• Proceedings of the Korean Society of Applied Pharmacology
• /
• 2001.11a
• /
• pp.102-102
• /
• 2001

Taurine (2-ethaneaminosulfonic acid) is one of the major intracellular ${\beta}$ -amino acids in mammals and is required for a number of biological processes including membrane stabilization, osmoregulation, antioxidation, detoxification, modulation of calcium flux and neurornodulation. The taurine transporter (TAUT) which contains 12 hydrophobic membrane-spanning domains has been cloned from dog kidney, rat brain, mouse brain, human thyroid, placenta and retina. In this study, The TAUT cDNA from the human intestinal epithelial cell, HT-29 was cloned and sequenced. Reverse-transcription polymerase chain reaction (RT-PCR) was performed to amplify partial cDNA encoding human intestinal TAUT. The coding region of the PCR product was 732 bp long. The primers were designed to encode highly conserved amino acid sequences near the transmembrane domains III (IPYFIFLF) and Ⅵ (KYKYNSYR) both in human and mouse. The TAUT cDNA amplified was ligated into the pGEX 4T-1 expression vector. The resulting sequence of human intestinal TAUT cDNA (Accession number of NCBI Genebank is AF346763) was identical to the sequences of the TAUTs previously determined in the human placenta and retina except 3 base pairs from that of the reported human thyroid. TAUT specific antibodies were generated to use them as biological tools in the studies of the biological role of TAUT. Peptides of 149-162 amino acid residue (14 amino acids) of the TAUT were synthesized. The synthetic peptide used in this study was LFQSFQKELPWAHC. This region was chosen not only to avoid putative glycosylation sites but also to exclude regions of known homology with GABA transporters in the extracellular hydrophilic domains. The synthetic peptide, TAUT-1 was conjugated with carrier protein, kehole lympet hemocyanin (KLH) to use as an antigen. When used for immunization on a rabbit to produce polyclonal antiserum, the conjugates elicited high -titered specific anti-TAUT-1 antibodies, which reacted well with the ovalbumin (OVA) conjugated peptides in ELISA. The KLH-conjugated peptide was also used as immunizing antigen in BALB/c mice to produce TAUT specific monoclonal antibodies. From the culture supernatant of the hybridoma, the specificity of anti-TAUT-1 monoclonal antibodies was confirmed by ELISA. Further applications of more tools in TAUT expression analysis will be performed such as western blotting and flow cytometry.

• Toxicological Research
• /
• v.9 no.1
• /
• pp.125-132
• /
• 1993

Antigenic potential of recombinant human growth hormone (LBD-007), a newly developed drug for growth hormone deficiency, was investigated in mice and guinea pigs. 1. Mice showed production of antibodies against LBD-009 (1.5IU/kg) with aluminum hydroxide gel(alum) as an adjuvant, Judaged by the heterologous anaphylaxis (PCA) test using rats. On the other hand, antibodies against ovalbumin (OVA) inoculated with alum were definitely detected. 2. In the studies with guinea pigs, both the inoculation of LBD-009 (0.15IU/kg-1.5IU/kg) only and of LBD-009 with complete Freund's adjuvant (CFA) as an adjuvant did produce weak positive reactions in homologous passive cutaneous anaphylaxis (PCA). On the other hand, the inoculation of ovalbumin with complete Freund's adjuvant (CFA) produced positive reaction in PCA.

• Journal of Microbiology and Biotechnology
• /
• v.25 no.2
• /
• pp.268-273
• /
• 2015

The fluorescent-antibody-to-membrane-antigen (FAMA) test is regarded as the "gold standard" to detect protective antibodies to varicella-zoster virus (VZV) because of its high sensitivity and specificity. Because the classic FAMA test uses an infectious virus for detection of antibodies to VZV, it is labor-intensive, and also requires special equipment for handling the virus. For this reason, we attempted to develop a simple and safe FAMA assay. Because VZV glycoprotein E (gE) is one of the major VZV glycoproteins, we used the gE protein for the FAMA test (gE FAMA). Here, we demonstrate that overexpression of gE in HEK293T cells can be used to measure antibodies in human serum, and that gE FAMA titers are closely correlated with gpEIA ELISA data. These results indicate that our gE FAMA test has the potential to measure antibodies to VZV.

• Proceedings of the Korean Society of Applied Pharmacology
• /
• 1994.11a
• /
• pp.61-67
• /
• 1994

One fundamental problem in developing an AIDS vaccine is antigenic variation of HIV. Despite a substantial induced immune response in gp120-immunized monkeys and humans, high titers of V3-directed type specific neutralizing antibodies may not be sufficient to neutralize continuously emerging new isolates. Several studies analyzing anti-gp120 antibodies in HIV-infected individuals have clearly indicated that most broadly neutralizing antibodies are directed to conformation-dependent epitopes. Therefore, it seems important to evaluate the potential efficacy of candidate gp120 vaccines at inducing such antibodies, that might be potentially protective against multiple HIV strains. One concern in the development of any recombinant protein as a vaccine is its stability when mixed with an adjuvant. This could be a particularly important factor for recombinant gp120, given the conformational nature of its major, broadly neutralizing, epitopes. We hypothesized that gp120 complexed with recombinant CD4 could stabilize the conformation-dependent epitopes and effectively deliver these epitopes to the immune system. In this study, a soluble gp120-CD4 complex in Syntex Adjuvant Formulation was tested in mice to analyze the anti-gp120 antibody response. With the aim of defining the fine specificity and neutalizing activities of the immune response, 17Mabs were generated and characterized. The studies indicate that the gp120-CD4 complex elicits neutralizing anti-gp120 antibodies, most of which are directed to the conformation dependent epitopes.

• Pediatric Infection and Vaccine
• /
• v.12 no.2
• /
• pp.108-113
• /
• 2005

Purpose : Human herpsevirus 8(HHV-8), a gamma herpsevirus, was initially identified from Kaposi sarcoma(KS) lesions and has been known to be associated with several malignancies including Kaposi sarcoma. HHV-8 seroprevalence is variable by different geographic areas and populations. The prevalence of HHV 8 infection in Korean children is unclear. So, we investigated the prevalence of HHV-8 specific antibodies in healthy children in Seoul, Korea. Methods : Sera were obtained from 112 children(age 1~15 years, 64 males and 48 females) who visited our hospital for routine health checkup and used for investigating sero-prevalence of anti-HHV-8 antibodies. An indirect immunofluorescent assay was used to detect the IgG antibodies to the lytic viral antigen(Biotrin, Dublin, Ireland). A peptide mix ELISA kit was used to detect the IgG antibodies to peptides specific for HHV-8 open reading frame (ORF)(Biotrin, Dublin, Ireland). Results : Of 112 children, 4 children younger than 6 years of age were seropositive to HHV-8[all 4(3.5%) were positive by IFA and 2(1.8%) were positive by ELISA]. Conclusion : These results suggest that the prevalence of antibody to HHV 8 in children in Korea is very low.

• IMMUNE NETWORK
• /
• v.2 no.2
• /
• pp.102-108
• /
• 2002

Background: This study was aimed to differentiate two forms of CTLA-4 (CD152) in activated peripheral blood lymphocyte and clarify the mechanism how cytoplasmic form of this molecule is targeted to cell surface. Methods: For this purpose we generated 2 different anti-human CD152 peptide antibodies and 5 different N'-terminal deletion mutant CTLA4Ig fusion proteins and carried out a series of Western blot and ELISA analyses. Antipeptide antibodies made in this study were anti-CTLA4pB and anti-CTLA4pN. The former recognized a region on extracellular single V-like domain and the latter recognized N'-terminal sequence of leader domain of human CD152. Results: In Western blot, the former antibody recognized recombinant human CTLA4Ig fusion protein as an antigen. And this recognition was completely blocked by preincubating antipeptide antibody with the peptide used for the antibody generation at the peptide concentration of 200 ug/ml. These antibodies were recognized human CD152 as a cytoplasmic sequestered- and a membrane bound- forms in phytohemagglutinin (PHA)-stimulated peripheral blood lymphocyte (PBL). These two forms of CD152 were further differentiated by using anti-CTLA4pN and anti-CTLA4pB antibodies such that former recognized cytosolic form only while latter recognized both cytoplasmic- and membraneforms of this molecule. Furthermore, in a transfection expression study of 5 different N'-terminal deletion mutant CTLA4Ig, mutated proteins were secreted out from transfected cell surface only when more than 6 amino acids from N'-terminal were deleted. Conclusion: Our results implies that cytosolic form of CTLA-4 has leader sequence while membrane form of this molecule does not. And also suggested is that at least N'-terminal 6 amino acid residues of human CTLA-4 are required for regulation of targeting this molecule from cytosolic- to membrane- area of activated human peripheral blood T lymphocyte.

• Clinical and Experimental Reproductive Medicine
• /
• v.12 no.1
• /
• pp.41-46
• /
• 1985

Various immunoserologic and cellular immunity techniques have been used to explore the presence of antisperm antibodies in the serum and seminal plasma of male patients and in the blood and genital fluid of infertile women. Several recent comparative investigations using various assays to detect and quantitate levels of antibody to human spermatozoa have produced widely varying results. So the first WHO workshop on iso- and autonatibodies to human spermatozoa in 1974 tried to establish some unification in the techniques used. The purpose of this study is to compare the results of two methods-the Kibrick macro-agglutination test and the Isojima micro-immobilization test-using the same test materials based on recommandation from WHO workshop. The results are as follows: 1. Twenty normal controls showed negative reactions in all the 2 tests. Out of 25 patients, the positive sera were noted in 15 (60%) on the Kibrick test and 13 (51%) on the Isojima test. 2. Twelve (48%) out of 25 patients showed positive reactions in the two tests, and 16 (64%) out of 25 patients showed positive reaction in one or more tests. 3. The titers of the antisperm antibodies on the Kibrick test was higher than that on the Isojima test. Therefore, it seems to be possible to increase the chances of detection of the antisperm antibodies, if two tests are imployed.

• Microbiology and Biotechnology Letters
• /
• v.27 no.4
• /
• pp.344-348
• /
• 1999

A competitive enzyme-linked immunosorbent assay(ELISA) for the detection of antibodies to Treponema pallidum(T.pallidum) was developed and evaluated. T.apllidum lysate was immobilized on the surface of microplate wells and horseradish peroxidase labeled human anti-T.pallidum lysate was immobilized on the surface of microplate wells and horseradish peroxidase labeled human anti-T.pallidum was prepared and used as a tracer. The performance of the competitive ELISA was evaluated by using different specimens. The competitive ELISA showed a sensitivity of 100% in a performance panel consisting of serum and plasma with anti-T.pallidum reactivity ranging from negative to strong positive by FTA-ABS test system and 120 plasma samples positive by TPHA. The specificity of the competitive ELISA was 100% in 1,200 plasma samples collected from healthy seronegative blood donors. These results suggest that the competitive ELISA provides an excellent assay method for the detection of antibodies to T.pallidum, and may be particularly useful for serological blood screening of syphilis.

• The Journal of the Korean Society for Microbiology
• /
• v.19 no.1
• /
• pp.79-83
• /
• 1984

To determine the existence of anti-HLA antibodies finally in 220 human placental extracts to be proved negative antiserum by previous anti-HLA A,B,C antibody screening procedure, the present study was performed by fractionation of immunoglobulins using saturated ammonium sulfate and by simple batch method on DEAE cellulose. Thereafter using known 150 T-lymphocyte panels, complement-dependent microlymphocytotoxicity test was performed to observe the existence of anti-HLA antibodies and the degree of the antibody response of the concentrates. The following results were obtained: 1. Of total 141 placental sera concentrated 45 cases(31.9%) were showed significant anti-HLA A,B,C antibody response after concentration(Excellent, 19(13.5%), Good, 3(2.1%), Weak, 23(16.3%)). 2. Anti-HLA specificities of placental sera obtained after concentration were A2, A24, B13, B27, B44, B51, CN1, C7. 3. A new type C new-1 anti-HLA antibody that is only expressed in Korean people, was obtained. 4. 79 placental sera purrified by simple batch method using DEAE cellulose were showed negative anti-HLA antibody responses.

• Korean Journal of Veterinary Service
• /
• v.46 no.4
• /
• pp.335-338
• /
• 2023

The COVID-19 pandemic, triggered by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has not only impacted human health on a global scale but also raised concerns about the vulnerability of a wide array of animals that are in close contact with humans. Particularly, the potential for infection and the subsequent immune response in domestic pets such as dogs and cats remain largely unexplored under natural living conditions. In this study, we have undertaken the task of detecting and tracking the presence of antibodies against SARS-CoV-2 in a small cohort of household pets-specifically, two dogs and two cats. Employing techniques such as the indirect ELISA and plaque reduction neutralization tests, we observed that the neutralizing antibodies against SARS-CoV-2 in these animals were maintained for a duration of up to six months following their initial positive test result. This duration mirrors the antibody response documented in human cases of COVID-19, suggesting a comparable post-infection immune response timeline between humans and these domestic animals.