• Title/Summary/Keyword: ELISA IgM

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Detection of Toxoplasma antigens and antibodies in mice infected with different strains of Toxoplasma gonnii (톡소포자충의 충주에 따른 항원과 항체의 검출 시기 및 양상)

  • 이영하;김재영
    • Parasites, Hosts and Diseases
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    • v.33 no.3
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    • pp.201-210
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    • 1995
  • This study aims to assess the possible strain-dependent variations in detection of ToxopLosmn antigens and antibodies. The virulent RH strain or avirulent Beverley strain of T gondii were injected into mice, intraperitoneally, and their antigens, antibodies and parasites were identified from the blood or tissues: liver, brain and spleen by ELISA, Western blot and PCR. In mice infected with RH strain, circulating antigens and parasitemia were first detected from 2 days after infection, and ToxopIasma DNA were found in the blood, liver, brain and spleen from 3 days after infection. It was impossible to detect specific IgM and IgG antibodies to T gondij and any specific band was not found by Western blot. In mice infected with Beverley strain, circulating antigens were detected between day 10 and day 35. The Toxoplusma DNA was found in the blood and liver from day 15 until day 60, and in the brain from day 20. But Toxoplosma DNA in the spleen were mainly detected between day 10 and day 30. The IgM antibodies were first appeared on day 10 post-infection, and were noted obviously increased between day 15 and 25. The IgG antibodies were first detected on day 15, and showed progressively increased titers. The antibody binding bands were specific according to infection period. Sera from mice infected with Beverley strain reacted mainly with the antigen of 27.5-kDa and 32.5-kDa. In conclusion, mice infected with RH strain revealed Toxoplosma antigens strongly, but not antibodies. However. mice infected with Beverley strain revealed both the Toxoplasma antigens and antibodies. The present results showed that immune responses are different between avirulent and virulent T gonnii.

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Usefulness of Salivary and Serum Auto-antibodies Against Tumor Biomarkers HER2 and MUC1 in Breast Cancer Screening

  • Laidi, Fatna;Bouziane, Amal;Errachid, Abdelhamid;Zaoui, Fatima
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.1
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    • pp.335-339
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    • 2016
  • Background: The aim of this work was to investigate if serum and salivary auto-antibodies, isotypes IgG and IgM, against HER2 and MUC1 tandem repeat fragments could play a role in breast cancer screening. Materials and Methods: Our case-control study was conducted in breast cancer patients, in early stages (n=29), at the gynecology service, Maternity Souissi Hospital, Rabat, Morocco and healthy woman (n=31). Salivary and serum auto-antibodies against HER2 and MUC1 (tandem repeat) were assessed by enzyme-linked immunosorbent assay (ELISA) and compared between patients and healthy women using the Mann-Whitney U test. A P-value <0.05 was considered to be statistically significant. Results: Our data showed higher expression of all serum and salivary autoantibodies in patients as compared to healthy women p<0.05. However, serum IgM anti-MUC1 expression did not show a significant difference between cases and controls (p=0.79). Similarly, salivary IgG anti-HER2 expression did not differ (p=0.15). The correlation between the different isotypes of antibodies revealed that the highest correlation was between salivary IgG anti-HER2 and salivary IgG anti-MUC1(r=0.65). In fact, we have found in saliva the correlation between autoantibodies anti-MUC1 and anti-HER2 more important than in serum (r=0.59 and r=0. 55). However, the correlation between serum and saliva values for all antibodies was weak. Conclusions: Autoantibodies against HER2 and MUC1 may provide a useful approach in breast cancer screening when using both serum and saliva values.

Impacts of Host Immunization on the Translocation of Intestinal Bacteria and Growth Performance in Weanling Piglets

  • Moon, H.K.;Hanz, In K.;Gentry, J.L.;Parmentier, H.K.;Schrama, J.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.2
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    • pp.180-185
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    • 1999
  • Effects of host immunization on bacterial translocation and growth performance in weanling piglets were studied. Twenty four barrows were assigned to one of two immunization treatments: Control group (CON: immunized with placebo) or Immunization group [IMMU: immunized with Antigen cocktail; Keyhole limpet hemocyanin (KLH), Ovalbumin (OA), and Tetanus toxoid (TT)]. On d0, piglets were weaned and intramuscularly immunized with 2 ml of placebo or Antigen cocktail, respectively. Antigen-specific Ig titers were determined by ELISA (Enzyme Linked ImmunoSorbent Assay). Ig titers to E. coli-derived lipopolysaccharides (LPS) were measured as the indicator of bacterial translocation. Ig titers to LPS were higher (p<0.10, 0.05 or 0.01) in CON group before immunization (d0), but the difference disappeared with time and IgA titers to LPS became higher (p<0.05) in IMMU group on d39. In IMMU group, IgG titers to LPS from d28 onwards showed positive correlations (p<0.10, 0.05, 0.01 or 0.001) with IgG titers to KLH from d11 onwards and with IgM titers to KLH from d7 onwards. Generally, growth performance was negatively related to IgG titers to LPS. Average daily gain for d28 to d35 showed negative correlations (p<0.10, 0.05, or 0.01) with IgG titers to LPS on d28 onwards in immunization group. These results reveal some evidences that host immunization might facilitate bacterial translocation and high humoral immune responses to LPS are negatively related with the growth performance.

Influence of Environmental Living Standards on Helicobacter pylori Infection in Korean Elementary School Children (서울 지역 초등학생의 생활환경과 Helicobacter pylori 양성률)

  • Kim, Je-Woo;Kim, Hyo-Shin;Chung, Ki-Sup
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.4 no.1
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    • pp.10-17
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    • 2001
  • Purpose: We measured anti-H. pylori IgG in Korean elementary school children living in Shinchon area of Seoul, Korea to evaluate the influence of environmental living standards on H. pylori infection. Methods: IgG antibodies to H. pylori were measured in plasma using a commercial ELISA kit (GAP IgG Helicobacter pylori, Bio-Rad Laboratories Inc., Hercules, CA, USA). Information on environmental status such as place of birth, parental income, type of housing, number of persons in the household, parents' occupation, family history of peptic ulcer disease and gastric cancer was obtained. Statistical analysis was done by Chi-square and logistic regression test using SPSS $7.0^{TM}$ for Windows. Results: Study subjects consisted of 571 children, and the age distribution ranged from 6.0 to 13.6 years with a mean of $9.6{\pm}1.8$ years. Male-to-female ratio was 1.1:1. The seropositive rates of H. pylori infection ranged from 10.4% in children aged 6 years to 30.9% in 12 year-old group, overall 16.8%. The prevalence of H. pylori infection progressively increased with age, but there was no significant difference in seropositive rates among children in different age groups (p=0.06). Seropositive rates of anti-H. pylori IgG on the basis of gender, place of birth, parental income, type of housing, parents' occupation, family history of peptic ulcer disease and gastric cancer showed no statistically significant difference. Interestingly, however, seropositive rate of anti-H. pylori IgG showed statistical significance in relation to number of persons in the household (p=0.003; Odds ratio 1.50 by logistic regression test). Conclusion: These results suggest that number of persons in the household is the most important factor among environmental living standards, and that risk of H. pylori infection increases by increment of 1.5 times as the number of persons in the household increases by one.

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Daily Amperometric Monitoring of Immunoglobulin E in a Mouse Whole Blood: Model of Ovalbumin Induced Asthma

  • Lee, Ju Kyung;Yoon, Sung-hoon;Kim, Sang Hee
    • Journal of the Korean Electrochemical Society
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    • v.25 no.1
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    • pp.13-21
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    • 2022
  • There is an increasing interest in monitoring of specific biomarker for determining progression of a disease or efficacy of a treatment. Conventional method for quantification of specific biomarkers as enzyme linked immunosorbent assay (ELISA) has high material costs, long incubation periods, requires large volume of samples and involves special instruments, which necessitates clinical samples to be sent to a lab. This paper reports on the development of an electrochemical biosensor to measure total immunoglobulin E (IgE), a marker of asthma disease that varies with age, gender, and disease in concentrations from 0.3-1000 ng/mL with consuming 20 µL volume of whole blood sample. The sensor provides rapid, accurate, easy, point-of-care measurement of IgE, also, sequential monitoring of total IgE with ovalbumin (OVA) induced mice is another application of sensor. Taken together, these results provide an alternative way for detection of biomarkers in whole blood with low volumes and long-term ex-vivo assessments for understanding the progression of a disease.

Characterization of Mouse B Lymphoma Cells (CH12F3-2A) for the Study of IgA Isotype Switching (IgA Isotype Switching 연구를 위한 마우스 B Lymphoma Cell (CH12F3-2A)의 특성 연구)

  • Jang, Young-Saeng;Choi, Seo-Hyeun;Park, Seok-Rae;Kim, Hyun-A;Park, Jae-Bong;Kim, Pyeung-Hyeun
    • IMMUNE NETWORK
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    • v.4 no.4
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    • pp.216-223
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    • 2004
  • Background: It is well known that IgA isotype switching is induced by $TGF-{\beta}1$. LPS-activated mouse normal B cells well differentiate into IgA secreting plasma cells under the influence of $TGF-{\beta}1$. Nevertheless, there are lots of difficulties in studying normal B cells in detail because it is not simple to obtain highly purified B cells, showing low reproducibility and transfection efficacy, moreover impossible to keep continuous culture. To overcome these obstacles, it is desperately needed to develop B cell line which acts like normal B cells. In the present study, we investigated whether CH12F3-2A lymphoma cells are appropriate for studying IgA isotype switching event. Methods: CH12F3-2A B cell line was treated with LPS and $TGF-{\beta}1$, then levels of germ-line (GL) transcripts were measured by RT-PCR, and $GL{\alpha}$ promoter activity was measured by luciferase assay. In addition, membrane IgA (mIgA) expression and IgA secretion were determined by FACS and ELISA, respectively. Results: $TGF-{\beta}1$, regardless of the presence of LPS, increased level of $GL{\alpha}$ transcripts but not $GL{\gamma}2b$ transcripts. However, IgA secretion was increased dramatically by co-stimulation of LPS and $TGF-{\beta}1$. Both mIgA and IgA secretion in the presence of $TGF-{\beta}1$ were further increased by over-expression of Smad3/4. Finally, $GL{\alpha}$ promoter activity was increased by $TGF-{\beta}1$. Conclusion: CH12F3-2A cell line acts quite similarly to the normal B cells which have been previously reported regarding IgA expression. Thus, CH12F3-2A lymphoma cell line appears to be adequate for the investigation of the mechanism(s) of IgA isotype switching at the cellular and molecular levels.

Evaluation of IgG4 Subclass Antibody Detection by Peptide-Based ELISA for the Diagnosis of Human Paragonimiasis Heterotrema

  • Intapan, Pewpan M.;Sanpool, Oranuch;Janwan, Penchom;Laummaunwai, Porntip;Morakote, Nimit;Kong, Yoon;Maleewong, Wanchai
    • Parasites, Hosts and Diseases
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    • v.51 no.6
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    • pp.763-766
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    • 2013
  • A synthetic peptide was prepared based on the antigenic region of Paragonimus westermani pre-procathepsin L, and its applicability for immunodiagnosis for human paragonimiasis (due to Paragonimus heterotremus) was tested using an ELISA to detect IgG4 antibodies in the sera of patients. Sera from other helminthiases, tuberculosis, and healthy volunteers were used as the references. This peptide-based assay system gave sensitivity, specificity, accuracy, and positive and negative predictive values of 100%, 94.6%, 96.2%, 100%, and 88.9%, respectively. Cross reactivity was frequently seen against the sera of fascioliasis (75%) and hookworm infections (50%). Since differential diagnosis between paragonimiasis and fascioliasis can be easily done by clinical presentation and fascioliasis serology, this cross reaction is not a serious problem. Sera from patients with other parasitoses (0-25%) rarely responded to this synthetic antigen. This synthetic peptide antigen seems to be useful for development of a standardized diagnostic system for paragonimiasis.

Effects of Natural Honeybee (Apis mellifera ligustica) Venom Treatment on the Humoral Immune Response in Pigs (Beevenom 처리가 돼지의 체내 면역반응에 미치는 효과)

  • 조성구;김경수;이석천
    • Journal of Animal Science and Technology
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    • v.48 no.6
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    • pp.933-942
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    • 2006
  • This experiment was carried out to investigate effects of honeybee venom treatment on the humoral immune response in pigs. Corresponding author : S. K. Cho, Dept. of Animal Sci. Chung-Buk National University, Kaesin-dong, Cheongju, 361-763, Korea. phone : 043-261-2551. E-mail : deercho@chungbuk.ac.kr To investigate effects of natural honeybee venom on the concentration of immunoglobulin G, A, and M, 20 piglets(LY×D) from 3 sows were allocated into two groups bee venom-treated group(10 piglets) and non-treated control(10 piglets). Natural honeybee venom was treated at 0, 3, 6 days after birth and the acupoints were Hai-men(ST-25), Du-kou(CV-8) and Jiao-chao(GV-1) points at 0, 3 days after birth and the regions of castration and tail amputation point at 6 days. Control group was injected 1㎖ of saline to the same site. Concentrations of IgG, A, and M were measured with immunoturbidimetric method at 0, 3, 7, 14, and 21 days after treatment. To investigate the effect of bee venom on the production of antibodies against hog cholera and atrophic rhinitis vaccines that were used as indicator antigens, 40 piglets(LYxD) from 5 sows were grouped as bee venom-treated group (20 piglets) and control group(20 piglets). Natural honeybee venom was treated at 0, 3days(castration, tail amputation) and 21days after birth. The acupoints were Hai-men(ST-25), Du-kou(CV-8) and Jiao-chao (GV-1) points at 0 day, the regions of castration and tail ampution at 3 days and Jiao-chao(GV-1) and Bai-hui(GV-20) points at 21days after birth(weaning). Control group was injected 1ml of saline to the same site. Atrophic rhinitis vaccine was injected twice at 24 and 44 days after birth and hog cholera vaccine was also injected twice at 44 and 64 days after birth. Antibody titers against Bordetella bronchiseptica and hog cholera virus were measured by using tube agglutination and ELISA tests at 24, 34, 44, 54 and 74 days after birth. Concentrations of IgG of treated group were 339.52, 366.48, 296.52, 242.06 and 219.06mg/dl at 0, 3, 7, 14 and 21 days after birth, respectively. In contrast, concentrations of IgG in control group were respectively 347.10, 334.14, 243.28, 205.18 and 191.58mg/dl during same periods with treated group. Concentrations of IgG at 0 day was not significantly different between the treated group and control group but treated group were significantly increased by 10.28% at 3 days after birth (P<0.02), 21.88% at 7 days after birth(P<0.01), 18.0% at 14 days after birth(P<0.07) and 14.3% at 21 days after birth(P<0.01). Concentrations of IgA and Ig M were not significantly different. Antibody titers against hog cholera virus were significantly increased by 57.0% at 24 days after birth(P<0.03), 74.6% at 34 days after birth (P<0.006), 48.6% at 44 days after birth(P<0.017), 45.0% at 54 days after birth(P<0.16) and 44.4% at 74 days after birth (P<0.006) in bee venom treated group in comparison with control group. Antibody titers against the Bordetella bronchiseptica was significantly increased in Beevenom treated group as 9.1% (P<0.32) at 24days, 39.7% (P<0.002) at 34days, 31.9% (P<0.02) at 44days, 33.4% (P<0.01) at 54days and 57.3% (P<0.007) at 74 days after birth when compared with those of control group pigs. Collecting together, the results in this study showed that immune responses were increased by treatment of natural honeybee venom to pigs. These results suggested that the treatment of bee venom could be used effectively for the increase of productivity in livestock industry.

Induction of a systemic IgG and secretory IgA responses in mice by peroral immunization with uropathogenic Escherichia coli adhesin protein coupled to cholera toxin A2B subunits

  • Lee, Yong-Hwa;Kim, Byung-Oh;Rhee, Dong-Kwon;Pyo, Suh-Kneung
    • Biomolecules & Therapeutics
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    • v.11 no.3
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    • pp.157-162
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    • 2003
  • The generation of secretory IgA antibodies(Abs) for specific immune protection of mucosal surfaces depends on stimulation of the mucosal immune system, but this is not effectively achieved by parenteral or even oral administration of most soluble antigens. Thus, to produce a possible vaccine antigen against urinary tract infections, the uropathogenic E. coli (UPEC) adhesin was genetically coupled to the ctxa2b gene and cloned into a pMAL-p2E expression vector. The chimeric construction of pMALfimHIctxa2b was then transformed into E. coli K-12 TB1 and its nucleotide sequence was verified. The chimeric protein was then purified by applying the affinity chromatography. The purified chimeric protein was confirmed by SDS-PAGE and western blotting using antibodies to the maltose binding protein (MBP) or the cholera toxin subunit B (CTXB), plus the N-terminal amino acid sequence was analyzed. The orderly-assembled chimeric protein was confirmed by a modified $G_{M1}$-ganglioside ELISA using antibodies to adhesin. The results indicate that the purified chimeric protein was an Adhesin/CTXA2B protein containing UPEC adhesin and the $G_{M1}$-ganglioside binding activity of CTXB. This study also demonstrate that peroral administration of this chimeric immunogen in mice elicited high level of secretory IgA and serum IgG Abs to the UPEC adhesin. The results suggest that the genetically linked CTXA2B acts as a useful mucosal adjuvant, and that the adhesin/CTXA2B chimeric protein might be a potential antigen for oral immunization against UPEC.

Diagnostic Efficacy of a Recombinant Cysteine Protease of Spirometra erinacei Larvae for Serodiagnosis of Sparganosis

  • Rahman, S.M. Mazidur;Kim, Jae-Hwan;Hong, Sung-Tae;Choi, Min-Ho
    • Parasites, Hosts and Diseases
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    • v.52 no.1
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    • pp.41-46
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    • 2014
  • The mature domain of a cysteine protease of Spirometra erinacei plerocercoid larva (i.e., sparganum) was expressed in Escherichia coli, and its value as an antigen for the serodiagnosis of sparganosis was investigated. The recombinant protein (rSepCp-1) has the molecular weight of 23.4 kDa, and strongly reacted with the sparganum positive human or mice sera but not with negative sera by immunoblotting. ELISA with rSepCp-1 protein or sparganum crude antigen (SeC) was evaluated for the serodiagnosis of sparganosis using patient's sera. The sensitivity and specificity of ELISA using rSepCp-1 protein were 95.0% (19/20) and 99.1% (111/112), respectively. In contrast, the sensitivity and specificity of ELISA with SeC were 100% (20/20) and 96.4% (108/112), respectively. Moreover, in experimentally infected mice, the sensitivity and specificity of both ELISA assays were 100% for the detection of anti-sparganum IgG. It is suggested that the rSepCp-1 protein-based ELISA could provide a highly sensitive and specific assay for the diagnosis of sparganosis.