Effect of Tunicamycin and 2-deoxy-D-glucose on the Cellular and Secreted Isoperoxidase Levels of Tobacco Callus

우구,이미영,김승수,Woo, Koo,Lee, Mi-Young,Kim, Soung-Soo Korean Society for Biochemistry and Molecular Biol 1987 한국생화학회지 Vol.20 No.1

담배 callus 배양시 tunicamycin(TM)과 2-deoxy-D-glucose(2-D-G)가 세포내 peroxidase와 배지로 분비되는 peroxidase의 활성과 isozyme pattern에 미치는 영향을 조사하였다. TM과 2-D-G 처리에 의하여 세포내 peroxidase의 비활성도는 전 배양기간동안 현저히 증가한 반면 배지로 분비되는 peroxidase의 비활성도는 배양초기를 제외하고는 점차 감소하였다. Isozyme patterns변화를 조사한 결과 M과 2-D-G처리에 의하여 세포내 peroxidase 중 glycoprotein인 cathodic isoperoxidase $C_4$ 가 뚜렷한 증가를 보인 반면 또 다른 glycoprotein인 anodic isoperoxidase $A_3$는 감소를 나타냈다. 한편 TM과 2-D-G 처리시 배지로 분비되는 peroxidase의 비활성도 감소는 주로 isoperoxidase $A_3$의 감소에 기인한다는 사실도 확인하였다. Anti-$A_3$ 혈청과 anti-$C_4$혈청을 사용하여 isoperoxidase $C_4$와 $A_3$의 양의 변화를 정량한 결과 비활성도 변화 및 pattern 변화와 일치하는 것으로 나타났다. The effects of tunicamycin(TM) and 2-deoxy-D-glucose(2-D-G) treatment on the specific activities and isozyme patterns of the cellular and secretory peroxidases of tobacco callus were investigated. The specific activities of the cellular peroxidases were increased by TM and 2-D-G treatment during entire culture periods, whereas the specific activities of the secretory peroxidases were decreased by TM and 2-D-G treatment. Comparisons of the isoperoxidase patterns of TM treated groups and 2-D-G treated groups with those of control groups showed that both TM and 2-D-G cause the significant elevation of the activities of cathodic isoperoxidases, especially isoperoxidase $C_4$ which is a glycoprotein. But the decrease of the specific activities of the secretory peroxidases by TM and 2-D-G was mainly due to the lowered level of anodic isoperoxidase $A_3$ which is also a glycoprotein. Immunoprecipitation with anti-$A_3$ antibody and anti-$C_4$ antibody were carried out to examine the changes in the synthesis and secretion of these isozymes due to the treatment of the callus with the glycosylation inhibitors. These results suggested that the increase of cellular $C_4$ activity was due to the increased amount of $C_4$ proteins and the decrease of the activity of secretory isoperoxidase $A_3$ was caused by the decrease of $A_3$ proteins in the medium in the presence of the glycosylation inhibitors.

Tunicamycin 과 2 - deoxy - D - glucose 가 Cellular Peroxidase 와 Secretory Peroxidase 의 생성에 미치는 영향

우구,이미영,김승수 ( Koo Woo,Mi Young Lee,Soung Soo Kim ) 생화학분자생물학회 1987 BMB Reports Vol.20 No.1

The effects of tunicamycin(TM) and 2-deoxy-D-glucose(2-D-G) treatment on the specific activities and isozyme patterns of the cellular and secretory peroxidases of tobacco callus were investigated. The specific activities of the cellular peroxidases were increased by TM and 2-D-G treatment during entire culture periods, whereas the specific activities of the secretory peroxidases were decreased by TM and 2-D-G treatment. Comparisons of the isoperoxidase patterns of TM treated groups and 2-D-G treated groups with those of control groups showed that both TM and 2-D-G cause the significant elevation of the activities of cathodic isoperoxidases, especially isoperoxidase C₄ which is a glycoprotein. But the decrease of the specific activities of the secretory peroxidases by TM and 2-D-G was mainly due to the lowered level of anodic isoperoxidase A₃ which is also a glycoprotein. Immunoprecipitation with anti-A₃antibody and anti-C₄ antibody were carried out to examine the changes in the synthesis and secretion of these isozymes due to the treatment of the callus with the glycosylation inhibitors. These results suggested that the increase of cellular C₄ activity was due to the increased amount of C₄ proteins and the decrease of the activity of secretory isoperoxidase A₃ was caused by the decrease of A₃ proteins in the medium in the presence of the glycosylation inhibitors.

Tunicamycin 과 2 - deoxy - D - glucose 가 Cellular Peroxidase 와 Secretory Peroxidase 의 생성에 미치는 영향

이미영,김승수,우구 생화학분자생물학회 1992 BMB Reports Vol.14 No.2

The effects of tunicamycin(TM) and 2-deoxy-D-glucose(2-D-G) treatment on the specific activities and isozyme patterns of the cellular and secretory peroxidases of tobacco callus were investigated. The specific activities of the cellular peroxidases were increased by TM and 2-D-G treatment during entire culture periods, whereas the specific activities of the secretory peroxidases were decreased by TM and 2-D-G treatment. Comparisons of the isoperoxidase patterns of TM treated groups and 2-D-G treated groups with those of control groups showed that both TM and 2-D-G cause the significant elevation of the activities of cathodic isoperoxidases, especially isoperoxidase C₄ which is a glycoprotein. But the decrease of the specific activities of the secretory peroxidases by TM and 2-D-G was mainly due to the lowered level of anodic isoperoxidase A₃ which is also a glycoprotein. Immunoprecipitation with anti-A₃antibody and anti-C₄ antibody were carried out to examine the changes in the synthesis and secretion of these isozymes due to the treatment of the callus with the glycosylation inhibitors. These results suggested that the increase of cellular C₄ activity was due to the increased amount of C₄ proteins and the decrease of the activity of secretory isoperoxidase A₃ was caused by the decrease of A₃ proteins in the medium in the presence of the glycosylation inhibitors.

47계대 Oka주 약독화 생백신 접종 후 2년간 임상 및 면역성의 추적 연구

강진한,김종현,허재균,우구,Kang, Jin Han,Kim, Jong Hyun,Hur, Jae Kyun,Woo, Koo 대한소아감염학회 2000 Pediatric Infection and Vaccine Vol.7 No.1

목 적 : 국내 건강한 소아에서 47계대 배양 Oka주 수두 약독화 생백신을 접종한 후 단기간 내의 면역원성과 안전성에 관한 연구를 시행한 다음 이들 접종아의 장기간에 걸친 접종효과와 면역성 유지에 관한 연구계획에 따라 일차적으로 접종 2년내에 수두 발생 여부와 발생시 임상적 특성을 확인하고 접종 2년 후 수두에 관한 면역상태의 평가와 잠복감염의 활성 여부를 파악하고자 하였다. 대상 및 방법 : 1997년 4월에서 8월 사이 가톨릭의대 소아과에서 시행하였던 47계대 Oka주 수두 약독화 생백신의 단기간 면역원성과 안전성의 연구 대상아 99명을 본 연구의 지속적인 동일 대상으로 하였다. 연구 대상아를 2년간 지속적으로 추적하여 수두의 발생여부를 확인하였다. 접종아중 수두에 이환 되었을 경우 발병기간, 발열을 포함한 피부병변의 정도, 합병증의 발생 여부, 치료투약의 여부 등을 파악하여 임상적 평가를 실시하고, 동시에 발생의 역학 상태를 파악하였다. 그리고 이 연구 기간 동안에 연구 대상아에서 잠복감염에 따른 대상포진의 발생을 동시에 관찰하였다. 또한 접종 2년후에 EIA와 FAMA의 면역학적 검사를 통하여 수두 대상포진 바이러스 IgG 항체와 수두 대상포진 바이러스의 세포막 항원에 대한 항체를 측정하여 접종 2년 후의 면역상태를 평가하였다. 그리고 이 결과를 통하여 2년 전에 확인하였던 접종 1개월 후 면역상태와 비교하였다. 결 과 : 연구대상자 99명을 2년간 추적한 결과 6명에서 수두가 발생되었다. 이들은 3일에서 5일 정도의 짧은 임상 경과를 보였고 2례에서 발열이 있었고 전 예에서 발진의 정도는 수포의 수나 확산 정도가 매우 미약하였으며 합병증을 보인 경우는 없었다. 그리고 치료를 위한 투약을 받은 경우는 없었고 전 예에서 가족내 또는 집단 발생 후 발생되었다. 2년 후 항체검사가 가능하였던 46명에서 EIA에 의한 수두 대상포진 바이러스 IgG 항체를 측정한 결과 1례를 제외한 전 예에서 양성을 보였으며 높은 항체가를 유지되었음을 알 수 있었고, FAMA에 의한 수두 대상포진 바이러스 세포막 항원에 대한 항체가는 전 예에서 양성을 보였다. 이들 추적 항체검사가 가능하였던 46명에서 접종 1개월 후 항체가와 2년 후 항체가를 비교한 결과 수두 대상포진 바이러스 IgG 항체와 수두 대상포진 바이러스 세포막 항원에 대한 항체는 모두 접종 2년 후에 높게 유지됨을 알 수 있었다. 그러나 전 예에서 연구기간 동안 대상포진의 발생은 관찰되지 않았다. 결 론 : 47계대 배양 Oka주 수두 약독화 생백신 접종 후 2년간 추적한 결과 접종아에서 집단 발생과 더불어 소수에서 수두가 발생되었으나 그 정도는 매우 미약하여 임상적으로 큰 문제가 없고 접종 2년 후에도 수두 대상포진 바이러스에 대한 면역상태는 잘 유지되고 있음을 확인하였다. Purpose : We previously reported the short-term immunogenecity and safety of 47 passaged Oka strain live attenuated varicella vaccine in healthy children in 1997. Now, we conducted this two-year follow-up study to confirm the maintenance of immunity, the occurrence of natural varicella infection and the activation of vaccine induced latent infection on the same vaccine. Methods : 99 children who had been immunized by 47 passaged Oka strain live attenuated varicella vaccine in 1997 were followed up by questionnaire, and 46 children out of study group were followed up serologically. They were asked to report any instance of varicella or herpes zoster since they had been immunized. If there was any evidence of varicella or herpes zoster, they should be clinically or serologically confirmed by doctor. Also, those patients' parents were asked to report any instance of varicella or herpes zoster in their family, playmate, kindergarten, school, or other settings. The immunity to VZV was confirmed by EIA and FAMA test. Results : 6 recipients developed breakthrough varicella after exposure to VZV in family, kindergarten and school during follow-up period. However, clinical features of those patients were very mild and self limited without therapy. And none of the recipients developed herpes zoster during this observation period. The results of EIA test showed that study subjects were all seropositive except one, and the antibody titers and GMT of FAMA test were seropositively maintained in all subjects. Statistically, the antibody titers of EIA and FAMA test confirmed two years after vaccination were higher than those results confirmed one month after vaccination. Conclusion : Our study results suggest that the immunity of 47 passaged Oka strain live attenuated varicella is well maintained until 2 years later after vaccination, and mild natural infection after exposure to VZV can be occurred with low rate. There were not developing zoster in study vaccine after vaccination for two-years.