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Cell-Based IL-15:IL-15Rα Secreting Vaccine as an Effective Therapy for CT26 Colon Cancer in Mice

  • Thi, Van Anh Do;Jeon, Hyung Min;Park, Sang Min;Lee, Hayyoung;Kim, Young Sang
    • Molecules and Cells
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    • v.42 no.12
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    • pp.869-883
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    • 2019
  • Interleukin (IL)-15 is an essential immune-modulator with high potential for use in cancer treatment. Natural IL-15 has a low biological potency because of its short half-life and difficulties in mass-production. IL-15Rα, a member of the IL-15 receptor complex, is famous for its high affinity to IL-15 and its ability to lengthen the half-life of IL-15. We have double-transfected IL-15 and its truncated receptor IL-15Rα into CT26 colon cancer cells to target them for intracellular assembly. The secreted IL-15:IL-15Rα complexes were confirmed in ELISA and Co-IP experiments. IL-15:IL-15Rα secreting clones showed a higher anti-tumor effect than IL-15 secreting clones. Furthermore, we also evaluated the vaccine and therapeutic efficacy of the whole cancer-cell vaccine using mitomycin C (MMC)-treated IL-15:IL-15Rα secreting CT26 clones. Three sets of experiments were evaluated; (1) therapeutics, (2) vaccination, and (3) long-term protection. Wild-type CT26-bearing mice treated with a single dose of MMC-inactivated secreted IL-15:IL-15Rα clones prolonged survival compared to the control group. Survival of MMC-inactivated IL-15:IL-15Rα clone-vaccinated mice (without any further adjuvant) exceeded up to 100%. This protection effect even lasted for at least three months after the immunization. Secreted IL-15:IL-15Rα clones challenging trigger anti-tumor response via CD4+ T, CD8+ T, and natural killer (NK) cell-dependent cytotoxicity. Our result suggested that cell-based vaccine secreting IL-15:IL-15Rα, may offer the new tools for immunotherapy to treat cancer.

A Study on the production of Monoclonal Antibodies against Rhodotrodula rubra (효모양진균의 단크론 생성)

  • Ryeom K.;Kim S. C.;Lee J. H.
    • Journal of environmental and Sanitary engineering
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    • v.4 no.1 s.6
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    • pp.17-28
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    • 1989
  • Having studied the production of monoclonal antibodies for developing a diagnosis medicine which shall be detected by a high-sensitivity test by using Rhodotorula rubra as a fungi-host which had been extracted through biochemical tests and follow-up examinations on Yeast-like fungi obtained from pulmonary tissues of pulmonary tuberculosis patients who had been in Kong ju National Tuberculosis Hospital from Jun. to Dec. in 1987, I. have gained such results as follows: 1. The fusion rate was influenced by feeder cell layers, cell density and time required to the cell fusion with cells in myelona subculture. 2. The fusion rate did not show any significant difference when the cell was applyed with two molecular weights, i.e., 1500 and 4000, of polyethylene glycol. 3. Fused cells after the addition of HAT selection media were bright and round, whereas unfused myelona cells and spleen cells were shrunk and granulated. 4. The cell fusion rate turned out to be about $57.2\%$(150 wells / 264 wells). 5. $10\%$(15 wells / 150 wells) of the positive reaction was detected in monoclonal antibody screening. 6. The titer which had reacted positively to Rhodotorula rubra fungal-host was 800 times in density after the gradual dilution of the produced monoclonal antibodies with Indirect ELISA method. 7. The Strongest specific reaction came out after the peroxidase labelled anti-human Immunogobulin had been applyed to Rhodotorula rubra for activating its nature after making drift with Carbonate-bicarbonate buffer (pH 9.6) and drying completely.

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Lobaric Acid Inhibits VCAM-1 Expression in TNF-α-Stimulated Vascular Smooth Muscle Cells via Modulation of NF-κB and MAPK Signaling Pathways

  • Kwon, Ii-Seul;Yim, Joung-Han;Lee, Hong-Kum;Pyo, Suhkneung
    • Biomolecules & Therapeutics
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    • v.24 no.1
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    • pp.25-32
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    • 2016
  • Lichens have been known to possess multiple biological activities, including anti-proliferative and anti-inflammatory activities. Vascular cell adhesion molecule-1 (VCAM-1) may play a role in the development of atherosclerosis. Hence, VCAM-1 is a possible therapeutic target in the treatment of the inflammatory disease. However, the effect of lobaric acid on VCAM-1 has not yet been investigated and characterized. For this study, we examined the effect of lobaric acid on the inhibition of VCAM-1 in tumor necrosis factor-alpha (TNF-${\alpha}$)-stimulated mouse vascular smooth muscle cells. Western blot and ELISA showed that the increased expression of VCAM-1 by TNF-${\alpha}$ was significantly suppressed by the pre-treatment of lobaric acid ($0.1-10{\mu}g/ml$) for 2 h. Lobaric acid abrogated TNF-${\alpha}$-induced NF-${\kappa}B$ activity through preventing the degradation of $I{\kappa}B$ and phosphorylation of extracellular signal-regulated kinases (ERK), c-Jun N-terminal kinases (JNK), and p38 mitogen activated protein (MAP) kinase. Lobaric acid also inhibited the expression of TNF-${\alpha}$ receptor 1 (TNF-R1). Overall, our results suggest that lobaric acid inhibited VCAM-1 expression through the inhibition of p38, ERK, JNK and NF-${\kappa}B$ signaling pathways, and downregulation of TNF-R1 expression. Therefore, it is implicated that lobaric acid may suppress inflammation by altering the physiology of the atherosclerotic lesion.

Zoledronate(Zometa(R))inhibits the formation of osteoblast in rat osteoblastic cell line UMR-106 (Zoledronate이 UMR-106 세포의 증식과 조골세포 형성에 미치는 영향)

  • Jeong, Ki-Hoon;Ryu, Dong-Mok;Jee, Yu-Jin;Lee, Deok-Won;Lee, Hyun-Woo
    • The Journal of the Korean dental association
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    • v.46 no.10
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    • pp.623-632
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    • 2008
  • Purpose : The purpose of this study is to identify the effect of zoledronate(Zometa(R)), which is most common nitrogen containing bisphosphonate, on survival, proliferation, and differentiation of osteoblast. Material & Methods: Twenty four cell culture plates containing essential medium were seeded with UMR-106 cell lines, at density of 5 x $10^4 cells per plates. Each plates were incubated with 5% $CO^2 incubator at $37^{\circ}C$. Starting from 2 days after incubation, cell culture medias were replaced, and added with osteogenesis induction media and 0, 0.01, 0.1, 0.5, 1, $3\muM$ of zoledronate(Zometa(R)), every 2 days, for 12 days. Control group was plates not added with zoledronate($0\muM$), and experiment group were plates added with different concentration of zoledronates(0, 0.01, 0.1, 0.5, 1, $3\muM$). Mature osteoblasts were identified with Alizarine Red staining, and protein samples were collected. Optical density was determined at wavelength of 405nm with ELISA reader. For viability analysis, cells were harvested and incubated with propidium iodide, and analysed with flow cytometry. Western blot technique was used to analyse Runx2 protein of osteoblast. Results : Secretion of bone matrix decreased as zoledronate concentration increased, and zoledronate did not effect survival rate of UMR-106 cells when measured with flow cytometer. Expression of Runx2 protein was inhibited as zoledronate concentration increased. Conclusion : From the results, we were able to identify that increase of zoledronate concentration inhibited differentiation of UMR-106 cell to osteoblast, without effecting quantity or survival rate.

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Effects of herbal mixture on blood glucose and lipid metabolism in type 2 diabetic mellitus mouse (천연물 복합제가 제 2형 당뇨 마우스에서 혈당 및 지질대사에 미치는 영향)

  • Seo, Dong-hyo;Joo, In-Hwan;Kim, Dong-Hee
    • Journal of Haehwa Medicine
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    • v.29 no.2
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    • pp.22-29
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    • 2020
  • Objectives: This study aims to investigate the effects of diabetes mellitus care mixture (DCM) on blood glucose and lipid metabolism in type 2 diabetic mellitus mice. DCM consisted of lagerstroemia speciose, allium hookeri, momordica charantia, amaranthus tricolor, and boesenbergia rotunda, which have been proven to have antidiabetic properties. Methods: In this study, we researched the effects of DCM in type 2 diabetic mellitus mice. C57BLKS/J mouse groups had no treatment, db/db mouse randomly assigned to 2 groups, and treated with distilled water and DCM (200 mg/kg/day). Blood glucose levels and body weight were checked every week. After 4 weeks of treatment, liver function indicators (AST, ALT, and LDH) and lipid metabolites (triglyceride, total cholesterol, LDL-cholesterol, HDL-cholesterol) were measured with a biochemistry analyzer. Diabetic factors (insulin, resistin, and leptin) were measured with ELISA. Results: DCM was decreased blood glucose, diabetic factors, liver function indicators, triglyceride, total cholesterol, and LDL-cholesterol significantly. Also, HDL-cholesterol was significantly increased in DCM group. The bodyweight of DCM group decreased but, no significant difference with the control group. DCM may have the potential to improved diabetes mellitus by regulating blood glucose levels and diabetic factors. Also protecting from diabetic complications by adjusting liver function indicators and lipid metabolites. Conclusions: These results suggest that DCM to be used as an oriental medicine for diabetes, the results of clinical trials are needed.

Studies on the pathogenic Escherichia coli isolated from piglets with diarrhea in Chonnam (전남 지방의 설사 자돈에서 분리된 병원성 대장균에 관한 연구)

  • 김자숙;박영춘;정인호;오은희;박석준;고홍범
    • Korean Journal of Veterinary Service
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    • v.19 no.2
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    • pp.139-153
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    • 1996
  • Porcine E coli infection is a disease caused by Enterotoxin produced by Enterotoxigenic Escherichia coli(ETEC). Enteric colibacillosis has become an economically important disease in pigs as a result of increasing intensification of farrowing management. The present study undertaken to obtain the antibiotic sensitivity and distribution of serogroups and pili producibility test of ETEC from E. coli isolates in Chonnam. The results obtained were as follows. 1. A total of 71 isolates identified as E, coli employing IMViC system from rectal specimens of 54 piglets with diarrhea. 2. In antibiotic sensitivity test, isolates showed high sensitivity to AN, CM, Fox, GM, but resistance to EM, NA TC. 3. The distribution of 25 Isolates of serogroups were 0141:K85(11.3%), 08:K87(8.5%), 064:K (5.6%), 0138:K8l (4.2%), 0139 :K82(2.8%), 0157:K88ac(1.4%) and 0149:K9l (1.4%). 4. MRHA of guinea pig erythrocytes was detected in 8 out of 25OK serotypes and 9 out of 46 unidentified serotypes. MRHA titers of serotypes showed from 64 to 128 in 0141: K85, 2 in 0138:K8l and no titers in 0139:K82. 5. The production of heat labile enterotoxin of ETEC was detect 39 out of 52 isolates showed $\beta$-hemolysin, 7 out of 52 isolates showed ${\gamma}$-hemolysin and 6 out of 52 isolates showed ${\gamma}$-hemolysin by $GM_1$ganglioside ELISA. The distribution of LT toxin were in 12 isolate showed $\beta$-hemolysin, 2 isolates showed ${\alpha}$-hemolysin and 3 isolates showed ${\gamma}$-hemolysin in 25 OK serotypes.

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Comparative evaluation of the murine immune responses to Salmonella enterica serovars Enteritidis, Gallinarum and Typhimurium infection (Salmonella enterica serovars Enteritidis, Gallinarum 및 Typhimurium의 마우스 면역반응의 비교평가)

  • Kim, Kiju;Kim, Dooree;Sun, Jisun;Park, Soyeon;Cho, Youngjae;Ko, Hyun-Jeong;Joo, Hong-Gu;Hahn, Tae-Wook
    • Korean Journal of Veterinary Research
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    • v.53 no.2
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    • pp.95-101
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    • 2013
  • The study was carried out to evaluate and compare the immune responses in mice experimentally infected with either wild-type or isogenic mutants of Salmonella enterica serovars Enteritidis (SE), Salmonella Typhimurium (ST) and Gallinarum (SG). The mutant strains were constructed by allelic replacement of some virulence-associated genes in the wild-type strains. Seven-week-old female BALB/c mice were orally or intraperitoneally inoculated by injecting bacterial suspension. To evaluate the immune responses, enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunospot (ELISPOT) assay were conducted with serum and fecal samples. As a result, the mice group infected orally with the SE mutant strain showed the highest level of specific IgA-secreting splenocytes, compared to the other groups. The peritoneally injected groups showed the greater levels of IgG1 than the orally injected groups, which was in a good agreement with the previous studies. In addition, the mutant infected groups had the similar secretion levels of antibodies with the wild-type infected groups. These results demonstrated that the SE mutant strain elicited humoral immune response as much as wild-type, implying that it can be useful as a delivery vehicle as well as a candidate of a live attenuated vaccine.

Alteration of Porcine Serum Albumin Levels in Pork Meat by Marination in Kiwi or Pineapple Juice and Subsequent Pan Broiling

  • Moon, Sung-Sil;Kim, Dongwook;Kim, Il-Suk;Ham, Jun-Sang;Park, Beom-Young;Jang, Aera
    • Food Science of Animal Resources
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    • v.34 no.3
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    • pp.355-361
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    • 2014
  • This study was conducted to evaluate the changes in porcine serum albumin (PSA), a major allergen, which occur when raw pork ham is marinated with kiwi or pineapple juice, and/or when the ham is pan broiled at $300^{\circ}C$ for 4 min after marination. In this study, raw pork ham was soaked for 4 h or 8 h in marinades containing commercial marinating sauce only, commercial marinating sauce and 7% kiwi juice, or commercial marinating sauce and 7% pineapple juice. When the meat was marinated and then pan-broiled, pork ham meat protein was significantly denatured and hydrolyzed, and the level of PSA in the meat was significantly reduced. The PSA contents of pork broiled without marination, pork that had been marinated in commercial marinating sauce alone, pork that had been marinated in commercial marinating sauce with kiwi juice, and pork that had been marinated in commercial marinating sauce with pineapple juice, were 95.4, 43.3, 14.3, and 5.4 ng/mL, respectively (p<0.05). Marinating with pineapple juice was more effective than marinating with kiwi juice; and marination for 8 h was more effective than marinating for 4 h. These results indicate that the level of PSA in pork ham is effectively reduced, when the meat is first marinated in sauces that contain kiwi or pineapple extracts for 8 h, rather than 4 h, and then cooked. Further study is needed to determine whether marinated pork meat reduces allergenicity in vivo, as well.

Effects of Lycii fructus and Astragalus membranaceus Mixed Extracts on Immunomodulators and Prevention of Diabetic Cataract and Retinopathy in Streptozotocin-induced Diabetes Rat Model (Streptozotocin으로 유발한 당뇨병성 쥐에서의 당뇨병성 백내장과 망막병증에 대한 구기자와 황기 혼합 추출물 등의 면역 조절 및 예방 효과)

  • Jeon, Yun-Hui;Moon, Jun-Woong;Kweon, Hyuk-Jung;Jeoung, Young-Jun;An, Chi-Sun;Jin, Hai-Lan;Hur, Sun-Jin;Lim, Beong-Ou
    • Korean Journal of Medicinal Crop Science
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    • v.18 no.1
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    • pp.15-21
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    • 2010
  • This study was conducted to investigate the effects of Lycii fructus and Astragalus membranaceus mixed extracts on immunomodulators and prevention in a streptozotocin-induced diabetes rat model. A total of 28 male rats were divided into four dietary groups and fed a commercial diet (A), commercial diet plus induced diabetes by a streptozotocin (STZ) injection (B), induced diabetes by STZ plus medicinal crop extracts(I&$H^{(R)}$) diet (C), and medicinal crop extracts (I&$H^{(R)}$) diet (D). Immunoblotting analyses revealed cytokine expression, and ELISA analyses revealed immunoglobulin E and nitric oxide production. As a results, the tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and inducible nitric oxide synthase (iNOS) as a inflammatory cytokine were decreased. Interleukin-6 (IL-6) and signal transducer and activation of transcription 3 (STAT3) cytokine related in diabetes expression through JAK/STAT3 pathway were also decreased. Furthermore, immunoglobulin E and nitric oxide production were decreased in the serum and lens, respectively. These results suggest that Lycii fructus and Astragalus membranaceus mixed extracts provide positive effects on immunomodulators and prevention in diabetes and eye disease complications.

A study on the Effects of Organism Vigor Information Solution to the Growth of Tumor-Derived Animal Cell (생물활성정보수가 종양 유래 동물세포 성장에 미치는 영향에 관한 고찰)

  • Jang, M.Y.;Lee, K.S.;Shin, J.W.;Mun, C.W.
    • Journal of Biomedical Engineering Research
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    • v.32 no.3
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    • pp.185-190
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    • 2011
  • The purpose of this study is to investigate the influence of organism vigor information solution(OVIS) on the animal cell growth and to set up a condition for cell culture. We investigated the reactions on the MG-63 and MCF-7 cell line in mixture culture media with various amount of REVIEW solution, which is an example among various OVIS. DMEM-HG was used as basic media. The concentration range of the mixture was limited from 0% to 15%. MTT assay is used for cell viability test. The cell was incubated for 14days and the MTT assay was performed on day 1, 3, 7, 10 and 14 throughout the experiment. We used the ELISA reader to measure the Optical Density(O.D) at 595 nm wavelength filter. MCF-7 was linearly proliferated according to culture time and concentrations of OVIS. On the other hand, MG-63 cells were measured the highest O.D value at 12%. The growth rates of both cells cultured in mixed culture media with OVIS are much higher than those in only basic media, DMEMHG, after 14days. It was confirmed that cell cultured at OVIS grows rapidly at certain period although cells showed a negative effect in initial stage.