• KSBB Journal
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• 제19권1호
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• pp.88-92
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• 2004

장티푸스 협막 다당체와 일본 뇌염 바이러스로 구성되어있는 혼합백신을 제조하여 마우스에서 면역성을 측정하였다. 혼합 항원 백신은 단일 항원을 투여한 경우보다 장티푸스와 일본 뇌염에 대해서 더 높은 IgG 항체 형성을 나타내었다. Aluminium hydroxide를 adjuvant로 첨가한 경우에도 첨가하지 않은 대조군에 비하며 모두 IgG 항체 형성이 증가하는 것으로 나타났다. 일본뇌염의 경우 중화항체 값 측정 시험에서도 혼합 항원백신은 일본뇌염 단독 항원 시에 비교해서 더 많은 중화항체 값을 형성하는 결과를 나타내었다. 본 실험에서 장티푸스 협막 다당체와 일본뇌염 바이러스의 혼합백신은 서로 다른 성분의 항원간에 나타날 수 있는 항체 형성에 대한 masking effect가 발생하지 않으며 오히려 synergic effect를 나타낸다는 사실을 보여주었다. 본 연구에서는 접종시기가 유사한 두 항원의 혼합 백신으로의 사용가능성을 높여주고 최근에 백신 산업에서 종류가 다른 여러 백신들을 혼합하여 다가백신을 제조 생산하는 추세의 산업화에 큰 기대효과를 제공할 수 있으리라 사려된다.

• 대한수의학회지
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• 제37권3호
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• pp.555-568
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• 1997

Fowl typhoid caused by Salmonella gallinarum has increased dramatically since 1992 and has caused a great economic losses in chicken industry by characterizing with high mortality. In these studies, we investigated the immunogenicity and protectivity in chickens which were immunized with outer membrane protein(OMP) extracted from isolates of S gallinarum against challenge with live microorganism. Outer membrane proteins were composed of various sizes of molecular weight including 14K, 22K, 31K, 36K, 40K and 55K and the most of them responded strongly against rabbit antisera in immunoblot analysis. The chickens vaccinated with OMP or vaccinated with whole-cell combined with OMP($200{\mu}g$/chickens) complex showed higher delayed type hypersensitivity(DTH) response than that of whole-cell vaccinated group. The protective rates of OMP or whole-cell combined with OMP complex group against challenge of S gallinarum were higher (above 75%) than those (45~50%) of whole-cell vaccinated group. All vaccines were safe and the body weight-gains of all vaccinated groups were not significantly different (p<0.05) from those of nonvaccinated control group. In vitro tests, OMP stimulated both the proliferation of lymphocytes and T-lymphocytes, and OMP-induced lymphocyte proliferation was higher in the cells of the immunized chickens with OMP than in those from the control chickens.

• Journal of Microbiology and Biotechnology
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• 제7권6호
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• pp.423-428
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• 1997

Vibrio vulnificus is a halophilic gram-negative human pathogen, which affects people with underlying liver diseases or a suppressed immune system, often leading to primary septicemia with a mortality rate of higher than 60%. In an effort to develop an oral vaccine against V. vulnificus infection, we prepared a whole cell killed vaccine of V. vulnificus on a large scale and compared the immunogenicity and protective efficacy of the vaccine administered in three formulation forms in rabbits. Since V. vulnificus O-antigen serotypes 1, 2, 3, 4, 5, and 7 account for more than 95% of clinical isolates, we prepared cell lysates from these six serotype strains and mixed in equal amounts for a vaccine. The vaccine was administered to rabbits intramuscularly (i.m.), orally as granules or as enteric-coated granules. In rabbits, all three formulation forms elicited a high level of serum IgG antibody reactive not only to the six strains but also to other O-antigen serotypes 6, 8 and 9, indicating cross-reactivities among the strains. Immunotherapeutic efficacy of the antisera was also evaluated by a passive immunization assay, which revealed that the orally immunized antisera as well as the i.m. immunized antisera was protective against a subsequent lethal challenge of V. vulnificus. These data demonstrate that oral immunization with a V. vulnificus whole cell lysate vaccine induced a systemic immune response and suggest the feasibility of development of this vaccine preparation as an oral vaccine.

• Journal of Korean Medical Science
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• 제33권51호
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• pp.340.1-340.14
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• 2018

Background: Various pneumococcal vaccines have been evaluated for immunogenicity by opsonophagocytic assay (OPA). A multiplexed OPA (MOPA) for 13 pneumococcal serotypes was developed by Nahm and Burton, and expanded to 26 serotypes in 2012. The development of new conjugate vaccines with increased valence has necessitated expanded MOPAs to include these additional serotypes. In this study, we validated this expanded MOPA platform and applied to measure antibodies against 11 additional serotypes (2, 8, 9N, 10A, 11A, 12F, 15B, 17F, 20B, 22F, and 33F) in human sera. Methods: All materials, including serum, complement, bacterial master stocks, and HL-60 cells, were evaluated for assay optimization. Following optimization, the assay was validated for accuracy, specificity, and intra- and inter-assay precision with sera from adult donors following standard protocols. The assay was applied to evaluate functional antibodies of 42 sera immunized with 23-valent pneumococcal polysaccharide vaccine (PPV23). Results: The expanded MOPA platform was specific for all serotypes, with the exception of serotype 20. The assay results were highly correlated with those obtained from single-serotype OPA, indicating acceptable accuracy. The coefficients of variation were 7%-24% and 13%-39% in tests of intra- and inter-assay precision, respectively, using three quality-control samples. A MOPA that included 11 additional serotypes in the PPV23 was established and validated with respect to accuracy, specificity, and precision. The opsonic indices of immune sera were obtained using this validated assay. Conclusion: The expanded MOPA will be useful for evaluation of the immunogenicity of PPV23 and future conjugate vaccine formulations.

• Journal of Microbiology and Biotechnology
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• 제32권10호
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• pp.1335-1343
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• 2022

COVID-19 is an emerging disease that poses a severe threat to global public health. As such, there is an urgent demand for vaccines against SARS-CoV-2, the virus that causes COVID-19. Here, we describe a virus-like nanoparticle candidate vaccine against SARS-CoV-2 produced by an E. coli expression system. The fusion protein of a truncated ORF2-encoded protein of aa 439~608 (p170) from hepatitis E virus CCJD-517 and the receptor-binding domain of the spike protein from SARS-CoV-2 were expressed, purified and characterized. The antigenicity and immunogenicity of p170-RBD were evaluated in vitro and in Kunming mice. Our investigation revealed that p170-RBD self-assembled into approximately 24 nm virus-like particles, which could bind to serum from vaccinated people (p < 0.001) and receptors on cells. Immunization with p170-RBD induced the titer of IgG antibody vaccine increased from 14 days post-immunization and was significantly enhanced after a booster immunization at 28 dpi, ultimately reaching a peak level on 42 dpi with a titer of 4.97 log₁₀. Pseudovirus neutralization tests showed that the candidate vaccine induced a strong neutralizing antibody response in mice. In this research, we demonstrated that p170-RBD possesses strong antigenicity and immunogenicity and could be a potential candidate for use in future SARS-CoV-2 vaccine development.

• Journal of Veterinary Science
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• 제24권1호
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• pp.15.1-15.13
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• 2023

Background: Inactivated vaccines are limited in preventing foot-and-mouth disease (FMD) due to safety problems. Recombinant virus-like particles (VLPs) are an excellent candidate for a novel vaccine for preventing FMD, given that VLPs have similar immunogenicity as natural viruses and are replication- and infection-incompetent. Objectives: The 3C protease and P1 polyprotein of type O FMD virus (FDMV) was expressed in yeast Hansenula polymorpha to generate self-resembling VLPs, and the potential of recombinant VLPs as an FMD vaccine was evaluated. Methods: BALB/c mice were immunized with recombinant purified VLPs using CpG oligodeoxynucleotide and aluminum hydroxide gel as an adjuvant. Cytokines and lymphocytes from serum and spleen were analyzed by enzyme-linked immunosorbent assay, enzyme-linked immunospot assay, and flow cytometry. Results: The VLPs of FMD were purified successfully from yeast protein with a diameter of approximately 25 nm. The immunization of mice showed that animals produced high levels of FMDV antibodies and a higher level of antibodies for a longer time. In addition, higher levels of interferon-γ and CD4⁺ T cells were observed in mice immunized with VLPs. Conclusions: The expression of VLPs of FMD in H. polymorpha provides a novel strategy for the generation of the FMDV vaccine.

• IMMUNE NETWORK
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• 제23권6호
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• pp.44.1-44.16
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• 2023

Mesenchymal stem cells (MSCs) are effective in treating autoimmune diseases and managing various conditions, such as engraftment of allogeneic islets. Additionally, autologous and HLA-matched allogeneic MSCs can aid in the engraftment of human allogeneic kidneys with or without low doses of tacrolimus, respectively. However, HLA alloantigens are problematic because cell therapy uses more HLA-mismatched allogeneic cells than autologous for convenience and standardization. In particular, HLA-mismatched MSCs showed increased Ag-specific T/B cells and reduced viability faster than HLA-matched MSCs. In CRISPR/Cas9-based cell therapy, Cas9 induce T cell activation in the recipient's immune system. Interestingly, despite their immunogenicity being limited to the cells with foreign Ags, the accumulation of HLA alloantigen-sensitized T/B cells may lead to allograft rejection, suggesting that alloantigens may have a greater scope of adverse effects than foreign Ags. To avoid alloantigen recognition, the β2-microglobulin knockout (B2MKO) system, eliminating class-I MHC, was able to avoid rejection by alloreactive CD8 T cells compared to controls. Moreover, universal donor cells in which both B2M and Class II MHC transactivator (CIITA) were knocked out was more effective in avoiding immune rejection than single KO. However, B2MKO and CIITA KO system remain to be controlled and validated for adverse effects such as the development of tumorigenicity due to deficient Ag recognition by CD8 T and CD4 T cells, respectively. Overall, better HLA-matching or depletion of HLA alloantigens prior to cell therapy can reduce repetitive transplantation through the long-term survival of allogeneic cell therapy, which may be especially important for patients seeking allogeneic transplantation.

• IMMUNE NETWORK
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• 제23권2호
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• pp.16.1-16.19
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• 2023

Bacillus Calmette-Guerin (BCG) vaccine is the only licensed vaccine for tuberculosis (TB) prevention. Previously, our group demonstrated the vaccine potential of Rv0351 and Rv3628 against Mycobacterium tuberculosis (Mtb) infection by directing Th1-biased CD4⁺ T cells co-expressing IFN-γ, TNF-α, and IL-2 in the lungs. Here, we assessed immunogenicity and vaccine potential of the combined Ags (Rv0351/Rv3628) formulated in different adjuvants as subunit booster in BCG-primed mice against hypervirulent clinical Mtb strain K (Mtb K). Compared to BCG-only or subunit-only vaccine, BCG prime and subunit boost regimen exhibited significantly enhanced Th1 response. Next, we evaluated the immunogenicity to the combined Ags when formulated with four different types of monophosphoryl lipid A (MPL)-based adjuvants: 1) dimethyldioctadecylammonium bromide (DDA), MPL, and trehalose dicorynomycolate (TDM) in liposome form (DMT), 2) MPL and Poly I:C in liposome form (MP), 3) MPL, Poly I:C, and QS21 in liposome form (MPQ), and 4) MPL and Poly I:C in squalene emulsion form (MPS). MPQ and MPS displayed greater adjuvancity in Th1 induction than DMT or MP did. Especially, BCG prime and subunit-MPS boost regimen significantly reduced the bacterial loads and pulmonary inflammation against Mtb K infection when compared to BCG-only vaccine at a chronic stage of TB disease. Collectively, our findings highlighted the importance of adjuvant components and formulation to induce the enhanced protection with an optimal Th1 response.

• Pediatric Infection and Vaccine
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• 제19권1호
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• pp.1-11
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• 2012

목 적 : 인플루엔자는 소아암 환자에게 이환율과 사망률이 높은 질환이나 소아암 환자에 대한 예방 접종률은 낮은 상태이다. 본 연구에서는 소아암 환자를 대상으로 인플루엔자 예방접종의 면역원성과 부작용에 대한 평가를 시행하였다. 방 법 : 2009년 10월부터 12월까지 원자력의학원에서 인플루엔자 예방접종(SK influenza IX vaccine$^{(R)}$)을 받은 25명의 소아암 환자를 대상으로 연구를 시행했다. 예방접종일과 접종 후 30일 뒤 2회에 걸쳐 채혈하였고 혈구응집억제 항체가를 측정하였다. 백신의 면역원성은 접종 전과 접종 후 30일의 혈구응집억제 항체가 1:40 이상인 피험자 비율, 접종 후 30일의 항체 양전율, 접종 전과 접종 후 30일 사이의 GMT 증가 배수로 평가하였다. 결 과 : 본 연구대상자 중에서 심각한 예방접종관련 부작용을 경험한 대상자는 없었다. 접종 후 혈구응집억제 항체가 1:40 이상을 보인 피험자의 비율은 H1N1 항원에 대해 68%, H3N2 항원에 대해 40%, B 항원에 대해 36%였다. 항체양전율은 H1N1 항원에 대해 12%, H3N2 항원에 대해 16%, B 항원에 대해 20%였다. GMT 증가 배수는 H1N1 항원에 대해 0.9, H3N2 항원에 대해 1.2, B 항원에 대해 1.8이었다. 결 론 : 본 연구의 대상자들은 인플루엔자 백신에 대해 제한적인 면역반응을 보였으나 일부 대상자들에게서 항체 양전이 나타났고 심각한 예방접종관련 부작용이 없었던 점을 고려할 때 소아암환자를 대상으로 매년 정기적인 인플루엔자 예방접종이 추천된다.

• Clinical and Experimental Pediatrics
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• 제63권7호
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• pp.265-271
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• 2020

Background: Pneumococcal diseases among children aged <5 years worldwide are associated with high annual mortality rates. Purpose: This study aimed to evaluate the immunogenicity and safety of GBP411, a 12-valent pneumococcal conjugant vaccine, with a dosing schedule of 2 primary doses plus 1 booster dose (2p+1) in healthy infants. Methods: This randomized active-controlled (Prevnar 13) double-blind phase 2 trial enrolled healthy subjects aged 6-10 weeks. Three serum concentrations of pneumococcal serotype-specific immunoglobulin G (IgG) were evaluated using the pneumococcal serotype-specific pneumonia polysaccharide enzyme-linked immunosorbent assay at 1 month after the primary doses and before and 1 month after the booster dose. The pneumococcal serotype-specific IgG titer was evaluated using a multiplex opsonophagocytic assay in a subset of 15 subjects per group. Results: After administration of the primary doses, the proportion of subjects who achieved pneumococcal serotype-specific IgG concentrations of >0.35 ㎍/mL was lower for some serotypes in the GBP411 group than in the comparator group (6B: 20.83% vs. 39.22%, P=0.047 and 19A: 58.33% vs. 90.20%, P<0.001). However, after administration of the booster dose, >97% of the subjects in each group achieved IgG concentrations of ≥0.35 ㎍/mL for all 12 serotypes. Increased immunogenicity was observed for some serotypes that showed significant intergroup differences after administration of the primary doses but not after the booster dose. We also found no significant intergroup difference in the overall incidence of solicited local adverse events. Furthermore, the overall incidence of solicited systemic adverse events was significantly lower in the GBP411 group than in the comparator vaccine group (79.59% vs. 98.04%; P=0.003). Conclusion: The GBP411 vaccine with a dosing schedule of 2p+1 may be immunogenic and safe for healthy infants.