Human humoral responses to antigens of Mycobacterium tuberculosis: immunodominance of high-molecular-mass antigens

Laal, Suman; Samanich, Karen M.; Sonnenberg, Michael G.; Zolla‐Pazner, Susan; Phadtare, Jaising Marutrao; Belisle, John T.

doi:10.1128/cdli.4.1.49-56.1997

Cited by 62 publications

(25 citation statements)

References 40 publications

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“…Knowledge of serological markers for TB patients coinfected with HIV would facilitate the development of diagnostic tests for TB. A study designed to characterize the target of the antibody response revealed that an 88-kDa antigen from CFP reacted with 70% of sera from TB patients (23). This same 88-kDa antigen was able to detect antibody in 74% of sera from HIV-positive patients who later developed TB (22).…”

Section: Discussionmentioning

confidence: 99%

“…More recently, the reactivity of high-molecular-weight antigens with patient sera has been described. In particular, an 88-kDa antigen was found to elicit a strong antibody response in patients with TB (23). Most importantly, the 88-kDa antigen, which is present in M. tuberculosis culture filtrate proteins (CFP), also has been described as a surrogate marker for TB in HIV-seropositive individuals, but its peptide sequence has been elusive (22).…”

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confidence: 99%

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Mass Spectrometric Identification of Mtb81, a Novel Serological Marker for Tuberculosis

Hendrickson

Douglass

Reynolds

et al. 2000

J. Clin. Microbiol.

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We have used serological proteome analysis in conjunction with tandem mass spectrometry to identify and sequence a novel protein, Mtb81, which may be useful for the diagnosis of tuberculosis (TB), especially for patients coinfected with human immunodeficiency virus (HIV). Recombinant Mtb81 was tested by an enzymelinked immunosorbent assay to detect antibodies in 25 of 27 TB patients (92%) seropositive for HIV as well as in 38 of 67 individuals (57%) who were TB positive alone. No reactivity was observed in 11 of 11 individuals (100%) who were HIV seropositive alone. In addition, neither sera from purified protein derivative (PPD)-negative (0 of 29) nor sera from healthy (0 of 45) blood donors tested positive with Mtb81. Only 2 of 57 of PPD-positive individuals tested positive with Mtb81. Sera from individuals with smear-positive TB and seropositive for HIV but who had tested negative for TB in the 38-kDa antigen immunodiagnostic assay were also tested for reactivity against Mtb81, as were sera from individuals with lung cancer and pneumonia. Mtb81 reacted with 26 of 37 HIV ؉ TB ؉ sera (70%) in this group, compared to 2 of 37 (5%) that reacted with the 38-kDa antigen. Together, these results demonstrate that Mtb81 may be a promising complementary antigen for the serodiagnosis of TB. The 38-kDa antigen was expressed in E. coli by using a hexahistidine tag similar to that used for Mtb81 (see below). The TB lysate was prepared by alternately homogenizing and sonicating three 100-mg ampoules of dessicated M. tuberculosis H37Ru (Difco, Franklin Lakes, N.J.) three times in 25 ml of 10 mM Tris (pH 8) containing 2% Nonidet P-40. The mixture was spun for 10 min at 13,000 ϫ g. The supernatant was used as the TB lysate.Study population. Blood was drawn from all patients after informed consent was obtained. Serum samples were obtained from individuals who had pulmonary TB alone prior to treatment (culture and/or acid-fast bacillus smear positive; see Results) or who had documented coinfections with HIV, as evidenced by a positive HIV type 1 and 2 antibody ELISA. These included samples from Uganda and South Africa that were obtained from Milton Tam (Program for Appropriate Technology in Health, Seattle, Wash.). Additional HIV ϩ TB ϩ serum samples from South Africa and samples from patients with lung cancer and pneumonia (China) were obtained from Robert Cole (AMRAD-ICT, French's Forest, Australia). This second group of HIV ϩ TB ϩ samples was selected on the basis of their negativity for the 38-kDa antigen in the AMRAD-ICT rapid test but smear-and/or culture-positive results (6, 34). To further evaluate the specificity of the Mtb81 antigen, we obtained sera from individuals who were PPD positive (Ͼ10-mm reaction zone) (culture, clinically, and radiographically negative for TB) and PPD negative (King County Tuberculosis Clinic, Seattle, Wash.). Samples from healthy blood donors (United States) were obtained from Boston Biomedica (West Bridgewater, Mass.), and sera from individuals who were HIV seropositive alone were obtained fr...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Mass Spectrometric Identification of Mtb81, a Novel Serological Marker for Tuberculosis

Hendrickson

Douglass

Reynolds

et al. 2000

J. Clin. Microbiol.

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“…Attempts to define the immunologically active components within this fraction have led to the purification and characterization of several proteins including the 6-kDa ESAT6 (56), 24-kDa MPT64 (39,47), 30-to 31-kDa Ag85 complex (21,24,39), and 45-kDa MPT32 (15,39,48). A strong humoral response against CFPs has also been noted (29), and antigens such as the 45-kDa MPT32 (18), the 38-kDa PstS homolog (9), and an 88-kDa protein complex (29,49) hold promise as tools for serodiagnosis.…”

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confidence: 99%

Definition of Mycobacterium tuberculosis culture filtrate proteins by two-dimensional polyacrylamide gel electrophoresis, N-terminal amino acid sequencing, and electrospray mass spectrometry

1997

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A number of the culture filtrate proteins secreted by Mycobacterium tuberculosis are known to contribute to the immunology of tuberculosis and to possess enzymatic activities associated with pathogenicity. However, a complete analysis of the protein composition of this fraction has been lacking. By using two-dimensional polyacrylamide gel electrophoresis, detailed maps of the culture filtrate proteins of M. tuberculosis H37Rv were generated. In total, 205 protein spots were observed. The coupling of this electrophoretic technique with Western blot analysis allowed the identification and mapping of 32 proteins. Further molecular characterization of abundant proteins within this fraction was achieved by N-terminal amino acid sequencing and liquid chromatography-mass spectrometry. Eighteen proteins were subjected to N-group analysis; of these, only 10 could be sequenced by Edman degradation. Among the most interesting were a novel 52-kDa protein demonstrating significant homology to an ␣-hydroxysteroid dehydrogenase of Eubacterium sp. strain VPI 12708, a 25-kDa protein corresponding to open reading frame 28 of the M. tuberculosis cosmid MTCY1A11, and a 31-kDa protein exhibiting an amino acid sequence identical to that of antigen 85A and 85B. This latter product migrated with an isoelectric point between those of antigen 85A and 85C but did not react with the antibody specific for this complex, suggesting that there is a fourth member of the antigen 85 complex. Novel N-terminal amino acid sequences were obtained for three additional culture filtrate proteins; however, these did not yield significant homology to known protein sequences. A protein cluster of 85 to 88 kDa, recognized by the monoclonal antibodies IT-57 and IT-42 and known to react with sera from a large proportion of tuberculosis patients, was refractory to N-group analysis. Nevertheless, mass spectrometry of peptides obtained from one member of this complex identified it as the M. tuberculosis KatG catalase/peroxidase. Thus, the detailed mapping of M. tuberculosis proteins, combined with state-of-the-art analytical techniques such as mass spectrometry, provides a basis for further analysis and rapid identification of biologically relevant molecules.

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“…7 In several studies, positive antibody responses in patients with tuberculosis were found 60-90% with specificities of 91-100%. 8,9,10,11 The higher rate of seropositivity for the microscopy/culture positive TB patients compared to that for the microscopy/culture negative TB patients may be due to more bacillary load exposed to immune system leading to more antibody production. The cause of negative response among microscopy/culture positive patients of both pulmonary and extrapulmonary TB might be due to intracellular nature of the bacilli resulting in less or no exposure to the immune system, or it may be due to immunosuppressive condition of the patients.…”

Section: Western Blot Analysismentioning

confidence: 99%

Comparison of Western Blot Technique with Microscopy and Culture for Diagnosis of Tuberculosis

Gazi

Miah

Ahmed

et al. 2016

Bangladesh J Med Microbiol

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Diagnosis of tuberculosis is usually done by microscopy for AFB and other tests. But each test has different limitation. This study was carried out to compare western blot technique with microscopy and culture for diagnosis of tuberculosis. Sputum and other samples were collected from 112 TB patients for bacteriological diagnosis by microscopy and culture for AFB. Serum samples were collected from 112 TB patients and 50 control subjects for detection of antibody response by western blot. For western blot analysis, Mycobacterium tuberculosis sonicate antigen extract was fractionated by electrophoresis on polyacrylamide gel. Western blot analysis revealed four polypeptides against which antibody response of study population were observed. The sensitivity of western blot was 73.21% which was higher than that of microscopy (63.39%) and culture (57.14%) for diagnosis of tuberculosis. The specificity of western blot was 92%.Bangladesh J Med Microbiol 2007; 01 (01):21-24

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Human humoral responses to antigens of Mycobacterium tuberculosis: immunodominance of high-molecular-mass antigens

Cited by 62 publications

References 40 publications

Mass Spectrometric Identification of Mtb81, a Novel Serological Marker for Tuberculosis

Mass Spectrometric Identification of Mtb81, a Novel Serological Marker for Tuberculosis

Definition of Mycobacterium tuberculosis culture filtrate proteins by two-dimensional polyacrylamide gel electrophoresis, N-terminal amino acid sequencing, and electrospray mass spectrometry

Comparison of Western Blot Technique with Microscopy and Culture for Diagnosis of Tuberculosis

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