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CagL 재조합 단백질 접종후에 Mongolian gerbil에서 나타나는 Helicobacter pylori 감염에 대한 반응
박은정,장성일,최윤희,김진문,김애련,김지혜,우계형,유윤정,이성행,차정헌,Bak, Eun-Jung,Jang, Sung-Il,Choi, Yun-Hui,Kim, Jin-Moon,Kim, Ae-Ryun,Kim, Ji-Hye,Woo, Gye-Hyeong,Yoo, Yun-Jung,Lee, Sung-Haeng,Cha, Jeong-Heon 한국미생물학회 2012 미생물학회지 Vol.48 No.2
Helicobacter pylori는 만성 위염, 소화성 궤양, 위암의 중요한 역학적 인자중 하나이다. H. pylori의 독성인자중 CagL은 숙주 세포와 H. pylori의 제 4형 분비기관(Type 4 secretion system)을 연결하는 adhesin으로 작용하는 섬모 단백질로 H. pylori가 발병하는데 중요한 역할을 하는 것으로 알려져 있다. 이번 연구는 저빌에 H. pylori를 감염시킨 동물 모델을 이용하여 CagL 재조합 단백질을 면역화시켰을 때 나타나는 효과를 평가하였다. 재조합 CagL은 클론되었고, 과발현시켜 정제하여 준비하였다 저빌은 H. pylori 감염 대조군과 H. pylori 감염 CagL 재조합 단백질 접종군으로 분류하였고, 접종시 알루미늄 애쥬번트를 사용하였다. 일주일 간격으로 4회 근육내 접종하였고, 마지막 접종 일주일 후, 모든 저빌에 H. pylori 7.13 균주를 $1{\times}10^9\;bacteria/500{\mu}l$ 농도로 위내 투여하였다. H. pylori 감염 6주째 모든 저빌을 희생하여 혈청 IgG 반응평가를 위한 ELISA를 실시하였고, 위에서는 집락화된 H. pylori의 수평가, 병리조직학적 평가 및 사이토카인 유전자발현을 조사하였다. CagL 재조합 단백질접종 일주일 후부터 H. pylori 감염 CagL 재조합 단백질 접종군의 혈청내 IgG 항체형성이 유의적으로 증가하였다. 위에서의 집락화된 세균수는 두군의 차이가 없었다. 저빌 체중에 대한 위무게 비율는 H. pylori 감염 CagL 재조합 단백질 접종군이 유의적으로 감소하였으나 병리조직학적 평가에서는 유의적인 차이는 확인하지 못하였다. 위에서의 IL-$1{\beta}$와 KC (IL-8 homologues)의 유전자발현 정도도 두 군사이에 유의적인 차이는 없었다. 이번 결과는 CagL 재조합 단백질의 접종은 IgG 항체형성은 효과적으로 자극하였지만 면역화된 숙주에서 세균 집락화의 감소 및 병변형성의 방어까지는 유도하지 못한 것으로 나타났으며, 앞으로 H. pylori 감염에 대해 유효한 면역 반응 및 질병 방어 효과를 나타내기 위해서 CagL을 포함한 다른 종류의 재조합 항원 사용 및 보조적으로 전신 면역 및 점막 면역을 효과적으로 유도하기 위해 안정성있는 애쥬번트의 사용을 고려해야 할 것으로 사료된다. Helicobacter pylori is an important factor of chronic gastritis, digestive ulcer, and stomach cancer. CagL, a virulence factor of H. pylori, is well-known as a pilus protein which acts as adhesion to host cell and a component of Type 4 secretion system. In this study, we evaluated the protective response of recombinant CagL protein (rCagL) using Mongolian gerbil animal model for H. pylori infection. The cagL gene was cloned from 26695 H. pylori followed by over-expression and purification of the protein in E. coli. Mongolian gerbils were immunized with rCagL protein mixed with aluminum adjuvant via intramuscular injections once a week during 4 weeks. At a week after the last immunization, the Mongolian gerbils were administrated with H. pylori 7.13 strain into the stomach and sacrificed to measure antibody titer on rCagL by ELISA and bacterial colonization in the stomach, and to examine the histopathological changes and cytokine expression at 6 week after challenge. Antibody titers on recombinant protein were significantly increased from a week after the first immunization. There was no significant change of the number of bacterial colony between control group and immunized group. The relative stomach weight was significantly decreased in immunized group, but the significant change of histopathological assessment was not observed in the stomach. Cytokine expression such as IL-$1{\beta}$ and KC also was not significantly different between control and immunized groups. These results indicate that rCagL could effectively induce the formation of the specific IgG antibodies. However, bacterial colonization and histopathological lesions could not be inhibited by the immunization in the stomach, indicating not enough protection against H. pylori infection. We consider that along with CagL other adequate antigens could be needed stimulating immune response and inducing protective effects against gastric disease, and also a better adjuvant could be considered.
박은정,채찬희,Bak, Eun-jung,Chae, Chan-hee 대한수의학회 1995 大韓獸醫學會誌 Vol.35 No.2
Basal cell tumors from 53 dogs were examined histologically and classified as basal cell adenoma(n=44), granular basal cell adenoma(n=1), basal cell carcinoma(n=3), basosquamous cell adenoma(n=1), basosquamous carcinoma(n=4) on the basis of malignancy and squamous metaplasia. Female was twenty three and male was thirty. Affected dogs are usually 5.6 years and sex predilection have not been seen. None of tumor examined was metastasized into other organs. Distinct patterns identified in the basal cell adenoma are solid(n=15), cystic(n=13), adenoid(n=8), medusa(n=5) and ribbon(n=3). Solid basal cell adenoma is common type in benign basal cell adenoma. Only one neoplasm was granular basal cell adenoma which was characterized by eccentric nucleus and abundant granular cytoplasm. Basal cell carcinoma showed anaplastic appearance histologically and had atypical basaloid epithelial cells and multinucleated giant cells with moderate number of mitotic figures. Both basosquamous cell adenoma and carcinoma had squamous metaplasia.
박은정,김대용,김재훈,권오경,Bak, Eun-jung,Kim, Dae-yong,Kim, Jae-hoon,Kweon, Oh-hyeong 대한수의학회 1996 大韓獸醫學會誌 Vol.36 No.2
A case of necrotizing meningoencephalitis was described in a 3-year-old female pug dog. Significant histopathological changes were confined to the brain. Histopathologically, brain lesion of the pug dog was characterized by (1) severe lymphocyte, plasma cell and macrophage infiltration in the meninges and gray and white matter of the cerebral hemisphere, (2) perivascular mononuclear cell cuffing and (3) focal polioencephalomalacia in the gray matter of cerebrum. Replicate sections of the brain and lung were negative for fungal infection, canine distemper and toxoplasmosis on PAS stain and immunohistochemistry, respectively. Based on clinical signs, histopathological and immunohistochemical studies, this case was diagnosed as necrotizing meningoencephalitis in pug dog that had not been preriously described in Korea.
국내 분리 토끼출혈병 바이러스(RHDVa)를 감염시킨 토끼 간장에서의 경시적인 초미세구조 변화와 apoptosis
박중원,전지은,박은정,김한,이명헌,황의경,김재훈,이중복,우계형,Park, Jung-Won,Chun, Ji-Eun,Bak, Eun-Jung,Kim, Han,Lee, Myeong-Heon,Hwang, Eui-Kyung,Kim, Jae-Hoon,Lee, Chung-Bok,Woo, Gye-Hyeong 대한수의학회 2013 大韓獸醫學會誌 Vol.53 No.1
In this study, to understand the pathogenesis of new rabbit hemorrhagic disease virus (RHDVa) serotype, we carried out to administrate RHDVa to rabbits, and to examine sequential electron microscopic changes and relationship between pathogenesis and apoptosis. TUNEL-positive cells began to be observed from 24 hours after inoculation (HAI) and the number of positive cells was slightly increased with the course of time. Whereas marked increase of positive cells was seen in the liver from the rabbits died acutely. Typical viral particles with cup-like projections and a diameter of 30~40 nm were detected in homogenized liver samples and tissues at 36 and 48, and 48 HAI, respectively. Ultrastructurally, glycogen deposition was observed from the first stage of hepatocellular degeneration by RHDVa infection and then, swelling and disruption of cristae of mitochondria by viral particles, swelling of smooth endoplasmic reticulum, vacuoles and vesicles were detected. Condensation, margination and fragmentation of chromatin were observed in degenerative hepatocytes at 36 and 48 HAI, indicating apoptotic bodies. These data offer that hepatocytic apoptosis by RHDV infection could be closely related with mitochondrial impairment in the hepatocytes.
이동은 ( Dong Eun Lee ),김지혜 ( Ji Hye Kim ),신동하 ( Dong Ha Shin ),차정헌 ( Jeong Heon Cha ),박은정 ( Eun Jung Bak ),유윤정 ( Yun Jung Yoo ) 한국치위생과학회 2015 치위생과학회지 Vol.15 No.3
Odontoclasts and osteoclasts resorb tooth root and alveolar bone, respectively. Many studies have focused on osteoclast formation in periodontitis, but effect of periodontitis on odontoclast formation is not clearly clarified. In this study, we observed formation of odontoclasts as well as osteoclasts in rats with ligature-induced periodontitis. To induce periodontitis, ligatures were placed around the first molars in left mandibles of rats. Rats were sacrificed at days 1, 3, and 10 after ligation. After tartrate resistant acid phosphatase (TRAP) staining in mandible section, the number of TRAP-positive odontoclasts and osteoclasts were histologically counted along the root and the alveolar bone surfaces of tooth, respectively. Odontoclasts increased until day 10 in mesial and furcation root surface, but did not increase in distal root surface. When compared odontoclast formation to osteoclast formation in mesial surface, osteoclasts peaked at day 3, and then decreased gradually, whereas odontoclasts were continuously increased until day 10. The number of odontoclasts was lower than that of osteoclasts before and after periodontitis induction. These indicate that periodontitis increased formation of odontoclasts as well as osteoclasts, but odontoclast formation occurs slower and weaker than that of osteoclasts.
개 유선종양의 병리학적 관찰 및 유선 혼합종양의 연골기원에 관한 면역조직화학적 연구
김용백,서일복,김재훈,박은정,김대용,한정희,Kim, Yong-baek,Seo, Il-bok,Kim, Jae-hoon,Bak, Eun-jung,Kim, Dae-yong,Han, Jeong-hee 대한수의학회 1997 大韓獸醫學會誌 Vol.37 No.4
Sixteen mammary gland tumors were collected from Seoul National University and Kangwon National University. The average age of the bitches with mammary gland tumor was 10 years. Total 17(60.7%) out of 28 tumor masses observed in 4th and 5th glands. Classification of these tumors according to Hampe and Misdorp were simple adenoma, complex adenoma, benign mixed tumor, papillary adenocarcinoma, solid adenocarcinoma and malignant mixed tumor. Immunohistochemical reaction of the intermediate filaments against normal canine mammary gland showed as followed; anti-cytokeratin 18 was strong and anti-cytokeratin 14 was moderate to the luminal epithelium. Anti-cytokeratin 14 and anti-pancytokeratin to the myoepithelium were showed strong, but anti-vimentin was weak in reactivity. Anti-vimentin to the interstitial cells was represented strong reactivity. The origin of cartilage in mixed tumor of canine mammary gland was studied immunohistochemically with antibodies against intermediate filament. In mammary gland mixed tumors, cartilage tumor tissues were surrounded with the irregularly demarcated three zones composed of adjacent star shaped cells in myxoid areas, proliferative spindle shaped cells and basal located proliferated cells. From basal proliferated cells to star shaped cells, the immunohistochemical reactivity of myoepithelium specific anti-pancytokeratin was decreased gradually and the reactivity of interstitial cell specific anti-vimentin was increased gradually. Based on these immunohistochemical staining patterns, we suggested that the origin of cartilagenous components in canine mammary gland mixed tumor is most likely to the proliferation and metaplsia of myoepithelium.
새로운 국내 분리 토끼출혈병바이러스(RHDVa)를 감염시킨 토끼에서의 경시적인 병리학적 변화와 조직 내 바이러스 항원 분포
박중원,전지은,양동군,박은정,김한,이명헌,황의경,이중복,우계형,Park, Jung-Won,Chun, Ji-Eun,Yang, Dong-Kun,Bak, Eun-Jung,Kim, Han,Lee, Myeong-Heon,Hwang, Eui-Kyung,Lee, Chung-Bok,Woo, Gye-Hyeong 대한수의학회 2012 大韓獸醫學會誌 Vol.52 No.2
Rabbit hemorrhagic disease is a highly acute and fatal viral disease caused by rabbit hemorrhagic disease virus (RHDV). Since first outbreak in Korea 1987, RHDV has been continually affected in the country, but the pattern of outbreak seem to be changed. In this study, to understand the pathogenesis of the new RHDVa serotype, we therefore carried out to inoculate RHDVa to rabbits, and to examine the sequential histopathologic changes and viral distribution. Macroscopically, various sized dark red or white spots or appearance were observed in the liver, lung, kidney uterus and ureter. In euhanized rabbits, significant pathologic findings such as infiltration of heterophils and mononuclear cells were observed at 24 hours after inoculation (HAI), and these were sequentially extended periportal to centrilobular area. However, in dead rabbits, severe hepatic degeneration and/or necrosis with relatively weak inflammatory responses were observed. RHDV antigens began to detect in liver, spleen, and lung from 12 HAI by PCR. Immunohistochemically, RHDV positive cells were seen in only liver from 24 HAI, and the degree of immunogen reactivity was stronger in dead rabbits than in euthanized ones. In conclusion, RHDVa caused the subacute or chronic infection accompanying low mortality and moderate to severe inflammatory reaction in rabbits, suggesting the possibility that RHD could become endemic.