Project description:We previously reported that a synthetic Nod1 ligand, FK565, induced coronary arteritis in mice similar to Kawasaki disease. However, the molecular mechanisms underlying this site-specific inflammation have remained elusive. In this study, we found that CD11c+MHC class II+ cells accumulated in the heart of FK565-treated mice prior to arteritis development. We used microarray analysis to detail gene expression of CD11c+MHC class II+ cells. To obtain gene expression profile of CD11c+MHC class II+ cells, we isolated these cells from hearts of FK565-treated mice. Briefly, female mice at 8weeks age were administered 500 μg of FK565 subcutaneously at day 0 and day 3. At day6, murine hearts were removed and digested with collagenase. CD11c+MHC II+ cells were sorted as PI–CD45+Ly6G–NK1.1–CD11b+CD11c+MHC II+ using FACS Aria cell sorter (BD Biosciences). Sorted cells were subjected to RNA preparation. Two independent replicates from ten mice were made.
Project description:The immune response against tuberculosis relies, at least in part, on CD4+ T cells. Protective vaccines require the induction of antigen-specific CD4+ T cells via mycobacterial peptides presented by MHC class-II in infected macrophages. We have purified MHC class-I and MHC-II peptides and analysed them by mass spectrometry. We have successfully identified 97 mycobacterial peptides presented by MHC-II and 54 presented by MHC-I, from 76 and 41 antigens, respectively. The sequences of selected peptides were confirmed by spectral match validation and immunogenicity evaluated by IFN-gamma ELISpot against peripheral blood mononuclear cells from volunteers vaccinated with BCG, M.tb latently infected subjects or patients with tuberculosis disease. Three antigens were expressed in viral vectors, and evaluated as vaccine candidates alone or in combination in a murine aerosol M.tb challenge model. When delivered in combination, the three candidate vaccines conferred significant protection in the lungs and spleen compared with BCG alone, demonstrating proof-of-concept for this unbiased approach to identifying novel candidate antigens.
Project description:Over expression of MHC Class l protein in skeletal muscle causes myositis. Phenotype after expression in young mice is more severe. We performed gene expression profiling on young and adult mice after over expression of self MHC class l protein in skeletal muscle Muscle from young ( early) , adult (Late) and cntrol (control) mice , n=3 each group, was used for gene expression profiling
Project description:Over expression of MHC Class l protein in skeletal muscle causes myositis. Phenotype after expression in young mice is more severe. We performed gene expression profiling on young and adult mice after over expression of self MHC class l protein in skeletal muscle Overall design: Muscle from young ( early) , adult (Late) and cntrol (control) mice , n=3 each group, was used for gene expression profiling
Project description:To further understand the early molecular events in antibody mediated chronic inflammation of the lungs, we utilized a murine Obliterative Airway Disease (OAD) model of native lungs where Abs to MHC class I induces bronchiolar obliteration and fibrosis. This study identified a unique transcriptional profile with a set of genes that were stimulated while another set of genes that were repressed in anti MHC I treated lungs compared to that of isotype treated lungs. Lung transplantation (LTx) is an accepted clinical intervention for various intractable pulmonary dysfunctions. However, de novo Abs to mismatched donor HLA (DSA) predispose lung-grafts to chronic rejection, Bronchiolitis Obliterans Syndrome (BOS). The anti-MHC induced mouse OAD model, the only available preclinical model for study of early pathogenesis following MHC ligation, produces lesions analogous to BOS. Overall design: Induction of gene expression was measured following intrabronchial administration of anti-MHC class I (H-2Kb) Ab at 200μg/mouse. Samples were collected on days 1, 2, 3 and 6 post-Ab administration. Isotype matched control Ab was administered at an equivalent dose and gene expression profile in isotype treated group served as the base line expression and was compared with that of anti-H-2Kb treated group. A total of 24 samples were processed with two treatment groups (anti-H-2Kb, isotype), four time points (days 1, 2, 3 and 6) and three biological replicates.