• Journal of Veterinary Science
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• v.24 no.3
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• pp.37.1-37.14
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• 2023

Background: Toll-like receptor (TLR) agonists have been used as adjuvants to modulate immune responses in both animals and humans. Objectives: The objective of this study was to evaluate the combined effects of the TLR 4 agonist monophosphoryl lipid A (MPL) and the TLR 3 agonist polyinosinic:polycytidylic acid (Poly I:C) on equine peripheral blood mononuclear cells (PBMCs), monocyte-derived dendritic cells (MoDCs), and bone marrow-derived mesenchymal stromal cells (BM-MSCs). Methods: The PBMCs, MoDCs, and BM-MSCs collected from three mixed breed horses were treated with MPL, Poly I:C, and their combination. The mRNA expression of interferon gamma (IFN-γ), interleukin (IL)-1β, IL-4, IL-6, IL-8, IL-12p40, tumor necrosis factor alpha (TNF-α), vascular endothelial growth factor (VEGF), and monocyte chemoattractant protein-1 (MCP-1) was determined using real-time polymerase chain reaction. Results: The combination of MPL and Poly I:C significantly upregulated immunomodulatory responses in equine cells/ without cytotoxicity. The combination induced greater mRNA expression of pro-inflammatory cytokines IFN-γ and IL-6 than MPL or Poly I:C stimulation alone in PBMCs. In addition, the combination induced significantly higher mRNA expression of IL-1β, IL-6, and IL-12p40 in MoDCs, and IL-8, MCP-1, and VEGF in BM-MSCs compared to stimulation with a single TLR agonist. Conclusions: The combination of MPL and Poly I:C can be used as a potential adjuvant candidate for vaccines to aid in preventing infectious diseases in horses.

• Korean Journal of Plant Resources
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• v.36 no.4
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• pp.257-263
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• 2023

Autophagy contributes to enhancing the immune system (innate and adaptive immune system) against foreign pathogens. Autophagy of macrophages is used as a major indicator for developing vaccine adjuvants to increase the adaptive immune response. In this study, water extracts from Rosa acicularis leaves (RAL) increased the production of immunostimulatory mediators and phagocytic activity in RAW264.7 cells. RAL increased p62/SQSTM1 expression. Inhibition of TLR4, JNK, and PI3K/AKT blocked RAL-mediated increase of p62/SQSTM1. RAL activated JNK and PI3K/AKT signaling. RAL-mediated activations of JNK and PI3K/AKT signaling were reversed by TLR4 inhibition. Taken together, it is believed that RAL-mediated autophagy may be dependent on activating via TLR4-dependent activation of JNK and PI3K/AKT signaling in macrophages.

• IMMUNE NETWORK
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• v.20 no.4
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• pp.33.1-33.19
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• 2020

We tested how adjuvants effect in a cancer vaccine model using an epitope derived from an autoactivation loop of membrane-type protease serine protease 14 (PRSS14; loop metavaccine) in mouse mammary tumor virus (MMTV)-polyoma middle tumor-antigen (PyMT) system and in 2 other orthotopic mouse systems. Earlier, we reported that loop metavaccine effectively prevented progression and metastasis regardless of adjuvant types and TH types of hosts in tail-vein injection systems. However, the loop metavaccine with Freund's complete adjuvant (CFA) reduced cancer progression and metastasis while that with alum, to our surprise, were adversely affected in 3 tumor bearing mouse models. The amounts of loop peptide specific antibodies inversely correlated with tumor burden and metastasis, meanwhile both T_H1 and T_H2 isotypes were present regardless of host type and adjuvant. Tumor infiltrating myeloid cells such as eosinophil, monocyte, and neutrophil were asymmetrically distributed among 2 adjuvant groups with loop metavaccine. Systemic expression profiling using the lymph nodes of the differentially immunized MMTV-PyMT mouse revealed that adjuvant types, as well as loop metavaccine can change the immune signatures. Specifically, loop metavaccine itself induces T_H2 and T_H17 responses but reduces T_H1 and T_reg responses regardless of adjuvant type, whereas CFA but not alum increased follicular T_H response. Among the myeloid signatures, eosinophil was most distinct between CFA and alum. Survival analysis of breast cancer patients showed that eosinophil chemokines can be useful prognostic factors in PRSS14 positive patients. Based on these observations, we concluded that multiple immune parameters are to be considered when applying a vaccine strategy to cancer patients.

• Korean Journal of Poultry Science
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• v.36 no.3
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• pp.201-206
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• 2009

This study was carried out to investigate dietary effects of illite on humoral immune response against Salmonella typhimurium flagella in Hyline Brown laying hens. Total of twenty eight 36-week-old hens were divided into two groups; The first was fed commercial diet as control and the other was fed diet containing 2 % illite. Each group was divided into 2 sub-groups and then performed immunization of Salmonella flagella with different adjuvants which were Freund's adjuvant and croton oil. The rates of egg-production in all groups were normal range but no difference was found between illite-treated and untreated groups. The serum antibody titers of Freund's adjuvant-used subgroup in illite fed groups were significantly increased, especially from 6-to 9-week after $1^{st}$ immunization compared to those of control groups (p<0.05). And the antibody titers of croton oil-used subgroup in illite-fed groups were also significantly increased, especially at 4-, 6- and 7-week (p<0.05). The results demonstrated that the feeding illite stimulated the immune response against S. typhimurium flagella antigen in laying hens and suggest that the supplementation of illite to the poultry diets may support protective effects against bacterial infections such as Salmonellosis.

• Journal of fish pathology
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• v.5 no.1
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• pp.19-27
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• 1992

A formalized bacterin(FKC) of Edwardsiella tarda, conjugated with/without Freund's complete adjuvant(FCA), Freund's incomplete adjuvant(FIA) and potassium aluminium sulfate(PAS), was administered by intraperitoneal injection to tilapia respectively. The tilapia were reared under indoor recirculating filter system. There were agglutinin titer increment in the tilapia which were administered not only FKC bacterin but also adjuvanted vaccines. But the producing time of the highest level of antibody in the immunized tilapia was different among vaccine preparation, i. e. FKC+FCA group and PAS+FKC group are between the 2nd and the 4th week, and FKC alone group is the 2nd week. In the challenge experiment with $2.5{\times}10^7\;CFU/ml$ or $2.5{\times}10^8\;CFU/ml$ of E. tarda T1123 to the tilapia immunized with vaccines, RPS above sixty was recorded both challenge dose in the tilapia which were 3 weeks after immunization with FKC+FCA, PAS+FKC and FKC alone, and the former dose 8 weeks after immunization with FCA+FKC and PAS+FKC. There were some resistance enhancement against E. tarda in the tilapia which were injected with adjuvant alone than that of control. As the results, the FCA or PAS adjuvanted vaccine is effective to sustain the defensible period against edwardsiellosis.

• Food Science of Animal Resources
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• v.27 no.2
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• pp.222-227
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• 2007

In order to produce the antibody rich eggs against Salmonella gallinarum(S.G.) causing fowl typhoid, the productions of immunoglobulin in eggs were compared and examined with the feed additives, the variety of adjuvants in vaccines to layers, and the existence of additive antigens other than target microorganism. The examination of the average contents of specific IgY in immunized group by supplying hardwood charcoal showed that the group supplied with 0.5% hardwood charcoal had the highest contents, implying that the supply of hardwood charcoal promoted the production of specific IgY. Adjuvant appeared to have little effect on the average contents of total IgY, but specific IgY contents increased in the immunized group with Freund's adjuvant. Addition of BCG in adjuvant treatment increased specific IgY however, this feature was not seen in aluminum hydroxide treated group. Immunization at 15 week layers resulted in higher laying rate than immunization at 21 week and addition of hardwood charcoal in feed recovered laying rate. It was therefore, concluded that the feed supplement, such as hardwood charcoal followed by a proper immunization program concerning adjuvant, vaccination period and supplementary microorganism hastened the production of IgY.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.10
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• pp.1478-1485
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• 2017

Objective: The effects of vaccinating 18-day-old chicken embryos with the combination of recombinant Eimeria profilin plus Clostridium perfringens (C. perfringens) NetB proteins mixed in the Montanide IMS adjuvant on the chicken immune response to necrotic enteritis (NE) were investigated using an Eimeria maxima (E. maxima)/C. perfringens co-infection NE disease model that we previously developed. Methods: Eighteen-day-old broiler embryos were injected with $100{\mu}L$ of phosphate-buffered saline, profilin, profilin plus necrotic enteritis B-like (NetB), profilin plus NetB/Montanide adjuvant (IMS 106), and profilin plus Net-B/Montanide adjuvant (IMS 101). After post-hatch birds were challenged with our NE experimental disease model, body weights, intestinal lesions, serum antibody levels to NetB, and proinflammatory cytokine and chemokine mRNA levels in intestinal intraepithelial lymphocytes were measured. Results: Chickens in ovo vaccinated with recombinant profilin plus NetB proteins/IMS106 and recombinant profilin plus NetB proteins/IMS101 showed significantly increased body weight gains and reduced gut damages compared with the profilin-only group, respectively. Greater antibody response to NetB toxin were observed in the profilin plus NetB/IMS 106, and profilin plus NetB/IMS 101 groups compared with the other three vaccine/adjuvant groups. Finally, diminished levels of transcripts encoding for proinflammatory cytokines such as lipopolysaccharide-induced tumor necrosis $factor-{\alpha}$ factor, tumor necrosis factor superfamily 15, and interleukin-8 were observed in the intestinal lymphocytes of chickens in ovo injected with profilin plus NetB toxin in combination with IMS 106, and profilin plus NetB toxin in combination with IMS 101 compared with profilin protein alone bird. Conclusion: These results suggest that the Montanide IMS adjuvants potentiate host immunity to experimentally-induced avian NE when administered in ovo in conjunction with the profilin and NetB proteins, and may reduce disease pathology by attenuating the expression of proinflammatory cytokines and chemokines implicated in disease pathogenesis.

• The Korean Journal of Pesticide Science
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• v.10 no.3
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• pp.210-217
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• 2006

The study was carried out to select the effective adjuvant as a photostabilizer for acetamiprid 5% SL(soluble concentrate) and to find its optimum content by using chemical and biological methods. Adjuvants used for the study were KS1 and KS2. Photodegradation test, insecticidal efficacies and residual analysis for the study were investigated. Although photodegradation rate of acetamiprid SL was decreased by addition of KS1 and KS2, the difference between KS1 and KS2 was not significant. As KS2 showed better efficacies against the cotton aphid (Aphis gossypii) and higher amount of residue on pepper than KS1, KS2 was selected as a photostabilizer for acetamiprid SL. Acetamipid SL with KS2 showed lower photodegradation rate, better efficacies against the cotton aphid (Aphis gossypii) and higher amount of residue on pepper as content of KS2 in acetamiprid SL was decreased. Therefore, it was concluded that the optimum content of KS2 was 0.1%. These results have demonstrated that the selected adjuvant could be used to enhance insecticidal activity and reduce spay dose of insecticide by protecting its photodegradation.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.53 no.2
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• pp.224-232
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• 2008

The health benefits associated with tea consumption have resulted in the wide inclusion of green tea extracts in botanical dietary supplements, which are widely consumed as adjuvants for complementary and alternative medicines. Tea contains polyphenols such as catechins or flavan-3-ols including (-)-epicatechin (EC), (-)-epigallocatechin (EGC), (-)-epicatechin gallate (ECG), and (-)-epigallocatechin gallate (EGCG), as well as the alkaloid, caffeine. The contents of catechins and caffeine in green tea are considered as a standard of quality evaluation of green tea. Therefor, the purpose of this study was to investigate the most suitable HPLC condition for simultaneous determination of catechins and caffeine in green tea extracts. The efficient HPLC analytical condition of catechins and caffeine contained green tea extracts was developed. The gradient elution employed a $250\;mm\;{\times}\;4.6\;mm$ i.d. YMC-pak ODS-AM 303 column. The gradient system was used two mobile phases. A gradient elution was performed with mobile phase A, consisting of 0.1% aqueous phosphoric acid, and mobile phase B, comprising 100% MeOH, and delivered at a flow rate of 1 mL/min as follows: $0{\sim}25\;min$, 80% A; $26{\sim}50\;min$, $80{\sim}70%$ A; 51 min, 80% A. $51{\sim}55\;min$, 80% A. The UV detection wavelength was set at 280 nm. The limit of detection (LOD) for catechins and caffeine standards were under 50 ng/mL.

• Korean Journal of Veterinary Research
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• v.46 no.3
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• pp.215-224
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• 2006

In order to search the availability of the lectin extracted from Korean mistletoe(Viscum album coloratum) as an adjuvant for the avian vaccines, attempts were made to determine toxicity of the lectin in chicks and its immunostimulating activity on the inactivated vaccines against Newcastle disease virus(NDV). For the determination of toxicity, the lectin was injected into the thigh muscle of SPF chicks(Charles River) of 1-week-old and observed hematologically and pathologically. For the determination of immunostimulating effects, lectin-adjuvanted, inactivated NDV vaccines were injected into the thigh muscle of SPF chicks in the same age group. Sera of the chicks were examined for the hemagglutination-inhibition(HI) antibodies induced, their HI titers and reaction to the NDV antigens. The data were further compared with those from aluminum hydroxide [$Al(OH)_3$]-adjuvanted vaccines and vaccines without adjuvant, and the results are as follows. There were no significant changes observed in the values of RBC, WBC, Hb, PCV, MCV, MCH, MCHC, AST, ALT, BUN, creatinine and total proteins in the chicks administered with lectin of 1.1, 2.2 and $22.2{\mu}g/kg$ body weight, which means the lectin has no effects on blood values and functions of liver and kidney. In histopathologic observation, no lesions were observed in the brain, heart, liver, lung, spleen, kidney, thymus and bursa of Fabricius of the chicks administered with lectin of 1.1, 2.2 and $22.2{\mu}g/kg$ body weight. There were inflammatory lesions, such as congestion, hemorrhage, edema, infiltration of macrophages and coagulation necrosis observed in the thigh muscle of chicks administered with lectin of $22.2{\mu}g/kg$ body weight, whereas no changes were observed in 1.1 and $22.2{\mu}g/kg$ lectin administered chicks. In chicks immunized with lectin($4.4{\mu}g/kg$ of body weight)-adjuvanted B1, LaSota and Ulster 2C vaccines, HI titers in reciprocal values for $log_2$ were 1.8-2.2 at 1 week after vaccination, which was similar with those of 1.5-2.9 by $Al(OH)_3$-adjuvanted vaccines. The HI titers by the lectin-adjuvanted vaccines reached to 3.9-5.3 at 4 weeks, whereas those by the $Al(OH)_3$-adjuvanted vaccines were more high as 7.3-9.3. Meanwhile, the immunostimulating effects of the lectin were recognized while compared to the HI titers with 2.4-3.7 in chicks immunized with vaccines without adjuvants at 4 weeks after vaccination. The chicks immunized with lectin-adjuvanted vaccines were enough to resist challenges by Kyojeongwon strain, a very virulent NDV at 4 weeks after vaccination as well as chicks immunized with $Al(OH)_3$-adjuvanted vaccines. The HI titers by the lectin-adjuvanted vaccines reached to high level as 8.7-10.3 as those with 8.2-9.6 by the $Al(OH)_3$-adjuvanted vaccines at 6 weeks after vaccination, which may be the booster effects by the challenge virus. Antibodies specific to the HN and F antigens of NDV were observed in the sera of both chicks immunized with lectin-adjuvanted vaccines and $Al(OH)_3$-adjuvanted vaccines.