• 제목/요약/키워드: immunological method

검색결과 108건 처리시간 0.028초

Immunotherapy with methyl gallate, an inhibitor of Treg cell migration, enhances the anti-cancer effect of cisplatin therapy

  • Kim, Hyunseong;Lee, Gihyun;Sohn, Sung-Hwa;Lee, Chanju;Kwak, Jung Won;Bae, Hyunsu
    • The Korean Journal of Physiology and Pharmacology
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    • 제20권3호
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    • pp.261-268
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    • 2016
  • $Foxp3^+$ $CD25^+CD4^+$ regulatory T (Treg) cells are crucial for the maintenance of immunological self-tolerance and are abundant in tumors. Most of these cells are chemo-attracted to tumor tissues and suppress anti-tumor responses inside the tumor. Currently, several cancer immunotherapies targeting Treg cells are being clinically tested. Cisplatin is one of the most potent chemotherapy drugs widely used for cancer treatment. While cisplatin is a powerful drug for the treatment of multiple cancers, there are obstacles that limit its use, such as renal dysfunction and the development of cisplatin-resistant cancer cells after its use. To minimize these barriers, combinatorial therapies of cisplatin with other drugs have been developed and have proven to be more effective to treat cancer. In the present study, we evaluated the effect of the combination therapy using methyl gallate with cisplatin in EL4 murine lymphoma bearing C57BL/6 mice. The combinatorial therapy of methyl gallate and cisplatin showed stronger anti-cancer effects than methyl gallate or cisplatin as single treatments. In Treg cell-depleted mice, however, the effect of methyl gallate vanished. It was found that methyl gallate treatment inhibited Treg cell migration into the tumor regardless of cisplatin treatment. Additionally, in both the normal and cisplatin-treated tumor-bearing mice, there was no renal toxicity attributed to methyl gallate treatment. These findings suggest that methyl gallate treatment could be useful as an adjuvant method accompanied with cisplatin therapy.

한약(韓藥)을 이용(利用)한 항암(抗癌) 실험연구(實驗硏究)의 경향(傾向)에 관(關)한 고찰(考察) (A study of the tendency of anti-cancer experimental study using herbal medicines)

  • 김현아;임성우;이원철
    • 대한한방종양학회지
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    • 제4권1호
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    • pp.211-232
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    • 1998
  • Because there were lots of side effects and tolerances to the existing anticancer therapeutics, the experiment extracting the anticancer effect from medicinal herbs is in progress liviely. Therefore the purpose of this study were to research the tendency and the course of anticancer studies. To research the tendency of anticancer studies, medicinal herbs of fifty three experimental papers were analyzed and to examine the course of studies, anticancer papers in the medical world were used. The obtained results were as follows: Methods of herbal medicinal treatments were elimination the pathogenic factor(祛邪) and supporting healthy energy(扶正) method used. In this study, immediately tumor bearing and immune response were the most important point. The subject of immediately tumor bearing was not in the specific cancer but in the influence on the life span of general cencerous cells. In the experimental study of immune response, the effect on NK cell activity of medicinal herbs most studied. The combined usage of medicinal herbs and anticancer agent mostly intended to know whether it inhibits the tumor cell growth. The serum test and blood cell number test show if medicinal herbs inhibit side effect of anticancer agent. More than 80 percents of used medicinal herbs, there were anticancer activities. However anticancer experimental studies using medicinal herbs two weak points. The one, it was difficult to choose a prescription according to differentiation of symptoms and signs(辨證論) of the Oriental Medicine, because we put to the test not a man but a mouse. The other, as we observed the indirect effect of the whole physiological regulation caused by synergic effects of the complex prescription, we don't understand the detailed mechanism of the herb. Therefore, if the anticancer effect of the herb is proved the experiment, we should research the concrete medical action of medicinal herbs and immunological analysis of herbal medicines to the body.

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Novel Modification of Growth Medium Enables Efficient E. coli Expression and Simple Purification of an Endotoxin-Free Recombinant Murine Hsp70 Protein

  • Zachova, Katerinat;Krupka, Michal;Chamrad, Ivo;Belakova, Jana;Horynova, Milada;Weigl, Evzen;Sebela, Marek;Raska, Milan
    • Journal of Microbiology and Biotechnology
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    • 제19권7호
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    • pp.727-733
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    • 2009
  • Heat shock protein 70 kDa (hsp70), a molecular chaperone involved in folding of nascent proteins, has been studied for its ability to activate innate and specific immunity. High purity hsp70 preparation is generally required for immunization experiments, because endotoxins and other immunologically active contaminants may affect immune responses independently of hsp70. We have developed a novel modification of E. coli-expression medium that enabled a simple two-step production and purification method for endotoxin-free recombinant hsp70. During Ni-NTA-based affinity purification of hsp70, a contaminating protein from host E. coli cells, L-glutamine-n-fructose-6-phosphate aminotransferase (GFAT), was identified. By testing various compounds, supplementation of growth medium with a GFAT metabolite,N-acetylglucosamine, was found to reduce GFAT expression and increase the total hsp70 yield five times. The new protocol is based on column purification of His-tagged hsp70 protein produced by E. coli with the modified medium, followed by endotoxin removal by Triton X-114 extraction. This approach yielded hsp70 with high purity and minimal endotoxin contamination, making the final product acceptable for immunization experiments. In summary, a simple modification of growth medium allowed production of recombinant mouse hsp70 in high yield and purity, thus compatible with immunological studies. This protocol may be useful for production of other Histagged proteins expressed in E. coli.

녹각 추출액의 면역학적 특성에 관한 연구 (Studies on the Immunological Characteristic of Cervi cornu Extract)

  • 김현식;허인회;이상준;안형수
    • 약학회지
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    • 제38권6호
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    • pp.806-813
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    • 1994
  • These experiments were conducted to investigate the effects of Cervi cornu extract on lymphocyte blastogenesis in spleen, thymus, lymph node, born marrow cells of Balb/c mouse, haemagglutination reaction against sheep red blood cell (SRBC), plaque forming cell (PFC) assay against SRBC and IL-2 production. Lymphocyte blastogenesis was determined by $[^3H]-thymidine$ incorporation. According to the lymphcoyte blastogenesis test on the immune cell. Ceriv cornu extrat was showed a potent mitogenic activity on the spleen and lymph node cells, but had mild mitogenic activity on the thymus and born marrow cells. Mitogenic active component of Crevi cornu extract was identified to be materials where molecular weights are higher than 5,000 by membrane filteration method. Cervi cornu extrat was shown to increase mitogenic effect on the lipopolysaccharide (LPS)-stimulated spleen cells significantly, but decrease mitogenic effect on the Con A stimulated spleen cell at the concentration 0.3%, 1% and 3%. Ceriv cornu extract didn't show to be haemagglutination reaction and showed to inhibit the Con A-induced haemagglutination reaction against SREC. Result of SRBC-PEC test. Ceriv cornu extract significantly increase the number of PEC at the concentration of 0.1% and 1%. When IL-2 or IL-4 production was determined by proliferation of CTLL-2 cells. Ceriv cornu extract was not shown to stimulate the production of IL-2. From the above results, it is shown that Ceriv cornu extract increased antibody production by B cells, but nor IL-2 production by helper T cells.

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Ferric Sulfate를 이용한 유치의 치수절단술 (PRIMANY TOOTH PULPOTOMY USING FERRIC SULFATE)

  • 이상헌;이미나;이상훈
    • 대한소아치과학회지
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    • 제25권4호
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    • pp.843-848
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    • 1998
  • Pulpotomy is a frequently used treatment modality in primary teeth. It is method by which infected coronal pulp is removed while retaining vital radicular pulp. Since its introduction in 1930 by Sweet formocresol remains the most popular medicament for this treatment. However, despite its outstanding bactericidal properties, formocresol is known to cause adverse tissue reactions. Theoretically, formocresol disinfects and fixes radicular pulp and thus prevents infection and internal resorption. In reality, however, it leads to chronic inflammation and is sometimes responsible for failures through abscess formation and internal root resorption. Also, Myers et al., in 1978, reported on the systemic distribution of FC and other studies have followed with reports of its immunological, mutagenic and carcinogenic effects. Much effort has, therefore, focused on the development of alternative medicaments and techniques. Since its introduction in 19C, ferric sulfate proven itself as an effective hemostatic agent and is used as an astringent in dentistry. In 1988, Landau and Johnsen suggested ferric sulfate be used as a medicament in pulpotomy and many studies have focused on it to overcome the toxic effects of FC. Ferric sulfate acts through its ferric ion and iron ion, which react with blood protein leading to aggregation. The aggregated protein acts to plug the blood vessels, causing mechanical hemostasis. As blood clot formation is minimal, there is reduced inflammation of radicular pulp and enhanced healing. There are no reports regarding its systemic distribution. This is a report of cases treated by the author using pulpotomy with ferric sulfate.

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Serum Levels of Type 2 Chemokines in Lepromatous Leprosy Patients

  • Lew, Wook;Nakamura, Koichiro;Tada, Yayoi;Kwahck, Ho;Chang, Soo Kyoung;Tamaki, Kunihiko
    • IMMUNE NETWORK
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    • 제2권4호
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    • pp.223-226
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    • 2002
  • Background: The type 2 deviated immunological state is predominant in lepromatous leprosy. Erythema nodosum leprosum (ENL) is an immune-complex mediated reaction that typically occurs in lepromatous leprosy. To date, the serum levels of tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-2 receptor, IL-10, IL-$1{\beta}$, IL-1 receptor antagonist and monocyte chemoattractant protein-1 (MCP-1) were reported to be higher in lepromatous leprosy. TNF-${\alpha}$ is also known to be higher in ENL, which is reduced after thalidomide treatment. However the serum type 2 chemokine levels in lepromatous leprosy patients have not been reported. Methods: The serum levels of the type 2 chemokines such as thymus and activation-regulated chemokine (TARC), macrophage-derived chemokine (MDC) and eotaxin together with IL-12 and IL-10 in the sera from leprosy patients were detected using an enzyme-linked solvent assay (ELISA) method. Results: The Serum TARC, MDC, eotaxin, IL-10 and IL-12 levels in lepromatous leprosy patients were not significantly different from the normal control levels. The serum levels were not significantly different between the paucibacillary group and multibacillary group. The serum TARC or MDC levels in the ENL patients were more reduced after a treatment containing thalidomide. Conclusion: The type 2 chemokines are not related to the severity of lepromatous leprosy. The larger reducing effect of the TARC or MDC levels in ENL patients by a treatment containing thalidomide suggests the potential role of these chemokines in the development of ENL and the therapeutic mechanism of thalidomide.

백서 좌골신경의 동종이식후 조직반응에 관한 실험적 연구 (AN EXPERIMENTAL STUDY ON TISSUE REACTIONS OF ALLOGENEIC SCIATIC NERVE GRAFT IN RAT)

  • 정형배;임창준;이동근;서재덕
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제13권2호
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    • pp.203-216
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    • 1991
  • Nerve allografts as a bridge of regeneration is useful in the repair of peripheral nerve defect resulting from trauma, and leprosy. But immunological rejection and complicated scar formation is an unavoidable problem in the application of allogeneic nerves. This article is intended to study of the regeneration of allogeneic nerve grafts in rats with histopathologically, scanning electron microscopically. 24 adult male Sprague-Dawley rats were used as the experimental animals. A 2cm skin incision was made on the lateral aspects of limb, parallel to femur. Segments of sciatic nerve trunk taken from rats, 10mm was resected at the middle of the thigh, nerve graft was inserted between the ends of gaps with perineural and epineural suture method with 10-0 prolene. Obsrevation was made simultaneously at 3 day, 1, 2, 3, 4, 5, 6, 8 weeks after surgery. The results were as follows. 1. In light and electronic microscopic studies, marked degenerative change of the graft nerves were observed at 2 weeks after surgery. 2. After surgery, blood clot fromation was observed at 3 day, granualtion tissue formation was observed at 2 week, and fibrous tissue proliferation was observed at 3 week. 3. In change of nerve fiber, there were Wallerian degeneration at early stage, decrease in degeneration at 4 week but degeneration of myeline was continuded at 8 week. 4. At 4 week, schwann cells proliferate at its cut ends to join with the distal and proximal stump of the damaged nerve. 5. Fibrous scar tissues are formed at 2 weeks and increased progressively in 8 weeks, which was interrupted the regeneration of grafted nerve.

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Comparison of Serum Fucose Levels in Leukoplakia and Oral Cancer Patients

  • Rai, Narendra Prakash;Anekar, Jayaprasad;Shivaraja, Shankara YM;Divakar, Darshan Devang;Al Kheraif, Abdulaziz Abdullah;Ramakrishnaiah, Ravikumar;Sebastian, Roopa;Raj, AC;Al-Hazmi, Ali;Mustafa, habil Mohamed
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권17호
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    • pp.7497-7500
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    • 2015
  • Background: Tumor markers, designated as a broad group of substances produced by malignancies, could be in the form of biochemical substances, immunological substances, cell surface changes and genetic alterations. Cancer, a disorder of cellular behavior is characterized by alteration of serum glycoproteins. L-fucose, a hexose, which is the terminal sugar in most of the plasma glycoproteins, may be useful as a tumor marker for the detection, monitoring and prognostic assessment of malignancies. The aim of the study was to ascertain the role of serum fucose as a biomarker for early detection of oral cancer and to compare serum fucose levels in healthy controls, leukoplakia and oral cancer patients. Materials and Methods: The study included 60 (100.0%) subjects, who were grouped as 20 (33.3%) control subjects, 20 (33.3%) squamous cell carcinoma patients and 20 (33.3%) leukoplakia patients. Fucose estimation was done using UV-visible spectrophotometry based on the method as adopted by Winzler using cysteine reagent. The results were analyzed statistically using ANOVA with Bonferroni post hoc tests. Results: Results showed a high significance in serum fucose in oral squamous cell carcinoma (OSCC) and leukoplakia subjects compared to normal controls. There was a gradual increase in the values noted from control to leukoplakia and to squamous cell carcinoma. Conclusions: Estimation of serum fucose may be a reliable marker and can be used as an effective diagnostic biomarker in oral squamous cell carcinoma patients.

Effect of Diclazuril on the Bursa of Fabricius Morphology and SIgA Expression in Chickens Infected with Eimeria tenella

  • Zhou, Bian-hua;Liu, Li-li;Liu, Jeffrey;Yuan, Fu-wei;Tian, Er-jie;Wang, Hong-wei
    • Parasites, Hosts and Diseases
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    • 제53권6호
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    • pp.675-682
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    • 2015
  • The effects of diclazuril on the bursa of Fabricius (BF) structure and secretory IgA (SIgA) expression in chickens infected with Eimeria tenella were examined. The morphology of the BF was observed by hematoxylin and eosin staining, while ultrastructural changes were monitored by transmission electron microscopy. E. tenella infection caused the BF cell volumes to decrease, irregularly arranged, as well as, enlargement of the intercellular space. Diclazuril treatment alleviated the physical signs of damages associated with E. tenella infection. The SIgA expression in BF was analyzed by immunohistochemistry technique. The SIgA expression increased significantly by 350.4% (P<0.01) after E. tenella infection compared to the normal control group. With the treatment of diclazuril, the SIgA was relatively fewer in the cortex, and the expression level was significantly decreased by 46.7% (P<0.01) compared with the infected and untreated group. In conclusion, E. tenella infection in chickens induced obvious harmful changes in BF morphological structure and stimulated the expression of SIgA in the BF. Diclazuril treatment effectively alleviated the morphological changes. This result demonstrates a method to develop an immunological strategy in coccidiosis control.

현삼메탄올 추출물이 LPS로 유도된 Raw 264.7 cell에서의 $TNF-{\alpha},\;IL-1{\beta}$, IL-6, 및 nitric oxide 생성에 미치는 영향 (Inhibitory effect of Scrophulariae Radix extract on $TNF-{\alpha},\;IL-1{\beta}$, IL-6 and Nitric Oxide production in lipopolysaccharide - activated Raw 264.7 cells)

  • 변성희;양재하;김상찬
    • 대한본초학회지
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    • 제20권2호
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    • pp.7-16
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    • 2005
  • Objectives : Scrophulariae Radix (SRE) is commonly used in combination with other herbs as a nutrient and health strengthening agent, and to remove 'heat' and replenish vital essence. The water-based extract of this herb can lower blood pressure in both anesthetized and concious animals, and exhibits an anti-inflammatory activity. But, there is lack of studies regarding the effects of SRE on the immunological activities in molecular levels. The present study was conducted to evaluate the effect of SRE on the regulatory mechanism of cytokines and nitric oxide (NO) in Raw 264.7 cells. Method : After the treatment of Scrophulariae Radix methanol extract, cell viability was measured by MTT assay, NO production was monitored by measuring the nitrite content in culture medium. COX-2 and iNOS were determined by Immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. Results : Results provided evidence that SRE inhibited the production of nitrite and nitrate (NO), inducible nitric oxide synthase (iNOS), $interleukin-1{\beta}\;(IL-1{\beta})$ and interleukin-6 (IL-6), and the activation of phospholylation of inhibitor ${\kappa}B{\alpha}\;(p-I{\kappa}B{\alpha})$ in Raw 264.7 cells activated with lipopolysaccharide (LPS). Conclusion : These findings suggest that Scrophulariae Radix can produce anti-inflammatory effect, which may playa role in adjunctive therapy in Gram-negative bacterial infections.

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