Project description:Background : Poxviruses have been shown to be efficient vectors for the production of protective immune responses in host species but also in on host species. Myxoma virus belongs to poxviridae family. In order to evaluate the capacity of (MYXV) as a vaccine vector in ruminants, we invistigated its interactions with bone marrow-derived dendritic cells (BM-DC). DCs are the most potent antigen-presenting cells and play a crucial role during the priming and reactivation of antigen-specific immune responses . Following infection by a pathogen, the functional changes of DC are essential since priming and polarization of the immune response depend on these changes . Therefore a better understanding of the modifications in gene expression of proinflammatory cytokines, chemokines and stimulatory molecules expression may prove useful in predicting whether or not a vaccine vector will be effective. Moreover, it may help identifying modifications for improving its efficacy. Methodology/Principal Findings: We investigated in vitro the interactions between recombinant MYXV expressing GFP protein, SG33-GFP and ovine BM-DCs. To gain a global view of the gene remodelling induced by MYXV, we infected BM-DCs from three sheep with SG33-GFP at different multiplicity of infection (MOI) (0, 1, 3) during different time (0, 3, 8 hours). We measured the expression of 15K probes in BM-DCs after each kind of conditions with ovine Agilent microarrays. Furthermore, a selected number of genes were confirmed by RT-qPCR. MYXV infection induces a strong reprogramming of the cells leading to the expression of pro-inflammatory cytokines and mobilisation of type I IFN pathways, cellular death and features associated with the activation of the adaptive immune response. Conclusion/Significance: Microarray profiling of a poxvirus-DC interaction help to delineate some interesting features of a vector candidate in ovine, and pave the way for additional improvements of a promising platform. Keywords : Myxoma virus ; bone marrow-derived dendritic cells ; transcriptome ; sheep; vaccine 9 samples: 3 time points and 3 replicates of MOI=1
Project description:Background : Poxviruses have been shown to be efficient vectors for the production of protective immune responses in host species but also in on host species. Myxoma virus belongs to poxviridae family. In order to evaluate the capacity of (MYXV) as a vaccine vector in ruminants, we invistigated its interactions with bone marrow-derived dendritic cells (BM-DC). DCs are the most potent antigen-presenting cells and play a crucial role during the priming and reactivation of antigen-specific immune responses . Following infection by a pathogen, the functional changes of DC are essential since priming and polarization of the immune response depend on these changes . Therefore a better understanding of the modifications in gene expression of proinflammatory cytokines, chemokines and stimulatory molecules expression may prove useful in predicting whether or not a vaccine vector will be effective. Moreover, it may help identifying modifications for improving its efficacy. Methodology/Principal Findings: We investigated in vitro the interactions between recombinant MYXV expressing GFP protein, SG33-GFP and ovine BM-DCs. To gain a global view of the gene remodelling induced by MYXV, we infected BM-DCs from three sheep with SG33-GFP at different multiplicity of infection (MOI) (0, 1, 3) during different time (0, 3, 8 hours). We measured the expression of 15K probes in BM-DCs after each kind of conditions with ovine Agilent microarrays. Furthermore, a selected number of genes were confirmed by RT-qPCR. MYXV infection induces a strong reprogramming of the cells leading to the expression of pro-inflammatory cytokines and mobilisation of type I IFN pathways, cellular death and features associated with the activation of the adaptive immune response. Conclusion/Significance: Microarray profiling of a poxvirus-DC interaction help to delineate some interesting features of a vector candidate in ovine, and pave the way for additional improvements of a promising platform. Keywords : Myxoma virus ; bone marrow-derived dendritic cells ; transcriptome ; sheep; vaccine
Project description:Infection of mouse bone marrow Flts3L-derived dendritic cells (FL-DCs) with chimeric human T cell leukemia virus type 1 (HTLV-1) results in the upregulation as well as selective downregulation of various DC-specific genes. Mouse bone marrow Flt3L-derived dendritic cells were infected with chimeric HTLV-1 and analyzed for the expression of various DC-specific genes after 12 hours. The expression of various genes was compared to the control (non-infected) cells.
Project description:Infection of mouse bone marrow Flts3L-derived dendritic cells (FL-DCs) with chimeric human T cell leukemia virus type 1 (HTLV-1) results in the upregulation as well as selective downregulation of various DC-specific genes.
Project description:Antigen presenting cells (APC) play a central role in the host immune response to pathogen invasion by triggering early innate responses and facilitating the subsequent adaptive immune system for effective control and clearance of the infection. Accordingly, APC critically impact the outcome of an immune response. In vitro bone marrow (BM) derived APC facilitate studies aimed at dissecting their biology and function. BM derived cell cultures differentiated with granulocyte-macrophage colony stimulating factor (GM-CSF) consists of a heterogeneous mix of both dendritic cells (DC) and macrophages (MØ). Here we addressed the question of the impact of heterogeneous APC populations in their response to RNA viruses using the well characterised lymphocytic choriomeningitis virus (LCMV) infection model of BMDC populations. We compared LCMV infection and treatment with the dsRNA mimic poly (I:C)) to characterise the response of different subsets of BMDC using single cell transcriptomic analysis. Our results identified differences in the transcriptional response of BM derived APC to LCMV and poly (I:C). BM-DC were more effective at antigen presentation to T cells, whereas BM-MØ were efficient cytokine producers.
Project description:The purpose is to obtain samples for mRNA analysis in primary mouse myeloid dendritic cells infected with wild type West Nile virus (WNVMT) and mutant virus (WNVE218A). Mouse (C57Bl/6J) primary myeloid dendritic cells (D10 after seeding of leukocytes from bone marrow) were seeded at 2 x 10^5 cells per well of non-tc treated 12-well plate in 0.8 mL R10 DC media after infection at a multiplicity of infection of 500. Infected samples were collected in triplet; time-matched mocks were collected in triplet in parallel with infected samples. Time points: 1, 8, 12 and 24 h post-infection. There were two independent biological replicates of the entire procedure. This study (WDC011) is a biological replicate of another (WDC010; GSE74628).
Project description:The purpose is to obtain samples for mRNA analysis in primary mouse myeloid dendritic cells infected with wild type West Nile virus (WNVMT) and mutant virus (WNVE218A). Mouse (C57Bl/6J) primary amyeloid dendritic cells (D10 after seeding of leukocytes from bone marrow) were seeded at 2 x 10^5 cells per well of non-tc treated 12-well plate in 0.8 mL R10 DC media after infection at a multiplicity of infection of 500. Infected samples were collected in triplet; time-matched mocks were collected in triplet in parallel with infected samples. Time points: 1, 8, 12 and 24 h post-infection. There were two independent biological replicates of the entire procedure. This study (WDC010) is a biological replicate of another (WDC011; GSE75222).
Project description:Gene expression profiling of BM-derived dendritic cells reveals important mechanisms associated with predisposition to Staphylococcus infections
Project description:We cultured bone marrow derived dendritic cells from WT and CD11c KO mice. Then, a group of bone marrow dendritic cells were stimulated with LPS overnight. We obtained bone marrow derived dendritic cells with or without LPS stimulation and analyzed proteomics profiles.