Synthetic Peptides for the Immunodiagnosis of Human Diseases

Gómara, María J.; Haro, Isabel

doi:10.2174/092986707780059698

Cited by 89 publications

(74 citation statements)

References 87 publications

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“…1. Inexpensive and rapid point-of-care (POC) tests based on the detection of antibodies to antigens and/or epitopes selected by a careful analysis of their immunogenic potential in patients and controls have been devised for other diseases (1,13). However, a recent WHO-sponsored analysis of the performances of currently available commercial serological tests for TB diagnosis showed that none of these tests provide accurate and reliable results for either pulmonary or extrapulmonary TB patients (46,47,56).…”

Section: Discussionmentioning

confidence: 99%

LipC (Rv0220) Is an Immunogenic Cell Surface Esterase of Mycobacterium tuberculosis

et al. 2012

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We have reported previously the identification of novel proteins of Mycobacterium tuberculosis by the immunoscreening of an expression library of M. tuberculosis genomic DNA with sera obtained from M. tuberculosis-infected rabbits at 5 weeks postinfection. In this study, we report the further characterization of one of these antigens, LipC (Rv0220). LipC is annotated as a member of the Lip family based on the presence of the consensus motif "GXSXG" characteristic of esterases. Although predicted to be a cytoplasmic enzyme, we provide evidence that LipC is a cell surface protein that is present in both the cell wall and the capsule of M. tuberculosis. Consistent with this localization, LipC elicits strong humoral immune responses in both HIV-negative (HIV ؊ ) and HIV-positive (HIV ؉ ) tuberculosis (TB) patients. The absence of anti-LipC antibodies in sera from purified protein derivative-positive (PPD ؉ ) healthy subjects confirms its expression only during active M. tuberculosis infection. Epitope mapping of LipC identified 6 immunodominant epitopes, 5 of which map to the exposed surface of the modeled LipC protein. The recombinant LipC (rLipC) protein also elicits proinflammatory cytokine and chemokine responses from macrophages and pulmonary epithelial cells. rLipC can hydrolyze short-chain esters with the carbon chain containing 2 to 10 carbon atoms. Together, these studies demonstrate that LipC is a novel cell surface-associated esterase of M. tuberculosis that is highly immunogenic and elicits both antibodies and cytokines/chemokines. There are an estimated 9 ϫ 10 6 new cases of tuberculosis (TB) and 2 ϫ10 6 TB-related deaths every year (55). Infection with Mycobacterium tuberculosis is initiated with the inhalation of a droplet bearing bacteria, and it takes months or years to progress to clinical TB. During this progression from initial infection to clinical disease, the in vivo bacteria adapt to continuously changing environments by altering their gene expressions (31,34,48,50). Previous studies to delineate culture filtrate (CF) proteins of M. tuberculosis that are recognized by antibodies during the natural course of disease progression demonstrated that the repertoire of antigens enlarges with the progression of infection (34)(35)(36)41). Interestingly, several of the M. tuberculosis antigens (malate synthase, MPT51, and ESAT6, for example) that elicit immune responses during the early stages of active infection have also been demonstrated to play important roles in the host-pathogen interaction (19-21, 57).To identify additional antigenic proteins of M. tuberculosis that are expressed during the early stages of active infection, we used sera obtained from M. tuberculosis aerosol-infected rabbits that were bled at 4 to 5 weeks postinfection to screen an expression library of M. tuberculosis genomic DNA (44). Antibodies in these sera identified several proteins known to contribute to the infection and survival of M. tuberculosis (exported repetitive protein [ERP], KatG, and MtrA) as well as novel prote...

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Section: Discussionmentioning

confidence: 99%

LipC (Rv0220) Is an Immunogenic Cell Surface Esterase of Mycobacterium tuberculosis

et al. 2012

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“…According to World Health Organization estimates, ϳ9 ϫ 10 6 new cases of tuberculosis (TB) and ϳ2 ϫ 10 6 TB-related deaths occur every year. The major proportion (Ͼ90%) of these cases and deaths occur in low-income developing countries where the diagnosis of TB is based primarily on the examination of sputum smears for acid-fast bacilli.…”

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confidence: 99%

“…In recent years, antigen discovery has been accelerated by the development of high-throughput screening assays such as microarray screening that allows the rapid selection of target potential candidate antigens from large numbers of proteins (22,25 Several immunodiagnostic tests based on synthetic peptides derived from carefully selected highly antigenic proteins have been devised for viral, bacterial, and parasitic diseases in recent years (3,6,17). In the current studies, we have screened libraries of overlapping peptides of immunogenic proteins identified from our microarray studies to investigate the feasibility of using selected peptides rather than recombinant proteins to devise a diagnostic test for TB.…”

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confidence: 99%

Peptide-Based Antibody Detection for Tuberculosis Diagnosis

Shen

Behera

Bhalla

et al. 2009

Clin Vaccine Immunol

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Tuberculosis (TB) is a major cause of morbidity and mortality, especially in developing countries. Despite significant limitations, microscopy remains the cornerstone of the global TB control strategy. As the TB epidemic escalates, new diagnostic methods that are accurate and also economical and simple to manufacture and deploy are urgently needed. Although several promising antigens have been identified and evaluated in recent years, the reproducible production of high-quality recombinant mycobacterial proteins with minimal batch-to-batch variation is difficult, laborious, and expensive. To determine the feasibility of devising a synthetic peptide-based diagnostic test for TB, we have delineated the immunodominant epitopes of three candidate antigens, Ag85B, BfrB, and TrxC, that were previously identified to be immunogenic in TB patients. The results demonstrate that combinations of carefully selected synthetic peptides derived from highly immunogenic proteins can be the basis for devising an immunodiagnostic test for TB.According to World Health Organization estimates, ϳ9 ϫ 10 6 new cases of tuberculosis (TB) and ϳ2 ϫ 10 6 TB-related deaths occur every year. The major proportion (Ͼ90%) of these cases and deaths occur in low-income developing countries where the diagnosis of TB is based primarily on the examination of sputum smears for acid-fast bacilli. Although microscopy is highly specific, it has some serious limitations, such as the fact that is has a low and variable sensitivity, is time-consuming, and is tedious. The need for multiple visits to provide specimens and obtain results before treatment can be initiated also leads to high rates of dropout of infectious patients (30). A rapid and economical diagnostic test that can provide an accurate means for the identification of sputum smear-positive TB cases is a major diagnostic priority of TB control programs in countries where TB is endemic.Antibody detection-based diagnostic assays have been successfully devised for several infectious diseases (7,15,26,30), but efforts to develop a serodiagnostic test for TB have yielded disappointing results for several decades and no currently available commercial immunodiagnostic tests for TB provide high sensitivity and specificity (24). However, interest in developing these assays for the diagnosis of TB continues due to their adaptability to simple, rapid formats, such as the dipstick and lateral-flow cassette formats, which are extremely useful in the field, where a sophisticated laboratory infrastructure and highly trained personnel are not available (8,30).Our earlier studies of the humoral immune responses elicited at different stages of active TB in humans demonstrated that only a small subset of the Mycobacterium tuberculosis culture filtrate proteins is recognized by antibodies in both patients with early TB and patients with advanced TB and in the presence of human immunodeficiency virus (HIV) coinfection (14,(20)(21)(22)(23). Our earlier studies also identified M. tuberculosis malate synthase (Rv1837c) a...

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“…28 Compared with native antigens, synthetic peptides and recombinant proteins are considered superior for immunodiagnostic purposes, because they are well-defined and can be produced in large quantities. 29,30 However, the low diagnostic sensitivities and specificities of some recombinant proteins remain a significant challenge in the serodiagnosis of CE. 31 For example, 34 CE samples, which were positive with native Ag5, showed diagnostic sensitivities of 65% and 21% when tested with rAg5 and a 38-kDa subunit of rAg5, respectively.…”

Section: Discussionmentioning

confidence: 99%

Lateral Flow Test Using Echinococcus granulosus Native Antigen B and Comparison of IgG and IgG4 Dipsticks for Detection of Human Cystic Echinococcosis

Khalilpour¹,

Sadjjadi²,

Moghadam³

et al. 2014

The American Society of Tropical Medicine and Hygiene

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Abstract. Cystic echinococcosis (CE) caused by infection with Echinococcus granulosus is of major concern for humans in many parts of the world. Antigen B was prepared from E. granulosus hydatid fluid, and Western blots confirmed eight batches showing a band corresponding to the 8-/12-kDa subunit with positive serum and no lowmolecular mass band ( 15 kDa) with negative serum. The batches were pooled and used to prepare lateral flow immunoglobulin G4 (IgG4) and IgG dipsticks. Diagnostic sensitivity was determined using serum samples from 21 hydatidosis patients, and diagnostic specificity was established using sera from 17 individuals infected with other parasites and 15 healthy people. IgG4 dipstick had a diagnostic sensitivity of 95% (20 of 21) and a specificity of 100% (32 of 32). The IgG dipstick had a sensitivity of 100% (21 of 21) and a specificity of 87.5% (28 of 32). Thus, both IgG and IgG4 dipsticks had high sensitivities, but IgG4 had greater specificity for the diagnosis of human CE.

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Synthetic Peptides for the Immunodiagnosis of Human Diseases

Cited by 89 publications

References 87 publications

LipC (Rv0220) Is an Immunogenic Cell Surface Esterase of Mycobacterium tuberculosis

LipC (Rv0220) Is an Immunogenic Cell Surface Esterase of Mycobacterium tuberculosis

Peptide-Based Antibody Detection for Tuberculosis Diagnosis

Lateral Flow Test Using Echinococcus granulosus Native Antigen B and Comparison of IgG and IgG4 Dipsticks for Detection of Human Cystic Echinococcosis

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