2.Study on the location of membrane and detection of antibody in the sera of genus-specific antigen LipL41s in patients with Leptospira interrogans.
Ye HU ; Zong-Qi GUO ; Bai-Li SUN ; Ping YANG ; Jie YAN
Chinese Journal of Epidemiology 2007;28(8):776-780
OBJECTIVETo determine the location on outer envelope and natural antibody response and types of genus-specific lipoprotein antigen LipL41s in patients with Leptospira interrogans.
METHODSMicroscope agglutination test (MAT) was used to examine leptospirosis patients' serum samples from Sichuan area, China. Ni-NTA affinity chromatography was performed to extract the target recombinant rLipL41/1 and rLipL41/2 products that expressed under inducement of IPTG. Western blot assay was performed to detect the immunoreactivity between the sera from the patients infected with different serogroups of L. interrogans and rLipL41s. Immune aurosol electron microscopy was selected to locate the position of LipL41s on leptospiral envelope. ELISA based on rLipL41s was established to confirm the level and types of specific antibody.
RESULTSL. interrogans serogroup icterohaemorrhagiae remained to be the most dominant leptospiral serogroup in Sichuan area. All the sera from patients infected with different serogroups of L. interrogans could efficiently recognize the LipL41s which were the protein molecular that located on the external surface of leptospiral envelope. In the 156 serum samples from MAT positive leptospirosis patients, the positive rates for rLipL41/1 or rLipL41/2 specific IgM appeared to be 84.6%-87.8% and 78.2%-83.3%, respectively, while for rLipL41/1 or rLipL41/2 specific IgG they were 69.2%-81.4% and 75.0%-80.1%, respectively.
CONCLUSIONLipL41s were the leptospiral superficial protein antigen of L. interrogans. Both the LipL41/1 and LipL41/2 could induce serum antibodies IgM and IgG with extensive antigenic-cross reaction during natural infection of L. interrogans in general populations. Hence, rLipL41/1 or rLipL41/2 could be used as the antigen candidate for developing universal genetic engineering vaccine and detection kit.
Antibodies, Bacterial ; blood ; immunology ; Antigens, Bacterial ; metabolism ; Bacterial Outer Membrane Proteins ; metabolism ; Cell Membrane ; metabolism ; Cross Reactions ; Humans ; Immunoglobulin G ; blood ; immunology ; Immunoglobulin M ; blood ; immunology ; Leptospira interrogans ; metabolism ; ultrastructure ; Leptospirosis ; immunology ; Species Specificity
3.Effects of autologous sera on immediate and late skin reaction to the house dust mite in atopic individuals.
Dong Ho NAHM ; Jung Won PARK ; Chein So HONG
Yonsei Medical Journal 1995;36(2):137-145
To evaluate the in vivo effect of autologous serum including antibodies to house dust mite in atopic individuals, we observed the immediate (15 mins) and late (6 hours) skin reactions (ISR, LSR) on intradermal (ID) test of serially diluted Dermatophagoides farinae antigens (DFa, Allergopharma, Germany) mixed with autologous sera (DFa-S) and diluent alone (DFa-D). We tested 34 DFa-skin reactive atopic individuals including 12 asthmatics (BA), 8 asthmatics on immunotherapy with DFa (IT), and 14 healthy atopic controls (AC). We observed complete inhibition of ISR in the lowest allergen dose of DFa-S in 7 (58.3%) of 12 BA, 3 (37.5%) of 8 IT, and 2 (14.3%) of 14 AC. In BA, the inhibition of ISR was more frequent than AC (p< 0.05). We observed larger late reactions in half of LSR positive cases on ID test by DFa-S than by DFa-D (> or = 1.5 X size; accentuation of LSR). Accentuation of LSR were shown more frequently by DFa mixed with larger amount of serum (25% in 1:1 mix; 80% in 1:3 mix, p< 0.05). But there were no differences of DFa-specific IgE and IgG subclass antibodies regardless of the inhibition of ISR or the accentuation of LSR. In conclusion, some autologous sera from DFa-sensitive individuals showed the inhibition of ISR and the accentuation of LSR on DFa-ID test.
Animal
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*Blood Physiology
;
Dermatitis, Atopic/blood/*immunology
;
Human
;
Hypersensitivity, Delayed/*immunology
;
Hypersensitivity, Immediate/*immunology
;
Immunoglobulin E/metabolism
;
Immunoglobulin G/metabolism
;
Intradermal Tests
;
Mites/*immunology
;
Skin/*immunology
;
Support, Non-U.S. Gov't
4.Allergic Asthma and Rhinitis Caused by Household Rabbit Exposure: Identification of Serum-Specific IgE and Its Allergens.
Jeong Hee CHOI ; Hyun Mi KIM ; Hae Sim PARK
Journal of Korean Medical Science 2007;22(5):820-824
Although rabbits are common domestic pets, severe respiratory allergic reactions to rabbits in households are unusual. Ory c 1, a 17-kDa glycoprotein found in saliva and fur, has previously been identified as a major rabbit allergen. In this report, we describe the cases of three patients with rabbit allergy who presented with asthma and/or rhinitis while living in households with detectable levels of serum-specific IgE and major IgE binding components. Three patients with rabbit allergy and 18 unexposed nonatopic healthy controls were enrolled. Enzyme-linked immunosorbent assays (ELISA) for serum-specific IgE and IgG4 to rabbit epithelium and inhibition ELISA were performed followed by sodium dodecye sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and IgE immunoblotting. All three patients with rabbit allergy had high serum-specific IgE antibody levels compared with controls. The results of the inhibition ELISA showed significant inhibition with the addition of rabbit epithelium, whereas no significant inhibition was noted with the addition of cat and dog epithelia. Two IgE-binding components with molecular weights of 16 kDa and 67.5 kDa were identified by IgE immunoblotting. In conclusion, rabbit exposure may induce IgE-mediated bronchial asthma and/or rhinitis in domestic settings.
Adolescent
;
Adult
;
Allergens/*blood
;
Animals
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Asthma/*immunology/metabolism
;
Electrophoresis, Polyacrylamide Gel
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Enzyme-Linked Immunosorbent Assay/methods
;
Female
;
Humans
;
Hypersensitivity/*immunology/metabolism
;
Hypersensitivity, Immediate/immunology
;
Immunoblotting
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Immunoglobulin E/*blood/*chemistry
;
Immunoglobulin G/chemistry
;
Male
;
Rabbits
;
Rhinitis/*immunology/metabolism
5.Evaluation of a Novel Array-Based Toxoplasma, Rubella, Cytomegalovirus, and Herpes Simplex Virus IgG Enzyme Linked Immunosorbent Assay and Its Comparison with Virion/Serion Enzyme Linked Immunosorbent Assays.
Dongsheng WU ; Yuanjian WU ; Liuhong WANG ; Weidong XU ; Qiao ZHONG
Annals of Laboratory Medicine 2014;34(1):38-42
BACKGROUND: The dramatic increase in use of the IgG test for toxoplasma, rubella, cytomegalovirus (CMV), and herpes simplex virus (HSV) [TORCH] has led to the requirement for a high-efficiency method that can be used in the clinical laboratory. This study aimed to compare the results of BGI-Array ELISA TORCH IgG (BGI-GBI, China) screening method to those of Virion/Serion TORCH IgG ELISA (Virion/Serion, Germany). METHODS: Serum specimens (n=400) submitted for routine IgG testing by Virion/Serion ELISA were also tested using the BGI-Array ELISA method. The agreements of these two kinds of method were analyzed by kappa-coefficients calculation. RESULTS: Following repeat testing, the BGI-Array ELISA TORCH IgG assays demonstrated agreements of 99.5% (398/400 specimens), 98% (392/400 specimens), 99% (396/400 specimens), and 99.5% (398/400 specimens), respectively. The BGI-Array ELISA IgG assays provided results comparable to Virion/Serion ELISA results, with kappa-coefficients showing near-perfect agreement for the HSV (kappa=0.87), rubella (kappa=0.92) and CMV (kappa=0.93) and substantial agreement for the toxoplasma (kappa=0.80) IgG assays. The use of the BGI-Array ELISA TORCH IgG assays could reduce the turnaround time (1.5 hr vs. 5 hr by Virion/Serion ELISA for 100 specimens) and were easy to use. CONCLUSIONS: BGI-Array ELISA TORCH IgG shows a good agreement with Virion/Serion ELISA methods and is suitable for clinical application.
Antibodies, Viral/blood
;
Cytomegalovirus/immunology/*metabolism
;
*Enzyme-Linked Immunosorbent Assay
;
Humans
;
Immunoglobulin G/*analysis/blood
;
Protozoan Infections/diagnosis
;
Reagent Kits, Diagnostic
;
Rubella virus/immunology/*metabolism
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Sensitivity and Specificity
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Simplexvirus/immunology/*metabolism
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Toxoplasma/immunology/*metabolism
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Virion/*immunology/metabolism
;
Virus Diseases/diagnosis
6.Congenital cytomegalovirus infection in Korean population with very high prevalence of maternal Immunity.
Young Mo SOHN ; Kook In PARK ; Chul LEE ; Dong Gwan HAN ; Won Young LEE
Journal of Korean Medical Science 1992;7(1):47-51
In order to asses congenital cytomegalovirus (CMV) infection in Korea, five hundred and seventy five pregnant women (mean age 29.5 +/- 3.8 yrs., mean gestational age at test 37.5 +/- 6.7 weeks) visiting the prenatal clinic at Severance Hospital, Seoul, Korea were studied. CMV IgG antibody was present in 96% (552/575) and IgM antibody was present in 0.7% (4/575) of the pregnant women by the third trimester. Four of 445 cord sera were positive for CMV IgM antibody (0.9%). Urine samples from 514 newborns were tested for the evaluation of congenital CMV infection. Six (1.2%) of 514 newborns excreted CMV in their urine. All the congenitally infected infants had subclinical involvement at birth and during the 12 months of the follow-up period. These results indicate that Korean pregnant women were highly immunized against CMV by the third trimester. Furthermore this study suggests that the rate of congenital CMV infection is relatively as high as rates previously reported from other countries, although there is a very high prevalence of maternal immunity. The incidence of maternal primary infection during pregnancy seems to be rare and therefore most congenital infections in Korea might be following by maternal reactivation or reinfection.
Adult
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Cytomegalovirus Infections/congenital/*epidemiology/immunology/metabolism
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Female
;
Follow-Up Studies
;
Humans
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*Immunity, Maternally-Acquired
;
Immunoglobulin G/blood
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Immunoglobulin M/blood
;
Infant, Newborn
;
Korea/epidemiology
;
Pregnancy
7.Incidence and Seroprevalence of Hepatitis A Virus Infections among Young Korean Soldiers.
Cheol In KANG ; Chang Min CHOI ; Tae Sung PARK ; Dong Jun LEE ; Myoung Don OH ; Kang Won CHOE
Journal of Korean Medical Science 2007;22(3):546-548
This study was performed to determine the incidence and seroprevalence of hepatitis A virus (HAV) infections in young soldiers in the Republic of Korea Army. From January 2000 through December 2004, a total of 147 hepatitis A cases were reported to the Armed Forces Medical Command. The annual incidence rates were 7.4 per 100,000 persons in 2000, 1.6 in 2001, 4.4 in 2002, 9.8 in 2003, and 6.2 in 2004, based on the reported cases among approximately 500,000 soldiers. All patients were males with a median age of 21 yr (range, 19-27). The most common symptom was nausea (86.5%), and all patients had recovered without complications. In addition, in order to evaluate the seroprevalence of HAV infection in young adults, serum samples were obtained from randomly selected young subjects among those who had been admitted to the Armed Forces Capital Hospital from September 2005 to February 2006. A total of 200 subjects were enrolled in the study to analyze the anti-HAV immune status. The overall anti-HAV IgG seropositive rate was 2% (4/200, 95% CI, 0.60-5.21%). Given the changing epidemiology of the disease and the associated increase in morbidity, it was suggested that the routine HAV vaccination for Korean military personnel might be necessary.
Adult
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Hepatitis A/*blood/*epidemiology/immunology
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Hepatitis A Antibodies/immunology
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Hepatitis A virus/*metabolism
;
Humans
;
Immunoglobulin G/blood
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Incidence
;
Korea
;
Male
;
Military Personnel
;
Seroepidemiologic Studies
8.Influence of fusion protein of IBDV VP2 and chicken interleukin-2 on immune response in chicken.
Chen WANG ; Zhanqin ZHAO ; Chunjie ZHANG ; Yichen LIU ; Ke DING ; Yinju LI ; Xiangchao CHENG ; Puyan CHEN
Chinese Journal of Biotechnology 2010;26(4):476-482
In order to research immunogenicity of the recombinant rVP2-IL-2 fusion protein, we obtained the rVP2-IL-2 fusion protein using Pichia pastoris expression system, and then evaluated its potential to induce immune responses in chicken. The effect was determined in the form of protective anti-IBDV VP2 titers, antibodies (IgG1 and IgG2a), lymphocyte proliferation, the levels of interferon-gamma and interleukin-4 cytokines, and challenge experiment. Antibody titers and proliferation lymphocyte level suggested that the fusion protein could elicit specific humoral immune and cellular immune responses, antibody sub-type results indicated that the rVP2-IL-2 fusion protein induced secretion both of IgG1 and IgG2a. The seem result elicited from cytokines ELISA test, secretion of both of Th1 (gamma-IFN) and Th2 (IL-4) were induced by the rVP2-IL-2 fusion protein. Challenge experiment result shown that chicken immunized the rVP2-IL-2 fusion protein obtained 85% protection. These results confirm that the fusion protein enhances the protection against IBDV through both humoral and cell-mediated immunity, and thus could serve as a candidate for the development of IBDV subunit vaccine.
Animals
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Antibodies, Viral
;
biosynthesis
;
blood
;
Chickens
;
immunology
;
Immunoglobulin G
;
blood
;
Interleukin-2
;
biosynthesis
;
genetics
;
Pichia
;
genetics
;
metabolism
;
Recombinant Fusion Proteins
;
biosynthesis
;
genetics
;
immunology
;
Th1 Cells
;
immunology
;
Th2 Cells
;
immunology
;
Vaccines, Subunit
;
immunology
;
Viral Structural Proteins
;
biosynthesis
;
genetics
;
Viral Vaccines
;
immunology
9.A Study of Oxidative Stress Parameters in Anti-Helicobacter Pylorus Immunoglobulin G Positive and Negative Gastric Cancer Patients.
Tevfik NOYAN ; Huseyin GUDUCUOGLU ; Mahmut ILHAN
Yonsei Medical Journal 2009;50(5):677-682
PURPOSE: Helicobacter pylorus (HP) is a Gram-negative spiral-shaped microaerophilic bacterium, which colonizes in the gastric mucosa of humans. The gastric human pathogen HP causes chronic gastritis and ulcers, and has a strong relationship with gastric cancer. The aim of this study was to determine advanced oxidation protein products (AOPP) levels, activities of myeloperoxidase (MPO) and catalase (CAT) in two groups. MATERIALS AND METHODS: For this aim, one group included 30 patients with gastric cancer (Group 1) and the other included 30 subjects with non-gastric cancer and Anti-HP immunoglobulin (Ig) G antibody positive (group 2). Anti-HP IgG antibody test values were found as positive in fifty percent of group 1 and all of the group 2 patients. RESULTS: Significantly increased AOOP levels were found in group 1 (p < 0.05) compared to group 2. There were no significant differences between the groups in regard to activities of MPO and CAT. In addition, AOPP level, MPO and CAT activities were similar among the Anti-HP IgG positive and negative subgroups of group 1 patients. CONCLUSION: The result of this study indicated that gastric cancer patients were characterized by increased protein oxidation, whereas there was no significant difference in oxidative stress parameters and antioxidant enzyme activity between the Anti-HP IgG positive and negative gastric cancer patients.
Adult
;
Aged
;
Catalase/blood
;
Female
;
Helicobacter Infections/*immunology/metabolism
;
Helicobacter pylori/*immunology
;
Humans
;
Immunoglobulin G/*blood
;
Male
;
Middle Aged
;
Oxidation-Reduction
;
*Oxidative Stress
;
Peroxidase/blood
;
Stomach Neoplasms/blood/immunology/*metabolism
10.Enhancement of immune responses in rabbits with a targeted anti-caries DNA vaccine pGJA-P.
Rong JIA ; Ming-wen FAN ; Ji-hua GUO ; Zhuan BIAN ; Zhi CHEN ; Fei YU
Chinese Journal of Stomatology 2004;39(3):248-251
OBJECTIVETo detect the immunoreactivity of targeted fusion anti-caries DNA vaccine pGJA-P in vitro, and the ability to enhance the immune responses compared with the non-targeted fusion anti-caries DNA vaccine pGLUA-P.
METHODSThe CHO cells were transfected with pGJA-P and the expression of recombinant protein in cultured supernatants were detected using Western blotting. 5 to 6-month-old female Japanese rabbits were immunized with either pGJA-P or pGLUA-P via either intramuscular injection (i.m.) or intranasal route (i.n.). The sera and saliva were collected and the antibody responses were checked by ELISA. The effect of immune sera on the synthesis of water-insoluble glucan by glucosyltransferase of S. mutans was examined.
RESULTSThe expressed protein could response to specific anti-GTF antibody. The antibody responses in serum generated by pGJA-P via i.m. were significantly higher than those generated by pGLUA-P (P < 0.01). The antibody responses in saliva generated by pGJA-P via i.n. were significantly higher than those generated by pGLUA-P (P < 0.01). The higher mucosal antibody response induced by pGJA-P via i.m. compared with pGLUA-P (P < 0.01) was detected. The immune sera of rabbits immunized by pGJA-P via i.m. most significantly inhibited the synthesis of water-insoluble glucan by glucosyltransferase.
CONCLUSIONSThe recombinant protein expressed by pGJA-P had the immunoreactivity to anti-GTF antibody. pGJA-P could induce faster and higher specific mucosal SIgA antibody responses via i.n. or serum IgG antibody responses via i.m. compared with non-targeted DNA vaccine, pGLUA-P. High titres of specific mucosal antibodies were found in rabbits immunized with pGJA-P via i.m. The immune sera of rabbits immunized by pGJA-P via i.m. displayed the ability of inhibiting the synthesis of water-insoluble glucan by glucosyltransferase.
Animals ; Bacterial Proteins ; immunology ; CHO Cells ; Cricetinae ; Dental Caries ; prevention & control ; Female ; Glucosyltransferases ; immunology ; metabolism ; Immunoglobulin G ; blood ; Membrane Glycoproteins ; immunology ; Rabbits ; Recombinant Fusion Proteins ; administration & dosage ; immunology ; Streptococcus mutans ; immunology ; Transfection ; Vaccines, DNA ; administration & dosage ; immunology