into recipient mice for 2 consecutive days from the first day after tumor inoculation, as described [49]

into recipient mice for 2 consecutive days from the first day after tumor inoculation, as described [49]. experiments. Table S1. The inhibition ratio (%) was calculated by the following formula: inhibition ratio (%)= [(A-B)/A]100, where A is the average tumor weight of the control group, and B is the tumor weight of the treated group. Table S2. The mean tumor volume of all groups for each of the in vivo experiments. 1476-4598-13-179-S2.doc (41K) GUID:?20D2CCDE-E86A-429A-B791-E300E3F44EA9 Additional file 3: Figure S2 Body weight and histology of major organs. A, Body weight. Body weight was measured every 3 days during the entire treatment. B, Sections Bmpr2 of heart, liver, spleen and kidney were stained with H&E. There was no obvious histological difference between the groups (magnification, 200). 1476-4598-13-179-S3.tiff (16M) GUID:?80BC99D7-4CEA-40B0-8190-263CFB87737C Abstract Background A safe and effective adjuvant plays an important role in the development of a vaccine. However, adjuvants licensed for administration in humans remain limited. Here, for the first time, we developed a novel combination adjuvant alum-polysaccharide-HH2 (APH) with potent immunomodulating activities, consisting of alum, polysaccharide of and the synthetic cationic innate defense regulator peptide HH2. Methods The adjuvant effects of APH were examined using NY-ESO-1 protein-based vaccines in prophylactic and therapeutic models. We further decided the immunogenicity and anti-tumor effect of NY-ESO-1-APH (NAPH) vaccine using adoptive cellular/serum therapy in C57/B6 and nude mice. Cell-mediated and GW7604 antibody-mediated immune responses were evaluated. Results The APH complex significantly promoted antigen uptake, maturation and cross-presentation of dendritic cells and enhanced the secretion of TNF-, MCP-1 and IFN- by human peripheral blood mononuclear cells compared with individual components. Vaccination of NAPH resulted in significant tumor regression or delayed tumor progression in prophylactic and therapeutic models. In addition, passive serum/cellular therapy potently inhibited tumor growth of NY-ESO-1-B16. Mice treated with NAPH vaccine produced higher antibody titers and greater antibody-dependent/independent cellular cytotoxicity. Therefore, NAPH vaccination effectively stimulated innate immunity, and boosted both arms of the adaptive humoral and cellular immune responses to suppress tumorigenesis and growth of melanoma. Conclusions Our study revealed the potential application of APH complex as a novel immunomodulatory agent for vaccines against tumor refractory and growth. showed an increasing trend with an increasing concentration of HH2. Among the PS-HH2 complexes, 1:4 and 1:2 (wt/wt) ratios of PS: HH2 resulted in a most potent induction of TNF- expression (699??48 pg/ml and 651??104pg/ml, respectively) in vitro and independently showed a synergistic effect of 2.1??0.1 and 2??0.2, respectively (Additional file 1: Physique S1B). Similarly, 1:4 and 1:2 (wt/wt) ratios of PS: HH2 resulted in a most potent induction of MCP-1 expression (16064??2051 pg/ml and 13725??1480 pg/ml, respectively). There was no significant difference between the two groups (Additional file 1: Physique S1C). Since safety is as important as potency for an adjuvant, we further investigated the potential cytotoxic effect of the PS-HH2 complexes by monitoring the GW7604 release of hemoglobin from red blood cells and lactate dehydrogenase (LDH) from peripheral blood mononuclear cells (PBMCs). Our data showed that the tested PS-HH2 complexes resulted in minimal or no release of hemoglobin or LDH from red blood cells (RBCs) (Additional file 1: Physique S1D) and PBMCs (Additional file 1: Physique S1E), respectively. Thus, 1:2 (wt/wt) PS: HH2 formulation was chosen to further examine GW7604 its adjuvant effect for NY-ESO-1 below. NAPH vaccine inhibits tumor growth in a melanoma model In the prophylactic model (Physique?1A and Additional file 2: Tables S1 and S2), the NY-ESO-1-B16 melanoma grew rapidly in control mice, with mean tumor volume of approximately 1675 mm3 by Day 22. In contrast, the tumor growth was significantly inhibited in the NY-ESO-1-alum-PS (NAP) group ( 0.05) (Figure?3B). Sera obtained from the NAPH group showed substantial higher titer of IgG1 than that from other groups.