A bursal pentapeptide (BPP-I), a novel bursal-derived peptide, exhibits antiproliferation of tumor cell and immunomodulator activity.

The bursa of Fabricius (BF) is the central humoral immune organ unique to birds. Here, we isolated a novel bursal pentapeptide I (BPP-I), LGPGP, from BF. BPP-I could play inhibition effect on MCF-7 but not on CEF or Vero cell proliferation in vitro, and enhance antitumor factor p53 protein expression. Also, BPP-I stimulated antibody production in a dose-dependent manner in hybridoma cell. Furthermore, BPP-I could induce various immune responses in mice immunization experiments, including increase antibody production and cytokines IL-4 and IFN-γ level, and induce T-cell immunophenotyping. These results suggest that BPP-I is a potential immunomodulator of antitumor and immunity. The study could provide some novel insights on the probable candidate reagent for the antitumor and immune improvement.

Gene expression profiling of hybridoma cells after bursal-derived bioactive factor BP5 treatment.

Bursa of Fabricius is the acknowledged vital humoral immune system for B cell differentiation and antibody production. To study the molecular mechanism underlying the effect of bursal-derived BP5, we used gene microarray to analyze the genomic expression profiling of BP5-treated hybridoma cells. BP5 exhibited an immunomodulatory effect on antibody production in hybridoma cells and induced alterations in the gene expression profiles related to the immune-related biological processes, such as T cell activation and proliferation, B cell activation, B cell-mediated immunity, and cytokines cytokine production involved in immune response. In addition, 26 biological pathways associated with immunomodulatory functions were regulated in BP5-treated hybridoma cells, in which p53 signal pathway played an important role in antitumor. Among these regulated genes, 12 differentially expressed genes were verified by qRT-PCR. The activation of p53 activity by BP5 was further confirmed by p53 luciferase reporter assay and p53 expression. Our data revealed that bursal-derived BP5 could regulate various immune-related cellular processes, including antitumor factor p53 signal pathway, perhaps partially accounting for the reported immunomodulatory roles and novel antiproliferation on tumor cells functions of bursal-derived bioactive factor BP5.

The Roles of Bursal Nonapeptide (BP9) on AIV Vaccine Immune Response in Chick Immunization and on Avian Immature B Cell.

BACKGROUND: Bursa of Fabricius plays the vital functions on B cell development and antibody production in poultry. The bursal-derived peptide plays the essential roles on avian immature B cell development. OBJECTIVES: Here we explored the functions of the recently reported bursal nonapeptide (BP9) on the antibody production and the molecular basis of BP9 on avian immature B cell. METHODS: Chicken were twice immunized with Avian Influenza Virus (AIV) inactivated vaccine plus with BP9 at three dosages, respectively. On two weeks after the second immunization, sera samples were collected from all experimental groups to measure AIV-specific Agglutination Inhibition (HI) antibody titers. Also, on 7th day after the second immunization, spleen lymphocytes were isolated from the immunized chicken to detect the lymphocyte viabilities. DT40 cells were treated with BP9 from 0.02 to 2 µg/mL for 4 and 20h to detect sIgM mRNA levels, and total RNAs from BP9-treated DT40 cells were collected to investigate the gene expression profiles of DT40 cells, and to analyze the enriched pathways and functional biological processes. Finally, nine gene expressions were validated with quantitative PCR (qPCR). RESULTS: Our investigation proved the strong regulatory roles of BP9 on AIV-specific HI antibody titers and lymphocyte viabilities. BP9 promoted sIgM mRNA levels in DT40 cells, and upregulated 598 gene expressions and downregulated 395 gene expressions in DT40 cells with 0.2µg/mL BP9 treatment. Moreover, our findings verified the significantly enriched six pathways and various the biological functional processes of BP9 on avian immature B cell. Also, we found eight signaling pathways in the enriched biological processes of BP9-treated DT40 cells, and the expressions of nine selected genes with qPCR were identical to that of microarray data. CONCLUSION: BP9 promoted the antibody production in the 21-old-day chicken immunization, and stimulated the sIgM expression in DT40 cells. Furthermore, we analyzed the gene expression profile and immune-related biological processes of DT40 cells treated with BP9, which provided some new insights into the mechanism on immature B cell development, and provided important references for adjuvant development on vaccine improvement and clinical application.

The Immunological Functions of Muramyl Dipeptide Compound Adjuvant on Humoral, Cellular-mediated and Mucosal Immune Responses to PEDV Inactivated Vaccine in Mice.

BACKGROUND: The muramyl dipeptide compound adjuvant, CVC1303, was one new resigned adjuvant to PEDV inactivated vaccine. Exploring the effects of CVC1303 on the immune induction to PEDV vaccine was of vital importance to the clinical application. OBJECTIVES: Here we explored the functions of CVC1303 on the humoral, cellular and mucosal immune response to PEDV vaccine in mice immunization. METHODS: Mice were twice subcutaneously injected with PEDV vaccine including high, medium and low dosages CVC1303, respectively. On 30th day after the second immunization, sera samples were collected from the immunized mice to measure PEDV-specific IgG and IgG subclasses levels, and lymphocytes were isolated to detect T cell subtype and intracellular IL-4 and IL-6 cytokine productions, and the expressions of co-stimulatory molecule on dendritic cells in the immunized mice. Small intestinal and lung washings were collected on 30th and 47th day after the second immunization to measure PEDV-specific IgA levels, and SP immunohistochemical method staining was employed to analyze the deviations of IgA+ positive cells in the small intestinal of the immunized mice. RESULTS: Our investigation proved the strong regulatory roles of CVC1303 on PEDV-specific IgG and IgG1 antibody and cytokines productions, and the significant increased CD3+CD4+T cells subpopulation and expressions of co-stimulatory molecules on dendritic cells in the immunized mice. Moreover, our findings verified the significantly enhanced PEDV-specific IgA antibody titers in small intestinal and lung in the mice immunized with PEDV vaccine and CVC1303. CONCLUSION: Compound adjuvant CVC1303 could effectively improve the PEDV-specific immune responses and mucosal immune, which provided an experimental basis for the further clinical application of new adjuvant CVC1303 and the development of improvement on the mucosal immune response.