Thirteen lipids were elected as specific markers for Zika virus infection (Brazilian strain), which were identified as putatively linked to the intracellular mechanism of viral replication and/or cell recognition. Our findings bring biochemical information that may translate into useful targets for breaking the transmission cycle1110CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPSem informaçãoSem informação2011/50400-0; 2015/06809-1; 2014/00084-2; 2014/00302-0; 2013/11343-

A Lipidomics Approach in the Characterization of Zika-Infected Mosquito Cells: Potential Targets for Breaking the Transmission Cycle

<div><p>Recent outbreaks of Zika virus in Oceania and Latin America, accompanied by unexpected clinical complications, made this infection a global public health concern. This virus has tropism to neural tissue, leading to microcephaly in newborns in a significant proportion of infected mothers. The clinical relevance of this infection, the difficulty to perform accurate diagnosis and the small amount of data in literature indicate the necessity of studies on Zika infection in order to characterize new biomarkers of this infection and to establish new targets for viral control in vertebrates and invertebrate vectors. Thus, this study aims at establishing a lipidomics profile of infected mosquito cells compared to a control group to define potential targets for viral control in mosquitoes. Thirteen lipids were elected as specific markers for Zika virus infection (Brazilian strain), which were identified as putatively linked to the intracellular mechanism of viral replication and/or cell recognition. Our findings bring biochemical information that may translate into useful targets for breaking the transmission cycle.</p></div

Scores plot between the first two principal components (PCs) selected.

<p>The explained variances are 33.5% for component 1 and 52.5% for component two.</p

Chemical images from MALDI-MSI showing the comparison between infected and uninfected cells.

<p>It is visually possible to assess the difference in lipid composition of ZIKV-specific characterized biomarkers.</p

Lipid markers elect by PLD-DA VIP score ≥ 1.5 and elucidated by MS/MS for <i>Aedes albopictus</i> C6/36 uninfected cells (Control group).

<p>Lipid markers elect by PLD-DA VIP score ≥ 1.5 and elucidated by MS/MS for <i>Aedes albopictus</i> C6/36 uninfected cells (Control group).</p

Semi-quantitative analysis of characteristic lipids showed that ZIKV-infected cells.

<p>The bars representing confidence interval of 95% (**** p<0.0001)</p