Project description:Next-Generation-Sequencing (NGS) technologies have led to important improvement in the detection of new or unrecognized infective agents, related to infectious diseases. In this context, NGS high-throughput technology can be used to achieve a comprehensive and unbiased sequencing of the nucleic acids present in a clinical sample (i.e. tissues). Metagenomic shotgun sequencing has emerged as powerful high-throughput approaches to analyze and survey microbial composition in the field of infectious diseases. By directly sequencing millions of nucleic acid molecules in a sample and matching the sequences to those available in databases, pathogens of an infectious disease can be inferred. Despite the large amount of metagenomic shotgun data produced, there is a lack of a comprehensive and easy-use pipeline for data analysis that avoid annoying and complicated bioinformatics steps. Here we present HOME-BIO, a modular and exhaustive pipeline for analysis of biological entity estimation, specific designed for shotgun sequenced clinical samples. HOME-BIO analysis provides comprehensive taxonomy classification by querying different source database and carry out main steps in metagenomic investigation. HOME-BIO is a powerful tool in the hand of biologist without computational experience, which are focused on metagenomic analysis. Its easy-to-use intrinsic characteristic allows users to simply import raw sequenced reads file and obtain taxonomy profile of their samples.
Project description:The female reproductive tract is one of the major mucosal invasion site of HIV-1. This site has been neglected in previous HIV-1 vaccine studies. Immune responses in the female reproductive tract after systemic vaccination remain to be characterized. Using a modified vaccinia virus Ankara (MVA) as a vaccine model, we characterized specific immune responses in all compartments of the female reproductive tract (FRT) of non-human primates after systemic vaccination. Memory T cells were preferentially found in the lower tract (vagina and cervix), whereas antigen-presenting cells and innate lymphoid cells were mainly located in the upper tract (uterus and fallopian tubes). This compartmentalisation of immune cells in the FRT was supported by transcriptomic analyses and correlation network. Polyfunctional MVA-specific CD8+ T cells were detected in the blood, lymph nodes, vagina, cervix, uterus and fallopian tubes. Anti-MVA IgG and IgA were detected in cervicovaginal fluid after a second vaccine dose. Systemic vaccination with an MVA vector thus elicits cellular and antibody responses in the female reproductive tract.
Project description:The vertebrate nuclear hormone receptor steroidogenic factor 1 (SF1; NR5A1) controls reproductive development and regulates the transcription of steroid-modifying cytochrome P450 genes. We find that the SF1-related Drosophila nuclear hormone receptor HR39 is also essential for sexual development. In Hr39 mutant females, the sperm-storing spermathecae and glandular parovaria are absent or defective, causing sterility. Our results indicate that spermathecae and parovaria secrete reproductive tract proteins required for sperm maturation and function, like the mammalian epididymis and female reproductive tract. Hr39 controls the expression of specific cytochrome P450 genes and is required in females both to activate spermathecal secretion and repress male-specific courtship genes such as takeout. Thus, a pathway that, in vertebrates, controls sex-specific steroid hormone production, also mediates reproductive functions in an invertebrate. Our findings suggest that Drosophila can be used to model more aspects of mammalian reproductive biology than previously believed. Experiment Overall Design: Wild type and Hr39(04443) Spermathecae, Wild type and Hr39(04443) Reproductive Tract
Project description:The female’s reproductive tract is exposed directly to the male’s ejaculate, making it a hotspot for mating-induced responses shortly after mating. In Drosophila melanogaster, changes in the reproductive tract are essential to optimize fertilization. To detect the earliest gene regulatory events that underlie these changes, we measured transcript abundances using RNA-seq and microRNA-seq of reproductive tracts of unmated females and females collected within 10-15 minutes after the end of mating, either to a wildtype male or to a male with defective BMP signaling in secondary cells of the accessory gland, which influences the composition of the male’s ejaculate. We observed transcript abundance changes for genes with roles in tissue morphogenesis, wound healing, the immune response and metabolism. Strikingly, predicted targets of microRNAs that respond to mating are enriched for overlapping functions, suggesting that mating-induced changes are in part regulated by microRNAs. Most of the differentially expressed RNAs are upregulated in response to mating, while most of the differentially expressed microRNAs are downregulated. This pattern suggests a response of activation and de-repression of gene programs that switch the reproductive tract to a “mated” state, rather than a repression of virgin-specific programs. Male genotype did not influence transcript levels, indicating that the earliest transcriptomic responses in the reproductive tract are not dependent on ejaculate components that require BMP signaling in secondary cells. Our results shed light on the molecular changes that accompany very early responses to mating and present candidate genes and microRNAs that can be further examined for their participation in alterations of the reproductive tract microenvironment in response to signals from the male.
Project description:In internally fertilizing species, the first direct molecular contact between the male and female gene products occurs within the female reproductive tract – a highly complex and rapidly evolving structure in many species. While molecular interactions between sperm and oocyte have received a lot of attention, efforts to gain insight into the crosstalk happening during the sperm cells journey to the site of fertilization have been limited, especially in mammals. The interactions between sperm cells and the female reproductive tract not only facilitate sperm guiding, storage, and protection against introduced pathogens but also comprise a sophisticated mechanism of female-mediated sperm selection. Molecular interaction partners that are involved directly in sperm assessment and selection, however, are difficult to pinpoint within the massive immunological response happening in the female reproductive tract following copulation. While we have some knowledge of sperm surface proteins interacting with the components of the female reproductive tract, the female counterparts of these interactions are still largely unknown. We characterized the murine female transcriptional response to semen following copulation. We mated males and females from different strains and used strain-specific SNPs to distinguish semen-derived from female reproductive tract-derived transcripts. We specifically focused on the different subdivisions of the female reproductive tract, with the aim to identify female molecules directly involved in sperm assessment. Our results show a large number of differentially expressed genes in uterus, uterotubal junction and oviduct compared to unmated control and significant differences in differential expression between the subsections.