• Parasites, Hosts and Diseases
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• 제34권4호
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• pp.255-258
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• 1996

닭에 있어서 닭와포자충의 재감염에 대한 방어기전을 이해하기 위하여 장기간에 걸쳐 혈청IgG항체가를 경시적으로 조사하였다. 2일령의 SPF 병아리(Dekalb-Warren, Sex-Sal-Link)에 $5{\;}{\times}{\;}10^5$개 그리고 IgG가가 정상으로 복귀한 다음 10일인 340일령에 $1{\;}{\times}{\;}10^7$개의 닭와포자충의 오오시스트를 경구투여하고 나서 채혈하여 간점형광항체법을 이용하여 혈청IgG 항체가를 측정하였다. IgG 가는 감염후 3일부터 38일까지는 대조군과 차이가 없었으나 45일부터 1:21.33으로 상승하기 시작하여 120일에 1:512로 최고치에 이른다음 점점 떨어져 330일에 1:4로 정상치에 이르렀다. 도전 감염후 7, 14, 28일에 각각 1:16, 1:512, 1:1.024로서 점점 높아져 정점에 이른다음 급격히 떨어져 63일에 정상으로 복귀하였다. 한편, 대조군의 항체가는 모두 1:2~1:4의 범위에 있었으며 이 원충의 도전감염전 및 후에 있어서 분변으로부터 집충시켜 제작한 Kinyoun 항산염색표본에서도 오오시스트를 전혀 검출할 수 없었다. 분변내의 오오시스트의 소실 시기와 최초 감염에 있어서 IgG가의 피이크는 일치하지 않았다.

• Biomolecules & Therapeutics
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• 제25권5호
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• pp.535-544
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• 2017

Carnosol is a phenolic antioxidant present in rosemary (Rosmarinus officinalis). It is known for anti-inflammatory effects, analgesic activity and anti-cancer effects. However, no study has been dedicated yet to its effect on atopic dermatitis (AD). Here, we show that carnosol effectively inhibited LPS-induced nitric oxide (NO) generation and expression of inflammatory marker proteins (iNOS and COX-2) in RAW 264.7 cells. In addition, carnosol effectively inhibits the phosphorylation of STAT3 and DNA binding activity in RAW 264.7 cells. Pull down assay and docking model analysis showed that carnosol directly binds to the DNA binding domain (DBD) of STAT3. We next examined the anti-atopic activity of carnosol ($0.05{\mu}g/cm^2$) using 5% Phthalic anhydride (PA)-induced AD model in HR1 mice. Carnosol treatment significantly reduced 5% PA-induced AD like skin inflammation in skin tissues compared with control mice. Moreover, carnosol treatment inhibits the expression of iNOS and COX-2 in skin tissue. In addition, the levels of $TNF-{\alpha}$, $IL-1{\beta}$, and Immunoglobulin-E in blood serum was significantly decreased in carnosol treated mice compared with those of 5% PA treated group. Furthermore, the activation of STAT3 in skin tissue was decreased in carnosol treated mice compared with control mice. In conclusion, these findings suggest that carnosol exhibited a potential anti-AD activity by inhibiting pro-inflammatory mediators through suppression of STAT3 activation via direct binding to DBD of STAT3.

• Journal of Microbiology and Biotechnology
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• 제11권6호
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• pp.1093-1098
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• 2001

The gene coding for a Lepidoptera-specific insecticidal crystalline (or control) protein (ICP), recognized as cryIAc, from Bacillus thuringiensis subsp. kurstaki HD-73, was cloned into the vector pBluscript ll SK-, and then transformed in Escherichia coli $DH5{\alpha}$. The clone was named EBtIAc and the chimeric phagemid, as pEBtIAc. Hyperexpression of CryIAc protoxin was observed in the extract of the culture of E. coli harboring pEBtIAc. Crystalline protoxin was purified by differential solubility. It was dissolved in alkaline pH, and exposed to trypsin to be activated. The molecular weights of the pro- and activated toxins on SDS-PAGE were estimated to be ca. 130 kDa and 60 kDa, respectively. The toxicity was tested by force-feeding larvae of gypsi moth (Lymantria diapar) with trypsinized protoxin. Using the batch of biologically active form of the toxin as an immunogen, anti-CryIAc antiserum was raised in a New Zealand white rabbit. Immunoglobulin G was fractionated from the seam by Protein-A sepharose affinity chromatography. Immunoreactivity of the antibody was examined by dot and Westerns blottings. It has been found that the anti- CryIAc antibody recognized the purified toxin at a level below a nanogram in terms of quantity. Using the antibody some of Bt-corns were able to be differentiated from tons of corn kernels which were imported from America as forage crops.

• Journal of Microbiology and Biotechnology
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• 제21권5호
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• pp.529-535
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• 2011

Single-chain variable fragment (scFv) is a fusion protein of the variable regions of the heavy (VH) and light (VL) chains of immunoglobulin, connected with a short linker peptide of 10 to about 20 amino acids. In this study, the scFv of a monoclonal antibody against the third domain of human CD4 was cloned from OKT4 hybridoma cells using the phage display technique and produced in E. coli. The expression, production, and purification of anti-CD4 scFv were tested using SDS-PAGE and Western blot, and the specificity of anti-CD4 scFv was examined using ELISA. A 31 kDa recombinant anti-CD4 scFv was expressed and produced in bacteria, which was confirmed by SDS-PAGE and Western blot assays. Sequence analysis proved the ScFv structure of the construct. It was able to bind to CD4 in quality ELISA assay. The canonical structure of anti-CD4 scFv antibody was obtained using the SWISS_MODEL bioinformatics tool for comparing with the scFv general structure. To the best of our knowledge, this is the first report for generating scFv against human CD4 antigen. Engineered anti-CD4 scFv could be used in immunological studies, including fluorochrome conjugation, bispecific antibody production, bifunctional protein synthesis, and other genetic engineering manipulations. Since the binding site of our product is domain 3 (D3) of the CD4 molecule and different from the CD4 immunological main domain, including D1 and D2, further studies are needed to evaluate the anti-CD4 scFv potential for diagnostic and therapeutic applications.

• 한국약용작물학회지
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• 제15권2호
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• pp.132-137
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• 2007

Inhibitory effect of Scutellaria baicalensis ethanol extracts (SR) on chemical mediator release and immunoglobulin (Ig) production from Sprague-Dawley rats originated cells as type I allergic reaction was examined. SR showed concentration-dependent inhibition on basal and concanavalin A (ConA)-stimulated Ig production. In the mesenteric lymph node lymphocytes, the inhibitory effect of SR on the IgE production in the presence of Con A was stronger than these on IgA and IgG production. Moreover, tumor necrosis factor-alpha $(TNF-{\alpha})$ production-inhibiting effect of SR in the presence ConA was observed. However, SR did not affect the production of $interferon-{\gamma}$. SR also inhibited histamine release from the peritoneal exudate cells stimulated with a calcium ionophore A23187. In the case of leukotriene B4, SR markedly inhibited it at the concentration of 100 mg/ml. From these results, ethanol extracts obtained from Scutellaria baicalensis may have an anti-allergic effect on the intestinal system of rats.

• Preventive Nutrition and Food Science
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• 제22권1호
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• pp.9-15
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• 2017

Vaccinium angustifolium, commonly known as the lowbush blueberry, is a rich source of flavonoids, with which various human physiological activities have been associated. The present study focuses on the investigation of the effect of the methanolic extract of V. angustifolium root extract (VAE) on high affinity immunoglobulin E receptor ($Fc{\varepsilon}RI$) ${\alpha}$ chain antibody (CRA-1)-induced allergic reaction in human basophilic KU812F cells. The total phenolic content of VAE was found to be $170{\pm}1.9mg$ gallic acid equivalents/g. Flow cytometry analysis revealed that the cell surface expression of $Fc{\varepsilon}RI$ was suppressed in a concentration-dependent manner upon culture with VAE. Reverse-transcriptase polymerase chain reaction analysis showed that the mRNA level of the $Fc{\varepsilon}RI$ ${\alpha}$ chain was reduced in a concentration-dependent manner as a result of VAE treatment. Western blot analysis revealed that the protein expression of $Fc{\varepsilon}RI$ and the phosphorylation of extracellular signal-regulated kinases (ERK) 1/2 were concentration-dependently inhibited by VAE. We determined that VAE inhibited anti-CRA-1-induced histamine release, in addition to the elevation of intracellular calcium concentration ([$Ca^{2+}$]i), in a concentration-dependent manner. These results indicate that VAE may exert an anti-allergic effect via the inhibition of calcium influx and histamine release, which occurs as a result of the down-regulation of $Fc{\varepsilon}RI$ expression through inhibition of ERK 1/2 activation.

• Preventive Nutrition and Food Science
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• 제11권1호
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• pp.54-60
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• 2006

The immunomodulatory effect of a new herbal preparation, HemoHIM, on the recovery from leukopenia induced by cyclophosphamide treatment was investigated. The HemoHIM was made up with an addition of the ethanol-insoluble fraction to the total water extract of Angelica Radix, Cnidii Rhizoma and Paeonia Radix. Daily oral administration of 100 mg/kg BW or 500 mg/kg BW HemoHIM accelerated the recovery from cyclophosphamide-induced leukopenia. HemoHIM increased the number of leukocytes and lymphocytes in the peripheral blood when compared with the cyclophosphamide-treated control. Moreover, the suppressed natural killer (NK) cell activity and interferon $(IFN)-{\gamma}$ secretion in the cyclophosphamide-treated mice were restored by the administration of HemoHIM. HemoHIM significantly reduced the abnormally heightened ratio of interleukin $(IL)-4/IFN-{\gamma}$ and immunoglobulin (Ig)E/IgG2a in the cyclophosphamide-treated mice. These results suggest that HemoHIM accelerates the recovery from leukopenia and alleviates the imbalanced T helper (Th)l/Th2 responses in the cyclophosphamide-treated mice. Additionally, HemoHIM was found to stimulate normal splenocytes to secrete not only Thl type cytokines such as $IFN-{\gamma}$ and IL-2, but also Th2 type cytokine IL-4. In conclusion, our results show that HemoHIM certainly has an influence on the balanced recovery of immune cells and the activation of their activities in the cyclophosphamide-treated mice.

• 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.

• 식물조직배양학회지
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• 제21권6호
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• pp.353-356
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• 1994

사람 B형 간염 바이러스의 pre-S2 표면항원에 결합하는 키메라 항체 유전자(카파 및 감마사슬의 cDNA클론)를 식물체에서 발현시키기 위해 식물체 발현벡터인 pBKS-1에 XbaI 자리를 이용하여 클로닝하였다. 이들 유전자를 포함하는 대장균의 플라스미드 핵산을 추출하여 아그로박테리움에 형질전환 시켰다. 다음 담배의 조직절편과 아그로박테리움을 공동배양함으로써 담배의 형질 전환을 시도하였다. 카나마이신이 포함된 신초유기배지에서 나온 신초를 시료로 하여 Western blot을 실시함으로써 이들 유전자가 형질전환 담배에서 안정하게 발현됨을 확인하였다.

• Pediatric Infection and Vaccine
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• 제17권2호
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• pp.177-181
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• 2010

KD의 급성 열성기에 드물지만 MAS가 발생할 수 있다. 저자들은 19개월 여아의 KD 치료 중에 발생된 MAS를 경험하였다. 환아는 IVIG과 steroid 치료를 받았지만, 발열과 간비종대는 지속되었고 골수생검에서 혈구탐식구증이 관찰되었다. KD에 동반된 MAS를 인지하여 적기에 치료하기 위해서는 임상양상과 치료반응을 토대로, MAS의 가능성을 고려하는 것이 필요하다.