Project description:Ectromelia virus (ECTV) has emerged as a valuable model for investigating the host-orthopoxvirus relationship as it relates to pathogenesis and the immune response. ECTV causes mousepox in most strains of mice, including BALB/c and DBA/2, and these are therefore classified as susceptible mice. Conversely, C57BL/6 and certain 129 strains display limited pathology and a very low mortality, and are thus classified as resistant. To understand the host genetic factors of different mouse strains in response to ECTV infection, we carried out a microarray analysis using Affymetrix Gene-Chip Mouse Genome Arrays of spleen tissues from BALB/c and C57BL/6 mice at 3 and 10 days post-ECTV infection. Differential Expression of Genes (DEGs) analyses revealed distinct differences in the gene profiles of resistant and susceptible mice infected with ECTV. Gene ontology and KEGG pathway analysis showed that the DEGs of susceptible mice were involved mainly in immunity, apoptosis, spliceosomes and cancer-related pathways, while the DEGs of resistant mice were largely involved in MAPK signaling and leukocyte transendothelial migration. This suggests that the susceptible BALB/c mice have a stronger response than the resistant C57BL/6 mice to ECTV infection. The BALB/c mice showed a strong induction of interferon-induced genes (ISGs), including guanylate binding proteins (GBPs), myxovirus resistance protein (Mx) GTPases, oligoadenylate synthetase (OAS) and IFN-induced protein with tetratricopeptide repeats (IFIT) family proteins, while the C57BL/6 mice upregulated more genes related to metabolic pathways. This suggests that the susceptible BALB/c mice have a stronger response than the resistant C57BL/6 mice to ECTV infection.
Project description:Ectromelia virus (ECTV) has emerged as a valuable model for investigating the host-orthopoxvirus relationship as it relates to pathogenesis and the immune response. We analyzed the transcriptional signatures of BALB/3T3 cells at different times after infection with Ectromelia virus. Mouse Genome 430 2.0 arrays were used to analyze global changes in gene transcripts to generate a pool of genes that was a fold change cut-off of ≥ 1.5 or ≤ 0.5 in infected samples versus non-infected samples. Gene ontology and KEGG pathway analysis showed that the DEGs were involved mainly in immunity, apoptosis, spliceosomes, MAPK signaling and cancer-related pathways.
Project description:Inflammatory monocytes (iMO) and B-cells are the main targets of the poxvirus ectromelia virus (ECTV) in the lymph nodes of mice and play distinct roles in surviving the infection. Infected and bystander iMO control ECTV’s systemic spread, preventing early death, while B-cells make antibodies that eliminate ECTV. Our work demonstrates that within an infected animal that survives ECTV infection, intrinsic and bystander infection of iMO and B-cells differentially control the transcription of genes important for immune cell function and, perhaps, cell identity. Bystander cells upregulate metabolism, antigen presentation, and interferon-stimulated genes. Infected cells downregulate many cell-type-specific genes and upregulate transcripts typical of non-immune cells. Bys and Inf iMO non-redundantly contribute to the cytokine milieu and the interferon response. Furthermore, we uncovered how IFN-I or IFN-γ signaling differentially regulates immune pathways in Inf and Bys iMO and, that at steady state, IFN-I primes iMO for rapid IFN-I production and antigen presentation.
Project description:Orthopoxviruses, such as variola and Mpox, encode a myriad of immunomodulatory proteins to promote pathogenesis. One notable family is the B22 family of proteins, which are highly conserved surface glycoproteins that potently inhibit T cell activation in vitro and ex vivo. However, there has been limited work investigating the impact of B22 proteins on both CD4+ and CD8+ T cell responses in vivo. Therefore, we used ectromelia (ECTV) as a natural host-pathogen model to investigate the impact of its B22 protein, C15, on T cell responses in vivo. To broadly investigate the impact of C15 on CD8+ effector (CD62L-CD44+) T cell development, we performed bulk RNA-seq on splenic CD8+ effector T cells 7 days post-infection with either WT ECTV or ECTV∆C15.
Project description:Orthopoxviruses, such as variola and Mpox, encode a myriad of immunomodulatory proteins to promote pathogenesis. One notable family is the B22 family of proteins, which are highly conserved surface glycoproteins that potently inhibit T cell activation in vitro and ex vivo. However, there has been limited work investigating the impact of B22 proteins on both CD4+ and CD8+ T cell responses in vivo. Therefore, we used ectromelia (ECTV) as a natural host-pathogen model to investigate the impact of its B22 protein, C15, on T cell responses in vivo. To broadly investigate the impact of C15 on CD4+ effector (CD62L-CD44+) T cell development, we performed bulk RNA-seq on splenic CD4+ effector T cells 7 days post-infection with either WT ECTV or ECTV∆C15.
Project description:Bacterial persister cells are phenotypic variants of regular cells that are tolerant to antibiotics. Analysis of clinical isolates of M. tuberculosis showed that strains vary substantially in their tolerance to antibiotics. The level of persisters was very high is some isolates, suggesting that these are hip mutants. We investigated gene expression differences in eight clinical isolates, four of which we characterized as high-persister strains and four as low-persister, or regular, strains. Comparison of gene expression patterns may provide clues as to the genetic mechanisms underlying persister formation.
Project description:Orthopoxviruses, such as variola and Mpox, encode a myriad of immunomodulatory proteins to promote pathogenesis. One notable family is the B22 family of proteins, which are highly conserved surface glycoproteins that potently inhibit T cell activation in vitro and ex vivo. However, there has been limited work investigating the impact of B22 proteins on both CD4+ and CD8+ T cell responses in vivo. Therefore, we used ectromelia (ECTV) as a natural host-pathogen model to investigate the impact of its B22 protein, C15, on T cell responses in vivo. To broadly investigate the impact of C15 on CD8+ effector (CD62L-CD44+) T cell development during the contraction phase, we performed bulk RNA-seq on splenic CD8+ effector T cells 10 days post-infection with either WT ECTV or ECTV∆C15.