• 동의생리병리학회지
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• 제19권5호
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• pp.1243-1250
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• 2005

This study was performed to investigate the effect of oral administration of deer horn against the asthma. Deer horn improves body metabolism and strengthens overall health, especially in elderly persons and young children. Additionally, it stimulates sexual function in females and can stimulate wound healing. Asthma was induced to Balblc mouse by i.p. injection and aerosol immunization with ovalbumin. It was observed the change of the eosinophil number in the BALF. Concentrations of IL-4, IL-5 in BALF and splenocyte were assessed by ELISA, IgG and IgE from serum were calculated by same method. The number of eosinophil in BALF was not significantly changed in deer horn group compared with control group. Concentration of IL-4 in BALF was significantly decreased in deer horn group compared with control group. Levels of IL-5 from BALF and splenocyte were significantly decreased in deer horn group compared with control group, respectively. Concentrations of IgE and IgG in serum were significantly decreased in deer horn group compared with control group, separately. We found that the effect of deer horn extract in asthma was implicated in reductions of IL-4, IL-5 released from Th2 cell, and decreases of IgG, IgE from plasma cell. These findings suggest that deer horn extract can produce anti-asthmatic effect, which may play a role in allergen-induced asthma therapy.

• IMMUNE NETWORK
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• 제22권6호
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• pp.50.1-50.19
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• 2022

Autoreactive B cells are not entirely deleted, but some remain as immunocompetent or anergic B cells. Although the persistence of autoreactive B cells as anergic cells has been shown in transgenic mouse models with the expression of B cell receptor (BCR) reactive to engineered self-antigen, the characterization of naturally occurring anergic B cells is important to identify them and understand their contribution to immune regulation or autoimmune diseases. We report here that a low-level expression of CD138 in the splenic B cells marks naturally arising anergic B cells, not plasma cells. The CD138^int B cells consisted of IgM^lowIgD^high follicular (FO) B cells and transitional 3 B cells in homeostatic conditions. The CD138^int FO B cells showed an anergic gene expression profile shared with that of monoclonal anergic B cells expressing engineered BCRs and the gene expression profile was different from those of plasma cells, age-associated B cells, or germinal center B cells. The anergic state of the CD138^int FO B cells was confirmed by attenuated Ca²⁺ response and failure to upregulate CD69 upon BCR engagement with anti-IgM, anti-IgD, anti-Igκ, or anti-IgG. The BCR repertoire of the CD138^int FO B cells was distinct from that of the CD138^- FO B cells and included some class-switched B cells with low-level somatic mutations. These findings demonstrate the presence of polyclonal anergic B cells in the normal mice that are characterized by low-level expression of CD138, IgM downregulation, reduced Ca²⁺ and CD69 responses upon BCR engagement, and distinct BCR repertoire.

• Toxicological Research
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• 제22권4호
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• pp.357-364
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• 2006

This study was undertaken to develop a reliable mice model demonstrating similar immunologic phenomena as human atopic dermatitis characterized with predominance of type-2 immune response. BALB/C mice and NC/Nga mice were sensitized twice with $100{\mu}l$ of 1% 2,4-dinitrochlorobenzene (DNCB) or vehicle (acetone : olive oil=4:1 mixture) in a week and challenged twice with $100{\mu}l$ of 0.2% DNCB or the vehicle at the following week. Mice were sacrificed at 19 days following the second DNCB or vehicle challenge for NC/Nga mice and at 28 days following the second DNCB or vehicle challenge for BALB/c mice. Upregulation of plasma 1gE, a hallmark of atopic dermatitis occurrence, was evident in the plasma obtained 4 day after the second DNCB challenge from BALB/c mice (approximately 4-fold) and NC/Nga mice (approximately 6-fold) treated with DNCB in comparison with that of the vehicle treated-control mice, and remain higher $3{\sim}4$ week after the second challenge. Ratio of plasma IgG1 versus IgG2a concentration was significantly higher in the mice treated with DNCB than the control mice, which also implies the skewed type-2 reactivity in vivo. Ratio of interleukin-4 versus interferon gamma produced in the splenic T cell culture supernatants was approximately 3-fold higher in the both strains of mice treated with DNCB than their control mice, respectively. The DNCB-treated mice demonstrated atopic dermatitis-like skin legions characterized with erythma, scaling, and hemorrhage, which was not observed with the control mice. Scratching on face or dorsal area was significantly more frequent (approximately 25-fold) in the DNCB-treated mice than the control at next day of the second DNCB challenge, and scratching frequency remains higher (approximately 4-fold) in the mice treated with DNCB than the control at 14 day following the second DNCB challenge. Overall, the mice model developed through sensitization and challenge with DNCB may be useful for research on atopic dermatitis and development of treatment materials for atopic dermatitis.

• Asian-Australasian Journal of Animal Sciences
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• 제18권10호
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• pp.1498-1504
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• 2005

Competitive direct ELISA was developed to detect gentamicin residues. Mice immunized with gentamicin-keyhole limpet hemocyanin (KLH) conjugate developed good antiserum titers, which gradually increased with booster injections, indicating immunization was successfully processed. Monoclonal antibody against gentamicin was prepared using hybridoma cells cloned by limit dilution of fused cells. IgG was purified from ascites fluid of hybridoma cell-injected mice through ammonium sulfate precipitation and Sephadex G-25 gel filtration. After the gel filtration, fractions of high antibody titer were further purified through affinity chromatography on protein A/G column. Monoclonal antibody against gentamicin was confirmed as IgG1, which has kappa light chain. Cross-reactivities ($CR_{50}$) of gentamicin monoclonal antibody to other aminoglycosides (kanamycin, neomycin, and streptomycin) were less than 0.005%, indicating the monoclonal antibody was highly specific for gentamicin. Standard curve constructed through competitive direct ELISA showed measurement range (from 80 to 20% of B/$B_0$ ratio) of gentamicin was between 1 and 40 ng/ml, and 50% of B/$B_0$ ratio was about 4 ng/ml. The gentamicin concentration rapidly increased to 1,300 ng/ml after the intramuscular administration up to 2 h, then sharply decreased to less than 300 ng/ml after 4 h of withdrawal, during which the elimination half-life ($t_{1/2}$) of gentamicin in the rabbit plasma was estimated to be 1.8 h. Competitive direct ELISA method developed in this study using the prepared monoclonal antibody is highly sensitive for gentamicin, and could be useful for detecting gentamicin residues in plasma of live animals.

• Preventive Nutrition and Food Science
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• 제5권4호
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• pp.219-224
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• 2000

Dieary effects of sea tangle on immune functions were investigated in diabetic mice. Four groups of ICR mice weighing 33.36$\pm$1.01 g were fed either an AIN-76 diet only (control), or with additional sea tangle powder, sea tangle water extract, and alginate at the level of 13.6%, 4%, and 1%, respectively by weight. Cellulose was omitted in sea tangle powder and alginate diets. After 10 days of feeding respective experimental diets, all mice were made diabetic by five consecutive intramuscular injections of streptozotocin (40mg/kg body weight per day) and fed the diets for four more weeks. Plasma IgG concentrations but not those of IgM were significantly higher in mice fed sea tangle powder, extract or alginate than those on the control diet. Plasma TNF$\alpha$ levels were, however, lower in those fed sea tangle power or water extract than control and alginate fed groups. TNF$\alpha$ releases from macro phages isolated from four groups and cultured with 5 $\mu\textrm{g}$/mL LPS for 24 hors showed a similar tendency to the results of plasma concentrations in the respective groups, but IL-1$\beta$ releases were not different among four groups. Lymphocyte proliferation in response to LPS (10$\mu\textrm{g}$/mL) measured using splenocytes cultured for 3 days was highest in the alginate fed group (594$\pm$38%) and those of sea tangle powder (536$\pm$47%) and extract (547$\pm$34%) fed groups tended to be higher than the control (523$\pm$30%). It is concluded that sea tangle contains immunomodulatory components besides alginate that could enhance humoral immunity of itself. The immunomodulatory effects of sea tangle constituents is regarded as beneficial for diabetic subjects.

• Journal of Pharmaceutical Investigation
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• 제29권2호
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• pp.87-92
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• 1999

Drug delivery to the brain may be achieved by producing chimeric peptide, attaching the drug to protein 'vectors' which are transported into the brain from the blood by a receptor-mediated transcytosis through the blood-brain barrier (BBB). Since the BBB expresses high concentrations of transferrin receptor, and it was reported that anti-transferrin receptor mouse monoclonal antibody (OX26) undergoes transcytosis through the BBB, it is logical to assume that a drug delivery system via transferrin receptor-mediated transcytosis is a promising strategy. In the present study, therefore, we tested feasibility of several OX26 based vectors for the brain delivery of a model drug. Avidin-based delivery vectors such as OX26-streptavidin (OX26-SA), OX26-neutralite avidin (OX26-NLA) were chemically synthesized vectors and OX26 immunoglobulin G 3 type $C_{H}3$ fusion avidin $(OX26\;IgG3C_H3-AV)$ was genetically engineered. To improve the efficiency of producing chimeric peptide, we used avidin-biotin technology. Pharmacokinetics of $[^3H]biotin$ bound to OX26-SA, OX26-NLA and $OX26\;IgG3C_H3-AV$ was determined by intravenous injection technique, and their stabilities in plasma were analyzed using HPLC. The brain delivery of $[^3H]biotin$ bound to OX26-SA, OX26-NLA and OX26\;$IgG3C_{H}3-AV$ (expressed as %ID/g brain) was $0.22{\pm}0.01$, $0.18{\pm}0.01$ and $0.25{\pm}0.09$, respectively. The areas under the plasma concentration versus time curve (AUC) for OX26-SA, OX26-NLA, $OX26\;IgG3C_H3-AV$ from time zero to 60 min were $209{\pm}10$, $195{\pm}9$, $134{\pm}29\;%ID\;min/ml$ respectively and their total clearances $(CL_{tot})$ were $1.00{\pm}0.09$, $1.08{\pm}0.07$ and $1.54{\pm}0.29\;ml/min/kg$, espectively. These results showed that these vectors possess preferable pharmaceutical (e.g., resonable stability) and pharmacokinetics (e.g., significant brain uptake and enhanced AUC) for brain delivery. Therefore, these vectors may be broadly useful in the brain delivery of drugs that are not transported into the brain to a significant extent.

• Toxicological Research
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• 제18권2호
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• pp.205-213
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• 2002

This study was undertaken to develop a subacute murine model for predicting occurrence of systemic anaphylaxis and to evaluate efficacy of various immunological parameters as the monitoring indices for the occurrence of anaphyalxis. The murine anaphyalxis model was developed through intraperitoneally sensitizing 100 $\mu\textrm{g}$ ovalbumin (OVA) in the presence of 1 mg alum and 300 ng cholera toxin twice a week interval followed by challenging 500 $\mu\textrm{g}$. OVA intravenously. Typical anaphylaxis symptoms were demonstrated at the both BALB/c mice, a strain prone to type-2 response, and C57BL/6 mice. a strain prone to type-1 response. Level of plasma histamine was approximately 50-fold or 30-fold higher in the mice sensitized with OVA than the mice sensitized with alum plus cholera toxin or the saline-treated mice after OVA challenge, respectively. Sensitization and challenge with OVA significantly enhanced plasma leukotriene $B_4$ level but not IgE levels in comparison with the control mice, which indicated the role of leukotriene $B_4$ for progression of anaphyalxis. Furthermore, among mice suffered from anaphylaxis, levels of OVA-specific IgGl were significantly higher in the BALB/c mice than in the C57BL/6 mice, which implied the genetic susceptibility for the induction of systemic anaphylaxis. Conclusively, simultaneous evaluation of histamine, leukotriene $B_4$, and allergen-specific IgG isotype may serve as more efficient monitoring tool for vaccine-related anaphyalxis.

• Journal of Chest Surgery
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• 제37권5호
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• pp.391-400
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• 2004

이종간 장기이식은 즉각적이고 비가역적 초급성 거부 반응을 초래하는데 자연항체인 IgM이 주원인으로 알려져 있다. 따라서 항체를 제거하는 것이 초급성 거부반응을 감소시키는 하나의 방법이 될 수 있다. 이에 연구자들은 돼지\$\longrightarrow$ 개 이소 심장이식 모델에서 자연항체를 제거하기 위해 원심분리 후 혈장분반술(postcentrifugal plasmapheresis-PCPP)을 사용하고 수혜견의 항체 변화와 이종 심장이식 시 생존시간을 관찰하였다. 대상 및 방법: 10∼20 kg의 이계 교배시킨 돼지를 공여돈으로, 25∼30 kg의 잡종개를 수혜견으로 사용하였다. 실험군에 해당하는 수혜견은 COBE TPE plasmapheresis device(COBE Laboratories, Lakewood. CO)로 PCPP를 시행하였다. 심장이식 2일 전과 이식 당일 PCPP를 진행하였다. 대조군은 PCPP를 수행하지 않았다. 그룹 1에서는 PCPP를 1 plasma-volume(PV)을 시행하고, 2일 후 2 PV를 실시하였다. 그룹 2는 2 PV를 시행하고 2일 후 2 PV로 PCPP를 실시하였다. 돼지의 심장은 이소심장 이식법으로 수혜견의 복강내 신하(infrarenal) 대동맥과 하대정맥에 이식하였다. 수혜견 혈장내의 총 IgM과 IgG의 분석은 ELISA방법을 이용하였다. 실험에서 얻은 시료로 혈청 albumin, 전해질, 보체 활성도와 응고 인자의 변화를 관찰하였다. 또한 이식된 심장의 초급성 거부 반응을 확인하기 위해 이식 거부반응을 경험한 심장 절편을 조직 병리학적으로 관찰하였다. 결과: PCPP 방법으로 처리한 그룹 2에서의 혈장내의 총 IsM과 IgG의 제거율은 각각 95.7$\pm$1.2%, 80.5$\pm$2.4%를 나타냈다. 또한 각 그룹에서 혈청 albumin 농도가 감소하는 것을 관찰하였는데 그룹 1에서는 2.8에서 1.4g/㎗, 그룹 2에서는 3.0에서 1.5 g/㎗의 감소를 보였다. 보체 활성도는 PCPP를 시행하면 감소하였다가 PCPP시행 후 약 24시간 후면 정상으로 회복되었다. 2 PV로 2회 시행한 PCPP에서의 보체 활성도는 시행 전의 10% 수준까지 감소되었다. PCPP가 혈액응고인자에 미치는 영향을 보면 2 PV의 PCPP 시행으로 fibrinogen의 농도가 시행 전의 20% 수준 이하로 떨어졌으나 이 효과는 24시간 이내에 회복되었고 antithrombin III도 비슷한 양상을 보였으나 fibrinogen보다 소폭의 감소를 보였다. PT, aPTT는 PCPP도중에 간혹 연장되는 경우가 있었으나 늘 일정한 결과는 아니었으며 이식 후에 측정 가능 범위 이상으로 연장되었다. D-dimer의 경우 PCPP 시행 동안에는 검출되지 않았으나 이식 후 10분부터 검출되어 지속되었다. PCPP를 수행하지 않은 그룹 0 수혜견에 이식한 심장의 생존시간은 5분이었으나 PCPP에 의해 자연항체를 제거 후 이식된 심장의 생존시간은 90분으로 현저한 증가를 보였다. 또한 조직 병리학적으로도 이식 거부에 대한 특징이 그룹 0에 비해 그룹 2에서 지연됨을 확인하였다. 결론: PCPP는 수혜견의 자연항체를 효과적으로 제거하여 초급성 거부반응을 감소시킴으로써 결국 돼지의 이종 이식 전 심장의 생존시간을 증가시켰다.

• Asian-Australasian Journal of Animal Sciences
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• 제33권5호
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• pp.722-731
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• 2020

Objective: Two experiments were conducted using 28 healthy multiparous sows to evaluate the oxidative stress status and reproductive performance of sows during gestation and lactation under different thermal environments. Methods: Fourteen multiparous sows were used in Exp. 1 under a high thermal environment, and the other 14 multiparous sows were used in Exp. 2 under a moderate thermal environment. In both experiments, reproductive performances of sows were recorded. Plasma samples were collected on d 35, 60, 90, and 109 of gestation, and d 1 and 18 of lactation for malondialdehyde, protein carbonyls, 8-hydroxy-deoxyguanosine, immunoglobulin g (IgG), and IgM analysis. Results: For sows in Exp. 1, plasma malondialdehyde concentration on d 109 of gestation tended to be greater (p<0.05) than it on d 18 of lactation. Plasma concentration of protein carbonyl on d 109 of gestation was the greatest (p<0.05) compared with all the other days. Plasma concentrations of 8-hydroxy-deoxyguanosine on d 109 of gestation was greater (p<0.05) than d 18 of lactation in Exp. 1. For sows in Exp. 2, there was no difference of malondialdehyde and protein carbonyl concentration during gestation and lactation. In both Exp. 1 and 2, litter size and litter weight were found to be negatively correlated with oxidative stress indicators. Conclusion: Sows under a high thermal environment had increased oxidative stress during late gestation indicating that increased oxidative damage to lipid, protein, and DNA could be one of the contributing factors for reduced reproductive performance of sows in this environment. This study indicates the importance of providing a moderate thermal environment to gestating and lactating sows to minimize the increase of oxidative stress during late gestation which can impair reproductive outcomes.

• IMMUNE NETWORK
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• 제15권1호
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• pp.27-36
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• 2015

In the present study, we investigated the protection conferred by a live attenuated Salmonella enterica serovar Typhimurium (ST) strain against Salmonella Typhimurium, Salmonella Gallinarum (SG), and Salmonella Enteritidis (SE) infection in layer chickens. Birds were orally primed with the attenuated ST strain at 7 days of age and then boosted at 4 weeks post prime immunization (PPI). Sequential monitoring of plasma IgG and mucosal secretory IgA (sIgA) levels revealed that inoculation with ST induced a significant antibody response to antigens against ST, SE, and SG. Moreover, significant lymphoproliferative responses to the 3 Salmonella serovars were observed in the immunized group. We also investigated protection against virulent ST, SE, and SG strain challenge. Upon virulent SG challenge, the immunized group showed significantly reduced mortality compared to the non-immunized group. The reduced persistence of the virulent ST and SE challenge strains in the liver, spleen, and cecal tissues of the immunized group suggests that immunization with the attenuated ST strain may not only protect against ST infection but can also confer cross protection against SE and SG infection.