Project description:Oral mucosa has a potential to maintain immunological homeostasis since little severe inflammation happens in the continuous antigen exposure in the oral cavity. SLIT applied for the allergen-specific immune suppression is performed by repeated antigen (Rpt Ag)-painting to sublingual mucosa (SLM). Among a series of immune cells at submucosa, heterogeneous DC/MF subsets contribute to the orchestration of mucosal immune responses. Previously, We found that Rpt Ag-painting to SLM exhausted typical CD11c+ DCs and induced round-type Macrophage-like CD206hi cells. In this study, we defined three major DC/Macrophage fraction in SLM after Rpt Ag-painting by the expressions of several surface molecules. Fraction 1 (Fr-1): CD206- CD11c+ F4/80lo; Fraction 2 (Fr-2): CD206- CD11c- F4/80+ and Fraction 3 (Fr-3): CD206hi CD11clo F4/80+. Microarray analyses revealed that total of 7282 up-regulated and 7389 down-regulated genes in Fr-3 were enriched compared with Fr-2, respectively. Genes in Fr-3 preferentially expressed molecules related to homeostatic process and negative regulation of T cell activation. Furthermore, new B7 family of immune checkpoint molecules express significantly higher on Fr-3. When we checked the expressions of genes related to M2 MF, Fr-3 cells preferentially express fizz1, aldh1a1 and aldh1a2 but not Ym-1 and arginase-1 compared to in vivo induced M2-like cells. These results indicate that Fr-3 has distinct features compared with M2 like cells and may has T cell regulation potential. Functional investigation imply that CD206hi cell dominant status induces suppression of Ag-specific T cell responses. In conclusion, these results suggest that CD206hi cells from SLM involved in immune tolerogenic inducement by expressing new B7 family molecules and modulate T cell immune responses.
Project description:Hemagglutinin of the influenza virus is the main external glycoprotein. This very immunogenic protein is the target of the most anti-influenza vaccines. DNA vaccines are new alternative to conventional inactivated ones. Four DNA vaccines were tested. Each tested variant was based on the pCI vector with nucleotide sequence encoding hemagglutinin from A/swan/Poland/305-135V08/2006 (H5N1, clade 2.2). In K3/pCI, GK/pCI and HAneo/pCI the different optimization algorithms of hemagglutinin encoding sequence without amino acids change were tested. In 3NF/pCI the NFkappaB binding sites flanking the expression cassette were included in order to improve the nuclear transfer. Comparative transcriptome analysis of mice vaccinated the following vaccine HAneo/pCI,K3/pCI, GK/pCI or 3NF/pCI versus empty vector demonstrated minor changes in genes expression pattern. Most genes were expressed on the similar level in the vaccinated individuals and in the control mice. Small number of genes in particular variants showed the expression different than in the control mice. In general, the identified genes with the changed expression included some genes involved in metabolic processes and none of them seem to induce any undesirable pathways nor disease.