• Title/Summary/Keyword: 2-site ELISA

Search Result 39, Processing Time 0.024 seconds

A STUDY ON SYSTEMIC AND LOCAL PRODUCTION OF IMMUNOGLOBULINS IN HUMAN PERIAPICAL CYSTS (치근단 낭종환자의 혈청 및 낭종내의 항체수준에 관한 연구)

  • Park, Kye-Yang;Lim, Sung-Sam
    • Restorative Dentistry and Endodontics
    • /
    • v.17 no.1
    • /
    • pp.222-234
    • /
    • 1992
  • The purpose of this study was to evaluate the systemic and local production of immunoglobulins and their levels in patients with periapical cysts using Enzyme - Linked Immunosorbent Assay. Streptococcus sanguis, Bacteroides gingivalis, and Bacteroides intermedius were grown for use as antigen and they were harvested by centrifugation. The patients were divided into two groups: patients of periapical cysts and normal control. 5 patients of each group were selected and their blood were obtained via intravenous puncture prior to surgical operation. Sera were prepared by centrifugation of each blood samples. Cyst fluid were aspirated from cystic cavity and cyst wall were excised at operation. Control tissue were also excised at extraction site of impacted wisdom teeth from normal control. Each tissue was prepared by homogenization and centrifugation. Then antibodies of each sample were measured by modified ELISA. The following results were obtained: 1. Serum IgG and IgM levels were not significantly different between patients with periapical cyst and normal control. 2. IgG and IgM levels of cyst fluid to Bacteroides gingivalis and Bacteroides intermedius were significantly higher than those of serum of patients with periapical cyst, but there was no significant difference to Streptococcus sanguis. 3. IgG and IgM levels of cyst wall to Bacteroides gingivalis and Bacteroides intermedius were significantly higher than those of control tissue, but there was no significant difference to Streptococcus sanguis. 4. IgG and IgM levels in cyst fluid and IgG levels in cyst wall were highest to Bacteroides gingivalis, and IgM levels in cyst wall were highest to Bacteroides intermedius.

  • PDF

Construction and Characterization of an Anti-Hepatitis B Virus preS1 Humanized Antibody that Binds to the Essential Receptor Binding Site

  • Wi, Jimin;Jeong, Mun Sik;Hong, Hyo Jeong
    • Journal of Microbiology and Biotechnology
    • /
    • v.27 no.7
    • /
    • pp.1336-1344
    • /
    • 2017
  • Hepatitis B virus (HBV) is a major cause of liver cirrhosis and hepatocellular carcinoma. With recent identification of HBV receptor, inhibition of virus entry has become a promising concept in the development of new antiviral drugs. To date, 10 HBV genotypes (A-J) have been defined. We previously generated two murine anti-preS1 monoclonal antibodies (mAbs), KR359 and KR127, that recognize amino acids (aa) 19-26 and 37-45, respectively, in the receptor binding site (aa 13-58, genotype C). Each mAb exhibited virus neutralizing activity in vitro, and a humanized version of KR127 effectively neutralized HBV infection in chimpanzees. In the present study, we constructed a humanized version (HzKR359-1) of KR359 whose antigen binding activity is 4.4-fold higher than that of KR359, as assessed by competitive ELISA, and produced recombinant preS1 antigens (aa 1-60) of different genotypes to investigate the binding capacities of HzKR359-1 and a humanized version (HzKR127-3.2) of KR127 to the 10 HBV genotypes. The results indicate that HzKR359-1 can bind to five genotypes (A, B, C, H, and J), and HzKR127-3.2 can also bind to five genotypes (A, C, D, G, and I). The combination of these two antibodies can bind to eight genotypes (A-D, G-J), and to genotype C additively. Considering that genotypes A-D are common, whereas genotypes E and F are occasionally represented in small patient population, the combination of these two antibodies might block the entry of most virus genotypes and thus broadly neutralize HBV infection.

ADSORPTION OF HUMAN RECOMBINANT FIBRONECTIN TO TITANIUM IN VITRO

  • Kim H.J.;Chang I.T.;Koak J.Y.;Heo S.J.;Yim S.A.;Jang J.H.
    • The Journal of Korean Academy of Prosthodontics
    • /
    • v.42 no.3
    • /
    • pp.301-306
    • /
    • 2004
  • Statement of problem. Fibronectin mediates its biological effects by binding to integrins on cell membranes through a consensus site including the Arg-Gly-Asp (RGD) sequence within tenth type III module. Purpose. The purpose of our study was to investigate the adsorption affinity of human recombinant fibronectin peptide (hFNIII 9-10) to titanium and to investigate the effect of the surrounding ionic composition on the adsorption process. Material and methods. As for evaluating the affinity of hFNIII 9-10 to Ti, titanium disks were incubated in 40, 80 and $120{\mu}g/ml$ hFNIII 9-10 solution at $37^{\circ}C$ overnight, repectively. As for evaluating the effect of surrounding ionic concentration, hFNIII 9-10 was dissolved in distilled water, phosphate buffered saline and RPMI 1640. Optical density (O.D.) was measured in ELISA reader. Results. The results were as follows; 1. The adsorption of hFNIII 9-10 showed significantly highest mean optical density (O.D.) value in $80{\mu}g/ml$. 2. The difference of ionic composition in DW, PBS and RPMI did not influence the adsorption amount of hFNIII 9-10.

PRODUCTION OF INTERLEUKIN-8 AND MONOCYTE CHEMOATTRACTANT PROTEIN-1 BY PERIPHERAL BLOOD MONOCYTES STIMULATED WITH MICROBES (세균에 노출된 말초혈액 단핵구에서 Interleukin-8과 Monocyte Chemoattractant Protein-1의 형성에 관한 연구)

  • Lee, Jung-Min;Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
    • /
    • v.25 no.2
    • /
    • pp.212-218
    • /
    • 2000
  • The recruitment of leukocytes to a site of inflammation is dependent on a complex interplay of a number of cytokines. Monocyte chemoattractant protein-1 (MCP-1) is a potent chemoattractant for monocytes, whereas interleukin-8 (IL-8) has chemotactic activity for neutrophils, lymphocytes, and basophils. The purpose of this study was to determine the effects of several microbes found in infected root canal systems on the production of inflammatoy cytokines, interleukin 8 and monocyte chemoattractant protein-1 from human peripheral blood mononuclear cells (PBMC). Monocytes isolated from peripheral blood were stimulated by group A streptococci (GAS, ATCC 19615), Enterococcus faecalis (ATCC 29212), Streptococcus mutans (ATCC 10449), Streptococcus sanguis (clinical isolate), and Candida albicans (ATCC 90029) respectively. Each of these bacteria induced dose-dependent induction in IL-8 and MCP-1 determined by ELISA. IL-8 production by each bacteria was decreased in the range of the microbe-to-PBMC ratios of 0.1-1.0. Group A streptococci was the week inducer of MCP-1 production. These results suggest that different oral pathogens induce specific dose-dependent patterns of cytokine release. Such patterns may provide a means of control of the type of immune celles particularly with regard to inflammatory leukocyte recruitment.

  • PDF

CA 15-3 (Mucin-1) and Physiological Characteristics of Breast Cancer from Lahore, Pakistan

  • Begum, Mumtaz;Karim, Sajjad;Malik, Arif;Khurshid, Rukhshan;Asif, Muhammad;Salim, Asmat;Nagra, Saeed Ahmed;Zaheer, Ahmad;Iqbal, Zafar;Abuzenadah, Adel Mohammed;Alqahtani, Mohammed Hussain;Rasool, Mahmood
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.13 no.10
    • /
    • pp.5257-5261
    • /
    • 2012
  • Background: High incidence of breast cancer and its fatal effect has reached an alarming stage across the globe, including the third world countries. Many factors have been reported to be associated with the development of breast cancer but detailed structural and functional information is missing. CA 15-3 is one of the known potential tumor marker of breast cancer; however little is known about structure and functional site of this protein. Present study aims to investigate the functional role of CA 15-3 in breast cancer, especially in development and metastasis. Material and Methods: Hundred female breast cancer patients confirmed by histopathological reports were included in the study. Their physiological characters were recorded in a performa. Enzyme linked immunosorbent assay (ELISA) technique was used to estimate serum CA 15-3 level. Immunohistochemistry was done for estrogen (ER), progesterone (PR) and Her2/neu receptors expression. Results: The study revealed the details of physiological characteristics of female breast cancer. Mean age was $37.72{\pm}5.99$ and $55.05{\pm}7.28$ years and serum CA 15-3 (MUC1) level was $60.47{\pm}8.59$ and $63.17{\pm}4.58$ U/ml in pre and post-menopause respectively, and both groups of women had sedentary life style. Their receptor status especially of progesterone, estrogen and HER-2/neu were positive in 50% of premenopausal women and 65% of postmenopausal women. Conclusion: There are multiple physiological factors promoting breast cancer. High serum CA 15-3 level and hormonal imbalance of ER, PR and Her2/neu appears to be the main cause of breast cancer. It may be possible that the functional sites of these proteins may be altered which may increase the chances of metastasis in breast cancer.

The Effect of Sonicated Extracts of Treponema Denticola and Treponema Lecithinolyticum on the Cytokine Secretion and Matrix Metalloproteinase Activation of Gingival Fibroblast (Treponema denticola와 Treponema lecithinolyticum의 분쇄액이 치은섬유아세포의 Cytokine 분비 및 Matrix metalloproteinase 활성에 미치는 영향)

  • Suh, Hye-Yuhn;Choi, Bong-Kyu;Choi, Seong-Ho;Cho, Kyoo-Sung;Kim, Chong-Kwan;Chai, Jung-Kiu
    • Journal of Periodontal and Implant Science
    • /
    • v.29 no.4
    • /
    • pp.979-995
    • /
    • 1999
  • This study was investigated to observe the effect of Treponema denticola cell sonicates(TDC) and Treponema lecithinolyticum cell sonicates(TLC) on cytokine secretion and matix metalloproteinase-2(MMP-2) activation of cultured human gingival fibroblast. Several experiments were performed including $IL-1{\beta}$, IL-6 ELISA for the effect on the $IL-1{\beta}$, IL-6 secretion of human gingival fibroblast. Also gelatinase zymography and gelatin dissolubility test for the activation of MMP-2 secreted by gingival fibroblast. The results were as follows. 1. The effect of TDC and TLC on IL-6 secretion of human gingival fibroblast showed statistically significant increase of IL-6 secretion in the TDC and TLC treated group compared to no treatment group(p<0.05) . 2. The amount of $IL-1{\beta}$ secretion was below the lower limit and there was no difference in the $IL-1{\beta}$ secretion of gingival fibroblast between TDC, TLC treated group and no treatment group. 3. The active form of pro MMP-2 with 72 kDa molecular weight was activated in both TDC and TLC treated group and clear band was appeared at 62kDa site on the zymography. 4. Gelatin dissolubility of MMP-2 secreted by gingival fibroblast was higher in TDC and TLC treated group compared to no treatment group(p<0.05). 5. In the TDC treated group, serine protease of T. denticola affect gelatin dissolubility. But in the TLC treated group gelatin was degraded by only MMP secreted by gingival fibroblast. Regarding to the above results, TDC and TLC have an effect on the IL-6 secretion increase of human gingival fibroblast and appears to activate pro MMP-2 which degrades collagen.

  • PDF

Expression and Function of Calcineurin in Inflammatory Arthritis (류마티스 관절염에서 칼시뉴린의 발현과 기능)

  • Park, Bo-Hyoung;Yoo, Seung-Ah;Hong, Kyung-Hee;Hyoung, Bok-Jin;Hwang, Yu-Na;Cho, Chul-Soo;Park, Won;Kim, Wan-Uk
    • IMMUNE NETWORK
    • /
    • v.6 no.1
    • /
    • pp.33-42
    • /
    • 2006
  • Background: Calcineurin plays a crucial role in T cell activation, cell growth, apoptosis, and angiogenesis, and its over-expression has been implicated in the pathogenesis of cardiomyopathy and stroke. However, the expression and function of calcineurin in the pathologic lesion of chronic inflammatory diseases, like rheumatoid synovium, remain to be defined. This study was aimed to determine the role of calcineurin in inflammatory arthritis and investigate the expression and function of calcineurin in the rheumatoid synovium and synoviocytes, the actual site of chronic inflammation. Methods: Immuno-histochemical staining using specific antibody to calcineurin was perfomed in the synovium of rheumatoid arthritis (RA). Fibroblast-like synoviocytes (FLS) from RA and osteoarthritis (OA) patients were isolated from RA and OA patients, and cultured with IL-1${\beta}$ and TNF-${\alpha}$ in the presence or absence of cyclosporin A, a calcineurin inhibitor. The calcineurin expression was assessed by phosphatase assay and Western blotting analysis. IL-6, -10, -17, matrix metalloproteinase (MMP)-1, -2, -3, and -9 released into the culture supernatants were measured by ELISA. After transfection with GFP-Cabin 1 gene into synoviocytes, the levels of IL-6 and MMPs were measured by ELISA. Results: Calcineurin was highly expressed in the lining layer of synovium and cultured synoviocytes of RA patients. The elevated calcineurin activity in the rheumatoid synoviocytes was triggered by proin flammatory cytokines such as IL-1${\beta}$ and TNF-${\alpha}$. In contrast, IL-10, an anti-inflammatory cytokine, failed to increase the calcineurin activity. The targeted inhibition of calcineurin by the over-expression of Cabin 1, a natural calcineurin antagonist, inhibited the production of IL-6 and MMP-2 by rheumatoid synoviocytes in a similar manner to the calcineurin inhibitor, cyclosporin A. Conclusion: These data suggest that abnormal activation of calcineurin in the synoviocytes may contribute to the pathogenesis of chronic arthritis, and thus provide a potential target for controlling inflammatory arthritis.

New Alternate Host of Rice stripe virus - 'Deulmuksae' (벼 줄무의잎마름병의 새로운 중간기주 '들묵새')

  • Yoon, Young-Nam;Lee, Bong-Choon;Jung, Ji-Hun;Kim, Jung-In;Hwang, Jae-Bok;Kim, Chang-Seok;Hong, Sung-Jun;Kang, Hang-Won;Song, Suk-Bo;Hong, Yeun-Gyu;Park, Sung-Tae;Lee, Key-Woon
    • Research in Plant Disease
    • /
    • v.15 no.2
    • /
    • pp.63-67
    • /
    • 2009
  • Rice stripe virus (RSV) is one of Tenuivirus Group, which is carried by small brown planthopper. There is an outbreak of RSV in South Korea at 20Ot, and 2007. The infection caused by RSV had been investigated on weeds around the rice cultivated areas 13 region and 26 site including Jeonbuk Buan and Chungnam Seocheon. There have a doubt as to alternate host of RSV is total 15 Family and 50 Species including Gramineae 24 species of Duksaepul (Alopecurus aequalis), H. sativum var. vulgare etc.. There is identified the infected RSV in Festuca myuros, Alopecurus aequalis, Hordeum sativum var. vulgare, Trisetum bifidum, Echinochloa crus-galli var. crus-galli, Digitaria ciliaris among this species. Deulmuksae is the overwintering exotic weed which sprout in Autumn and wither in Spring and commonly growed as green manure crop or cover crop. In order to identify the infection rate furthermore, 111 samples which were collected at Buan Gyehwa-myeon region, and 50 samples from Seocheon Maseo-myeon in June, 2008, were ELISA tested. The results are 32 positives from Buan, 28.8% infection rate, 8 positives from Seocheon 16.0% infection rate. RSV infection of Deulmuksae is not reported currently, and follow report first describes the Deulmuksae as an alternate host of RSV.

Effect of IRES Controlled Reporter Gene on Screening and Production of Recombinant Human EPO Proteins from Cultured CHO Cells

  • Lee Hyun Gi;Park Jin-Ki;Kim Sung-Woo;Ko Eun-Mi;Kim Byoung-Ju;Jo Su-Jin;Byun Sung-June;Yang Boh-Suk;Chang Won-Kyong;Lee Hoon-Taek;Lee Poong-Yeon
    • Reproductive and Developmental Biology
    • /
    • v.30 no.2
    • /
    • pp.81-85
    • /
    • 2006
  • This study was conducted to examine the effect of IRES controlled reporter gene on screening and production of recombinant human erythropoietin (EPO) proteins from cultured CHO cells. The cDNA was cloned for EPO from human liver cDNA Using site-directed mutagenesis, we generated recombinant human EPO (rhEPO) with two additional N-glycosylations (Novel erythropoiesis-stimulating protein: NESP). Wild type hEPO and NESP were cloned into expression vectors with GFP reporter gene under regulatory control of CMV promoter and IRES so that the vectors could express both rhEPO and GFP. The expression vectors were transfected to cultured CHO-K1 cells. Under microscopy, expression of GFP was visible. Using supernatant of the culture, ELISA assay, immunocytochemistry and in vitro assay using EPO dependant cell line were performed to estimate biological activity to compare the production characteristics (secretion levels, etc.) between rhEPO and NESP. The activity of NESP protein, obtained by mutagenesis, was described and compared with its rhEPO counterpart produced under same conditions. Although NESP had less secretion level in CHO cell line, the biological activity of NESP was greater than that of rhEPO. These results are consistent with previous researches. We also demonstrated that rhEPO and GFP proteins expressed simultaneously from transfected CHO cell line. Therefore we conclude that use of GFP reporter gene under IRES control could be used to screen and produce rhEPO in cultured CHO cells.

The relationship between the variants in the 5'-untranslated regions of equine chorionic gonadotropin genes and serum equine chorionic gonadotropin levels

  • Liu, ShuQin;Lian, Song;Yang, YunZhou;Fu, ChunZheng;Ma, HongYing;Xiong, ZhiYao;Ling, Yao;Zhao, ChunJiang
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.30 no.12
    • /
    • pp.1679-1683
    • /
    • 2017
  • Objective: An experiment was conducted to study the association between the single nucleotide polymorphisms (SNPs) in 5'-untranslated regions (5'-UTR) of equine chorionic gonadotropin (eCG) genes and the serum eCG levels. Methods: SNPs in 5'-UTR of eCG genes were screened across 10 horse breeds, including 7 Chinese indigenous breeds and 3 imported breeds using iPLEX chemistry, and the association between the serum eCG levels of 174 pregnant Da'an mares and their serum eCG levels (determined with ELISA) was analyzed. Results: Four SNPs were identified in the 5'-UTR of the $eCG{\alpha}$ gene, and one of them was unique in the indigenous breeds. There were 2 SNPs detected at the 5' end of the $eCG{\beta}$ subunit gene, and one of them was only found in the Chinese breeds. The SNP g.39948246T>C at the 5'-UTR of $eCG{\alpha}$ was associated significantly with eCG levels of 75-day pregnant mare serum (p<0.05) in Da'an mares. Prediction analysis on binding sites of transcription factors showed that the g.39948246T>C mutation causes appearance of the specific binding site of hepatocyte nuclear factor 3 forkhead homolog 2 (HFH-2), which is a transcriptional repressor belonging to the forkhead protein family of transcription factors. Conclusion: The SNP g.39948246T>C at the 5'-UTR of $eCG{\alpha}$ is associated with eCG levels of 75-day pregnant mare serum (p<0.05).