• IMMUNE NETWORK
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• 제12권1호
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• pp.27-32
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• 2012

Background: Toll-like receptors (TLRs) have been extensively studied in recent years. However, functions of these molecules in murine B cell biology are largely unknown. A TLR4 stimulant, LPS is well known as a powerful polyclonal activator for murine B cells. Methods: In this study, we explored the effect of a murine TLR9 stimulant, M6-395 (a synthetic CpG ODNs) on B cell proliferation and Ig production. Results: First, M6-395 was much more potent than LPS in augmenting B cell proliferation. As for Ig expression, M6-395 facilitated the expression of both TGF-${\beta}1$-induced germ line transcript ${\alpha}$ ($GLT{\alpha}$) and IL-4-induced $GLT{\gamma}1$ as levels as those by LPS and Pam3CSK4 (TLR1/2 agonist) : a certain Ig GLT expression is regarded as an indicative of the corresponding isotype switching recombination. However, IgA and IgG1 secretion patterns were quite different--these Ig isotype secretions by M6-395 were much less than those by LPS and Pam3CSK4. Moreover, the increase of IgA and IgG1 production by LPS and Pam3CSK4 was virtually abrogated by M6-395. The same was true for the secretion of IgG3. We found that this unexpected phenomena provoked by M6-395 is attributed, at least in part, to its excessive mitogenic nature. Conclusion: Taken together, these results suggest that M6-395 can act as a murine polyclonal activator but its strong mitogenic activity is unfavorable to Ig isotype switching.

• 대한핵의학회지
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• 제26권1호
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• pp.116-123
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• 1992

$^{123}I$ has ideal half life of 13 hours, suitable 159 keV gamma energy for imaging, and easy labeling methods. In Korea Cancer Center Hospital, $^{123}I$ has been produced by MC-50 cyclotron. The purpose of this study is looking for good labeling condition of $^{123}I$ and $^{99m}Tc$ to nonspecific human polyclonal IgG, and comparing these with $^{67}Ga-citrate$ in the abscess bearing mice. Human polyclonal nonspecific IgG was labeled with 0.2 M phosphate buffer added $^{123}I$ by chloramine T method. Human polyclonat nonspecific IgG was labeled with $^{99m}Tc-gluconate$ after activation with $\beta-mercaptoethanol$. In the abscess bearing mice, the radioactivity in the abscess was higher in 24 hours than 6 hours after injection. In the abscess, $^{123}I$ nonspecific IgG had higher uptake than $^{99m}Tc-IgG\;or\;^{67}Ga-citrate$. There was no significant difference in absecess uptake of $^{123}I-IgG$ among 24, 72, 120 hours abscess age. Further clinical researches with $^{123}I-nonspecific$ IgG, and other immunoscintigraphies using $^{123}I$ are expected.

• Journal of Gastric Cancer
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• 제1권3호
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• pp.129-135
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• 2001

Purpose: Dysplasia or flat adenoma of the stomach is regarded as a precancerous lesion. However, the frequency and the evolutionary process of malignant transformation of gastric dysplasia are still debated. In order to see whether the lesion was a monoclonal or a polyclonal proliferation, clonality was assayed by X-linked HUMARA polymorphism. Materials and Methods: DNA was extracted from the paraffin-embedded tissue of 16 consecutive cases of endoscopic biopsy, eight of which supplied both dysplastic and nondysplastic tissue for comparison. HUMARA was amplified by PCR with or without pretreatment with methylationsensitive restriction enzyme, HpaII. The amplification products were electrophoresed on polyacrylamide gel and silver-stained. Results: Among the 16 cases, 13 cases were informative and 3 cases noninformative. Of the 13 cases, one case showed skewed lyonization, rendering 12 cases to be analyzed further. A monoclonal band pattern was noted in 2 cases, and a polyclonal band pattern in 10 cases. A review of the histopathologies of the monoclonal and the polyclonal cases did not reveal features discriminating the two groups. Conclusion: These results suggest that gastric dysplasia is a disease entity heterogeneous in the genetic level, and many cases may be non-neoplastic.

• Parasites, Hosts and Diseases
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• 제42권1호
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• pp.35-40
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• 2004

The nfa 1 gene was cloned from a cDNA library of pathogenic Naegleria fowleri by immunoscreening; it consisted of 360 bp and produced a 13.1 kDa recombinant protein (rNfa1) that showed the pseudopodia-specific localization by immunocytochemistry in the previous study. Based on the idea that the pseudopodia-specific Nfa1 protein mentioned above seems to be involved in the pathogenicity of N. fowleri, we observed the effect of an anti-Nfa1 antibody on the proliferation of N. fowleri trophozoites and the cytotoxicity of N. fowleri trophozoites on the target cells. The proliferation of N. fowleri trophozoites was inhibited after being treated with an anti-Nfa1 polycional antibody in a dose-dependent manner for 48 hrs. By a light microscope, CHO cells co-cultured with N. fowleri trophozoites (group I) for 48 hrs showed severe morphological destruction. On the contrary, CHO cells co-cultured with N. fowleri trophozoites and anti-Nfa1 polyclonal antibody (1:100 dilution) (group II) showed less destruction. In the LDH release assay results, group I showed 50.6% cytotoxicity, and group II showed 39.3%. Consequently, addition of an anti-Nfa1 polyclonal antibody produced a decreasing effect of in vitro cytotoxicity of N. fowleri in a dose-dependent manner.

• 한국식품위생안전성학회지
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• 제13권2호
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• pp.129-134
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• 1998

Nivalenol(NIV)의 검출을 위한 효소면역측정법(ELISA)을 개발하기 위하여 tetraacetyl nivalenol(Ac4-NIV)에 대한 다클론항체를 생산하고 그 조건을 확립하였다. Ac4-NIV-hemisuccinate를 bovine serum albumin에 공유결합 시킨 Ac4-NIV-HS-BSA를 Freund's adjuvant와 함께 수차례 토끼에 피하면역하였다. 가장 높은 항체가를 나타낸 항혈청으로부터 정제한 항체와 Ac4-NIV-HS-HRP conjugate를 이용하여 직접 경합 ELISA(cdELISA)를 확립하였다. 그 표준 곡선으로부터 Ac4-NIV의 검출 범위는 10~5,000 ng/ml(ppb)임을 알 수 있었다. 특이항체의 Ac4-NIV과 acetyl T-2에 대한 반응성은 각각 100, 70%였으나, NIV, deoxynivalenol, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, triacetyl deoxynivalenol, fusarenon-X, T-2에 대한 반응성은 0.1% 이하로 극히 미약하였다. NIV를 인위적으로 오염시킨 옥수수 시료를 70% acetonitrile 추출하고 acetylation 한 다음 cdELISA를 행하였을 때, 분석의 회수율은 100, 300, 1,000 ng/g(ppb)에서 각각 108, 143, and 70%(평균, 107%)로 나타났다.

• Journal of Animal Science and Technology
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• 제47권1호
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• pp.19-28
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• 2005

The objectives of the current study were to develop polyclonal antibodies in sheep against adipocyte plasma membrane(APM) proteins isolated from swine, to investigate tissue specificity, and to determine cytotoxic effects of antiserum on swine adipocytes. Plasma membrane proteins from adipocyte, brain, heart, kidney, liver, and spleen were isolated using a 32% sucrose gradient. Adult male sheep was immunized three times at three week interval with the purified swine APM proteins. Antiserum was taken from immunized sheep at 10, 12, and 14 days after the third immunization. Antiserum expressed strong reactivity with APM proteins determined by enzyme-linked immunosorbent assay(ELISA), and the reactivity could be detected at dilutions in excess of 1 : 81,000. Antiserum showed very low binding affinity with proteins isolated from brain, heart, kidney, liver, or spleen. Tissue specificity of the antiserum was reconfirmed by Western immunoblotting using anti-sheep immunoglobulin G•alkalinephosphatase conjugate as a secondary antibody. The reactivity of antiserum to the external surface of fixed swine adipocytes was confmned by an immunohistochemical technique using anti-sheep immunoglobulin G-FITC. Confluent swine adipocytes in culture were lysed by antiserum treatment and cytosolie lactate dehydrogenase(LDH) was released as a dose-dependent patterns while adipocytes treated with normal sheep serum maintained their integrity and expressed low level of LDH. These results implicate that fat contents in the pigs can be reduced by immunological methods.

• Applied Biological Chemistry
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• 제49권3호
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• pp.196-201
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• 2006

Vitellogenin은 어류의 난황 단백질의 전구체 물질로서 암컷 혈청에서 발견되는 단백질이며, 외부에서 에스트로겐이나 내분비계장애물질에 노출된 경우에는 수컷이나 미성숙한 암컷에서도 이의 합성이 촉진되는 것으로 보고되었다. 따라서 수컷에서 유도되는 vitellogenin은 외인성 에스트로겐 물질에 노출되었음을 암시하는 중요한 지표로 인정되고 있다. Vitellogenin에 대한 항체를 생산하기 위하여 잉어의 vitellogenin 서열 중의 일부분에 대한 peptide를 합성한 후 그 합성 peptide에 대한 항체를 제작하였다. 그리고 $17{\beta}$-estradiol을 주입한 잉어의 혈청에서 DE-52 이온 교환 크로마토그래피를 사용하여 vitellogenin을 정제한 후 이에 대한 항체를 제작하였다. 본 연구에서는 상기의 합성 vitellogenin peptide에 대하여 제작한 다클론 항체와 vitellogenin 단백질에 대한 다클론 항체가 추후 에스트로겐의 지표로 사용될 수 있는 지를 조사하고자 항체의 반응성을 조사하였다. 정제하여 얻은 vitellogenin에 대한 다클론 항체는 Western blotting 시 $17{\beta}$-estradiol을 주입한 잉어의 혈청과 암컷 잉어의 혈청에 있는 vitellogenin과 반응한 반면에 vitellogenin peptide에 대한 다클론 항체는 이들과 반응하지 않았다. 이는 공유결합적으로 많은 변형이 일어난 vitellogenin 단백질은 vitellogenin 합성 펩타이드에 대한 항체로는 검출되지 않음을 나타낸다.

• Biomolecules & Therapeutics
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• 제2권4호
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• pp.376-382
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• 1994

Repeated injections of low-doses of mercuric chloride in rats or mice induce polyclonal activation which includes the induction of anti-glomerular basement membrane (GBM) antibodies and circulating immune complex and it results in nephritis. Because this disease is autoimmune mediated disease resulted from immune dysfunction, immunomodulators are used to control the symptoms or to cure the disease. Irpex lacteus Fr. is a kind of new medicinal fungus. The polysaccharide fraction extracted from submerged fermentation of Irpex lacteus Fr. decreased the serum agglutinin, serolysin and IgM plaque forming cells in normal mice. The hitherto obtained clinical results suggested that it significantly improved the oligourea, edema, and hypertension in patients who have nephritis. To elucidate the action-mechanisms of Irpex lacteus Fr., we established the experimental model of HgCl$_2$induced polyclonal activation by intraperitoneal administrations of HgCl$_2$to mice. To assess the immunomodulating effect of Irpex lacteus fraction, we Investigated its effects on the mitogen induced proliferation and IgM PFC counts of splenic lymphocytes in mice during the treatment of HgCl$_2$. The Irpex lacteus polysaccharide reduced the abnormally increased mitogen induced Iymphocyte proliferation and IgM PFCs to almost normal levels. And the Irpex lacteus polysaccharides prevented the increasement of serum immunoglobulin level induced by HgCl$_2$. These data suggested that the Irpex lacteus polysaccharides might have the immunomodulating activity to prevent and /or improve the HgCl$_2$ induced autoimmune disease.

• Parasites, Hosts and Diseases
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• 제60권2호
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• pp.143-147
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• 2022

Acanthamoeba keratitis (AK) is a rare ocular disease, but it is a painful and sight-threatening infectious disease. Early diagnosis and adequate treatment are necessary to prevent serious complications. While AK is frequently diagnosis via several PCR assays or Acanthamoeba-specific antibodies, a more specific and effective diagnostic method is required. This study described the production of a polyclonal peptide antibody against the periplasmic binding protein (PBP) of A. castellanii and investigated its diagnostic potential. Western blot analysis showed that the PBP antibody specifically reacted with the cell lysates of A. castellanii. However, the PBP antibody did not interact with human corneal epithelial (HCE) cells and the other 3 major causative agents of keratitis. Immunocytochemistry (ICC) results revealed the specific detection of A. castellanii trophozoites and cysts by PBP antibodies when A. castellanii were co-cultured with HCE cells. PBP antibody specificity was further confirmed by co-culture of A. castellanii trophozoites with F. solani, S. aureus, and P. aeruginosa via ICC. The PBP antibody specifically reacted with the trophozoites and cysts of A. polyphaga, A. hatchetti, A. culbertsoni, A. royreba, and A. healyi, thus demonstrated its genus-specific nature. These results showed that the PBP polyclonal peptide antibody of A. castellanii could specifically detect several species of Acanthamoeba, contributing to the development of an effective antibody-based AK diagnostics.

• 한국발생생물학회지:발생과생식
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• 제9권2호
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• pp.135-142
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• 2005

무척추동물에서 척추동물에 이르기까지 대부분의 난생동물들의 난황 단백질의 전구체를 vitellogenin(VTG)이라 한다. 난생 척추동물에서 VTG는 간에서 합성되어 혈액을 통해 난세포로 전이된다. 암컷 어류는 정상적인 생식주기에서 난황단백전구체 형성이 시작되면 혈중 농도는 급격히 증가하게 된다. 수컷도 VTG의 유전자를 가지고 있기 때문에 낮은 수준의 내인성 에스트로겐으로 아주 적은 양의 단백질이 있을 수 있다. 그렇지만 수컷에 외인성에스트로겐 유사물질에 노출되면 그 양은 증가하게 된다. 따라서 어류에서 VTG는 내분비계 장애물질 효과를 조사하는 유용한 생물학적 추적자로 사용할 수 있다. 이 연구에서는 에스트로겐 유사물질에 오염된 지역에 서식하는 망둑어류의 혈중 VTG의 수준을 정량할 수 있는 방법을 제공하고자 한다. 이러한 목적을 위해 꾹저구(Chaenogobius annularis)의 VTG를 분리하고, 이에 대한 단클론 항체와 다클론 항체를 제작하여 혈중 VTG 수준을 정량할 수 있는 sandwich 경합 ELISA system을 만들었다. 난황 단백질에 대한 단클론 항체와 다클론 항체를 사용한 ELISA system에 대한 타당성 검토를 하였다. 순차적으로 희석한 암 컷의 혈청에 대한 흡광도 곡선은 VTG 표준농도의 곡선과 평행하였으나 수컷의 혈청에 대한 흡광도 곡선은 평행하지 않았다. 이 VTG에 대한 sandwich ELISA system으로 꾹저구(C. annularis)의 혈중 VTG의 수준을 정량뿐만 아니라 문절망둑(Acanthogobius flaviman)과 풀망둑(A. hasta)의 혈중 VTG 수준을 정량할 수 있다.