• Title/Summary/Keyword: IgM

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Plant-derived PAP proteins fused to immunoglobulin A and M Fc domains induce anti-prostate cancer immune response in mice

  • Yang Joo Kang;Deuk-Su Kim;Seyoung Kim;Young-Jin Seo;Kisung Ko
    • BMB Reports
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    • v.56 no.7
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    • pp.392-397
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    • 2023
  • In this study, recombinant Fc-fused Prostate acid phosphatase (PAP) proteins were produced in transgenic plants. PAP was fused to immunoglobulin (Ig) A and M Fc domain (PAP-IgA Fc and PAP-IgM Fc), which were tagged to the ER retention sequence KDEL to generate PAP-IgA FcK and PAP-IgM FcK. Agrobacterium-mediated transformation was performed to produce transgenic tobacco plants expressing four recombinant proteins. Genomic PCR and RT-PCR analyses confirmed the transgene insertion and mRNA transcription of PAP-IgA Fc, PAP-IgM Fc, PAP-IgA FcK, and PAP-IgM FcK in tobacco plant leaves. Western blot confirmed the expression of PAP-IgA Fc, PAP-IgM Fc, PAP-IgA FcK, and PAP-IgM FcK proteins. SEC-HPLC and Bio-TEM analyses were performed to confirm the size and shape of the plant-derived recombinant PAP-Fc fusion proteins. In mice experiments, the plant-derived IgA and IgM Fc fused proteins induced production of total IgGs including IgG1 against PAP. This result suggests that IgA and IgM Fc fusion can be applied to produce recombinant PAP proteins as a prostate cancer vaccine in plant expression system.

Developmental Changes of Serum IgA, IgG and IgM Concentrations in Broiler Chicks - III. Isolation of IgM and Developmental Changes of Serum IgM Levels (육계의 혈청중 면역글로부린(IgA, IgG, IgM)농도의 발육시기별 변화상 - III. IgM 분리 및 발육시기별 농도수준)

  • 김정우;이민호;김춘수;김상희;박근식
    • Korean Journal of Poultry Science
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    • v.21 no.3
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    • pp.175-182
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    • 1994
  • An experiment was conducted to establish a large scale production method of anti-serum against chicken IgM and to profile the developmental changes of serum IgM levels during the feeding period(from hatching to 7 weeks of age) in broiler chicks. Blood samples were taken from Hubbard chicken at the age of hatching, three days of age, and weekly thereafter till to 7 weeks of age. The pure IgM was isolated from ammonium sulfate treated chicken serum by both sephadex G-200 and sepharose CL-6B chromatography. The breaking-through peak containing IgM appeared from the fraction 26 to 28. These fractions consisted mainly of IgM when tested by anti-chicken IgM(Nordic, Netherlands). Immunized with the heavy chain of this purified IgM, the rabbit immune sera(anti-chicken IgM) were formed a reaction only with the purified chicken IgM. The quantitative assay of serum IgM were carried by RID method. The optimal time for diffusion was 14 hours and the coefficient of determination($R^{2}$) for regression equation of standard curve was 0.992. It was observed that IgM concentrations were the highest at hatching(3.23 mg /mL), after that decreased gradually. From 2 to 5 weeks of age the levels unchanged(2.0 ~ 2.3mg /mL), and gradually decreased to 7 weeks of age(1.3 mg /mL).

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Anti-IgE mAb Suppresses Systemic Anaphylaxis through the Inhibitory IgG Receptor Fc ${\gamma}$ RIIb in Mice - Interaction between Anti-IgE and Fc ${\gamma}$ RIIb -

  • Kang, Nam-In;Jin, Zhe-Wu;Lee, Hern-Ku
    • IMMUNE NETWORK
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    • v.7 no.3
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    • pp.141-148
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    • 2007
  • Background: Anti-IgE mAb which binds circulating but not receptor-bound IgE has been shown to be effective in treatment for asthma and other allergic diseases. However, the mechanisms by which anti-IgE mAb influences the pathophysiological responses are remained to be illustrated. This study was undertaken to examine the therapeutic efficacy of non-anaphylactogenic anti-mouse IgE mAb using murine models of IgE-induced systemic fatal anaphylaxis. Methods: Active systemic anaphylaxis was induced by either penicillin V(Pen V) or OVA and passive systemic anaphylaxis was induced by either anaphylactogenic anti-mouse IgE or a mixture of anti-chicken gamma globulin (CGG) IgG1 mAb and CGG. The binding of the Fc portion of anti-IgE to CHO-stable cell line expressing mouse Fc ${\gamma}$ RIIb was examined using flow cytometry. Fc fragments of anti-IgE mAb were prepared using papain digestion. The expression of phosphatases in lungs were assessed by Western blotting and immunohistochemistry. Results: Anti-IgE mAb prevented IgE- and IgG-induced active and passive systemic fatal reactions. In both types of anaphylaxis, anti-IgE mAb suppressed antigen-specific IgE responses, but not those of IgG. Anti-IgE mAb neither prevented anaphylaxis nor suppressed the IgE response in Fc ${\gamma}$ RIIb-deficient mice. The Fc portion of anti-IgE mAb was bound to murine Fc ${\gamma}$ RIIb gene-transfected CHO cells and inhibited systemic anaphylaxis. Anti-IgE mAb blocked the anaphylaxis-induced downregulation of Fc ${\gamma}$ RIIb-associated phosphatases such as src homology 2 domain-containing inositol 5-phosphatase (SHIP) and phosphatase and tensin homologue deleted on chromosome ten (PTEN). Conclusion: Anti-IgE mAb prevented anaphylaxis by delivering nonspecific inhibitory signals through the inhibitory IgG receptor, Fc ${\gamma}$ RIIb, rather than targeting IgE.

Electrophysiological features and prognosis of peripheral neuropathy associated with IgM monoclonal gammopathy: a single-center analysis in South Korea

  • Sooyoung Kim;Bit Na Lee;Seung Woo Kim;Ha Young Shin
    • Annals of Clinical Neurophysiology
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    • v.25 no.2
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    • pp.84-92
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    • 2023
  • Background: Clinical spectrum of immunoglobulin M (IgM) monoclonal gammopathy varies from IgM monoclonal gammopathy of unknown significance (IgM-MGUS) to hematological malignancies. We evaluated the clinical features, electrophysiological characteristics, and prognosis of patients with peripheral neuropathy associated with IgM monoclonal gammopathy (PN-IgM MG). Methods: We retrospectively evaluated 25 patients with PN-IgM MG. Peripheral neuropathy was classified as axonal, demyelinating, or undetermined, based on electrophysiological studies. We classified the enrolled patients into the IgM-MGUS and malignancy groups, and compared the clinical and electrophysiological features between the groups. Results: Fifteen patients had IgM-MGUS and 10 had hematologic malignancies (Waldenström's macroglobulinemia: two and B-cell non-Hodgkin's lymphoma: eight). In the electrophysiological evaluation, the nerve conduction study (NCS) criteria for demyelination were met in 86.7% of the IgM-MGUS group and 10.0% of the malignancy group. In particular, the distal latencies of the motor NCS in the IgM-MGUS group were significantly prolonged compared to those in the malignancy group (median, 9.1 ± 5.1 [IgM-MGUS], 4.2 ± 1.3 [malignancy], p = 0.003; ulnar, 5.4 ± 1.9 [IgM-MGUS], 2.9 ± 0.9 [malignancy], p = 0.001; fibular, 9.3 ± 5.1 [IgM-MGUS], 3.8 ± 0.3 [malignancy], p = 0.01; P-posterior tibial, 8.3 ± 5.4 [IgM-MGUS], 4.4 ± 1.0 [malignancy], p = 0.04). Overall treatment responses were significantly worse in the malignancy group than in the IgM-MGUS group (p = 0.004), and the modified Rankin Scale score at the last visit was higher in the malignancy group than in the IgM-MGUS group (2.0 ± 1.1 [IgM-MGUS], 4.2 ± 1.7 [malignancy], p = 0.001), although there was no significant difference at the initial assessment. Conclusions: The risk of hematological malignancy should be carefully assessed in patients with PN-IgM MG without electrophysiological demyelination features.

Serologic Tests of IgG and IgM Antibodies and IgG Avidity for Diagnosis of Ocular Toxoplasmosis

  • Rahimi-Esboei, Bahman;Zarei, Mohammad;Mohebali, Mehdi;Valian, Hossein Keshavarz;Shojaee, Saeedeh;Mahmoudzadeh, Raziyeh;Salabati, Mirataollah
    • Parasites, Hosts and Diseases
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    • v.56 no.2
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    • pp.147-152
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    • 2018
  • This prospective study was aimed to detect acute and chronic ocular toxoplasmosis by comparison of anti-Toxoplasma gondii IgM and IgG antibody levels and IgG avidity test. One hundred and seventeen patients with ocular toxoplasmosis (OT) who referred to the Farabi Eye Hospital, Tehran, Iran were included in this study. Of the patients, 77 cases were positive for anti-T. gondii IgG, and 8 cases were positive for anti-T. gondii IgM. IgG avidity test revealed 11, 4, and 102 cases were low, intermediate, and high, respectively, and 6.8% and 9.4% of cases were positive for IgM and IgG avidity tests, respectively (P=0.632). Agreement (Kappa value) between paired tests IgG-IgM, IgG-IgG avidity, and IgM-IgG avidity was 0.080, 0.099, and 0.721, respectively (P<0.05). This study showed that conventional serologic tests (IgM and IgG levels) and IgG avidity correlate well each other and can be used to differentiate recent infections from old OT. It seems that reactivated old infections rather than recently acquired infections are majority of Iranian OT patients.

Cloning and Expression of Partial Japanese Flounder (Paralichthys olivaceus) IgD

  • Choi, Dae-Han;Jang, Han-Na;Ha, Dae-Mang;Kim, Jae-Wha;Oh, Chan-Ho;Choi, Sang-Hoon
    • BMB Reports
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    • v.40 no.4
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    • pp.459-466
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    • 2007
  • The cDNA sequence of the Japanese flounder (Paralychthys olivaceus) IgD has been previously reported (GenBank accession no. AB052658) and this was followed by the detection of IgD mRNA expression in some flounder organ tissues. However, it has not been determined whether the flounder IgD gene is virtually expressed into IgD protein. To characterize the flounder immunoglobulins utilized in elucidating the mechanism, evolution and diversity of the flounder immune system, antibodies specific to IgD and IgM were necessary. In the present study, partial flounder recombinant IgD (rIgD), IgM (rIgM) and the conserved regions of IgD and IgM (rCIg) were produced by cloning the cDNA sequence using isotype specific primers which were designed to produce unique fragments of IgD and IgM specific amino acid sequences. The production of recombinant Igs was ascertained by SDS-gel electrophoresis and immunoblot analysis using anti-T7$\cdot}$Taq antibody. The produced recombinant Igs were purified using affinity columns, and used as immunogens. Antibodies specific to the isotype of flounder Igs were generated by immunizing rabbits with rfIgs and the antibodies produced were identified by enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Specificities of the generated antibodies were evaluated by testing cross-reactivity between recombinant IgM and IgD. By ELISA, rabbit antibodies against the rfIgD fragment (anti-rfIgD) failed to recognize any kind of flounder serum Igs, whereas respective antibodies against rfCIg (anti-rfCIg) and rfIgM fragments (anti-rfIgM) reacted with serum Igs. Likewise, in immunoblot assays, though anti-rfIgD did not, both anti-rfCIg and anti-rfIgM bound with the ~85 kd flounder IgM heavy chain. By flow cytometry analysis, anti-rfCIg, anti-rfIgD and anti-rfIgM reacted with 6%, 3% and 6.5% of cells, respectively, suggesting that flounder IgD is not secreted in serum but expressed on flounder B-like cell surfaces as in mammals. Antibodies produced against recombinant flounder Igs could be used to develop sandwich assay systems for detecting flounder Igs and for further investigating the flounder immune system.

Comparison of Direct-labeling Method of Antibody with $^{99m}Tc$ and $^{188}Re$ (농양이식백서에서 $^{99m}Tc,\;^{188}Re$ 직접표지항체의 비교)

  • Choi, Tae-Hyun;Lim, Sang-Moo;Choi, Chang-Woon;Woo, Kwang-Sun;Chung, Wee-Sup;Lim, Soo-Jeong
    • The Korean Journal of Nuclear Medicine
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    • v.33 no.1
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    • pp.84-93
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    • 1999
  • Purpose: We investigated the direct labeling method of antibody with $^{99m}Tc$ and $^{188}Re$ and examined the stability and function of these labeled compounds in in vitro and in vivo. Materials and Methods: Disulfide bond of nonspecific human IgG was reduced to -SH group by 2-mercaptoethanol. Stannous ion was used to reduce $^{99m}Tc$ and $^{188}Re$. The stability of $^{99m}Tc$-IgG and $^{188}Re$-IgG was estimated upto 24 hrs. Biodistribution was evaluated in abscess bearing rats at 4 and 24 hr post-injection of $^{99m}Tc$ or $^{188}Re$ labeled IgG. Results: The number of -SH group per reduced IgG molecule was 2.34. The labeling yield of $^{99m}Tc$-IgG and $^{188}Re$-IgG were 90% and 95%, respectively The stability of $^{99m}Tc$-IgG at 1, 4, 6 and 24 hr was 91%, 83%, 78%, 7% and that of $^{188}Re$-IgG at 1, 4, 16 and 24 hr was 94%, 80%, 47%, 42%, respectively. At 4 hr post-injection of $^{99m}Tc$-IgG, high uptake was found on kidney, blood, stomach and abscess ($9.42{\pm}0.68,\;1.43{\pm}0.24,\;0.86{\pm}0.18,\;0.72{\pm}0.10$ %ID/g, respectively). The uptakes at 24 hr were kidney, abscess,.itomach, and blood in descending order. In case of $^{188}Re$-lgG, high uptake at 4 hr post injection appeared on kidney, blood, abscess and stomach ($3.92{\pm}0.62,\;1.32{\pm}0.08,\;0.88{\pm}0.01,\;0.26{\pm}0.06$, respectively). The uptakes at 24 hr were kidney, abscess, blood and stomach in descending order. The abscess to blood uptake ratio of $^{99m}Tc$-IgG was 0.5 at 4 hr and 2.02 at 24 hr and that of $^{188}Re$-IgG was 0.67 and 1.29. Conclusion: $^{99m}Tc$-IgG and $^{188}Re$-IgG canbe labeled efficiently with direct labeling method. However, $^{99m}Tc$-IgG and $^{188}Re$-IgG, labeled with direct method, was unstable. Further study is needed to enhance the stability of the antibody labeling.

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Developmental Changes of Serum IgA, IgG and IgM Concentrations in Broiler Chicks 1. Isolation of IgG and Developmental Changes of Serum IgG Levels (육용계의 혈청중 면역글로불린(IgA, IgG, IgM) 농도의 발육시기별 변화상 I. 혈청 IgG의 분리 및 발육시기별 농도 수준)

  • 김정우;이진언;김춘수;김상희;박근식
    • Korean Journal of Poultry Science
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    • v.20 no.2
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    • pp.65-72
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    • 1993
  • The experiment was conducted to estasblish the large scale production of anti-serum against chicken IgG and to profile the developmental changes of serum IgG levels during all the feeding period(from hatching to 7 weeks of age) in broiler chicks. Blood sample were taken from Hubbard chicken hi-daily or /and weekly during the experimental period. The pure IgG was isolated from ammonium sulfate treated chicken serum by ion exchange chromatography. The quantitative assay of serum IgG were carried by RID method. It was observed that IgG concentrations were showed the highest at hatching(4.25 $m\ell$/$m\ell$), after that dicreased rapidly, lowest at 2 weeks of age(0.81 $m\ell$/$m\ell$), and gradually increased to 7 weeks of age (2.48 $m\ell$/$m\ell$) There was no differences of IgG level between sex, but the IgG levels of male chicks dicreased more rapidly and increased earlier thereafter than those of females during the first two weeks of agg.

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Developmental Changes of Serum IgA, IgG and IgM Concentrations in Broiler Chicks - II. Isolation of IgA and Developmental Changes of Serum IgA Levels (육계의 혈청중 면역글로부린(IgA, IgG, IgM)농도의 발육시기별 변화상 - II. IgA 분리 및 발육시기별 농도수준)

  • 김정우;이민호;김춘수;김상희;박근식
    • Korean Journal of Poultry Science
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    • v.21 no.3
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    • pp.169-174
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    • 1994
  • An experiment was conducted to establish a large scale production method of anti-serum against chicken IgA and to profile the developmental changes of serum IgA levels during the feeding period(from hatching to 7 weeks of age) in broiler chicks. Blood samples were taken from Hubbard chicken at the age of hatching, 3 days of age, and weekly thereafter till to 7 weeks of age. The pure IgA was isolated from ammonium sulfate treated chicken bile juice by gel filtration chromatography ( Sepharose GL-6B) - The quantitative assay of serum IgA were carried by RID method. Developmental changes of serum IgA concentrations were 0.42 mg /mL at hatching, thereafter dicreased gradually, lowest at 1 week of age(0.17 mg /mL), and gradually increased to 7 weeks of age(2.73 mg /mL). There was no sexual difference in serum IgA level, but female chicks showed higher IgA levels than male chicks during the experimental period.

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Immunohistoehemical Observation on the Antigens Inducing IgG and IgM Antibodies against Sparganum (IgG와 IgM 항체를 유도하는 sparganum의 항원에 관한 면역조직화학적 및 전기영동에 의한 연구)

  • 김창환;최완성
    • Parasites, Hosts and Diseases
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    • v.29 no.4
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    • pp.339-354
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    • 1991
  • Localization and characterization of the antigenic components of sparganum which induced IgG and IsM antibodies in the host were studied by immunohistochemical techniques and SDS-PAGT and Western blotting. The antigen recognized by IgG antibody of rats or mice which were immunised by infection or injection of crude extracts of metacestodes of Spirometra erinacei, was located in the parenchyme of sparganum, especially at the cortex and around the calcareous corpuscles. The immunoreaction was demonstrated not only in the encysted fibrous wall of host but around the arterioles or venules in the connective tissue of host. The antigen recognized by IgM antibody of rats or mice was also observed in the parenchyme of sparganum and in the connective tissue of host. By 5∼20% gradient SDS-PAGE and EIBT, we detected antigenic components by IgG and 1gG antibodies of the rat or mouse immunized by infection or injection of crude extract of spargana. Twenty-three antigenic bands from crude extracts of spargana were recognized by IgG antibody and 15 components by IgM antibody of immunized rats. Out of the bands recognized by IgG and IgM antibodies, 15 were cross-reacted each other. Twenty components of eBlcretory-secretory proteins from spargana were recognized by IgG, and 5 components by IgM antibody of immunized rats. By IgG and IgM antibodies of immunized mice, 16 components of crude extracts were recognized by IgG antibody and 9 components by IgM antibody. Twenty components of excretory-secretory preparation were recognized by IgG antibody and 5 components by IgM antibody. Thirteen components of crude extracts were cross-reacted by IgG antibody of rats and mice.

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