• IMMUNE NETWORK
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• 제9권5호
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• pp.179-191
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• 2009

Background: The present study examines a hypothesis that short allergen-derived peptides may shift an Aspergillus fumigatus (Afu-) specific TH2 response towards a protective TH1. Five overlapping peptides (P1-P5) derived from Asp f1, a major allergen/antigen of Afu, were evaluated for prophylactic or therapeutic efficacy in BALB/c mice. Methods: To evaluate the prophylactic efficacy, peptides were intranasally administered to naive mice and challenged with Afu-allergens/antigens. For evaluation of therapeutic efficacy, the mice were sensitized with Afu-allergens/antigens followed by intranasal administration of peptides. The groups were compared for the levels of Afu-specific antibodies in sera and splenic cytokines evaluated by ELISA. Eosinophil peroxidase activity was examined in the lung cell suspensions and lung inflammation was assessed by histopathogy. Results: Peptides P1-, P2- and P3 decreased Afu-specific IgE (84.5~98.9%) and IgG antibodies (45.7~71.6%) in comparison with Afu-sensitized mice prophylactically. P1- and P2-treated ABPA mice showed decline in Afu-specific IgE (76.4~88%) and IgG antibodies (15~54%). Increased IgG2a/IgG1 and IFN-${\gamma}$/IL-4 ratios were observed. P1-P3 prophylactically and P1 therapeutically decreased IL-5 levels and eosinophil peroxidase activity. P1 decreased inflammatory cells' infiltration in lung tissue comparable to non-challenged control. Conclusion: Asp f1-derived peptide P1, prophylactically and therapeutically administered to Balb/c mice, is effective in regulating allergic response to allergens/antigens of Afu, and may be explored for immunotherapy of allergic aspergillosis in humans.

• Clinical and Experimental Pediatrics
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• 제48권5호
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• pp.506-511
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• 2005

목 적 : 폐구균 혈청형 6B와 6A는 폐구균 감염의 중요한 원인균이다. 6B 백신은 다당질 구조의 유사성으로 6A와 교차 면역반응을 일으키며 6B에 의해 생성된 6A에 대한 항체는 감염에서 방어 작용을 할 수 있다고 생각되고 있다. 이를 규명하기 위해 성인에게 폐구균 백신 접종 후 형성된 6B와 6A에 대한 항체의 옵소닌 작용 능력을 측정하여 교차 면역반응을 연구하였다. 방 법 : 건강한 성인 24명에게 혈청형 6B만 포함된 폐구균 다당질 백신을 접종하고 접종 전과 접종 한달 후 혈청에서 혈청형 6B와 6A에 대한 특이 항체의 옵소닌 작용 역가를 OPKA로 측정하였다. 결 과 : 6B에 대한 옵소닌 작용 역가는 백신 접종 전과 접종 후 모두 6A에 대한 역가보다 의미있게 높았다. 백신 혈청형인 6B에 대해서 뿐 아니라 교차 반응하는 혈청형인 6A에 대해 다당질 백신 접종 후 성인에서 옵소닌 작용 역가가 의미 있게 증가하였으므로 백신에 포함된 6B 다당질은 6A에 대해서 교차 방어 항체를 유도하였으나 모든 경우에 해당되지는 않았다. 1명의 성인에서 접종 후에도 계속적으로 6A에 대한 옵소닌 작용 역가가 측정되지 않았다. 결 론 : 백신에 포함된 폐구균 혈청형 6B에 대한 다당질은 혈청형 6A에 대해 대부분 교차 면역 반응을 일으켜 기능적 항체 형성을 유발하지만 드문 경우 이런 교차 면역 반응이 형성되지 않는 경우도 있다. 앞으로 이에 대한 연구가 소아와 노인 등의 다른 연령 군에서도 시행되어야 할 것이다. 또한 교차 방어 형성의 유무는 폐구균 백신의 임상 연구와 백신 사용 후 분리되는 폐구균의 혈청형 검사를 통한 임상 연구에서 직접적으로 연구되어야 할 것이다.

• 대한수의학회지
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• 제41권2호
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• pp.211-218
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• 2001

병원성 내장리슈만편모충(Leishmania donovani)에서 추출한 GP63 또는 LPG 항원을 liposome으로 캡슐화하고 보강제로서 BCG를 조합하여 DBA-2N 마우스에 면역접종을 한 후, 내장리슈만편모충의 병원성 amastigote를 접종하여 이들 물질의 방어면역 효과를 관찰하였다. 그 결과 GP63 과 LPG, 그리고 BCG를 모두 첨가하여 접종한 마우스의 간 조직에서 유의성 있는 내장리슈만편모충의 감소가 관찰되었으나 감소율은 27.4%에 불과하였다. 실험적 피부리슈만편모충증에 대하여 성공적인 방어면역성을 나타낸 GP63이 내장리슈만편모충 감염에 대하여 방어면역성을 상실한 원인을 분석하기 위한 실험에서 C3H 마우스에 GP63-GST 단백질과 BCG를 혼합하여 면역접종하고 내장리슈만편모충의 병원성 amastigote로 접종한 후, 혈청 내 특이항체와 비장세포에서의 감마인터페론 및 IL-5의 생산을 관찰하였다. 그 결과 GP63-GST와 BCG를 혼합하여 면역 접종한 마우스의 간 조직에서도 유의성 있는 amastigote의 감소는 관찰되지 않았다. 한편 이들 마우스의 비장세포에서는 BCG 만을 접종한 군에 비해 10배 이상의 감마인터페론과 3배의 IL-5가 생산되었다. 이와 같은 사실은 GP63-GST 단백질과 BCG를 혼합하여 접종한 마우스에서 Th1 및 Th2 타입 면역반응이 모두 활성화되었음을 시사하며, Th1 뿐만 아니라 Th2 타입 면역 반응도 함께 활성화된 것이 실험적 내장리슈만편모충 감염에 대한 방어면역에 실패한 원인 중의 하나일 것으로 사료되었다.

• 한국임상수의학회지
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• 제23권3호
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• pp.308-313
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• 2006

This study was performed to monitor the level of serum canine parvovirus(CPV) antibody titers in adult dogs throughout the Korea from January 2003 to April 2004. A total of clinically healthy 885 dogs between 1 year and 17 years old were included in this study. Serum antibody titers against CPV were measured by means of hemagglutination inhibition(HI) titers at the time dogs were brought to the hospital for revaccination. Most of dogs had been primarily vaccinated or previously revaccinated. Dogs were grouped by age, breed, sex, and primary vaccination and revaccination to determine whether these factors were associated with antibody titers. Serum CPV titers ${\geq}80$ were considered protective. Protective antibodies against CPV were present in 95.0% of the population. Breed, age, and primary vaccination and revaccination were not significantly associated with serum CPV antibody titers. But sex was significantly associated with CPV antibody titers. The results of this study have shown that there is a need to reconsider the annual revaccination strategy against CPV infection.

• Journal of Microbiology and Biotechnology
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• 제19권9호
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• pp.982-986
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• 2009

For the production of ghost bacteria vaccine to prevent the streptococcal disease in aquaculture fish species, a double cassettes vector was constructed and cloned in Escherichia coli DH5${\alpha}$. Ghost bacteria vaccine production from Escherichia coli DH5${\alpha}$/pHCE-InaN-GAPDH-Ghost 37 SDM (SIG) was maximized at a glucose concentration of 1 g/l, agitation of 300 rpm, and aeration of 1 vvm. The maximal efficiency of ghost bacteria formation was obtained at the mid-exponential phase ($OD_{600}=2.0$) with the concentration of 0.77 g/l for SIG. The molecular mass of GAPDH was detected at 67 kDa with the insoluble fraction, by SDS-PAGE and Western blot. The protective efficacy of ghost bacteria vaccine was evaluated by challenge test using olive flounder. The cumulative mortalities of the positive control, formalin-killed cell (FKC) vaccine, and SIG vaccine immunized groups were 91%, 74%, and 57%, respectively. These results suggest that SIG vaccine showed efficacy as a vaccine and had a higher potential to induce protective antibodies than did FKC vaccine.

• Biotechnology and Bioprocess Engineering:BBE
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• 제9권6호
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• pp.490-494
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• 2004

An efficacy test of PRP (polyribosylribitol phosphate)-TT (Tetanus toxoid) conjugate vaccines was carried out using BALB/c mice as an animal model by inoculating Haemophilus in­fluenzae type b (Hib) with a virulence enhancement factor (VEF). Three administrations of the conjugate vaccines at 2-week intervals elicited a significantly high level of PRP antibodies (P>0.0001). The protective activity of the PRP immunization was challenged with either Hib with iron dextran (Hib/) or with a combination of mucin and hemoglobin (Hibmh) as a VEF. The me­dium lethal dose $(LD_{50})$ for Hibmh and Hibiwas measured as 10 CFU (Colony Forming Unit) and $2.5{\times}10^{8}$ CFU respectively. Each immunized animal was challenged with five or ten times the $LD_{50}$ level of bacteria with a VEF. A significant difference in mortality between the immunized and control mice (P> 0.01) was observed with the Hibmh challenge inoculation but not with the Hibi challenge inoculation. These results show that a combination of mucin and hemoglobin was able to enhance the virulence of Hib in BALB/c mice to cause a lethal infection, thus suggesting that BALB/c mice introduced to this method can be an effective model animal for testing the protective efficacy of H. influenzae conjugate vaccines.

• IMMUNE NETWORK
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• 제5권2호
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• pp.68-77
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• 2005

Background: Costimulation is a critical process in Ag-specific immune responses. Both B7.1 and CD28 molecules have been reported to stimulate T cell responses during antigen presentation. Therefore, we tested whether Ag-specific immune responses as well as protective immunity are influenced by coinjecting with B7.1 and CD28 cDNAs in a mouse HSV-2 challenge model system. Methods: ELISA was used to detect levels of antibodies, cytokines and chemokines while thymidine incorporation assay was used to evaluate T cell proliferation levels. Results: Ag-specific antibody responses were enhanced by CD28 coinjection but not by B7.1 coinjection. Furthermore, CD28 coinjection increased IgG1 production to a significant level, as compared to pgD+pcDNA3, suggesting that CD28 drives Th2 type responses. In contrast, B7.1 coinjection showed the opposite, suggesting a Th1 bias. B7.1 coinjection also enhanced Ag-specific Th cell proliferative responses as well as production of Th1 type cytokines and chemokines significantly higher than pgD+pcDNA3. However, CD28 coinjection decreased Ag-specific Th cell proliferative responses as well as production of Th1 types of cytokines and chemokine significantly lower than pgD+pcDNA3. Only MCP-1 production was enhanced by CD28. B7.1 coimmunized animals exhibited an enhanced survival rate as well as decreased herpetic lesion formation, as compared to pgD+pcDNA3. In contrast, CD28 vaccinated animals exhibited decreased survival from lethal challenge. Conclusion: This study shows that B7.1 enhances protective Th1 type cellular immunity against HSV-2 challenge while CD28 drives a more detrimental Th2 type immunity against HSV-2 challenge, supporting an opposite role of B7.1 and CD28 in Ag-specific immune responses to a Th1 vs Th2 type.

• Journal of Microbiology and Biotechnology
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• 제27권8호
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• pp.1539-1548
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• 2017

Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen that commonly causes fatal infections in cystic fibrosis and burn patients as well as in patients who are hospitalized or have impaired immune systems. P. aeruginosa infections are difficult to treat owing to the high resistance of the pathogen to conventional antibiotics. Despite several efforts, no effective prophylactic vaccines against P. aeruginosa are currently available. In this study, we investigated the activity of the CIA06 adjuvant system, which is composed of alum and de-O-acylated lipooligosaccharide, on a P. aeruginosa outer membrane protein (OMP) antigen vaccine in mice. The results indicated that CIA06 significantly increased the antigen-specific IgG titers and opsonophagocytic activity of immune sera against P. aeruginosa. In addition, the antibodies induced by the CIA06-adjuvanted vaccine exhibited higher cross-reactivity with heterologous P. aeruginosa strains. Finally, mice immunized with the CIA06-adjuvanted vaccine were effectively protected from lethal P. aeruginosa challenge. Based on these data, we suggest that the CIA06 adjuvant system might be used to promote the immunogenicity and protective efficacy of the P. aeruginosa OMP vaccine.

• 한방안이비인후피부과학회지
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• 제29권3호
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• pp.14-26
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• 2016

Objectives : This study was performed to investigate the protective effects and mechanisms of Salvia miltiorrhizae Radix extract (SME) on endotoxin shock.Methods : We used two models; LPS-induced sepsis model for in vivo model, and murine peritoneal macrophages responses for in vitro. SME was administrated orally to mice. After 1 hr, LPS was injected intraperitoneally. Survival rate was checked each time per 12 hr for 5 days. Mice were sacrificed 3 hr after LPS injection, then blood samples and organs were harvested. Cytokines secretion was measured by ELISA. Organs tissues were observed with microscope. Murine peritoneal macrophages were cultured for 1 hr either in a medium alone or in a medium that contained SME, as indicated. Then, the cells were treated with LPS for 24 hr. mRNA levels of cytokines were measured by real-time RT-PCR. Cytokine levels in the supernatants were measured by ELISA. The amount of nitrite was measured by using the Griess method to evaluate NO production. The cell lysates were analysed by Western blotting using antibodies for iNOS and β-actin was used as an internal control to monitor equal protein loading.Results : SME improverd the survival rate of mice model. SME inhibited the secretion of inflammatory cytokines and organs damages on Endotoxin Shock model. SME suppressed cytokine expression, cytokine secretion,NO production, iNOS expression in LPS-induced murine peritoneal macrophages.Conclusions : The results suggest that SME has protective effects on endotoxin shock through suppression of inflammatory cytokines, organ damages, NO production and so on.

• 한국동물위생학회지
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• 제24권1호
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• pp.21-29
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• 2001

To compare of serum antibody titers using ELISA and HI, serum samples were collected from 100 breeders and their progeny 550 broilers. The breeders and broilers were vaccinated with infectious bronchitis(IB)- and Newcastle disease(ND)-viruses according to general vaccination program. The antibodies in serum samples against IB and ND viruses were detected by enzyme-linked immunosorbent assay(ELISA) using commercial ELISA kit and hemagglutination inhibition(HI) test. Geometric mean titer(GMT) of ELISA and In titers were monitored from 1-day-old to 35-day-old broilers and compared to those of breeder chickens. The antibody titers of breeders vaccinated with ]B virus showed 47,800, ELISA and 7.2, HI, respectively. Progeny chicks, 1-day-old, vaccinated with IBV showed high antibody titers than those of breed chickens. Those chicks were maintained protective antibody levels until 11-day-old. From 14-day-old, the antibody level decreased below protective levels. In ND, breeders serum antibody titers ELISA and Eiu were 30,200 GMT and 8.7 HI titer, respectively. On 1-day-old chicks, antibody levels was decreased to half in ELISA(16,270) compared with those of breeders, but In titers was 7.4. Progeny broilers, protective antibody level was maintained until 14- day-old by ELISA, but at 11-day-old by HI titers. After then, ND antibody titer was continuously decreased underdefense level. These result indicated that the ELISA method be more sensitive than HI titration to detect serum antibody level for IBV and NDV.