• BMB Reports
• /
• 제53권9호
• /
• pp.478-483
• /
• 2020

Kudoa septempunctata is a myxozoan parasite that causes food poisoning in individuals consuming olive flounder. The present study aimed to investigate the currently insufficiently elucidated early molecular mechanisms of inflammatory responses in the intestine owing to parasite ingestion. After Kudoa spores were isolated from olive flounder, HT29 cells were exposed to spores identified to be alive using SYTO-9 and propidium iodide staining or to antigens of Kudoa spores (KsAg). IL-1β, IL-8, TNF-α and NFKB1 expression and NF-κB activation were assessed using real-time PCR, cytokine array and western blotting. The immunofluorescence of FITC-conjugated lectins, results of ligand binding assays using Mincle-Fc and IgG-Fc, CLEC4E expressions in response to KsAg stimulation, and Mincle-dependent NF-κB activation were assessed to clarify the early immune-triggering mechanism. Inflammatory cytokines (IL-1β, GM-CSF and TNF-α), chemokines (IL-8, CCL2, CCL5 and CXCL1) and NF-κB activation (pNF-κB/NF-κB) in HT29 cells increased following stimulation by KsAg. The immunofluorescence results of spores and lectins (concanavalin A and wheat germ agglutinin) suggested the importance of Mincle in molecular recognition between Kudoa spores and intestinal cells. Practically, data for Mincle-Fc and KsAg binding affinity, CLEC4E mRNA expression, Mincle immunofluorescence staining and hMincle-dependent NF-κB activation demonstrated the involvement of Mincle in the early immune-triggering mechanism. The present study newly elucidated that the molecular recognition and immune-triggering mechanism of K. septempunctata are associated with Mincle on human intestinal epithelial cells.

• 대한본초학회지
• /
• 제23권2호
• /
• pp.111-121
• /
• 2008

Objectives : The present study was carried out to investigate the effects of Astragali radix on the asthma treatment. EAR-I is an extract of astragali radix cultured for one year and EAR-III is an extract of astragali radix cultured for three years. Methods : Two asthma-induced groups mice group was treated respectively with EAR- I and EAR- III. The other group was not. Each group was analyzed in vivo and in vitro experiments. Results : In asthma-induced mice by OVA treatment, the weight of lung, total cell number of leukocyte and eosinophil in BALF, both EAR- I and EAR- III treated groups were significantly decreased compared to control group. IL-4, IL-5, IL-13, histamine, IgE in BLAF of group treated with both EAR-I and EM-III were significantly decreased compared to control group. In FACS analyzing, granulocytes, $CD3e^-$/$CCR3^+$, $CD3^+$/$CD19^$, $CD3e^+$/$CD69^+$, $CD4^+$, $CD8^+$ and $GR-1^+$/$CD11b^+$ were significantly increased in asthma induced mice group by OVA treatment compared to control group, and decreased in group treated with both EAR- I and EAR-III. Conclusions : The present data proves that extract of astragali radix has an effect on the inhibiting asthma. Moreover, astragali radix cultured for three years was proved to be superior to the one cultured for one year.

• 한국동물위생학회지
• /
• 제43권2호
• /
• pp.89-97
• /
• 2020

This study was carried out to examine a novel inactivated Salmonella Typhimurium (S. Typhimurium) vaccine candidate for protection of mice against salmonellosis by immunization of BALB/c mice using various type adjuvant. The novel type-inactivated vaccine candidate was constructed by adding Chlorhexidine digluconate solution. BALB/c mice were divided into 6 groups of 15 mice apiece. The mice were intramuscularly (IM) primed at 6 weeks of age and were IM boosted 8 weeks of age. Groups A and B mice were injected with sterile phosphate-buffered saline as controls; group C mice were inoculated with 5×10⁸ cells/100 µL of formalin-inactivated S. Typhimurium cells and adjuvant ISA70; groups D~F mice were immunized with 5×10⁸ cells/100 µL of the inactivated vaccine candidate and adjuvant ISA70, adjuvant IMS1313 and adjuvant IMS1313 containing 30 ㎍/mL of GI24, respectively. All mice (except group A mice) were orally challenged with a virulent S. Typhimurium strain at 10 weeks of age. Mice from groups C-F had significantly increased IgG levels compared to control groups (A-B) mice. The levels of splenocyte IFN-γ and IL-4 in mice of all groups were measured by ELISA, resulting in increased immunity in group F mice compared to those of groups A-E mice. These data suggested that systemic and cell-mediated immune responses were highly induced by IM immunization with the vaccine candidate and adjuvant IMS1313 containing GI24. Furthermore, clinical signs such as death were observed in only 20% of group F mice after virulent Salmonella strain challenge, however, groups B and C (100%), and groups D and E (60%) mice died. This data suggested that mice immunized by intramuscular prime and booster with this vaccine candidate and adjuvant IMS1313 containing GI24 effectively protected mice from salmonellosis.

• Applied Microscopy
• /
• 제30권1호
• /
• pp.45-59
• /
• 2000

항암제로 널리 사용되는 cis-platin은 백금(platinum) 원소에 염소기와 암모니아기가 수평면의 cis-위치에 결합되어 있는 금속화합물이다. Cis-platin은 인체의 종양에 상당한 항암효과가 있어 악성 난소종양, 두경부의 악성상피종양, 방광암 및 자궁경부암에 효과가 있다고 알려져 있으나 이 약제는 세포의 DNA합성을 억제하는 기능이 이미 밝혀져 있다. 저자는 cis-platin을 가임기의 여성에 투여되었을 때 난관상피 세포를 구성하는 섬모세포의 섬모형성에도 필연적으로 손상 및 억제적인 작용이 있을 것으로 생각되어 섬모의 주요 구성분인 미세소관의 $\alpha-tubulin$과 cis-platin과의 관계를 추구하고자 하였다. 실험동물로는 건강한 체중 $150\sim200gm$의 자성흰쥐를 사용하였으며 estradiol benzoate를 4일간 매일 투여함으로써 섬모세포를 난관내에서 지속적인 활성을 유지시킨 뒤 cis-platin을 실험군의 복강내로 주사한 후 1일, 3일, 5일 및 7일 경과시에 각각 실험동물의 난관에서 상피세포내 $\alpha-tubulin$의 발현을 관찰하기 위해 mouse antirat $\alpha-tubulin$ monoclonal antibody와 2차 항체로 bio-tinylated goat anti-rat IgG를 각각 사용하여 면역조직 화학법을 시행한 후 광학현미경으로 관찰하였다. 또 일부조직은 전자현미경 조직절편을 제작하여 1차 항체로 mouse anti rat $\alpha-tubulin$ monoclonal antibody와 2차 항체로 직경 15nm의 금과립을 결합시킨 goat anti-mouse IgG를 사용하여 면역조직 반응을 시행하고 투과전자현미경으로 관찰하여 다음과 같은 결과를 얻었다. 1. Estradiol benzoate를 4일간 매일 일정량을 투여한 흰쥐의 난관섬모세포내 $\alpha-tubulin$의 면역반응은 estradiol 투여후 1일, 3일 및 5일군에서 강한 반응을 나타내었다. 2. Estradiol beozoate를 4일간 투여한 후 cia-platin을 투여한 흰쥐 난관섬모세포내 $\alpha-tubulin$ 반응은 cis-platin 투여 1일군과 3일군에서 약한 반응을 나타내었으나 제 5일군에서는 강한 반응으로 회복되었다. 3. Cis-platin투여한 후 제 1일 및 3일군의 흰쥐 난관 섬모세포내 $\alpha-tubulin$반응은 첨부세포질에서는 감소되었고 기저체, 섬모등에서는 $\alpha-tubulin$반응이 대조군과 비교하면 변동이 없었다. 이상의 결과로 미루어 난관섬모세포내 $\alpha-tubulin$은 cis-platin투여에 의해 감소되는 것으로 결론 지을 수 있었다.