• Allergy, Asthma & Immunology Research
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• 제10권6호
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• pp.698-715
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• 2018

Purpose: Hrd1 has recently emerged as a critical regulator of B-cells in autoimmune diseases. However, its role in the pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP) remains largely unexplored. This study aimed to examine Hrd1 expression and B-cell accumulation and their possible roles in CRSwNP. Methods: Quantitative real-time polymerase chain reaction, immunohistochemistry, enzyme-linked immunosorbent assay and Western blotting were used to assess gene and protein expression in nasal tissue extracts. Cells isolated from nasal tissues and peripheral blood mononuclear cells were characterized by flow cytometry. Local antibody production was measured in tissue extracts with a Bio-Plex assay. Additionally, changes in Hrd1 expression in response to specific inflammatory stimuli were measured in cultured dispersed polyp cells. Results: Nasal polyps (NPs) from patients with eosinophilic CRSwNP (ECRS) had increased levels of Hrd1, B-cells and plasma cells compared with NPs from patients with non-eosinophilic CRSwNP (non-ECRS) or other control subjects (P < 0.05). The average Hrd1 levels in B-cells in NPs from ECRS patients were significantly higher than those from non-ECRS patients and control subjects (P < 0.05). NPs also contained significantly increased levels of several antibody isotypes compared with normal controls (P < 0.05). Interestingly, Hrd1 expression in cultured polyp cells from ECRS patients, but not non-ECRS patients, was significantly increased by interleukin-$1{\beta}$, lipopolysaccharide and Poly(I:C) stimulation, and inhibited by dexamethasone treatment (P < 0.05). Conclusions: Differential Hrd1 expression and B-cell accumulation between the ECRS and non-ECRS subsets suggests that they can exhibit distinct pathogenic mechanisms and play important roles in NP.

• IMMUNE NETWORK
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• 제3권1호
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• pp.1-7
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• 2003

Type II collagen (CII), major component of hyaline cartilage, has been considered as an auto-antigen in rheumatoid arthritis (RA). However, the clinical and biological significances with regard to the CII autoimmunity need to be clarified in human RA. The presence of antibodies to CII has been identified in sera, synovial fluid, and cartilage of patients with RA. In our study, the increased titer of IgG anti-CII in sera was well correlated with C-reactive protein, suggesting that this antibody may reflect the inflammatory status of RA. The titer of anti-CII antibodies (anti-CII Abs) tended to be higher in early stages of diseases. In our extending study, among 997 patients with RA, 269 (27.0%) were positive for circulatory IgG antibody to CII, those levels were fluctuated over time. It is hard to assess the significant amount of T cell responses to CII and CII (255~274) in RA. By using a sensitive method of antigen specific mixed lymphocyte culture, we can detect the presence of CII-reactive T cells in peripheral blood mononuclear cells of RA patients. Sixty seven (46.9%) of 143 patients showed positive CII reactive T cell responses to CII or CII (255~274). The frequencies of CII reactive T cells were more prominent in inflamed synovial fluid (SF) than in peripheral blood. These T cells could be clonally expanded after consecutive stimulation of CII with feeding of autologous irradiated antigen presenting cells (APC). Moreover, the production of Th1-related cytokine, such as IFN-${\gamma}$, was strongly up-regulated by CII reactive T cells. These data suggest that T cells responding to CII, which are probably presenting the IFN-${\gamma}$ producing cells, may play an important role in the perpetuation of inflammatory process in RA. To evaluate the effector function of CII reactive T cells, we investigated the effect of CII reactive T cells and fibroblasts-like synoviocytes (FLS) interaction on the production of pro-inflammatory cytokines. When the CII reactive T cells were co-cultured with FLS, the production of IL-15 and TNF-${\alpha}$ from FLS were significantly increased (2 to 3 fold increase) and this increase was clearly presented in accord to the expansion of CII reactive T cells. In addition, the production of IFN-${\gamma}$ and IL-17, T cell derived cytokines, were also increased by the co-incubation of CII reactive T cells with FLS. We also examined the impact of CII reactive T cells on chemokines production. When FLS were co-cultured with CII stimulated T cells, the production of IL-8, MCP-1, and MIP-1${\alpha}$ were significantly enhanced. The increased production of these chemokines was strongly correlated with increase the frequency of CII reactive T cells. Conclusively, immune response to CII was frequently found in RA. Activated T cells in response to CII contributed to increase the production of proinflammatory cytokines and chemokines, which were critical for inflammatory responses in RA. The interaction of CII-reactive T cells with FLS further augmented this phenomenon. Taken together, our recent studies have suggested that autoimmunity to CII could play a crucial role not only in the initiation but amplification/perpetuation of inflammatory process in human RA.

• 대한치과교정학회지
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• 제33권6호
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• pp.453-463
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• 2003

본 연구는 MC3T3El 조골세포가 저산소증에 반응하여 유발될 수 있는 세포 고사조절 기전을 구명하고자 함에 목적이 있다. $2\%$ 저산소증의 조건하에서 MC3T3El 조골세포는 DNA 사다리 분절 헝성을 보였으며 형광성 염료인 Hoechst 33258로 염색된 핵 구조 형태 관찰시 시간이 지남에 따라 세포고사 현상을 관찰할 수 있었다 Pancaspase 억제제인 Z-VAD-FMK나 특정한 caspase-3 억제제인 Z-DEVD-CHO로 사전 처치하였을 경우에는 저산소증에 의한 DNA 사다리 분절형성이 농축에 비례하여 억제되었다. caspase-3류의 프로테아제(DEVDase) 활성 증가가 세포고사 중에 관찰되었으나 caspase-1 (YVADase)의 활성은 없었다. 어떤 caspase가 세포고사에 관여하는지를 확인하기 위하여 anti-caspase-3 또는 anti-caspase-6의 항체를 이용한 western blotting이 시행되었다. caspase-3의 활성산물에 해당하는 17-KDa단백질과 caspase-6의 활성산물인 20-KDa 단백질이 세포용해물에서 발생되었다. 또한 시간 경과와 더불어 caspase-6의 활동의 상징인 Lamin A의 분열을 일으켰으며, 사이토크롬 C를 cytosol로 방출하였다. 이로써 저산소증에 의한 조골세포의 고사 과정에 사이토크롬 C의 방출이 포함된 caspase의 활성이 관여한다는 것을 확인할 수 있었다.

• Asian-Australasian Journal of Animal Sciences
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• 제18권10호
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• pp.1445-1450
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• 2005

This experiment was conducted to evaluate the effect of feed withdrawal (F) and heat acclimatization (A) on malebroiler and -layer chickens responses to acute heat stress (AHS) at four weeks of age. Totals of ninety male chicks of broiler or layer type were randomly allocated into 30 pens of grower batteries with raised wire floors. Chicks were subjected to F and A three times a week through the first three weeks of age. At each time, feed withdrawal and heat acclimatization (T = $35^{\circ}C$) lasted for six and four hours, respectively. Feed consumption (FC), body weight (BW), and feed conversion ratio (FCR) were recorded weekly for broiler type chickens only. At four weeks of age, all groups of chickens were exposed to AHS (T = $39{\pm}1^{\circ}C$) for three hours. Before and after AHS challenge, body temperature (Tb), heterophil (H), and lymphocyte (L) counts were recorded, and H/L ratio was calculated. Antibody (Ab) response to sheep red blood cells (SRBC) was assessed from all treatments without being exposed to AHS. Group F of broiler-type chickens weighed less (p<0.05) compared to control group. Also, both A and F groups of broiler-type chickens consumed less (p<0.05) feed when compared to control group. Acute heat stress elevated Tb of all treatment groups, however the increase was more profound (p<0.001) in broiler chicks. Broiler chicks of both A and F groups showed a tendency to have higher (p = 0.08) Tb when compared to control group. Acute heat stress elevated (p<0.001) H/L ratio in both types of chickens. Broiler chicks maintained higher (p<0.001) H/L ratio. Both F and A groups reduced (p<0.01) the level of elevation in H/L ratio compared to control groups of both types of chickens. Neither A nor F group affected the Ab production in response to SRBC. However, there was a tendency towards higher Ab responses in F group when compared to other groups in both types of chickens. Results of the present study demonstrate that previous history of feed withdrawal or episodes of heat exposures improved chicks'physiological withstanding of AHS and a tendency to improved humoral immune response.

• 한국임상수의학회지
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• 제21권1호
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• pp.20-22
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• 2004

개의 아토피성 피부염의 주된 세균성 감염은 Staphylococcus intermedius에 의해 발생한다. 본 연구의 목적은 아토피성 피부염을 가지고 있는 환견의 혈청과 정상견의 혈청을 이용하여 체액성 면역반응을 유발하는 주된 세균단백질을 규명하는데 있다. 건국대학교 수의과대학 부속동물병원에 내원한 아토피성 피부염 및 표재성 세균성 농피증을 앓고 있는 환견의 혈청 및 정상견의 혈청을 분리하여 본 실험에 사용하였으며 아토피성 피부염을 가지고 있는 환견에서 S. intermedius 균을 분리하였다. Brain heart infusion 액체배지 조건 및 $37^{\circ}C$ 배양조건에서 호기상태로 증균시켰으며 증균후 포도상구균을 PBS완충액에 부유시킨후 원심분리하고 최종적으로 lysostaphin을 이용하여 세균단백질을 분리하였다. 수거한 세균단백질은 SDS-PAGE을 이용하여 단백질을 전기영동하였으며 영동후 nitrocellulose membrane을 이용하여 단백질을 이동시켰다. Western blot은 anti-dog-IgG, 아토피성 피부염의 혈청 및 정상혈청을 이용하였다. 결론적으로 아토피성 피부염의 혈청을 이용해 확인한 S. intermedius의 주요 세균단백질은 18, 31, 75, 및 110 kDa이였다. 현재의 연구결과로 볼 때 아토피성 피부염에 감염된 대부분의 환견은 S. intermedius의 세균단백질에 대한 체액성 면역반응이 유발된다고 생각된다.

• 대한수의학회지
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• 제34권2호
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• pp.307-313
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• 1994

We have previously shown that crude water extract of Euonymus alatus (EA) had strong prophylactic effect against chemically induced-and tumor cell implanted-cancer, and that the mechanisms responsible for its antitumor effects were due to nonspecific enhancement of the NK cell activities and the cell mediated immunity. However, it was unknown that any components of crude extract did work so, since it consisted of several components. In this paper, we fractionated the crude watar EA-extract into several fraction such as hexane-, ethylether-, ethyl acetate-, n-butanol- and water soluble-fraction, and screened the immune regulating activities of each fraction by the evaluation of lymphokine production and activated lymphocyte proliferation. As a result of the component fraction of EA-extract, it was found that n-butanol fraction was a potent immunostimulator, and the remained water soluble fraction also contained some stimulator, But, other fraction did not showed any remarkable effect. It is therefore suggested that EA-glycosides in n-butanol fraction may be new one of the potent biological response modifiers. The present study was also undertaken in an efforts to investigate the effects of elm-bark(EB, Ulmus clavidiana var japonica), which has been used for curing ulcer and inflammation as a folk medicine without any kind of experimental evidence to support this, on the cellular- and humoral-immune responses, lymphocyte function and NK cell activities in mice. Regardless of time and duration of EB-treatment, Arthus reaction and antibody response to SRBC were not modified by EB, but delayed hypersensitivity to SRBC was significantly enhanced only when EB was treated prior to SRBC-sensitization. EB slightly inhibited the proliferation responses of splenocytes to PHA-stimulation, but it significantly augmented the responses of these cells to S aureus Cowan 1 and Con A-activation, and these effects were manifested only when EB was added at culture initiation. EB did not influence Ig secretion of spleen cells but it significantly augmented the Con A-induced 1L 2 and MIF production of splenocytes. NK cell activities of splenocytes were markedly riled when effector cells were pretreated with EB and this augmentation was dine to the increase of binding affinity of effector cells to target cells and the target cell lytic activities of effector cells. These results led to the conclusion that EB triggers increase of cellular immune responses, such as delayed hypersensitivitiy, lymphokine production and NK cell activities. Also these results suggested that EB contains potent immune stimulants, which may provide the rational basis for their therapeutic use as one of the new biological response modifiers.

• 대한한방소아과학회지
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• 제21권2호
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• pp.69-88
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• 2007

Objectives In this study, we investigated the clinical effect of Imoyongsutang and Imoyongsutang plus Maduryong on the viscosity of mucin solution, the instantly type allergy, the delayed type allergy, the carbon clearance, the pulmonary thromboembolism for the lung damaged rats and mice. Methods The gastric mucin and incubation time, pulmonary thromboembolism induced by sodium arachidonic acid, the pulmonary thromboembolism induced by ADP, vascular permeability response, non inhibitory effects, the delayed type hypersensitivity response to picryl chloride, serum $Na^+$ level, $K^+$ and $Cl^-$ level, ${\alpha}-index$ in phagocytic activity were measured. Results 1. Both the solid extracts of Imoyongsutang and Imoyongsutang plus Maduryong gave some high significance results on the gastric mucin and incubation time on the viscosity of mucin solution in rats, and both groups had similar result. 2. Both the solid extracts of Imoyongsutang and Imoyongsutang plus Maduryong were revealed feeble effect on the pulmonary thromboembolism induced by sodium arachidonic acid in mice, and both groups had similar result. 3. Both the solid extracts of Imoyongsutang and Imoyongsutang plus Maduryong were revealed feeble effect on the pulmonary thromboembolism induced by ADP in mice, and after medication, the value was increased than the before one. 4. Both the solid extracts of Imoyongsutang and Imoyongsutang plus Maduryong were recognized. significance on vascular permeability response induced by histamine in rats. And the significance of the Imoyongsutang plus Maduryong is rather higher than that of Imoyongsutang. 5. The extract of Imoyongsutang recognized no significance symptoms on vascular permeability response induced by serotonin in rats, but the solid extract of Imoyongsutang plus Maduryong resulted recognized significance. 6. Both the solid extracts of Imoyongsutang and Imoyongsutang plus Maduryong were revealed non inhibitory effects on the 48 hour homologous PCA in rats provoked by the IgE-like antibody against the egg albumin. 7. Both the solid extracts of Imoyongsutang and Imoyongsutang plus Maduryong were remarkably revealed inhibitory effect on the delayed type hypersensitivity response to picryl chloride in mice. And the significance of the latter is rather higher than that of the former. 8. The solid extract of Imoyongsutang was revealed inhibitory eects on the delayed type hypersensitivity response to SRBC in mice, but thffe solid extract of Imoyongsutang plus Maduryong recognized significance. 9. The solid extract of Imoyongsutang was recognized significance on the lung TBA value of $O_3$ intoxicated rats, but the solid extract of Imoyongsutang plus Maduryong recognized no significance. 10. Both the solid extracts of Imoyongsutang and Imoyongsutang plus Maduryong was recognized significance on serum $Na^+$ level in $O_{3}-intoxicated$ rats. And the significance of the latter is rather higher than that of the former. 11. Both the solid extracts of Imoyongsutang and Imoyongsutang plus Maduryong were revealed non inhibitory effects on serum $K^+$ and $CL^-$ level $O_{3}-intoxicated$ Rats 12. The solid extracts Imoyongsutang was recognized significance on K-index in phagocytic activity in mice, but the solid extract of Imoymgsutang plus Maduryong recognized no significance. 13. The solid extract Imoyongsutang was recognized on significance on ${\alpha}-index$ in phagocytic activity in mice. but the solid extract of Imoyongsutang plus Maduryong recognized significance. Conclusions According to the above findings, it is suggested that the sold extract of Imoyongsutang and Imoyongsutang plus Maduryong were revealed effects on asthma cough or dyspnea caused by the abnormal rising of lung-allergy and throat discomfort so that they retain effectiveness on the instantly and delayed type allergy, the pulmonary thromboembolism and the lung damages in rats and mice.

• 생명과학회지
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• 제30권10호
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• pp.905-911
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• 2020

본 연구에서는 Keyhole limpet hemocyanin (KLH)에 대한 체액성 및 세포성 면역반응 유도에 대한 PAMAM dendrimer G4 (PAMAM)의 증강 효과를 조사하였다. PAMAM을 KLH와 2주 간격으로 2회 피하주사로 면역한 후, KLH에 대한 특이항체를 측정한 결과, KLH+PAMAM 면역 그룹은 KLH만을 단독으로 면역한 그룹에 약 30배이상 높은 유의한 항체가(IgG+IgA+IgM) 상승을 나타냈다. ELISA 분석에 의해 KLH 특이적인 면역글로부린의 isotype을 측정한 결과, PAMAM를 혼합하여 면역함으로서 IgG1, IgG2a, IgG2b, IgG3 및 IgM 항체의 역가가 유의하게 증가하는 것으로 확인되었다. 또한 면역 개시 7주째에 면역동물에 KLH 항원을 피하주사하고 swelling reaction을 통해 세포성 면역반응인 지연형 과민반응(DTH)을 측정한 결과, KLH+PAMAM으로 면역한 그룹에서 KLH만을 단독으로 면역한 그룹에 비해 높은 DTH 유도활성이 관찰되었다. 한편 면역동물의 비장세포를 취하여 in vitro에서 KLH로 재자극한 후 림프구 증식반응과 사이토카인 유도활성을 측정한 결과, PAMAM을 혼합하여 면역한 그룹에서 KLH 단독 면역 그룹에 비해 림프구의 증식반응에 유의하게 증가하였으며, Th1 type (IFN-γ)과 Th2 type (IL-4) 사이토카인의 생성도 모두 상승하는 것으로 확인되었다. 이상의 결과로부터 PAMAM dendrimer는 함께 투여된 항원물질에 의해 유도되는 세포성 면역과 체액성 면역을 상승시키는 활성이 있는 것으로 확인되었으며, 이는 PAMAM dendrimer가 면역 adjuvant로서 응용 가능한 소재임을 입증하는 것이다.

• Tuberculosis and Respiratory Diseases
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• 제52권5호
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• pp.441-452
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• 2002

연구배경 : 아포프토시스를 억제하는 단백질들은 암의 발생, 진행 및 치료 반응에 있어서 중요한 역할을 한다고 알려져 있다. bcl-2는 지금까지 가장 잘 알려진 항아포프토시스 단백질이고 최근에 survivin이라 하는 IAP군과 HSP 등이 새롭게 밝혀졌고 이들은 많은 암종에서 발견되었다. 이에 저자들은 비소세포폐암을 대상으로 survivin, HSP70, 그리고 bcl-2의 발현에 대해서 면역조직화학적 분석을 시행함으로써 임상적 특성과의 관련성을 알아보고자 하였다. 방 법 : 99예의 비소세포폐암 조직으로 변역조직화학적 염색을 시행하였으며 일차 항체로 anti-survivin rabbit polyclonal antibody, anti-HSP70 mouse monoclonal antibody, anti-Bcl-2 mouse monoclonal antibody를 이용하였다. 각각의 단백질 발현과 여러 임상적, 조직학적 지표들과의 연관성은 Chi-square test를 이용하여 비교하였다. 모든 통계적 분석은 SPSS software를 이용하였다. 결 과 : 99예 중 남녀 비는 78 : 21이었고 평균 연령은 56.1세였으며 조직학적 분류는 편평상피암이 42예로 가장 많았고, 병리학적 병기로는 IB기 가 30예로 가장 많았다. Survivin은 33예 (33.3%)에서 발현되었고 여성에서 발현률이 유의하게 높았고 비흡연자에서 발현률이 높은 경향을 보였으며 흡연자 중 흡연양이 증가할수록 발현률은 유의하게 감소하였다. 또한 재발된 환자군에서 survivin은 유의하게 높은 발현률을 보였다. HSP70은 총 99예 중 84예 (84.8%)에서 발현되어 3 가지 단백질 중 가장 높은 빈도를 보였으나 유의한 관련성을 보이는 임상 지표는 없었다. bcl-2는 18예 (18.2%)에서 발현되었고 bcl-2 발현군에서의 재발률이 유의하게 높았고 흡연양이 많을수록 발현률이 감소하는 경향을 보였으나 다른 임상 지표와는 관련성이 없었다. 각 단백질의 발현군과 비발현군 사이에 중간 생존기간을 비교하였으나 통계적 유의성은 없었다. 결 론 : 본 연구에서 survivin 발현은 비흡연자에서 높은 경향을 보이고 여성과 종양이 재발한 군에서 유의하게 높은 발현률을 보이고 bcl-2 발현군에서 재발률이 유의하게 높은 반면 HSP70의 발현은 임상적, 조직학적 지표들과 관련성이 없었다. 결론적으로 발암과정에 중요한 아포프토시스에 관여하는 survivin, HSP 그리고 bcl-2에 대한 보다 광범위한 연구가 필요할 것으로 생각된다.

• 대한치과교정학회지
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• 제24권2호
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• pp.303-317
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• 1994

치아이동시 발생되는 치근 흡수에 관련된 특이면역반응의 역할을 연구하기 위해 치근 흡수에 따른 치근항체의 역가 변화를 관찰하였다. 생후 2년된 성견 다섯마리를 실험동물로 사용하였다. closed coil spring을 사용하여 각 실험동물의 상악 6전치에 200-250gm의 함입력을 가하였으며, 하악 6전치를 발거하여 동종항원으로 사용하였다. 발거된 하악 전치의 상아질 부위를 분리하여 6M Guanidine-HCl-10% EDTA(pH 5.0)에서 해리시킨 다음 부유물 만을 투석하여 항원을 준비하였다. 치아를 함입시키기 전 1회 혈청을 채취하고 함입시키면서 매주 간격으로 11회 형철을 채취하였다. 실험 9주째 치근 흡수가 일어나고 있는 상악 6전치를 모두 발거하여 항원의 근원을 제거하였다. 혈청 내 치근항체의 역가는 ELISA(enzyme-linked immunosorbent assay)로 측정하였다. 항체 특이성에 대한 대조군으로 혈청과 치근 항원 그리고 혈청과 관련없는 박테리아 항원(Porphyro monas gingivalis 33277)을 섭씨 25도에서 3시간 반응시켜 모든 과정에서 항원-항체 반응을 측정하였다. 치근 흡수를 관찰하기 위하여 실험 전과 실험 후 한 달 간격으로 방사선 교합사진을 촬영하였고 9주째 발거한 치아의 치근단 부위를 입체현미경으로 관찰하였다. 치근항원에 대한 자가항원의 역가 변화를 측정하여 다음과 같은 결과를 얻었다. 자가항체의 역가는 치아가 함입되면서 즉시 감소하였다가 1주후 다시 증가하였으며 4주째 최고수준에 도달한 다음 다시 지속적으로 감소하였다. 흡수 중인 치근을 발거한 직후 급격히 증가하는 양상을 보였다. 치근 항원과 반응시킨 혈청내에서는 항원-항체 반응이 거의 나타나지 않았으며 관련없는 박테리아 항원과 반응을 시킨 혈청에서는 활성도가 동종 치근 항원에 대한 자가항체의 활성도와 비슷하게 나타났다. 방사선 교합사진 상에서는 육안으로 구별할 수 있는 차이가 거의 없었으나 입체현미경하에서는 치근단 부위의 다양한 흡수 양상을 관찰할 수 있었다.