• Title/Summary/Keyword: immunological methods

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Study on Antibody-enzyme Coupling and Enzyme Immunoassay Methods (효소-항체의 결합 및 효소면역측정 방법의 연구)

  • Jang Sean Il
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.3
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    • pp.874-879
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    • 2004
  • Alakaline phosphatase (ALP)- or horseradish peroxidase (HRP)-antibody conjugate was used frequently on the immunological detection methods such as enzyme-linked immunosobent assay (ELISA), immunobolt, immunohistochemistry. The classical enzyme-antibody coupling method by one-step (direction) injection of glutaraldehyde bring into being disadvantage such as low sensitivity of antigen detection because of homopolymers. This study was modified with the dialysis glutaraldehyde method to provide simple coupling through E-amino residues present in most protein. The dialysis glutaraldehyde coupling effects were better than the classical one-step glutaraldehyde injection in antigen detection of ELISA and immunobolt. Optimal dose of the dialysis glutaraldehyde solution was 0.10-0.25 %. This results suggest that the dialysis glutaraldehyde coupling method can readily applied to antigen detection of in vitro and in vivo.

The effect of prescriptions prepared by adding medicinal herbs(Rehmannia glutinosa Residue, Poria cocos Bark) and fermenting herbal materials based on formulas Iksuyoungjingo on immunological activity (익수영진고가미방의 지황박, 복령피 배합과 발효에 따른 제제가 면역활성에 미치는 영향)

  • Lee, Yu-Mi;Kim, Wang-In;Choi, Dong-Hee;Kim, Mi-Rae;Moon, Yang-Seon;Kim, Ji-Eun;Youn, Dae-Hwan;Son, Hong-Seok;Na, Chang-Su
    • Herbal Formula Science
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    • v.26 no.1
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    • pp.13-26
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    • 2018
  • Objectives : This study was carried out to enhance the activity of Iksuyoungjingo, which has the tonify Qi, nourish Yin effect of oriental medicine, and to study the effect of the prescriptions on the immunological activities. Methods : Immunosuppression was induced by methotrexate (MTX) 2 mg / kg, the experimental group was divided into IYGgami-I, IYGgami-I-F, IYGgami-II and IYGgami-II-F groups. Each prescription was administered with drinking water for 20 days, and body weight was measured every 5 days during this period. Leukocyte, $TNF-{\alpha}$, IL-2, IL-6, IgE, spleen weight and body weight were measured Results : In the changes of $TNF-{\alpha}$, IL-2 and IL-6 as pro-inflammatory elements, all of the experimental groups showed a significant increase compared to the control group. In the IgE changes, the IYGgami-I-F, IYGgami-II and IYGgami-II-F groups showed a significant decrease compared to the control group. In the changes of spleen weight, the IYGgami-II-F group showed a significant increase compared to the control group. In the changes of WBC and lymphocytes, the IYGgami-I-F group showed a significant increase compared to the control group. Conclusions : From the above results, it can be observed that the efficacy against immunity is exerted in all of the preparations, and it was confirmed that the efficacy was maintained constant even when utilizing the Rehmannia glutinosa Residue and Poria cocos Bark, and that a more beneficial effect can be exerted in the effectiveness when the fermentation is carried out.

Immunological benefits by ginseng through reciprocal regulation of Th17 and Treg cells during cyclosporine-induced immunosuppression

  • Heo, Seong Beom;Lim, Sun Woo;Jhun, Joo Yeon;Cho, Mi La;Chung, Byung Ha;Yang, Chul Woo
    • Journal of Ginseng Research
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    • v.40 no.1
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    • pp.18-27
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    • 2016
  • Background: It is not clear whether ginseng affects cyclosporine A (CsA)-induced desirable immunosuppressive action. In this study, we evaluated the immunological influence of combined treatment of ginseng with CsA. Methods: Using CD4+ T cells from mouse spleens stimulated with the T cell receptor (TCR) or allogeneic antigen-presenting cells (APCs), we examined the differentiation of naïve T cells into T helper 1 (Th1), Th2, Th17, and regulatory T cells (Tregs), and their cytokine production during treatment by Korean Red Ginseng extract (KRGE) and/or CsA. The influence of KRGE on the allogeneic T cell response was evaluated by mixed lymphocyte reaction (MLR). We also evaluated whether signal transducer and activator of transcription 3 (STAT3) and STAT5 are implicated in this regulation. Results: Under TCR stimulation, KRGE treatment did not affect the population of CD4+interferon gamma ($IFN{\gamma}$)+ and CD4+interleukin (IL)-4+ cells and their cytokine production compared with CsA alone. Under the Th17-polarizing condition, KRGE significantly reduced the number of CD4+IL-17+ cells and CD4+/phosphorylated STAT3 (p-STAT3)+ cells, but increased the number of CD4+CD25+forkhead box P3 (Foxp3)+ cells and CD4+/p-STAT5+ cells compared with CsA alone. In allogeneic APCs-stimulated CD4+ T cells, KRGE significantly decreased total allogeneic T cell proliferation. Consistent with the effects of TCR stimulation, KRGE reduced the number of CD4+IL-17+ cells and increased the number of CD4+CD25+Foxp3+ cells under the Th17-polarizing condition. Conclusion: KRGE has immunological benefits through the reciprocal regulation of Th17 and Treg cells during CsA-induced immunosuppression.

Effects of heat stress on growth performance, selected physiological and immunological parameters, caecal microflora, and meat quality in two broiler strains

  • Awad, Elmutaz Atta;Najaa, Muhamad;Zulaikha, Zainool Abidin;Zulkifli, Idrus;Soleimani, Abdoreza Farjam
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.5
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    • pp.778-787
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    • 2020
  • Objective: This study was conducted to investigate the effects of normal and heat stress environments on growth performance and, selected physiological and immunological parameters, caecal microflora and meat quality in Cobb 500 and Ross 308 broilers. Methods: One-hundred-and-twenty male broiler chicks from each strain (one-day-old) were randomly assigned in groups of 10 to 24 battery cages. Ambient temperature on day (d) 1 was set at 32℃ and gradually reduced to 23℃ on d 21. From d 22 to 35, equal numbers of birds from each strain were exposed to a temperature of either 23℃ throughout (normal) or 34℃ for 6 h (heat stress). Results: From d 1 to 21, strain had no effect (p>0.05) on feed intake (FI), body weight gain (BWG), or the feed conversion ratio (FCR). Except for creatine kinase, no strain×temperature interactions were observed for all the parameters measured. Regardless of strain, heat exposure significantly (p<0.05) reduced FI and BWG (d 22 to 35 and 1 to 35), immunoglobulin Y (IgY) and IgM, while increased FCR (d 22 to 35 and 1 to 35) and serum levels of glucose and acute phase proteins (APPs). Regardless of temperature, the Ross 308 birds had significantly (p<0.05) lower IgA and higher finisher and overall BWG compared to Cobb 500. Conclusion: The present study suggests that the detrimental effects of heat stress are consistent across commercial broiler strains because there were no significant strain×temperature interactions for growth performance, serum APPs and immunoglobulin responses, meat quality, and ceacal microflora population.

Comparison of immune response and HPLC analysis for combination of Aconiti Lateralis Radix Preparata and Glycyrrhizae Radix (법제 부자와 감초의 배합 비율에 대한 HPLC 분석 및 면역 활성 비교 연구)

  • Lee, Jin-Ah;Ha, Hye-Kyung;Jung, Da-Young;Seo, Chang-Seob;Lee, Ho-Young;Shin, Hyeun-Kyoo
    • The Korea Journal of Herbology
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    • v.25 no.4
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    • pp.23-29
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    • 2010
  • Objectives : To investigate the immunological activities, we evaluated the combination ratio of Aconiti Lateralis Radix Preparata and Glycyrrhizae Radix (AG) on murine macrophage cell line (RAW 264.7) and ovalbumin/aluminium (OVA/Alum)-immunized mice. Methods : The cellular proliferation and the production of nitric oxide were examined in a macrophage cell line, RAW 264.7 cells, in the presence of the combination ratio of Aconiti Lateralis Radix Preparata and Glycyrrhizae Radix. C57BL/6 mice were immunized intraperitonially with ovalbumin/aluminium ($100{\mu}g/200{\mu}g$) on day 1, 8, and 15. The combination ratio of Aconiti Lateralis Radix Preparata and Glycyrrhizae Radix (1 g/kg/day) was orally administrated for 3 weeks. On day 22, splenocyte and plasma were collected for mitogen-induced proliferation, lymphocyte subpopulation by flow cytometry and measurement of AST (Aspirate aminotransferase), ALT (Alanine aminotransferase), and antibodies (OVA-specific antibodies of the IgG, IgG1, and total IgM classes). Results : Aconiti Lateralis Radix Preparata treatment had no influence on immune responses. The proliferation and NO production of macrophage and proliferation of splenocyte were increased as the higher ratio of Glycrrhizae Radix. The proliferation of splenocyte, lymphocyte subpopulation and production of antibody (total IgM, OVA-specific IgG and OVA-specific IgG1) were increased as the higher ratio of Glycrrhizae Radix on OVA-immunzed mice. Conclusions : These results suggest that the higher ratio of Glycyrrhizae Radix can increase immunological activities such as NO production in RAW264.7 cells, splenocyte proliferation and immunoglobulin production in OVA-immunized mice.

Effects of Herbal Acupuncture of Clematis Mandshurica Maxim. at Sinsu-hyul (BL 23) on Adjuvant Arthritis in Rats.

  • Kim, Kap-Sung;Lee, Seung-Deok;Lee, Chang-Hwan
    • The Journal of Korean Medicine
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    • v.27 no.4
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    • pp.174-181
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    • 2006
  • Objective : Based on the immunological dysfunction in RA and the immunological feature of Clematis mandshurica Maxim. herbal acupuncture solution (CM-HAS), this study examined whether subcutaneous (s.c.) administration of CM-HAS has anti-inflammatory effects on adjuvant arthritis (AA) in the rat. Methods : Complete Freund's adjuvant was used to induce AA in rats. Secondary paw swelling of AA rats was measured, and polyarthritis index was scored. The administration of CM-HAS (2, 5, 10 mg/kg) to the Sinsu-hyul (BL 23) acupuncture point subcutaneously (s.c.) inhibited the inflammatory response and restored the weight of body and immune organs of AA rats. In this study. inhibitory effect of CM-HAS on cyclooxygenase-2 (COX-2) was evaluated. The plant material selected for this study has been used in Korean medicine for the treatment of various diseases that are considered as inflammatory in nature, e.g. asthma, arthritis, rheumatism, fever, edema, infections, snakebite and related inflammatory diseases. Results : There were significant secondary inflammatory reactions in AA rats, accompanied with the simultaneous decrease of body and immune organs weight. The administration of CM-HAS (2, 5, 10 mg/kg, s.c.) improved the above changes significantly. CM-HAS showed inhibitory activity against COX-2, which supports their traditional uses. In this study, aspirin and indomethacin were used to contrast with CM-HAS as the COX-2 inhibitors. Conclusion: The findings of this study may explain at least in part why CM-HAS has been traditionally used for the treatment of inflammatory conditions in traditional Korean and Chinese medicine.

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Effects of Acupuncture and Radix Astragali Aqua-acupuncture on Transcriptional Expression of Mouse Cytokines $IL-1{\beta}$ (현유혈의 침자극과 황기약침이 실험용 생쥐의 면역활성물질 $IL-1{\beta}$의 유전자 발현에 미치는 영향)

  • 손수곤;김종수;박원환
    • The Journal of Korean Medicine
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    • v.21 no.4
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    • pp.16-25
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    • 2000
  • Objective : Acupuncture and Radix Astragali aqua-acupuncture stimuli have long been used to cure human diseases. However, the exact physiological and biochemical mechanisms involved remain undiscovered. Thus, many attempts have been made to show the scientific mechanisms involved. The effects of acupuncture and Radix Astragali aqua-acupuncture, which was known to date, as follow; effective circulation of body blood system and proliferation of leucocytes. Methods : In this study, we have applied acupuncture and Radix Astragali aqua-acupuncture stimuli to mouse on Sinsuhyul, a stimulative point of oriental medicine, to see effects on the expression of cytokine $IL-1{\beta}$. Mice were treated with lipopolysaccharide(LPS) for inflammation induction and then reverse transcriptase-polymerase chain reaction (RT-PCR) using each primer set were performed to trace the amounts of mRNA. Results : 1. $IL-1{\beta}$ was not expressed in LPS-nontreated mice at 15 to 60 min after acupuncture-stimuli. However, expression occurred after 3hrs. 2. $IL-1{\beta}$ was specifically expressed in LPS-treated mice at 30 min after acupuncture-stimuli. 3. $IL-1{\beta}$ was expressed in LPS-nontreated mice at 30 min after Radix Astragali aqua-acupuncture stimuli, however, not expressed at 60, 180 min. 4. $IL-1{\beta}$ was gradually expressed in LPS-treated mice at 15 to 180 min after Radix Astragali aqua-acupuncture stimuli. Conclusions : $IL-1{\beta}$ in LPS-treated mice was more effective than that of LPS-nontreated mice. We are now in the process of elucidating the immunological action mechanism of acupuncture and Radix Astragali aqua-acupuncture stimuli. And cytokine $IL-1{\beta}$ can be used not only as a basis of the effects of acupuncture and Radix Astragali aqua-acupuncture but also as a diagnosis guide through the immunological actions of those.

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Effect of Gupoongjeseuptang (GPJST) on DNCB (dinitrochlorobenzene)-induced Atopic Dermatitis-like Model NC/Nga Mice (구풍제습탕(驅風除濕湯)이 DNCB로 유도된 NC/Nga mice의 아토피 피부염에 미치는 영향)

  • Yoon, Jae-Eun;Kim, Yun-Hee;Han, Jae-Kyung;Kim, Yun-Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.22 no.3
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    • pp.105-137
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    • 2008
  • Objectives : The purpose of this study is to investigate the effect of Gupoongjeseuptang (GPJST) on atopic dermatitis by in vivo experiment using NC/Nga atopic dermatitis mouse, which has histological and clinical similarities to the atopic dermatitis of human. Methods : To investigate the effect of GPJST on atopic dermatifis, we evaluated atopic dermatitis-like skin lesions by clinical skin index and analyzed immunological parameters in peripheral blood mononuclear cells (PBMCs), splenocytes, draining lymph node (DLN) and performed skin histology in ears and dorsal skin of atopic dermatitis-like skin NC/Nga mouse in vivo. Results : In vivo, clinical skin severity score were significantly lower in GPJST group than control group. IgE, IL-6, $TNF-{\alpha}$, IgG1, IgM, IgG2a and IgG2b levels in serum decreased remarkably in GPJST group than control group. Also, total absolute number of $CD3^+CD69^+$, and $CCR3^+$ cells recovered as normal in PBMCs and $CD3^+$, $CD3^+CD69^+$ decreased significantly compared with control group in isolated DLN from NC/Nga mouse and total absolute number of $CD11b^+Gr-1^+$, $CCR3^+CD3^+$ in dorsal skin of NC/Nga mouse decreased by GPJST. We analyzed ear and neck-back skin after biopsy and dyeing by hematoxyline/eosin (H&E) and toluidine staining (mast cells marker) and obtained results that GPJST are very effective to histological symptoms (dermal and epidermal thickening, hyperkeratosis and inflammatory cell (CD4, $CCR3^+$) infiltration). Conclusions : This study demonstrates immunological activity of GPJST on atopic dermatitis-like model mice.

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Effects of Lobophytum crassum extract(MC-1) on Various Immunological Factors Related to Pathogenesis of Atopic Dermatitis in Dermatophagoides Pteronyssinus Treated NC/Nga mice (연산호 추출물(MC-1)이 아토피피부염 유발 NC/Nga 생쥐의 혈액내 면역 관련 인자에 미치는 영향)

  • Choi, Hak-Joo;Sim, Boo-Yong;Miyamoto, Tomofumi
    • The Korea Journal of Herbology
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    • v.30 no.1
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    • pp.95-101
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    • 2015
  • Objectives : The aim of this study was to confirm whether or not coral has a preventive effect on development of atopic dermatitis induced by house mite(dermatophagoides pteronyssinus) in NC/Nga mice. Methods : This study was undertaken by using a reliable Atopic dermatitis mouse model demonstrating similar immune response. Lobophytum crassum was administered orally to NC/Nga mouse for 3 weeks. In order to verify the effectiveness of Lobophytum crassum in atopic dermatitis treatment, its role in immune factors were observed in NC/Nga mice. Results : ALT, AST, BUN and creatine levels were all within in the normal ranges in MC-1 200 and 400 (mg/kg) treated groups, indicating no induced toxicity. MC-1 200 and 400 (mg/kg) groups decreased of atopic dermatitis skin manifestation in NC/Nga mouse of MC-1 200 and 400 (mg/kg) groups compared to that of the control group and decreased the ratio of WBC and lymphocyte in blood. Also, MC-1 200 and 400 (mg/kg) groups significant decreased the ratio of CD4+, CD8+, CD11b+/Gr1+, B220/CD23 and CD4/CD25 immune cell ratio in ALN. Finally MC-1 200 and 400 (mg/kg) groups significantly increased the ratio of CD4+, CD8+, B220/CD23 and CD4/CD25 immune cells in DLN. Conclusions : Theses results suggested that Lobophytum crassum has suppressive effects on aberrant and overactive immunological activities in dermatophagoides pteronyssinus-induced dermatitis mice of NC/Nga.

Studies on Esterase of Pieris rapae L. II. Biochemical Properties and Immunological Studies (배추흰나비(Pieris rapae L.)의 esterase에 관한 연구 II. 생화학적 특성 및 면역학적 연구)

  • 박철호;김학열;여성문
    • The Korean Journal of Zoology
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    • v.33 no.3
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    • pp.337-345
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    • 1990
  • The properties of three esterases (E2, E6 and E11) which were previously purified from Pieris rapae L. were determined and physiological role of E6 was inferred using immunological methods. Based on inhibitor study, all of the purified esterases were found to be carboxylesterases (EC 3.1.1.1). The Km values for E2, E6 and E11 were determined to be 6.89 X 10-$^4$M. 3.19 $\times$ l0-$^4$M and 3.69 X 10-$^4$M, respectively. The molecular weights of E2, E6 and E11 were estimated to be 42 KD, 81 KD and 174 KD, respectively. The isoelectric points of E2, E6 and E11 were estimated to be pH 5.54, pH 5.89 and pH 6.50, respectively. The concentration of E6 during development was highest at the late 5th instar larval stage and that according to organs at the same stage was highest in midgut. These results suggest that E6 might be a hydrolase involved in the digestion of dietary lipids.

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