scholarly journals Mycobacterium tuberculosis HBHA Protein Reacts Strongly with the Serum Immunoglobulin M of Tuberculosis Patients

2006 ◽  
Vol 13 (8) ◽  
pp. 869-875 ◽  
Author(s):  
A-Rum Shin ◽  
Kil-Soo Lee ◽  
Ji-Sook Lee ◽  
Su-Young Kim ◽  
Chang-Hwa Song ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Immunoblot Analysis ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Heparin Binding ◽  
Igg Antibodies ◽  
Humoral Immune ◽  
Igm Antibodies ◽  
Humoral Immune Responses ◽  
Mycobacterial Antigens

ABSTRACT Identification and characterization of serologically active mycobacterial antigens are prerequisites for the development of diagnostic reagents. We examined the humoral immune responses of active tuberculosis (TB) patients against Triton-soluble proteins extracted from Mycobacterium tuberculosis by immunoblotting. A 29-kDa protein reacted with immunoglobulin M (IgM) in the pooled sera of the patients, and its N-terminal amino acid sequence matched that of the heparin-binding hemagglutinin (HBHA). Recombinant full-length HBHA was expressed in Escherichia coli (rEC-HBHA) and M. smegmatis (rMS-HBHA). In immunoblot analysis, the IgM antibodies of the TB patients reacted strongly with rMS-HBHA but not with rEC-HBHA, whereas the IgG antibodies of these patients reacted weakly with both recombinant HBHA proteins. In enzyme-linked immunosorbent assay analysis using rMS-HBHA and 85B as antigens, the mean levels and sensitivities of the anti-HBHA IgM antibodies of the TB patients were significantly higher than those of the anti-antigen 85B IgM antibodies, while the IgG antibodies showed the opposite results. Of interest in this respect, the pooled sera from the TB patients that contained anti-HBHA IgM antibodies neutralized the entry of M. tuberculosis into epithelial cells. These findings suggest that IgM antibody to HBHA may play a role in protection against extrapulmonary dissemination.

scholarly journals Humoral Immune Responses of Tuberculosis Patients in Brazil Indicate Recognition of Mycobacterium tuberculosis MPT-51 and GlcB

2008 ◽  
Vol 15 (3) ◽  
pp. 579-581 ◽  
Author(s):  
Cristina Melo Cardoso Almeida ◽  
Arioldo C. Vasconcelos ◽  
André Kipnis ◽  
Ana Lúcia Andrade ◽  
Ana Paula Junqueira-Kipnis
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Immune Responses ◽  
Immunosorbent Assay ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Tuberculosis Patients ◽  
Humoral Immune ◽  
Humoral Immune Responses ◽  
Humoral Responses

ABSTRACT The humoral responses to recombinant MPT-51 and GlcB was determined by using an enzyme-linked immunosorbent assay. Levels of immunoglobulin M (IgM) against MPT-51 and IgG against GlcB were higher among tuberculosis (TB) patients than among control individuals. When the MPT-51 and GlcB assays were combined, 90.8% specificity and 75.5% sensitivity were observed. MPT-51 and GlcB were recognized in the humoral responses of Brazilian TB patients.

scholarly journals Diagnosis of Eastern Equine Encephalomyelitis Virus Infection in Horses by Immunoglobulin M and G Capture Enzyme-Linked Immunosorbent Assay

1994 ◽  
Vol 6 (1) ◽  
pp. 34-38 ◽  
Author(s):  
Sudhir P. Sahu ◽  
Arnold D. Alstad ◽  
Douglas D. Pedersen ◽  
James E. Pearson
Keyword(s):  
Virus Isolation ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibodies ◽  
Serum Samples ◽  
Capture Elisa ◽  
Igm Antibodies ◽  
Viral Antigens ◽  
Encephalomyelitis Virus ◽  
Eastern Equine Encephalomyelitis

Immunoglobulin M (IgM) and G (IgG) capture enzyme-linked immunosorbent assays (ELISAs) were used as possible adjuncts to hemagglutination inhibition (HI) and virus neutralization (VN) tests to differentiate between reaction to recent exposure to eastern equine encephalomyelitis (EEE) virus and those due to prior vaccination. Serum samples were evaluated by the IgM-capture ELISA, and the results were compared with those of HI and VN tests. Of 381 serum samples, 51% (195 samples) were positive by HI test (≥1:40) and 54% (205 samples) were positive by VN test (≥ 1:10), but only 35% (132 samples) were positive by IgM-capture ELISA (≥ 1:100). With only a few exceptions, the sera with IgG ELISA titers had a VN titer of ≥1: 100. When EEE virus isolation and serology were compared, the EEE cases were divided into three categories: 1) peracute cases- the serum was negative for EEE IgM and IgG by the ELISA, negative for VN antibody, but HI antibody positive; 2) acute cases-IgM and HI antibody positive but negative for IgG and VN antibody; and 3) transitional cases–positive for IgM and IgG antibodies, HI titers of 1:40–1:160, and VN titers of ≥ 1: 100. IgM antibodies of EEE virus were monospecific and did not cross-react with western or Venezuelan equine encephalomyelitis viral antigens by the ELISA. Isolation of EEE virus from tissue or detection of IgM antibodies to EEE virus in a single serum sample is evidence of recent EEE infection and differentiated serum from previously vaccinated horses. However, the presence of IgM in the absence of virus isolation could also be due to a recent viral vaccination.

scholarly journals Early Antibody Responses to Experimental Mycobacterium bovis Infection of Cattle

2006 ◽  
Vol 13 (6) ◽  
pp. 648-654 ◽  
Author(s):  
W. R. Waters ◽  
M. V. Palmer ◽  
T. C. Thacker ◽  
J. P. Bannantine ◽  
H. M. Vordermeier ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Skin Testing ◽  
Rapid Test ◽  
Kinetic Studies ◽  
Immunoblot Analysis ◽  
Immunoglobulin M ◽  
Serum Samples ◽  
Mycobacterial Antigens ◽  
New Generation ◽  
And Control

ABSTRACT Bovine tuberculosis persists as a costly zoonotic disease in numerous countries despite extensive eradication and control efforts. Sequential serum samples obtained from Mycobacterium bovis-infected cattle were evaluated for seroreactivity to mycobacterial antigens. Animals received M. bovis by aerosol, intratonsil, intranasal, or intratracheal inoculation. Assays included the multiantigen print immunoassay for determination of antigen recognition patterns, immunoblot analysis for sensitive kinetic studies, and the VetTB STAT-PAK test, a novel, rapid test based on lateral-flow technology. Responses to MPB83 were detected for all M. bovis-infected animals regardless of the route or strain of M. bovis used for inoculation. Other less commonly recognized antigens included ESAT-6, CFP-10, and MPB70. Responses to MPB83 were detectable as early as 4 weeks after inoculation, were boosted upon injection of purified protein derivatives for skin testing, and persisted throughout the course of each of the four challenge studies. MPB83-specific immunoglobulin M (IgM) was detected prior to MPB83-specific IgG detection; however, early IgM responses rapidly waned, suggesting a benefit of tests that detect both IgM- and IgG-specific antibodies. The VetTB STAT-PAK test detected responses in sera from 60% (15/25) of the animals by 7 weeks after challenge and detected responses in 96% (24/25) of the animals by 18 weeks. These findings demonstrate the potential for new-generation antibody-based tests for the early detection of M. bovis infection in cattle.

scholarly journals Purified Hexameric Epstein-Barr Virus-Encoded BARF1 Protein for Measuring Anti-BARF1 Antibody Responses in Nasopharyngeal Carcinoma Patients

2010 ◽  
Vol 18 (2) ◽  
pp. 298-304 ◽  
Author(s):  
E. K. Hoebe ◽  
S. H. Hutajulu ◽  
J. van Beek ◽  
S. J. Stevens ◽  
D. K. Paramita ◽  
...  
Keyword(s):  
Nasopharyngeal Carcinoma ◽  
Epstein Barr Virus ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Humoral Immune ◽  
Barr Virus ◽  
Humoral Immune Responses ◽  
Human Sera ◽  
Epstein Barr ◽  
Humoral Responses

ABSTRACTWHO type III nasopharyngeal carcinoma (NPC) is highly prevalent in Indonesia and 100% associated with Epstein-Barr virus (EBV). NPC tumor cells express viral proteins, including BARF1, which is secreted and is considered to have oncogenic and immune-modulating properties. Recently, we found conserved mutations in the BARF1 gene in NPC isolates. This study describes the expression and purification of NPC-derived BARF1 and analyzes humoral immune responses against prototype BARF1 (B95-8) and purified native hexameric BARF1 in sera of Indonesian NPC patients (n= 155) compared to healthy EBV-positive (n= 56) and EBV-negative (n= 16) individuals. BARF1 (B95-8) expressed inEscherichia coliand baculovirus, as well as BARF1-derived peptides, did not react with IgG or IgA antibodies in NPC. Purified native hexameric BARF1 protein isolated from culture medium was used in enzyme-linked immunosorbent assay (ELISA) and revealed relatively weak IgG and IgA responses in human sera, although it had strong antibody responses to other EBV proteins. Higher IgG reactivity was found in NPC patients (P= 0.015) than in regional Indonesian controls or EBV-negative individuals (P< 0.001). IgA responses to native BARF1 were marginal. NPC sera with the highest IgG responses to hexameric BARF1 in ELISA showed detectable reactivity with denatured BARF1 by immunoblotting. In conclusion, BARF1 has low immunogenicity for humoral responses and requires native conformation for antibody binding. The presence of antibodies against native BARF1 in the blood of NPC patients provides evidence that the protein is expressed and secreted as a hexameric protein in NPC patients.

scholarly journals Preliminary Serological Study of Helicobacter pylori Infection in Some University Students

2020 ◽  
Vol 6 ◽  
pp. 1
Author(s):  
Abdullah A Mahrazi ◽  
Mohammad A Khibrani ◽  
Khatib S Ismail ◽  
Emad Abada ◽  
◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
University Students ◽  
Large Scale ◽  
Confirmatory Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Blood Collection ◽  
Igg Antibodies ◽  
Igm Antibodies ◽  
H Pylori

Helicobacter pylori has been associated with peptic ulcer and gastric carcinoma. This study aimed to find the seroprevalence of H. pylori infection in some male students of Jazan University, Saudi Arabia. Twenty students were enrolled in the study (n = 20). Informed consent was obtained from the students. About 2 ml blood was collected intravenously in Improvacuter® evacuated blood collection tubes. The blood was allowed to clot at room temperature. The serum was collected and stored at –20°C for further use. The separated serum was used to detect IgG and IgM antibodies by Enzyme Linked Immunosorbent Assay (ELISA) against H. pylori for the in vitro diagnosis. A total of 11 (55.00%) students tested positive for IgG antibodies against H. pylori indicating previous infection. All the samples tested negative for IgM antibodies against H. pylori indicating no active infection. The seroprevalance of IgG antibodies against H. pylori was found to be very high in some male university students and is a cause of concern regarding their health. Obesity (p < 0.05; Value statistically significant), stress and bad eating habits, eating out, drinking carbonated beverages, and eating spicy food were some of the factors found to be associated with IgG seropositive students. The students were counseled and were instructed to undergo a confirmatory test and get medical intervention. Further large-scale studies need to be performed to plan action against this disease causing organism and to improve the health of students.

scholarly journals Idiotypes of anti-Ia antibodies. I. Expression of the 14-4-4S idiotype in humoral immune responses.

1981 ◽  
Vol 154 (2) ◽  
pp. 397-409 ◽  
Author(s):  
S L Epstein ◽  
K Ozato ◽  
J A Bluestone ◽  
D H Sachs
Keyword(s):  
Immune Responses ◽  
Immunosorbent Assay ◽  
Heavy Chain ◽  
Antigenic Specificity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Absorption Analysis ◽  
Specific Enzyme ◽  
Cellular Receptors ◽  
Humoral Immune ◽  
Humoral Immune Responses

The idiotype of a mouse monoclonal anti-I-E antibody, 14-4-4S, has been studied using a heterologous anti-idiotypic reagent. This antibody recognizes Ia. 7, an antigenic specificity present in all strains expressing a product of the I-E subregion. Expression of the 14-4-4S idiotype in humoral immune responses was analyzed by an idiotype-specific enzyme-linked immunosorbent assay system. The idiotype was readily detectable in C3H.SW anti-C3H alloantisera, the same immunization combination from which the hybridoma was derived. Absorption analysis demonstrated the anti-I-E specificity of the idiotype-positive molecules in these alloantisera. Penetrance of idiotype expression was high among individual C3H.SW immune mice (9 of 10 tested). To examine genetic requirements for idiotype expression, an immunization was performed using as responders CWB mice, congenic with C3H.SW but differing at the heavy chain allotype loci. Immune sera of individual CWB mice contained very little or no idiotype, demonstrating that levels of idiotype expression are influenced by allotype-linked genes, although the influence of other genes has not been ruled. The 14-4-4S idiotype therefore represents a shared idiotype of anti-Ia antibodies and provides opportunities for analysis of the idiotypes of cellular receptors for the corresponding Ia antigen.

Immunodiagnosis and molecular validation of Toxoplasma gondii infection among patients with end-stage renal disease undergoing haemodialysis

Parasitology ◽  
2019 ◽  
Vol 146 (13) ◽  
pp. 1683-1689 ◽  
Author(s):  
Zahra Arab-Mazar ◽  
Shirzad Fallahi ◽  
Davood Yadegarynia ◽  
Amirreza Javadi Mamaghani ◽  
Seyyed Javad Seyyed Tabaei ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Renal Disease ◽  
End Stage Renal Disease ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Controls ◽  
Blood Samples ◽  
Igm Antibodies ◽  
Stage Renal Disease ◽  
End Stage

AbstractInfection is a significant cause of morbidity and mortality in patients with chronic kidney disease, especially who were under dialysis due to their depressed immunity. Toxoplasma gondii is a ubiquitous parasite that causes severe manifestations in immunocompromised patients. This case-control study was conducted to the immunodiagnosis and molecular validation of T. gondii infection among patients with end-stage renal disease undergoing haemodialysis. The study population consisted of 260 haemodialysis patients and 259 healthy controls referred to the main dialysis centres of Tehran, Iran during 2016. Anti-T. gondii immunoglobulin G (IgG) and immunoglobulin M (IgM) antibodies were assessed using enzyme-linked immunosorbent assay. As well, the T. gondii genomic DNA in whole blood samples of IgM-positive patients and healthy controls was evaluated using GRA6-polymerase chain reaction (PCR) and SAG1-loop-mediated isothermal amplification (LAMP) assays. The anti-T. gondii IgG and IgM antibodies were detected in 175 (67.3%) and 18 (7%) of haemodialysis patients and 122 (47%) and 4 (1.5%) of controls, respectively. Two of the 18 blood samples from IgM-positive patients and none of the IgM-positive control subjects were positive by GRA6-PCR. Whereas, nine and two blood samples of IgM-positive patients and controls were positive for Toxoplasma DNA by a SAG1-LAMP technique respectively. The seropositivity of the Toxoplasma IgM antibody was significantly different between haemodialysis patients and healthy controls which was confirmed by PCR and LAMP. The higher prevalence of T. gondii infection in haemodialysis patients compared with the controls proposes that these patients can be a group at risk for toxoplasmosis and screening for toxoplasmosis before dialysis is necessary for the patients.

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