ABSTRACT: Intestinal microbial communities and Holdemanella isolated from HIV pos, neg men who have sex with men increase frequencies of lamina propria CCR5+ CD4 T cells
Project description:Intestinal microbial communities and Holdemanella biformis isolated from HIV+/− men who have sex with men increase frequencies of lamina propria CCR5+ CD4+ T cells
Project description:This RNA seq experiment was designed to identify a gene signature of a CCR5-expressing subset of human intestinal Th17-cells. T helper-cells from peripheral blood of healthy donors and from the lamina propria of Crohn’s Disease patients were FACS-purified according to the expression of the chemokine receptors CCR6, CCR5 and CXCR3. Four CCR6+Th- subsets were isolated according to CXCR3 and CCR5 expression: two CXCR3- Th17 subsets (CCR5-: “cTh17” or CCR5+: “pTh17”) and two subsets of CXCR3+ Th1/17-cells (CCR5+ or CCR5-)
Project description:Mammals differ more than 100-fold in maximum lifespan. Here, we conducted comparative transcriptomics on 26 species with diverse lifespans. We identified thousands of genes with expression levels negatively or positively correlated with a species' maximum lifespan (Neg- or Pos-MLS genes). Neg-MLS genes are primarily involved in energy metabolism and inflammation. Pos-MLS genes show enrichment in DNA repair, microtubule organization, and RNA transport. Expression of Neg- and Pos-MLS genes is modulated by interventions, including mTOR and PI3K inhibition. Regulatory networks analysis showed that Neg-MLS genes are under circadian regulation possibly to avoid persistent high expression, whereas Pos-MLS genes are targets of master pluripotency regulators OCT4 and NANOG and are upregulated during somatic cell reprogramming. Pos-MLS genes are highly expressed during embryogenesis but significantly downregulated after birth. This work provides targets for anti-aging interventions by defining pathways correlating with longevity across mammals and uncovering circadian and pluripotency networks as central regulators of longevity.
Project description:Using Gfi1b conditional mice, deletion of gfi1b in the hematopietic system was induced by injecting MxCre tg Gfi1bfl/fl mice with pIpC. 30 days after injection, Cd150 pos, Cd 48 neg, Lin neg Sca and c-kit pos stem cells were sortrted from Gfi1bfl/fl and Mxcre tg Gfi1bfl/fl mice and analysed. We used the mouse Affymetrix Gene ST Array.
Project description:Cellular composition and responsiveness of immune system evolve upon ageing and are influenced by biological sex. CD4+ T cells from women living with HIV exhibit decreased viral replication ex vivo compared to men. We thus hypothesized that these findings could be recapitulated in vitro, and infected primary CD4+ T cells with HIV-based vectors pseudotyped with VSV-G or HIV envelopes. We used cells isolated from twenty donors to interrogate the effect of sex and age on permissiveness over a six-day activation kinetics. Our data identified an increased permissiveness to HIV between 24 and 72h post-stimulation. Sex- and age-based analysis at these time points showed increased susceptibility to HIV of cells isolated from males and from donors over 50 years old, respectively. Parallel assessment of surface marker expression revealed higher frequencies of activation markers (CD69, CD25, HLA-DR) and immune check point inhibitors (PD-1 and CTLA-4 in cells from highly permissive donors. Furthermore, positive correlations were identified between CD69, PD-1 and CTLA-4 expression kinetics and HIV expression kinetics. Cell population heterogeneity was assessed by single-cell RNA-Seq analysis and no cell subtype enrichment was identified according to sex. Finally, transcriptomic analyses further highlighted the role of activation in those differences with enriched activation and cell cycle gene sets in men and in older women cells. Altogether, this study brought further evidence about individual features affecting HIV replication at the cellular level and should be considered in latency reactivation studies for HIV cure.
Project description:Cellular composition and responsiveness of immune system evolve upon ageing and are influenced by biological sex. CD4+ T cells from women living with HIV exhibit decreased viral replication ex vivo compared to men. We thus hypothesized that these findings could be recapitulated in vitro, and infected primary CD4+ T cells with HIV-based vectors pseudotyped with VSV-G or HIV envelopes. We used cells isolated from twenty donors to interrogate the effect of sex and age on permissiveness over a six-day activation kinetics. Our data identified an increased permissiveness to HIV between 24 and 72h post-stimulation. Sex- and age-based analysis at these time points showed increased susceptibility to HIV of cells isolated from males and from donors over 50 years old, respectively. Parallel assessment of surface marker expression revealed higher frequencies of activation markers (CD69, CD25, HLA-DR) and immune check point inhibitors (PD-1 and CTLA-4 in cells from highly permissive donors. Furthermore, positive correlations were identified between CD69, PD-1 and CTLA-4 expression kinetics and HIV expression kinetics. Cell population heterogeneity was assessed by single-cell RNA-Seq analysis and no cell subtype enrichment was identified according to sex. Finally, transcriptomic analyses further highlighted the role of activation in those differences with enriched activation and cell cycle gene sets in men and in older women cells. Altogether, this study brought further evidence about individual features affecting HIV replication at the cellular level and should be considered in latency reactivation studies for HIV cure.
Project description:CCR5 is the main HIV co-receptor. We aimed to (1) compare CCR5 expression on immune cells between people living with HIV (PLHIV) using combination antiretroviral therapy (cART) and HIV-uninfected controls, (2) relate CCR5 expression to viral reservoir size and (3) assess detereminants of CCR5 expression. Percentages of CCR5 positive cells (%) and CCR5 mean fluorescence intensity (MFI) assessed by flow cytometry in monocytes and lymphocyte subsets were correlated to host factors, HIV-1 cell-associated (CA)-RNA and CA-DNA, plasma inflammation markers and metabolites.
Project description:We sorted Group 3 MB xenografts tumors by PRTG surface expression and performed single-cell RNA sequencing on the PRTG-pos and PRTG-neg fractions.
Project description:Using Gfi1b conditional mice, deletion of gfi1b in the hematopietic system was induced by injecting MxCre tg Gfi1bfl/fl mice with pIpC. 30 days after injection, Cd150 pos, Cd 48 neg, Lin neg Sca and c-kit pos stem cells were sortrted from Gfi1bfl/fl and Mxcre tg Gfi1bfl/fl mice and analysed. We used the mouse Affymetrix Gene ST Array. The study should determine whether loss of Gfi1 alters the gene expression pattern in the hematopietic stem cells.
Project description:To further development of our gene expression approach to CD300a deficiency on dendritic cells (DCs) in colonic lamina propria, we have employed whole genome microarray expression profiling as a discovery platform to identify genes with the potential to distinguish CD300a deficiency on DCs in colonic lamina propria from those of WT mice. Colonic lamina propria DCs were obtained by cell sorter from WT and CD300a deficient mice raised under SPF and GF condition. Expression of Ifnb1 was significantly higher in CD300a deficient DCs, quantified in the same RNA samples by real-time PCR. Gene expression in WT and CD300a colonic lamina propria DCs raised under SPF and GF conditions were measured. Colonic lamina propria cells were obtained from 5 mice in each conditions. Takara-Bio