• Title/Summary/Keyword: yIgG

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Separation of Immunoglobulin G from Colostrum by Reverse Micelles of Cationic Surfactant (양이온 계면활성제로 형성된 역미셀을 이용한 초유 Immunoglobulin G의 분리)

  • 노경현;임지영
    • Food Science of Animal Resources
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    • v.24 no.1
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    • pp.80-85
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    • 2004
  • This study was carried out to separate immunoglobulin G(IgG) from colostrum using reverse micellar extraction of cationic surfactant and to suggest suitable extraction conditions. The reconstituted colostrum powder was solubilized into a reverse micellar phase containing CDAB(cetyldimethylethyl ammonium bromide) by mixing equal volume of the aqueous and organic phase with constant stirring. The solubilization of proteins from the aqueous to the organic phase was manipulated by pH and ionic strength of the aqueous phase and concentration of surfactant in the organic phase. Based on the SDS-PAGE and densitometry, about more than 90% of initial IgG was remained in the aqueous phase after reverse micellar extraction. Although the aqueous phase contained lactoferrin and bovine serum albumin as minor components, about 93% of the total protein was IgG. The efficient extraction was achieved by the reaction of sodium phosphate buffer(pH 8) containing 50 mM KCl and organic phase containing 100 mM CDAB. The separation of IgG using reverse micellar extraction was simple, highly efficient and easy to be scaled up.

Degradation of human immunoglobulins and cytotoxicity on HeLa cells by live Trichomonas vaginalis (질편모충 단백질분해효소의 세포독성 및 인체면역글로불린 분해능)

  • 민득영;류재숙
    • Parasites, Hosts and Diseases
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    • v.35 no.1
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    • pp.39-46
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    • 1997
  • The present study was undertaken to determine whether live T. uaginnlis degrades human secretory IgA, serum If and IgG molecules. Human immunoglobulins were exposed to live trophozoites, parasite Iysate, and excretory-secretory product (ESP) of T ucginnlis. To determine the fragmentation of immunoglobulins, the reaction sample was subjected to SDS-PAGE and EITB, and peroxidase conjugated antihuman IgA and IgG were used as probes. Live trophozoites degraded secretory IgA, serum IgA and IgG, and degradation were pressed forward by the prolongation of the incubation time and by increasing the number of trichomonads respectively. Also the Iysates and ESP of trichomonads degraded IgA and IgG. The cysteine and serine proteinase inhibitors such as I-64, antipain, iodoacetic acid, iodoacetamide, TLCK reduced the ability of cleaving immunoglobulins. The proteinase activity and cytotoxicity of T. uaginnlis to HeLa cells were decreased when live T. vusinalis was treated with metallo-proteinase inhibitor as well as cysteine and serine proteinase inhibitors. These results suggest that proteinase secreted from live T ucginclis may play a part role in host pathogenesis by T. uosinnlis, and the cleaving ability of host immunoglobulins by the proteinase may contribute as a one of immune evasion mechanism for parasite survival in the host.

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IgG Western Blot for Confirmatory Diagnosis of Equivocal Cases of Toxoplasmosis by EIA-IgG and Fluorescent Antibody Test

  • Khammari, Imen;Saghrouni, Fatma;Yaacoub, Alia;Meksi, Sondoss Gaied;Ach, Hinda;Garma, Lamia;Fathallah, Akila;Said, Moncef Ben
    • Parasites, Hosts and Diseases
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    • v.51 no.4
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    • pp.485-488
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    • 2013
  • The performance values of available techniques used in serodiagnosis of toxoplasmosis are satisfactory but they raise problems of equivocal and discordant results for very low IgG titers. Recently marketed, LDBio-Toxo II IgG Western blot (IB) showed an excellent correlation with the dye test. We estimated the proportion of equivocal and discordant results between the enzyme immunoassay Platelia Toxo IgG (EIA-IgG) and fluorescent antibody test (FAT) and assessed the usefulness of the IB as a confirmatory test. Out of 2,136 sera collected from pregnant women, 1,644 (77.0%) tested unequivocally positive and 407 (19.0%) were negative in both EIA-IgG and FAT. The remaining 85 (4%) sera showed equivocal or discordant results. Among them, 73 (85.9%) were positive and 12 (14.1%) were negative in IB. Forty-one (89.1%) equivocal sera in EIA-IgG and 46 (86.8%) equivocal sera in FAT were positive in IB. Reducing the cut-off values of both screening techniques improved significantly their sensitivity in detecting very low IgG titers at the expense of their specificity. In conclusion, equivocal results in routine-used techniques and their discordance in determination of the immune status in pregnancy women were not uncommon. IB test appeard to be highly useful in these situations as a confirmatory technique.

Distribution of $^{123}I,\;^{99m}Tc-Human$ Polyclonal Nonspecific IgG and $^{67}Ga-Citrate$ in Abscess bearing Mice ($^{123}I,\;^{99m}Tc$ 사람 비특이 IgG 및 $^{67}Ga-Citrate$의 실험동물에서 염증병소 섭취율의 비교)

  • Lim, Sang-Moo;Woo, Kwang-Sun;Chung, Wee-Sup;Awh, Ok-Doo;Seo, Yong-Sup;Lee, Jong-Doo
    • The Korean Journal of Nuclear Medicine
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    • v.26 no.1
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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.

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Molecular Mechanism of Male Germ Cell Apoptosis after Busulfan Treatment

  • Kim, Jin-Hoi
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.63-65
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    • 2002
  • Identification of spermatogonial stem cell-specific surface molecules is important in understanding the molecular mechanisms underlying the maintenance and differentiation of these cells. We have found that spermatogonia from busulfan treated mice expressed an autoantigen that distinguishes between undifferentiated and differentiated spermatogonia. Four to six weeks after busulfan treatment, germ cells located in the basal compartment of seminiferous epithelium show isotype-specific IgG deposits that form due to autoimmunity. Before busulfan treatment, the level of testicular IgG was very low but IgG levels began to increase after week 4 and peaked at week 6. When cells from the busulfan treated testis were analyzed using laser scanning cytomeoy (LSC), the frequency of cells positive for IgG deposits, 6-integrin, and 1-integrin were 16.5${\pm}$3.8%, 11.8${\pm}$2.6%, and 9.0${\pm}$ 1.4%, respectively. Immunofluorescent staining suggested that most, if not all of the cells with IgG-deposits isolated from a laminin-coated dish, were also positive for a spermatogonial stem cell marker \ulcorner6-integrins as well as for a germ cell-specific marker TRA 98. We determined serum and intratesticular IgG levels and the soundness of seminiferous tubule basement membrane from busulfan treated mice using electron microscopy, in order to study the mechanism responsible for IgG deposits in spermatogonia. We found that the basement membranes of seminiferous tubules from busulfan treated mice were severely impaired when compared to those of normal adult, neonates and w/wv mice. Furthermore, new blood cells were observed in the surface of the damaged basement membrane along the seminiferous tubules. These results suggest that the IgG in spermatogonial stem cells accumulates from circulating blood through the impaired basement membranes induced by busulfan treatment. Taken together, our study suggests that IgG can be used as a new marker for undifferentiated spermatogonia cells.

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Specific IgG antibody responses in experimental cat metagonimiasis (실험적 요꼬가와흡충증에서의 특이 IgG 항체 반응)

  • Jo, Seung-Yeol;Kim, Seok-Il;Gang, Sin-Yeong
    • Parasites, Hosts and Diseases
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    • v.25 no.2
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    • pp.149-153
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    • 1987
  • In order to observe the feasibility of serologic diagnosis of metagonimiasis, saline extracts of metacercariae and 4-week old adults were prepared. Sera from 25 experimentally infected cats were collected from 3 days to 12 weeks after infection. Their levels of specific IgG antibody were measured by ELISA together with 3 sera from non-infected cats. Specific IgG antibody levels began to rise in 7 days after infection, reached their peak in 2-4 weeks and made a plateau thereafter. Cats infected with hundreds of adult worms showed minimal rise of the antibody level. Adult antigen was superior to metacercarial antigen in detecting the specific IgG antibody.

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The Change of IgG Antibody Titer to Measles, Mumps and Rubella According to Age (국내 홍역, 유행성이하선염 및 풍진 백신 접종 후 연령에 따른 항체양성율 변화)

  • Pang, Sung Joon;Choi, Kyong Min
    • Pediatric Infection and Vaccine
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    • v.18 no.2
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    • pp.117-123
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    • 2011
  • Purpose : We investigated the change of antibody titer to measles, mumps and rubella according to age after vaccination. Methods : The IgG antibody titers to measles, mumps and rubella were tested on the residual serum from patients aged 7-20 years old after routine laboratory testing in the hospital with informed consent from the parents. Results : Antibody to measles was present in 275 cases out of 408 cases with a positive rate of 67.4%, the mean IgG titer was 2.77${\pm}$1.18 Index. Antibody to mumps was present in 112 cases out of 408 cases with a positive rate of 27.5%, the mean IgG titer was 2.08${\pm}$1.29 Index. Antibody to rubella was present in 367 cases out of 408 cases with a positive rate of 90.0%, the mean IgG titer was 60.46${\pm}$63.47 IU/mL. Conclusion : It is important to maintain a high rate of vaccination coverage in order to prevent an outbreak of measles, mumps, or rubella. It is also important to stress the maintenance of vaccination records for further reference.

The alteration of the positive rate of cytomegalovirus IgG antibody among preschool period children (학령 전기 소아에서 거대세포바이러스 IgG 항체 양성률의 변동)

  • Seo, Se-Young;Park, Sang-Jun;Hwang, Ja-Young;Hahn, Seong-Hoon;Kim, So-Young;Kim, Hyun-Hee;Lee, Wonbae
    • Clinical and Experimental Pediatrics
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    • v.49 no.1
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    • pp.51-55
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    • 2006
  • Purpose : In order to evaluate the time of disappearance of cytomegalovirus(CMV) IgG antibodies from mothers, and the alteration of the positive rate of CMV IgG antibodies among preschool period children, we investigated the positive rate of CMV antibodies among preschool children. Methods : We studied 391 children who visited the Department of Pediatrics from March, 2001 to February, 2004. We measured the serum CMV IgG of 217 children and the serum CMV IgM of 358 children. Results : The positive rate of CMV IgG antibodies is 83.9 percent(the number of positive IgG children is 182 out of 217). The alteration of the positive rate is 92.9 percent in 0-3 months, 75.0 percent in 4-6 months and the nadir was 20.0 percent in 7-9 months. Then, the positive rate increased to 83.9 percent in 22-24 months. After 22 months, the positive rate was 92.1 percent(the number of positive IgG children was 105 out of 114). The positive rate of CMV IgM antibody by age is 3.3 percent in 0-1 months, 3.6 percent in 1-2 months, 10.5 percent in 2-3 months, 14.3 percent in 3-4 months, 14.3 percent in 4-5 months, and then the results of five children among 148 children were positive. The distribution was one in 22-23 months, one in 25-26 months, one in 27-28 months, one in 28-29 months, one in 40-41 months. We discovered IgM positive children succesively from birth to 5 months, but sporadically after 5 months. Conclusion : The CMV IgG from mothers has decreased since birth and the time of nadir is 7-9 months. But it increases to a mean value of 83.9 percent at 22-24 months because of perinatal or postnatal infections.

The Effect of Copper, Selenium and Vitamin E on the IgG Level and Growth Rate of Broiler Chicks (Copper, Selenium과 Vitamin E의 첨가 급여가 육용계의 IgG수준과 성장율에 미치는 효과)

  • 김정우;김춘수;김상희;박근식
    • Korean Journal of Poultry Science
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    • v.20 no.2
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    • pp.55-64
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    • 1993
  • The experiment was conducted to compare the effects of supplements of copper, vitamin I and selenium on growth and immune responses of broiler chicks fed cornsoy diets. The basal diet contained 21% crude protein, 2,800 kcal ME, 10 mg Vitamin E, 10 mg copper and 0.1 mg selenium per kg diet. Additions of the basal diet were copper (150mg and 250 mg/kg) or combination of vitamin I(200 mg/kg) and selenium(2 mg/kg). Serum immunoglobulin G(IgG) concentrations and body weight gain were determined weekly from hatching to 7 weeks of age. Additions of copper(150mg, 250mg) to the basal diet were showed, at the four weeks of age, 4.8% and 4.5% higher in body weight gain than that of control group, respectively. The active immune system of copper and (Vit. E+Se) treated groups developed one week earlier than control group(basal diet). Negative correlation between IgG concentration and body weight gain was showed at the period from hatching to three weeks of age and, thereafter, positive correlation were identified (p<0.01). Mortality rates were observed lower in all treated groups than that of control. In conclusion, the lower the levels of serum IgG, at the first two weeks of age, the lower in disease Infection and the higher in body weight gain.

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Busulfan-Induced IgG-Protein Complex of Germ Cells and Its Utility for Selection of Spermatogonial Stem Cells

  • 주학진;천영신;권득남;김진회
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.38-38
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    • 2001
  • Spermatogonial stem cells은 sperrnatogenesis에서 중요한 역할을 하며, 곡세정관의 기저막에 위치하고 있는 것으로 알려져 있다. 그러나, 그 동안 이 세포에 특이하게 발현되는 marker가 거의 알려져 있지 않아 spermatogonial stem cell의 연구에 많은 어려움을 가져왔다. 최근 일반적인 stem cell이 갖는 특성 중, 기저막과 상호작용을 하는 surface protein으로 integrin이 존재한다는 사실을 이용하여, anti-$\alpha$$_{6}$/ 또는 anti-$\beta$$_1$ integrin항체로 germ cell을 선발하여 정소에 이식한 결과, 높은 효율로 이식세포유래의 정자발생이 가능하다는 결과가 보고되었다 (Shinohara et al., 1999). 한편, 항암제의 일종인 busulfan을 마우스에 투여(40mg/kg)한 후 4-5주가 경과하면 세정관의 기저막에 위치하는 spermatogonia를 제외하고 대부분의 생식세포는 소멸한다 본 실험의 목적은 이러한 사실들을 이용하여 spermatogonial stem cell의 특성을 밝히고, 이 생식세포를 보다 간편하고 손쉽게 선발할 수 있는 시스템을 확립하는데 있다. Busulfan처리 후 5주가 경과된 마우스와 정상적인 13주령의 마우스 testis로부터 세포를 분리한 후 FITC-conjugated anti-IgG를 이용한 면역형광법으로 측정.분석한 결과, 형광표식된 세포비율이 대조군과 비교하여 busulfan을 처리한 경우에서 유의적인 증가를 보였다.(17$\pm$3.8%. 0.7$\pm$0.3% busulfan vs control). 또한, IgG와 결합한 단백질이 존재하는 이들 세포들은 곡세정관의 기저막을 따라 위치하며, 단백질과 복합체를 형성한 IgG는 anti-Ig $G_{2a}$와 반응하지 않는다는 사실을 관찰했다. 이러한 IgG 복합체를 형성한 세포들의 특성을 이용하여, IgG와 반응을 하지 않는 것으로 확인된 이차 항체인 an1i-Ig $G_{2}$와 일차 항체인 anti-$\alpha$$_{6}$ 또는 anti-$\beta$$_1$ integrin항체를 이용하여 측정.분석하였다. Busulfan을 처리한 마우스 정소에서 분리한 세포를 다시 laminin으로 코팅된 dish에서 선발.회수해서, anti-lgG, anti-$\alpha$$_{6}$ 또는 anti-$\beta$$_1$ integrin항체로 각각 표식된 세포비율을 비교하였다. Laminin으로부터 선발.회수한 세포에서는 IgG복합체가 $\alpha$$_{6}$ 또 는 $\beta$$_1$integrin과 거의 같은 수준에서 높은 비율로 표식되었다. 결론적으로, busulfan에 의해 유도된 IgG와 결합가능한 단백질은 $\alpha$$_{6}$$\beta$$_1$ integrin과 마찬가지로 immunoglobulin G를 이용하여 spermatogonial stem cell의 선발을 가능하게 했다. 따라서, busulfan처리시 IgG는 미분화된 정조세포의 선발을 위한 하나의 marker로서 사용가능함을 시사한다.다.

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