• Title/Summary/Keyword: B3 antibody

Search Result 513, Processing Time 0.03 seconds

Suppressive Effects of Chenilyeomgamibang (CGB) and Chenggihaedok-san (CHS) on TNCB(trinitrochlorobenzene)-induced Atopic Dermtitis NC/Nga Mice Model (TNCB로 유발한 아토피피부염 생쥐 모델에서 천일염가미방(天日鹽加味方)과 청기해독산(淸肌解毒散)의 병용 효과)

  • Lee, Kyoung-Mee;Gim, Seon-Bin;Choi, Hak-Ju;Choi, Jeong-June;Roh, Seong-Soo;Kim, Dong-Hee
    • The Korea Journal of Herbology
    • /
    • v.24 no.4
    • /
    • pp.215-224
    • /
    • 2009
  • Objectives : Atopic dermatitis is a chronic inflammatory disease characterized by typically distributed eczematous skin lesion with pruritus, lichenification and dry skin. In this study, we performed to assess the therapeutic effects of co-treatment of Chenilyeomgamibang (CGB) and Chenggihaedok-san (CHS, C&C) on the TNCB(trinitrochlorobenzene)-induced atopic dermatitis in NC/Nga mice, characterized by the onset of atopic dermatitis along with an increase the number of inflammatory cells and dysregulation of Th2 cytokines. Methods : Defined amount of CGB was sprayed on mice skin and CHS was simultaneously orally administrated to TNCB treated NC/Nga mice for 5 weeks. The immune cell types were caracterized by flow cytometry using each specific antibody. The amount of Th2 cytokines in serum and splenocytes culture supernatant were measured by ELISA. Results : Administration of C&C significantly reduced clinical dermatitis severity including pruritus, edema, eczematous and erythema. Histological findings indicated that the thickening of epidermis/dermis and dermal infiltration of inflammatory cells were dramatically reduced. Flow cytometry analysis showed that infiltrated immune cell numbers of CCR3+, B220+/IgE+, Gr-1+/CD11b+, and CD117+ were significantly reduced in C&C-treated dorsal skin lesion. Furthermore, T cell composition rate in PBMC was also dramatically decreased by the treatment. C&C greatly down-regulated production of Th2 cytokines including IL-4, IL-5, IL-13 in the serum. The down- regulatory effects of C&C on these Th2 cytokines production were also detected in CD3/ CD28 activated splenocytes. Conclusions : These results indicated that C&C is a plausible therapeutic agent for treatment of atopic dermatitis through regulating the Th2 skewed immune system.

Differentiation of Dopaminergic and Cholinergic Neurons from Mesenchymal-like Stem Cells Derived from the Adipose Tissue (사람 지방 유래 중간엽 줄기세포의 도파민성 및 콜린성 신경세포분화)

  • Hong, In-Kyung;Jeong, Na-Hee;Kim, Ju-Ran;Do, Byung-Rok;Kim, Hea-Kwon;Kang, Sung-Goo
    • Development and Reproduction
    • /
    • v.12 no.1
    • /
    • pp.31-39
    • /
    • 2008
  • Neural tissue has limited intrinsic capacity of repair after injury, and the identification of alternate sources of neural stem cells has broad clinical potential. We isolated mesechymal-like stem cells from human adipose tissues (AT-MSCs), and studied on transdifferentiation-promoting conditions in neural cells. Dopaminergic and cholinergic neuron induction of AT-MSCs was also studied. Neural differentiation was induced by adding bFGF, EGF, dimethyl sulphoxide (DMSO) and butylated hydroxyanisole(BHA) in N2 Medium and N2 supplement. The immunoreactive cells for $\beta$-tubulin III, a neuron-specific marker, GFAP, an astrocyte marker, or Gal-C, an oligodendrocyte marker, were found. AT-MSCs treated with bFGF, SHH and FGF8 were differentiatied into dopaminergic neurons that were immunopositive for TH antibody. Differentiation of MSCs to cholinergic neurons was induced by combined treatment with basic fibroblast growth factor (bFGF), retinoic acid (RA) and sonic hedgehog (Shh). AT-MSCs treated with DMSO and BHA rapidly assumed the morphology of multipolar neurons. Both immunocytochemistry and RT-PCR analysis indicated that the expression of a number of neural markers including neuro D1, $\beta$-tubulin III, GFAP and nestinwas markedly elevated during this acute differentiation. While the stem cell markers such as SCF, C-kit, and Stat-3 were not expressed after preinduction medium culture, we confirmed the differentiation of dopaminergic and cholinergic neurons by TH/$\beta$-tubulin III or ChAT/ $\beta$-tubulin III positive cells. Conclusively, AT-MSCs can be differentiated into dopaminergic and cholinergic neuronsand these findings suggest that AT-MSCs are alternative cell source of treatment for neurodegenerative diseases.

  • PDF

New Whitening Agent From Pimpinella brachycarpa (참나물추출물의 멜라닌 생성저해 효과)

  • Kim, Jin-Hwa;Sim, Gwan-Sub;Lee, Dong-Hwan;Lee, Geun-Soo;Lee, Bum-Chun;Pyo, Hyeong-Bae
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • v.33 no.3
    • /
    • pp.203-208
    • /
    • 2007
  • To develop a new whitening agent for cosmetics from natural products, Pimpinella brachcarpa was selected for its inhibitory effect on melanogenesis in B16 melanoma cells. Crude ethanolic extract of P. brachycarpa and its four fractions-hexane, ethyl acetate(EtOAc), butanol and aqueous were evaluated for antioxidative effects and tyrosinase inhibitory activity. To elucidate the mechanism of active compounds of P. brachycarpa, we investigated the changes in protein level of tyrosinase, TRP-1 and TRP-2 using Western blotting and the changes in mRNA level of tyrosinase using RT-PCR technique. Following UV irradiation, expression of ET-1 in HaCaT keratinocytes was measured by quantitative enzyme immunoassay(EIA) using human ET-1 antibody. Crude ethanolic extract of P. brachycarpa and its four fractions-hexane, EtOAc, butanol and aqueous had free radical scavenging effect by 87.2, 2.5, 97.2, 80.5, 49.8% at 100 ${\mu}g/mL$ and tyrosinase inhibitory effect by 18.3, 15.1, 55.4, 13.1, 0 % at 100 ${\mu}g/mL$. P. brachycarpa EtOAc fraction significantly inhibited melanin production in B16 melanoma cells. Treatment with P. brachycarpa extract for 72 h suppressed the biosynthesis of melanin up to 58 % at 100 ${\mu}g/mL$. Especially, the EtOAc fraction of P. brachycarpa reduced the tyrosinase activity and tyrosinase expression in B16 melanoma cells in a dose-dependent manner. mRNA levels of tyrosinase and TRP-1 were markedly reduced by the EtOAc fraction of P. brachycarpa. Moreover, at the concentrations of $12.5{\sim}50{\mu}g/mL$ of the fraction, the production of UV-induced ET-1 in HaCaT keratinocytes(24 h after 8 $mJ/cm^2$ UVB irradiation) was reduced about 40%(p<0.05). P. brachycarpa could be used as a new natural skin-whitening agent due to the inhibitory effect of on melanin biosynthesis and endothelin-1 expression.

Comparison of Split versus Subunit Seasonal Influenza Vaccine in Korean Children over 3 to under 18 Years of Age

  • Kang, Seah;Kim, Dong Ho;Eun, Byung Wook;Kim, Nam Hee;Kang, Eun Kyeong;Lee, Byong Sop;Kim, Yun-Kyung
    • Pediatric Infection and Vaccine
    • /
    • v.26 no.3
    • /
    • pp.161-169
    • /
    • 2019
  • Purpose: This study was conducted to compare immunogenicities and reactogenicities of the trivalent inactivated subunit influenza vaccine and split influenza vaccine in Korean children and adolescents. Methods: In total, 202 healthy children aged 36 months to <18 years were enrolled at six hospitals in Korea from October to December 2008. The subjects were vaccinated with either the split or subunit influenza vaccine. The hemagglutinin inhibition antibody titers against the H1N1, H3N2, and B virus strains were measured, and the seroconversion rates, seroprotection rates, and geometric mean titers were calculated. All subjects were observed for local and systemic reactions. Results: Both the split and subunit vaccine groups had similar seroprotection rates against all strains (95.9%, 94.9%, 96.9% vs. 96.0%, 90.9%, and 87.9%). In children aged 36 to <72 months, the seroprotection rates were similar between the two vaccine groups. In children aged 72 months to <18 years, both vaccines showed high seroprotection rates against the H1N1, H3N2, and B strain (98.4%, 98.4%, 98.4% vs. 97.0%, 95.5%, and 91.0%), but showed relatively low seroconversion rates (39.1%, 73.4%, 35.9% vs. 34.3%, 55.2%, and 38.8%). There were more local and systemic reactions in the split vaccine group than in the subunit vaccine group; however, no serious adverse reactions were observed in both groups. Conclusions: Both the split and subunit vaccines showed acceptable immunogenicity in all age groups. There were no serious adverse events with both vaccines.

Measurement of the Levels of IgG Subclasses Reactive to Salmonella typhi in the Sera of Patients with Typhoid Fever (장티푸스환자의 혈청내 Salmonella typhi에 대한 IgG subclass항체의 분포)

  • Kim, Young-Jung;Hwang, Eung-Soo;Kang, Jae-Seung;Cha, Chang-Yong;Chang, Woo-Hyun;Kim, Yoon-Won;Cho, Min-Ki;Min, Chang-Hong
    • The Journal of the Korean Society for Microbiology
    • /
    • v.21 no.4
    • /
    • pp.447-453
    • /
    • 1986
  • To diagnose the typhoid fever rapidly and accurately in clinically suspected patients, the levels of IgG subclass antibody were measured by enzyme-linked immunosorbent assay(ELISA). With symptom, blood culture and agglutination test, tested persons were categorized into 6 groups as typhoid fever, FUO, paratyphi A or B, other bacterial infctions, cancers, and control. ELISA was performed on the polyvinyl chloride plates coated with killed whole cell($10^8\;cell/ml$) of S. typhi 0901W by poly-L-lysine applied as binding substance (and polyvinyl chloride as solid phase). The distribution of the level of IgG subclass antibodies in each group was analyzed and compared with other groups. The results obtained were summarized as follow: 1. The optimal dilution of the sera from patients with typhoid fever was 1:160, and those of the sheep anti-human IgG subclass and the peroxidase conjugated rabbit anti-sheep IgG were 1:4000 and 1:5000, respectively. 2. The absorbance levels of IgG subclass in the sera of typhoid fever patients were as follows; a) IgG1 value is $0.439{\pm}0.110$ b) IgG2 value is $0.416{\pm}0.165$ c) IgG3 value is $0.449{\pm}0.145$ d) IgG4 value is $0.525{\pm}0.154$ IgG subclass levels in the sera of typhoid patients were much higher than in control group and patient with paratyphi A or B as well as other infectious diseases. The sensitivity and the specificity in differential diagnosis of typhoid fever and other febrile diseases were 92% and 79% in the assay of IgG1 respectively, whereas those in the assay of IgG2 were 97% and 72%, respectively (above absorbance 0.3). 3. The absorbance levels of IgG subclass in the serial sera of typhiod fever patients tend to decrease to the level of absorbance 0.3 in 10 months from the onset of illness. 4. The order of absorbance levels of IgG subclass in the serum of each group were typhoid fever, paratyphi A or B, other infectious diseases, control and cancer. 5. For the serodiagnosis of typhoid fever against other febrile diseases, the sensitivity and the specificity in the assay of IgG2 activity were 76% and 93% in absorbance 0.4, respectively. 6. In the distribution of the level of each IgG subclass in the sera of FUO patients which were suspected of typhoid fever, the positive rate was ranged from 36% to 82%. This suggest that more than 50% of FUO patients are caused by S. typhi.

  • PDF

Growth Performance, Humoral Immune Response and Carcass Characteristics of Broiler Chickens Fed Alkali Processed Karanj Cake Incorporated Diet Supplemented with Methionine

  • Panda, K.;Sastry, V.R.B.;Mandal, A.B.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.18 no.5
    • /
    • pp.677-681
    • /
    • 2005
  • A study was conducted to see the effect of dietary incorporation of alkali (1.5% NaOH, w/w) processed solvent extracted karanj cake (SKC) supplemented with methionine on growth performance, humoral immune response and carcass characteristics of broiler chickens from 0 to 8 weeks of age. One hundred and twenty, day- old broiler chicks were wing banded, vaccinated against Marek' disease and distributed in a completely randomized design (CRD) into 3 groups of 40 chicks each, which was further replicated to 4 and fed on diet containing soybean meal and those of test groups were fed diets containing alkali (1.5% NaOH) treated SKC partially replacing soybean meal nitrogen of reference diet (12.5%) without or with supplementation of methionine (0.2%). Individual body weight of chicks and replicate-wise feed intakes were recorded at weekly intervals throughout the experimental period. Feed consumption from 1 to 14, 28, 42 and 56 d of age was recorded for each replicate and feed conversion efficiency (weight gain/feed intake) for the respective period was calculated. Mortality was monitored on daily basis. On 28$^{th}$ day of experimental feeding, two birds of each replicate in each dietary group (8 birds/diet) were inoculated with 0.1 ml of a 1.0% suspension of sheep red blood cells (SRBC) and the antibody titre (log 2) was measured after 5 days by the microtitre haemmagglutination procedure. After 42 days of experimental feeding, a retention study of 4 days (43-47 d) duration was conducted on all birds to determine the retention of various nutrients such as DM, N, Ca, P and GE. On 43$^{rd}$ day of experimental feeding, one representative bird from each replicate of a dietary treatment (4/dietary group) was sacrificed, after fasting for two hours with free access to water, through cervical dislocation to observe the weight of dressed carcass, primal cuts (breast, thigh, drumstick, back, neck and wing), giblet (liver, heart and gizzard), abdominal fat and digestive organs. The body weight gain of chicks fed reference diet and those fed diet incorporated with NaOH treated SKC (12.5% replacement) with or without methionine supplementation was comparable during 0 to 4 weeks of age. However, dietary incorporation of alkali processed SKC replacing 12.5% nitrogen moiety of soybean meal resulted in growth retardation, subsequently as evidenced by significantly (p<0.05) lowered body weight gain during 0 to 6 weeks of age in birds fed diet incorporated with alkali processed SKC at 6.43% without methionine as compared to those supplemented with methionine or reference diet. Dietary incorporation of alkali (1.5% NaOH) processed SKC replacing 12.5% of soybean meal nitrogen in the diet of broiler chickens had no adverse effect on feed conversion ratio during all the weeks of experimental feeding. The humoral immune response (HIR) as measured by the antibody titre in response to SRBC inoculation was comparable among all the dietary groups. No significant difference in the intake and retention of DM, N, Ca, P or GE was noted among the chicks fed reference and alkali processed SKC incorporated diets with or without methionine supplementation. None of the carcass traits varied significantly due to dietary variations, except the percent weight of liver and giblet. The percent liver weight was significantly (p<0.05) higher in the birds fed diet incorporated with alkali processed SKC as compared to that in other two groups. Thus solvent extracted karanj cake could be incorporated after alkali (1.5% NaOH, w/w) processing at an enhanced level of 6.43%, replacing 12.5% of soybean meal nitrogen, in the broiler diets up to 4 weeks of age, beyond which the observed growth depression on this diet could be alleviated by 0.2% methionine supplementation.

The Expression of neuronal Nitric Oxide Synthase in Reinnervated Recurrent Laryngeal Nerve (흰쥐에서 편측 반회후두신경 재지배 후 neuronal Nitric Oxide Synthase(nNOS)의 발현과 후두기능회복과의 관계)

  • 정성민;김성숙;조윤희;구태완;박수경;신유리
    • Journal of the Korean Society of Laryngology, Phoniatrics and Logopedics
    • /
    • v.12 no.1
    • /
    • pp.46-54
    • /
    • 2001
  • Background and Objectives : Nitric oxide(NO) is a short-lived molecule with messenger and cytotoxic functions in nervous, cardiovascular, and immune systems. Among the three distinct NOS isoforms, the neuronal isoform is expressed in small, discrete neuronal populations of CNS and PNS. Axonal injury in adult animals results in a dramatic NOS up-regulation in many types of central and peripheral neurons which normally lack the enzyme or express it only at very low levels. In previous study, we confirmed the efficacy of PEMS on the early functional recovery in rats with surgically transected and reanastomosed recurrent laryngeal nerve. Therefore, after we obtained functionally recovered rats using PEMS in this study, we studied to evaluate the expression of nNOS through the analysis of the difference between functional recovery group and non-recovery group in the recurrent laryngeal nerve. Materials and Method : Using 84 healthy male Sprague-Dawley rats, transections and primary anastomosis were performed on their left recurrent laryngeal nerves. Rats were then randomly assigned to 2 groups. The rats in group A(n=42) received PEMS by placing them in custom cages equipped with Helm-holz coils(3 hr/day, 5 days/wk, for 12 wk). The rats in group B(n=42) were handled the same way as the group A, except that they did not receive PEMS. Laryngovideoendoscopy was performed before and after surgery and followed up weekly. Laryngeal EMG was obtained in both PCA and TA muscles. Immunohistochemisty staining using monoclonal anti-neuronal nitric oxide synthase(nNOS) antibody was performed to detect nNOS in recurrent laryngeal nerve and nodose ganglion. Results : 20 rats(63%) in group A and 5 rats(17%) in the group B showed recovery of vocal fo1d motion. The number of NOS-positive cells was increased in functionally-recovered rats. NOS-staining intensity was reduced 12 weeks after nerve injury. The difference between PEMS group and non-stimulated group was not found. Conclusion : This study shows that nNOS may exert a beneficial effect on nerve regeneration and functional repair.

  • PDF

All-trans Retinoic Acid Induced Differentiation of Rat Mammary Epithelial Cells Cultured in Serum-free Medium

  • Ki, Min-Hyo;Paik, Kee-Joo;Lee, Ji-Hyeon;Chung, Hae-Young;Lee, Kyung-Hee;Kim, Kyu-Won;Kim, Nam-Deuk
    • Archives of Pharmacal Research
    • /
    • v.21 no.3
    • /
    • pp.298-304
    • /
    • 1998
  • Retinoids are applied to not only cancer prevention but also cancer chemotherapy by stimulating differentiation of cells. We studied differentiation inducing effect of all-trans retinoic acid (ATRA) by studying proportion of high dense fractions of stem-like cells and the size of S phase fraction in cell cycle. From mammary organoids obtained from 7- to 8-week old F344 female rat mammary gland, we cultured rat mammary epithelial cells (RMEC) and treated physiological doses of $10^{-6}$, $10^{-7}$, and $10^{-8}$ M ATRA from the first day and then cultured for 4, 7, and 14 days. After that, immunostaining was performed using peanut agglutinin (PNA) and anti-Thy-1.1 monoclonal antibody (Thy-1.1) that can be used as markers of differentiation. We separated four different cell subpopulations by flow cytometry: cells negative to both reagents (B-), PNA-positive cells (PNA+), Thy-1.1-positive cells (Thy-1.1+), and cells positive to both reagents (B+). We observed continuous decreases of high dense fractions of stem-like cells (PNA+ subpopulations) for 14 days and as much decreases as high doses of ATRA, which were thought to be proportional to doses of ATRA. We labeled RMEC with bromodeoxyuridine and investigated cell cycle fractions that went through S phase. We observed a tendency of decrease of S phase fraction with time in culture, which, is thought to be related to continuous decreases of PNA+ subpopulations and inhibitory role of ATRA on cell cycle. These results suggest that physiological doses of ATRA could stimulate differentiation of RMEC and convert stem-like RMEC to differentiated cells in SFM for a relatively long period of 14 days.

  • PDF

Expression of Nucleocapsid Protein Gene of Maaji Virus and Use of the Protein as an Immunodiagnostic Antigen of Hemorrhagic Fever with Renal Syndrome (마지바이러스 Nucleocapsid Protein 유전자의 발현과 신증후 출혈열 진단용 항원으로의 이용)

  • Lee, Pyung-Woo;Kim, Yun-Cheol;Paik, Woo-Hyun
    • The Journal of Korean Society of Virology
    • /
    • v.26 no.1
    • /
    • pp.77-90
    • /
    • 1996
  • Nucleocapsid protein (NP)which exists in the particle of hantavirus and surrounds the viral RNA genome is one of the major structural proteins and plays role of antigen to elicit the antibody detected predorminantly right after infection of the virus in the patients of hemorragic fever with renal syndrome (HFRS)or experimental animals. NP is important target antigen in serological diagnostic system of HFRS utilizing whole antigens from the native virus particle, such as IFA, ELISA and Western blotting. Therefore, the preparation of this protein in the level of higher quantity and purity is desirasble for developed dianosis of the disease. The purpose of this study is the cloning of NP gene which exists in the S genome segment of Maaji (MAA) virus and expression of the gene to obtain qualified, genetically engineered NP to be utilized as an immunodiagnostic antigen. First of all, for the purpose of amplifing the MAA-NP gene by PCR, the specific primers were built from the known nucleotide sequence of Hantaan viral NP gene. The viral cDNA of the NP gene was synthesized by using the primers and RNase $H^-$ AMV reverse transcriptase. Thereafter, using this cDNA as a template, the NP gene was amplified specifically by Taq DNA polymrerase. The pT7blue (R)T-overhang vector systems were used for cloning of the amplified NP gene. The expression system was consisted of BL21 (DE3)pLysS and pET16b as a host and a plasmid repectively. Into Ndel site of pET16b, NP gene was ligated with cohesive end for the expression. Insertion of NP gene in the plasmid was confirmed by PCR and mini prep methods. For expression, IPTG was used and the expressed protein was characterized by Western blotting. The MAA-NP was expressed as the form of inclusion body (insoluble fraction)and the protein purified by affinity and metal chealating columns reacted specifically with the sera from patients of HFRS as to be tested by ELISA and Western blotting.

  • PDF

Monoclonal antibodies to recombinant Der p2, a major house dust mite allergen: specificity, epitope analysis and development of two-site capture ELISA

  • Yong, Tai-Soon;Lee, Sang-Mi;Park, Gab-Man;Lee, In-Yong;Ree, Han-Il;Kim, Kyung-Sup;Oh, Sang-Hwan;Park, Jung-Won;Hong, Chein-Soo
    • Parasites, Hosts and Diseases
    • /
    • v.37 no.3
    • /
    • pp.163-169
    • /
    • 1999
  • House dust mite allergens have been well established as sensitizing agents that are important in the induction of allergic diseases. In order to analyze epitopes of the allergen and to develop a quantitative method of the allergen exposure, monoclonal antibodies against a recombinant Der p 2 (rDer p 2), one of the major allergens of Dermatophogoides pteronyssinus, were produce. Four monoclonal antibodies produced wee species-specific and did not cross-react to the D. farinae crude extract. Two of the monoclonal antibodies were found to be IgG1 and the others were IgM. For the analysis of epitopes, a Der p 2 cDNA encoding 126 amino acids (aa) was dissected into three fragments with several overlapping peptides, A (aa residues 1-49), B (44-93), and C fragment (84-126). Three monoclonal antibodies showed reactivities to the recombinant B fragment and to the full-length rDer p 2, but one monoclonal antibody reacted only with the full-length rDer p 2. Two-site capture ELISA was developed using two different monoclonal antibodies for quantitating Der p2 in house dust. The sensitivity limit was 4ng/ml with rDer p2 and $8{\;}\mu\textrm{g}/ml$ with the d. pteronyssinus crude extract. The result suggested that the assay using monoclonal antibodies against rDer p2 could be useful for the environmental studies and for the standardization of mite allergen extracts.

  • PDF