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Item Activation of Dendritic Cells by Soypeptide Lunasin: Implication in Vaccine Adjuvant(Office of the Vice Chancellor for Research, 2014-04-11) Flores, Sarah; Dong, Melissa; Tung, Chun-Yu; Chang, Hua-ChenAdjuvants enhance the immunogenicity of vaccines and improve the immune responses. Although many adjuvants are currently used in research, FDA approved aluminum salt (Alum) remains the most often used in human vaccines. Alum is known to promote the Th2 immune response and enhance antibody production, but is less efficient on eliciting Th1 and CTL cellular responses. Thus, it is prudent to improve the effectiveness of current adjuvants or to develop a novel alternative adjuvant. We have recently identified lunasin, a seed peptide from soybeans, as a novel immune modulator. The objective is to define the effectiveness of lunasin peptide as an adjuvant that can enhance the protective immunity of vaccines. Our studies have revealed stimulatory effects of lunasin on dendritic cells (DCs) by regulating expression of a number of genes that are important for immune responses. Lunasin-treated human conventional DCs (cDCs) not only expressed elevated levels of co-stimulatory molecules (CD86) but also exhibited up-regulation of chemokines (CCL2, CCL3, CCL4) and cytokine (IL-1β). To determine the function of lunasin-treated cDCs, these cells were co-cultured with allogeneic human peripheral blood CD4+ T cells for 7 days in the mixed lymphocyte reaction. Lunasin-treated cDCs induced almost 2-fold higher proliferation of allogeneic CD4+ T cells when comparing with a sham treatment. To verify the in vivo effects, lunasin was administered into mice. Increased CD86 expression was found in cDCs from spleens of mice treated with lunasin. Furthermore, mice vaccinated with lunasin-adjuvanted ovalbumin (OVA) had reduced tumor growth following challenging with OVA-expressing A20 B-lymphoma cells. Taken together, our data suggest that lunasin may act as a vaccine adjuvant by targeting DCs to enhance and modulate the immune responses to antigens.Item Activation of Natural Killer Cell by Lunasin and Cytokine(Office of the Vice Chancellor for Research, 2014-04-11) Kyazike, Sharifah; Lewis, David; Tung, Chun-Yu; Han, Ling; Chang, Hua-ChenCancer immunotherapy is one of the emerging therapeutic strategies to harness the immune system to eradicate chemotherapy-resistant cancerous cells. NK cells can recognize and eliminate cancer cells before adaptive immunity is developed. Human NK cells can be divided into 2 major subsets based on their surface expression of CD56. NK cells with CD56 bright populations are major cytokine producers, while NK cells expressing CD56 dim have higher lytic activity. Due to the role of NK cells in cancer surveillance, any approach to enhance their activity may augment cancer treatment. We have recently shown that soypeptide Lunasin is a novel immune modulating agent that, together with cytokines, enhances IFN- γ and Granzyme B expression by NK cells. This synergism augments the natural cytotoxicity of NK cells against various tumors in vitro as well as in the xenograft model. The objective of this study is to evaluate the effects of Lunasin on antibody-dependent cellular cytotoxicity (ADCC) activity of NK cells against Rituximab-coated human B-lymphoma Raji cells. We also evaluated the expression of several markers involved in NK-mediated tumorcidal activity using flow cytometry. Together, these results suggest that Lunasin could enhance the efficacy of NK cell-based immunotherapy for cancer.Item Mechanisms of Gene Regulation by Soy Peptide Lunasin in Innate Immune Cells(Office of the Vice Chancellor for Research, 2015-04-17) Casiano-Rivera, Félix M.; Tung, Chun-Yu; Chang, Hua-ChenLunasin is a seed peptide containing 43 amino acids, originally isolated from soybeans. Recently, a novel function of lunasin was discovered, as it acts as an immune modulating agent regulating gene expression of various innate immune cells. Lunasin strongly activated the expression of genes encoding for type I interferon and inflammatory cytokines. Nonetheless, the molecular mechanisms of lunasin’s gene regulation are relatively unknown. Our current hypothesis states that lunasin is able to induce activation of various transcription factors, resulting in gene expression in immune cells. Human dendritic cells (DCs) or monocytes were purified from peripheral blood mononuclear cells (PBMCs) in order to determine the activation of transcription factors. Phosphorylation of STAT1 and NF-ĸB (p65) were evident in cells treated with lunasin using flow cytometry and Western blotting. The results will ultimately lead to the signaling pathways involved in gene expression regulated by lunasin in innate immune cells. By defining the signaling pathway of lunasin, we can have a better understanding of its application in immune modulation.