• 대한수의학회지
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• 제45권3호
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• pp.311-323
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• 2005

This study tested the effect of a trivalent (feline panleukopenia; FPV, feline viral rhinotracheitis; FHV, feline calicivirus infection; FCV) inactivated vaccine in cats. The vaccine was tested for the safety in guinea pigs, mice and cats. Also, it was tested for the efficacy in cats. The vaccine was inoculated to cats at 7~9 and 10~12 weeks of age (conventional schedule) and the serological response to vaccination was assessed and was compared to the unvaccinated group. All cats were bled by jugular venipuncture for FPV, FHV and FCV specific serological test (virus neutralizing antibody, VN) at 7~9, 10~12 and 13~15 weeks. After last bleeding, all cats were inoculated with each virus (FPV : orally $2ml\;10^{7.5}\;TCID_{50}/ml$, FHV : nasally $1ml\;10^{7.0}\;TCID_{50}/ml$ and FCV : nasally $1ml\;10^{7.0}\;TCID_{50}/ml$). The Vaccine verified excellent protective effect in guinea pigs, mice and cats. The VN antibody titers of the unvaccinated group cats against FPV, FHV and FCV were <2~16, on the other hand the vaccinated group cats were $512{\sim}{\geq}4096$, 64~1024 and 64~1024, respectively. When all cats were challenged with virulent viruses, the survival rates of the vaccinated group cats were over 80%, while the survival rates of the unvaccinated group cats were less 20%. The typical clinical signs were not observed in the vaccinated group cats, but the typical clinical signs and histopathological lesions were observed in the unvaccinated group cats. As the result of tests, the VN values obtained in this study appeared to be high enough to protect cats from viral challenges. The trivalent (FPV, FHV, and FCV) inactivated vaccine seemed to be very effective, for prevention of feline viral diseases (FPV, FHV, and FCV).

• 대한수의학회지
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• 제26권2호
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• pp.265-276
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• 1986

The bovine serum factor influencing the growth of swine testicle (ST) cell and the END effect of hog cholera SN test was studied. Throughout the experimental studies. following results were obtained and summarized. 1. Bovine whole serum of 16(76.2%) and 4(19.0%) samples out of 21 have shown a positive ST cell growth and the END effect, respectively. However, all of 21(100%) and 8(38.1%) samples out of 21 serum supernatant fractions, prepared from the bovine whole serum, have shown positive ST cell growth and END effect, respectively. 2. In the SDS-polyacrylamide gel electrophoretic analysis of the bovine whole serum and the supernatant fractions, ST cell growth inhibiting factor was proved present in globulin fraction and in whole gel plate as a diffusible component. 3. The END ineffective component present in the whole serum and its supernatant fraction was proved to be BVDV neutralizing antibody. 4. The difference of osmolarity, optical density, pH, degree of precipitant formation following heat cold treatment, A/G ratio as we11 as electrophoretic pattern and NDV SN index of the samples were not correlated to the degree of 57 cell growth and to the END effectiveness.

• Journal of Microbiology and Biotechnology
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• 제19권10호
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• pp.1271-1279
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• 2009

To develop low endotoxic and multi-immunogenic outer membrane vesicles (OMVs), a deletion mutant of the msbB gene in Salmonella enterica serovar Typhimurium (S. Typhimurium) was used as a source of low endotoxic OMV, and an expression vector of the canine parvovirus (CPV) VP2 epitope fused to the bacterial OmpA protein was constructed and transformed into the Salmonella ${\Delta}msbB$ mutant. In a lethality test, BALB/c mice injected intraperitoneally with the Salmonella ${\Delta}msbB$ mutant survived for 7 days, whereas mice injected intraperitoneally with the wild type survived for 3 days. Moreover, all mice inoculated orally with the ${\Delta}msbB$ mutant survived for 30 days, but 80% of mice inoculated orally with the wild type survived. The OmpA::CPV VP2 epitope fusion protein was expressed successfully and associated with the outer membrane and OMV fractions from the mutant S. Typhimurium transformed with the fusion protein-expressing vector. In immunogenicity tests, sera obtained from the mice immunized with either the Salmonella msbB mutant or its OMVs containing the OmpA::CPV VP2 epitope showed bactericidal activities against wild-type S. Typhimurium and contained specific antibodies to the CPV VP2 epitope. In the hemagglutination inhibition (HI) assay as a measurement of CPV-neutralizing activity in the immune sera, there was an 8-fold increase of HI titer in the OMV-immunized group compared with the control. These results suggested that the CPV-neutralizing antibody response was raised by immunization with OMV containing the OmpA::CPV VP2 epitope, as well as the protective immune response against S. Typhimurium in BALB/c mice.

• Journal of Microbiology
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• 제43권6호
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• pp.537-545
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• 2005

Even though neutralizing antibodies against the Hantaan virus (HTNV) has been proven to be critical against viral infections, the cellular immune responses to HTNV are also assumed to be important for viral clearance. In this report, we have examined the cellular and humoral immune responses against the HTNV nucleocapsid protein (NP) elicited by virus infection or DNA vaccination. To examine the cellular immune response against HTNV NP, we used $H-2K^b$ restricted T-cell epitopes of NP. The NP-specific $CD8^+$ T cell response was analyzed using a $^{51}Cr-release$ assay, intracellular cytokine staining assay, enzyme-linked immunospot assay and tetramer binding assay in C57BL/6 mice infected with HTNV. Using these methods, we found that HTNV infection elicited a strong NP-specific $CD8^+$ T cell response at eight days after infection. We also found that several different methods to check the NP-specific $CD8^+$ T cell response showed a very high correlation among analysis. In the case of DNA vaccination by plasmid encoding nucleocapsid gene, the NP-specific antibody response was elicited $2\~4$ weeks after immunization and maximized at $6\~8$ weeks. NP-specific $CD8^+$ T cell response reached its peak 3 weeks after immunization. In a challenge test with the recombinant vaccinia virus expressing NP (rVV-HTNV-N), the rVV-HTNV-N titers in DNA vaccinated mice were decreased about 100-fold compared to the negative control mice.

• Tuberculosis and Respiratory Diseases
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• 제43권3호
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• pp.348-358
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• 1996

연구배경 : Interleukin-5(IL-5)는 호산구 보이는 여러 질환들과 관련이 있으며 특히 알레르기성 천식에서 호산구의 침윤정도와 밀접한 관계가 있는 것으로 알려져 있다. 그러나 IL-2도 증상있는 천식환자의 기도에서 상승됨이 관찰되어 호산구 침윤정도와 상관관계가 있는 것이 밝혀졌다. IL-2가 호산구의 생존을 증가시키는 기전이 IL-5의 표현을 증가시킴으로써인지 또는 다른 경로를 통하여 작용하는 것인지 알기위해 다음과 같은 방법으로 호산구 생존에 미치는 IL-2의 영향을 관찰하였다. 방법 : 호산구증다증을 보인 환자의 말초혈액으로부터 호산구를 분리하여 trypan blue dye exclusion test를 이용하여 생존율을 측정하였으며 Randolp 용액을 이용하여 호산구를 계수하였다. 1) IL-2, IL-5 존재하의 호산구 생존율 및 IL-2와 anti IL-5 존재하의 호산구 생존율을 측정하였다. 2) IL-2 존재하의 말초혈액단핵구에서 IL-5 m-RNA 표현을 Reverse Transcription-Polymerase Chain Reaction(RT-PCR) 방법을 통하여 관찰하였다. 3) IL-2로 자극한 호산구의 IL-2 수용체 발현 증가를 유세포분석기(Flow cytometer)로 측정하였다. 결과 : 1) 호산구의 생존율은 IL-2 및 IL-5에 대한 농도의존성을 보이며 증가하였다. 2) IL-2에 의해 증가된 호산구의 생존율은 anti IL-5에 의해 억제되지 않았다. 3) IL-2로 자극된 말초혈액단핵구는 IL-5 m-RNA를 표현하지 않았다. 4) IL-2는 호산구에서 IL-$2{\alpha}$ 수용체의 표현을 증가시키며 IL-$2{\beta}$ 수용체의 표현은 변화가 없었다. 결론 : 사람에서는 IL-2는 IL-5 형성증가를 통하지 않고 호산구에 IL-2 수용체를 증가시킴으로써 호산구의 생존율을 증가시킨다.

• 한국가금학회지
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• 제39권2호
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• pp.113-120
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• 2012

본 연구를 통하여 Vero 세포에서 강독형 NDV인 Kr005주를 55대 연속 계대하여 Vero 세포에 적응된 Kr005/V 주를 작성하였다. Kr005/V 주는 Vero 세포에서 $10^{7.8}/mL$의 높은 바이러스 증식성을 보였으며, 감염세포는 cell rounding과 같은 CPE를 형성하였다. 또한 이열성(heat labile) NDV Kr005주와 달리 NDV Kr005/V 주는 내열성(thermostable)을 보였다. 계태아에서 평균 치사 시간(MDT)를 측정한 결과, 강독형인 Kr005주(49.6시간의 MDT)와 달리 Kr005/V 주는 120시간이상의 MDT를 보여 약독형 바이러스(lentogenic virus)로 분류되었다. Kr005/V 주의 HN 단백질과 F 단백질의 염기서열을 분석한 결과, HN 단백질의 443번째 아미노산에서만 변이(T에서 S)가 관찰되었다. Kr005/V 주와 Vero 세포를 사용하여 야외 닭 혈청을 대상으로 혈청중화 항체가를 측정한 결과, Kr005주와 CEF 세포를 사용하여 측정한 혈청중화 항체가와 97%의 상관성을 나타내었다.