RESUMEN
BACKGROUND: Host inflammation contributes to determine whether SARS-CoV-2 infection causes mild or life-threatening disease. Tools are needed for early risk assessment. METHODS: We studied in 111 COVID-19 patients prospectively followed at a single reference Hospital fifty-three potential biomarkers including alarmins, cytokines, adipocytokines and growth factors, humoral innate immune and neuroendocrine molecules and regulators of iron metabolism. Biomarkers at hospital admission together with age, degree of hypoxia, neutrophil to lymphocyte ratio (NLR), lactate dehydrogenase (LDH), C-reactive protein (CRP) and creatinine were analysed within a data-driven approach to classify patients with respect to survival and ICU outcomes. Classification and regression tree (CART) models were used to identify prognostic biomarkers. RESULTS: Among the fifty-three potential biomarkers, the classification tree analysis selected CXCL10 at hospital admission, in combination with NLR and time from onset, as the best predictor of ICU transfer (AUC [95% CI] = 0.8374 [0.6233-0.8435]), while it was selected alone to predict death (AUC [95% CI] = 0.7334 [0.7547-0.9201]). CXCL10 concentration abated in COVID-19 survivors after healing and discharge from the hospital. CONCLUSIONS: CXCL10 results from a data-driven analysis, that accounts for presence of confounding factors, as the most robust predictive biomarker of patient outcome in COVID-19.
Asunto(s)
COVID-19/diagnóstico , Quimiocina CXCL10/sangre , Enfermedad de la Arteria Coronaria/diagnóstico , Diabetes Mellitus/diagnóstico , Hipertensión/diagnóstico , Biomarcadores/sangre , Proteína C-Reactiva/metabolismo , COVID-19/sangre , COVID-19/inmunología , COVID-19/mortalidad , Comorbilidad , Enfermedad de la Arteria Coronaria/sangre , Enfermedad de la Arteria Coronaria/inmunología , Enfermedad de la Arteria Coronaria/mortalidad , Creatina/sangre , Diabetes Mellitus/sangre , Diabetes Mellitus/inmunología , Diabetes Mellitus/mortalidad , Femenino , Hospitalización , Humanos , Hipertensión/sangre , Hipertensión/inmunología , Hipertensión/mortalidad , Inmunidad Humoral , Inmunidad Innata , Inflamación , Unidades de Cuidados Intensivos , L-Lactato Deshidrogenasa/sangre , Recuento de Leucocitos , Linfocitos/inmunología , Linfocitos/patología , Masculino , Persona de Mediana Edad , Neutrófilos/inmunología , Neutrófilos/patología , Pronóstico , Estudios Prospectivos , Estudios Retrospectivos , SARS-CoV-2 , Índice de Severidad de la Enfermedad , Análisis de SupervivenciaAsunto(s)
COVID-19 , SARS-CoV-2 , Pruebas Diagnósticas de Rutina , Humanos , Técnicas de Diagnóstico Molecular , Nasofaringe , ARN Viral , SalivaRESUMEN
During the last year, mass screening campaigns have been carried out to identify immunological response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and establish a possible seroprevalence. The obtained results gained new importance with the beginning of the SARS-CoV-2 vaccination campaign, as the lack of doses has persuaded several countries to introduce different policies for individuals who had a history of COVID-19. Lateral flow assays (LFAs) may represent an affordable tool to support population screening in low-middle-income countries, where diagnostic tests are lacking and epidemiology is still widely unknown. However, LFAs have demonstrated a wide range of performance, and the question of which one could be more valuable in these settings still remains. We evaluated the performance of 11 LFAs in detecting SARS-CoV-2 infection, analyzing samples collected from 350 subjects. In addition, samples from 57 health care workers collected at 21 to 24 days after the first dose of the Pfizer-BioNTech vaccine were also evaluated. LFAs demonstrated a wide range of specificity (92.31% to 100%) and sensitivity (50% to 100%). The analysis of postvaccination samples was used to describe the most suitable tests to detect IgG response against S protein receptor binding domain (RBD). Tuberculosis (TB) therapy was identified as a potential factor affecting the specificity of LFAs. This analysis identified which LFAs represent a valuable tool not only for the detection of prior SARS-CoV-2 infection but also for the detection of IgG elicited in response to vaccination. These results demonstrated that different LFAs may have different applications and the possible risks of their use in high-TB-burden settings. IMPORTANCE Our study provides a fresh perspective on the possible employment of SARS-CoV-2 LFA antibody tests. We developed an in-depth, large-scale analysis comparing LFA performance to enzyme-linked immunosorbent assay (ELISA) and electrochemiluminescence immunoassay (ECLIA) and evaluating their sensitivity and specificity in identifying COVID-19 patients at different time points from symptom onset. Moreover, for the first time, we analyzed samples of patients undergoing treatment for endemic poverty-related diseases, especially tuberculosis, and we evaluated the impact of this therapy on test specificity in order to assess possible performance in TB high-burden countries.
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Anticuerpos Antivirales/sangre , Prueba Serológica para COVID-19/métodos , Vacunas contra la COVID-19/inmunología , SARS-CoV-2/inmunología , Glicoproteína de la Espiga del Coronavirus/inmunología , Adulto , Vacuna BNT162 , COVID-19/diagnóstico , Técnicas Electroquímicas , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Inmunoensayo/métodos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Masculino , Tamizaje Masivo/métodos , Pruebas en el Punto de Atención , Sensibilidad y Especificidad , Tuberculosis/diagnóstico , Adulto JovenRESUMEN
One hundred and twenty-two Mycobacterium chimaera strains isolated in Italy from cardiac surgery-related patients, cardiac surgery-unrelated patients and from heater-cooler units, were submitted to whole-genome sequencing and to subsequent SNP analysis. All but one strains isolated from cardiac surgery-related patients belonged to Subgroup 1.1 (19/23) or Subgroup 1.8 (3/23). Only 28 out of 79 strains isolated from heater-cooler units belonged to groupings other than 1.1 and 1.8. The strains isolated from cardiac surgery-unrelated patients were instead distributed across the phylogenetic tree. Our data, the first on isolates from Italy, are in agreement with a recent large genomic study suggesting a common source, represented by strains belonging to Subgroups 1.1 and 1.8, of cardiac surgery-related Mycobacterium chimaera infections. The strains belonging to groupings other than 1.1 and 1.8 isolated from heather-cooler units evidently resulted from contaminations at hospital level and had no share in the Mycobacterium chimaera outbreak. One Mycobacterium chimaera strain investigated in this study proved distant from every previously known Mycobacterium chimaera Groups (1, 2, 3 and 4) and we propose to assign to a novel group, named "Group 5".
Asunto(s)
Infección Hospitalaria/microbiología , Infecciones por Mycobacterium no Tuberculosas/genética , Infecciones por Mycobacterium/genética , Mycobacterium/aislamiento & purificación , Procedimientos Quirúrgicos Cardíacos/efectos adversos , Infección Hospitalaria/genética , Brotes de Enfermedades , Contaminación de Equipos , Femenino , Genómica , Humanos , Italia/epidemiología , Masculino , Mycobacterium/patogenicidad , Infecciones por Mycobacterium/epidemiología , Infecciones por Mycobacterium/microbiología , Infecciones por Mycobacterium no Tuberculosas/epidemiología , Infecciones por Mycobacterium no Tuberculosas/microbiología , Micobacterias no Tuberculosas/genética , Micobacterias no Tuberculosas/patogenicidad , Polimorfismo de Nucleótido Simple/genética , Microbiología del Agua , Secuenciación Completa del GenomaRESUMEN
Background: Interleukin (IL)-7 promotes the generation, expansion, and survival of memory T cells. Previous mouse and human studies showed that IL-7 can support immune cell reconstitution in lymphopenic conditions, expand tumor-reactive T cells for adoptive immunotherapy, and enhance effector cytokine expression by autoreactive T cells. Whether pathogen-reactive T cells also benefit from IL-7 exposure remains unknown. Methods: In this study, we investigated this issue in cultures of peripheral blood mononuclear cells (PBMCs) derived from patients infected with various endemic pathogens. After short-term exposure to IL-7, we measured PBMC responses to antigens derived from pathogens, such as Mycobacterium tuberculosis, Candida albicans, and cytomegalovirus, and to the superantigen Staphylococcus aureus enterotoxin B. Results: We found that IL-7 favored the expansion and, in some instances, the uncovering of pathogen-reactive CD4 T cells, by promoting pathogen-specific interferon-γ, IL-2, and tumor necrosis factor recall responses. Conclusions: Our findings indicate that IL-7 unveils and supports reactivation of pathogen-specific T cells with possible diagnostic, prognostic, and therapeutic significance of clinical value, especially in conditions of pathogen persistence and chronic infection.
Asunto(s)
Antígenos/inmunología , Linfocitos T CD4-Positivos/inmunología , Interleucina-7/inmunología , Anciano , Candida albicans/inmunología , Candidiasis/inmunología , Citomegalovirus/inmunología , Infecciones por Citomegalovirus/inmunología , Femenino , Humanos , Inmunoterapia Adoptiva/métodos , Interferón gamma/inmunología , Interleucina-2/inmunología , Leucocitos Mononucleares/inmunología , Activación de Linfocitos/inmunología , Masculino , Mycobacterium tuberculosis/inmunología , Tuberculosis/inmunología , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
A thorough phenotypic and genotypic analysis of 150 strains belonging to the Mycobacterium terrae complex resulted in the identification of a number of previously unreported sequevars (sqvs) within the species known to belong to the complex. For the species Mycobacterium arupense, three sqvs were detected in the 16S rRNA gene, six sqvs in the hsp65 gene and 15 sqvs in the rpoB gene; in Mycobacterium senuense two sqvs were present in each of the three genetic regions; in Mycobacterium kumamotonense four, two and nine sqvs were found, respectively, and in M. terrae three, four and six sqvs were found, respectively. The inappropriate inclusion of Mycobacterium triviale within the M. terrae complex was confirmed. The limited utility of biochemical tests and of mycolic acid analyses for the differentiation of the members of M. terrae complex was also confirmed. The survey allowed the recognition of three previously undescribed species that were characterized by unique sequences in the 16S rRNA, hsp65 and rpoB genes. Mycobacterium engbaekii sp. nov. (proposed previously 40 years ago but never validly published) was characterized by pink photochromogenic pigmentation and rapid growth; phylogenetically it was related to Mycobacterium hiberniae. The type strain of this species, of which eight strains were investigated, is ATCC 27353(T) (â=âDSM 45694(T)). A cluster of 24 strains was the basis for the description of Mycobacterium heraklionense sp. nov., which has an intermediate growth rate and is unpigmented; nitrate reductase activity is typically strong. Closely related to M. arupense with respect to the 16S rRNA gene, M. heraklionense sp. nov. could be clearly differentiated from the latter species in the other genetic regions investigated. The type strain is NCTC 13432(T) (â=âLMG 24735(T)â=âCECT 7509(T)). Mycobacterium longobardum sp. nov., represented in the study by seven strains, was characterized by a unique phylogenetic location within the M. terrae complex, clearly divergent from any other species. The type strain is DSM 45394(T) (â=âCCUG 58460(T)).
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Micobacterias no Tuberculosas/clasificación , Filogenia , Técnicas de Tipificación Bacteriana , ADN Bacteriano/genética , Genes Bacterianos , Datos de Secuencia Molecular , Ácidos Micólicos/análisis , Micobacterias no Tuberculosas/genética , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADNRESUMEN
PURPOSE: The purpose of this study is to evaluate the rate of Mycobacterium tuberculosis infection in uveitis patients using an ELISPOT-IFN-gamma (ELISPOT-MTP) assay and a tuberculin skin test (TST). METHODS: Fifty-three patients with suspicious tuberculous uveitis, seen at the Ocular Immunology and Uveitis Service, Scientific Institute San Raffaele, Milan, Italy, were compared with 233 healthy control subjects. All uveitis patients, together with healthy control subjects, underwent in-house ELISPOT-MTP assay and then the TST. RESULTS: None of the patients had signs of active tuberculosis. A total of 75.5% of uveitis patients showed positive TST reaction while 58.5% responded positively to ELISPOT-MTP. In healthy individuals, these responses were 30.5% and 25.3%, respectively (p<0.0001). In a different diagnosis subset, TST and ELISPOT positivity were, respectively, 80% and 50% in anterior uveitis; 75% and 50% in intermediate uveitis; 100% and 87.5% in serpiginous-like choroiditis; 90% and 80% in posterior uveitis; and 57.1% and 42.9% in panuveitis. Serpiginous-like choroiditis and posterior uveitis patients had a higher number of ELISPOT-MTP positive results and a higher grade of intensity of ELISPOT-MTP responses compared to healthy control subjects (p=0.0098). CONCLUSIONS: Our uveitis patients had higher M tuberculosis infection rate and grade of intensity response than healthy control subjects detected by ELISPOT-MTP. This response is statistically significant and higher in patients with serpiginous-like choroiditis and posterior uveitis.
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Ensayo de Immunospot Ligado a Enzimas , Mycobacterium tuberculosis/inmunología , Prueba de Tuberculina , Tuberculosis Ocular/diagnóstico , Uveítis/diagnóstico , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Antígenos Bacterianos/inmunología , Proteínas Bacterianas/inmunología , Niño , Femenino , Humanos , Masculino , Persona de Mediana Edad , Fragmentos de Péptidos/inmunología , Tuberculosis Ocular/inmunología , Uveítis/inmunología , Adulto JovenRESUMEN
We evaluate the performance of the GenoType® MTBDRsl (Hain Lifescience Nehren, Germany) for the detection of second-line resistant tuberculosis and we correlate the frequency of mutations to different Mycobacterium tuberculosis genotypes. We tested 175 strains and 59 clinical specimens interpreting the results according to the Standards for Reporting of Diagnostic Accuracy recommendations. All the strains were also investigated by spoligotyping and Mycobacterial Interspersed Repetitive Units-Variable Number of Tandem Repeats typing. The performances of the MTBDRsl in detecting resistance to fluoroquinolones (FQ), second-line injectable drugs (SLID), and ethambutol (EMB) on clinical isolates were similar (specificity â¼ 99%, sensitivity â¼ 70%, and positive predictive value (PPV) â¼ 99%). Of the 59 respiratory specimens, three samples were classified as "indeterminate". The specificity in detecting resistances was similar for FQs and EMB 100% (95% CI 92.7-100%) and 100% (95% CI 83.9-100%), respectively with a PPV of 100% (95% CI 64.6-100%) and 100% (95% CI 87.9-100%), respectively. Detection of SLID showed a specificity of 89.1% (95% CI 77.0-95.3%) and a PPV of 58.3% (95% CI 32.0-80.7%). Sensitivity for FQ-resistance detection was 100% (95% CI 64.6-100%), whereas for SLID and EMB it was 89.1% (95% CI 77.0-95.3%) and 86.1% (95% CI 71.3-93.9%), respectively. We detected a significant association between mutations in the rrs gene and Beijing lineage. The MTBDRsl can be used to "rule in" extensively drug-resistant strains of tuberculosis in a high risk group; the low sensitivity and negative predicted value (NPV) make confirmation by conventional drug susceptibility testing mandatory when mutations are not identified. NPV for SLID is higher in Beijing strains, showing that the predictive values of the molecular tests are related to the genetic background.
Asunto(s)
Técnicas de Genotipaje/métodos , Mycobacterium tuberculosis/aislamiento & purificación , Tuberculosis Resistente a Múltiples Medicamentos/diagnóstico , Antituberculosos/farmacología , Secuencia de Bases , Etambutol/farmacología , Fluoroquinolonas/farmacología , Variación Genética , Técnicas de Genotipaje/instrumentación , Técnicas de Genotipaje/normas , Humanos , Pruebas de Sensibilidad Microbiana/instrumentación , Pruebas de Sensibilidad Microbiana/métodos , Datos de Secuencia Molecular , Mutación , Mycobacterium tuberculosis/efectos de los fármacos , Mycobacterium tuberculosis/genética , Valor Predictivo de las Pruebas , Análisis de Secuencia de ADN/instrumentación , Análisis de Secuencia de ADN/métodos , Tuberculosis Resistente a Múltiples Medicamentos/tratamiento farmacológicoRESUMEN
In rheumatic patients candidate to anti-TNF-alpha treatment, there is an increased risk of developing tuberculosis (TB). The tuberculin skin test (TST), the standard diagnostic test for latent tuberculosis infection (LTBI), suffers low specificity and sensitivity. Here, we compared the performance characteristics of an in-house ELISPOT-IFN-gamma assay (using a restricted pool of Mycobacterium tuberculosis-specific peptides or MTP) to TST for the diagnosis of LTBI in 69 rheumatic patients candidate to anti-TNF-alpha treatment and in 60 healthy LTBI individuals. Among the 69 patients enrolled, 17 (25%) had a positive TST response and 15 (22%) a positive ELISPOT-MTP response. Among the patients with a positive TST result, eight had a positive and nine a negative ELISPOT-MTP response, whereas among the 49 patients with a negative TST result, 42 were ELISPOT-MTP negative, but seven (14%) were ELISPOT-MTP positive, with three indeterminate results. The agreement between the two tests was poor (k = 0.341, 95% CI = 0.060 to 0.622) and the test of symmetry was not significant (P = 0.8). Considering the ELISPOT assay, rheumatic patients had a reduced number of spot-forming cells after stimulation of lymphocytes with PHA or PPD when compared with healthy LTBI individuals. Thus, the ELISPOT-IFN-gamma assay performs better than the TST in recognizing patients with LTBI, on one hand reducing the number of patients submitted to isoniazid prophylaxis, and on the other hand, since the assay is less biased by immunosuppressive regimens than TST, recognizing LTBI patients among those with a negative TST response.
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Ensayo de Immunospot Ligado a Enzimas , Tuberculosis Latente/diagnóstico , Mycobacterium tuberculosis/inmunología , Enfermedades Reumáticas/complicaciones , Prueba de Tuberculina , Adulto , Anticuerpos Monoclonales/uso terapéutico , Femenino , Humanos , Tuberculosis Latente/complicaciones , Tuberculosis Latente/inmunología , Masculino , Persona de Mediana Edad , Enfermedades Reumáticas/terapia , Sensibilidad y Especificidad , Factor de Necrosis Tumoral alfa/antagonistas & inhibidoresRESUMEN
We investigated the distribution of natural killer (NK) cell subsets, their activating and inhibitory receptors, and their cytolytic potential, in primary human immunodeficiency virus (HIV)-infected (PHI) individuals at baseline and during 1 year of follow-up with or without antiretroviral therapy, and compared the results with those obtained in treatment-naïve, chronically HIV-infected (CHI) individuals, and HIV-seronegative (HN) healthy individuals. The proportion of the CD56(dim) and CD56(bright) subsets decreased with disease progression, whereas that of the CD56(-) CD16(+) subset increased. In the CD56(dim) subset, the proportion of cells with natural cytotoxicity receptors (NCRs) decreased with disease progression, and their cytolytic potential was reduced. Conversely, the CD56(bright) subset was characterized by a high proportion of NCR-positive, killer cell immunoglobulin-like receptor (KIR)-positive NKG2A(+) cells in both CHI and PHI individuals, which was associated with an increase in their cytolytic potential. During the 1 year of follow-up, the PHI individuals with high viraemia levels and low CD4(+) T-cell counts who received highly active antiretroviral therapy (HAART) had a similar proportion of NK subsets to CHI individuals, while patients with low viraemia levels and high CD4(+) T-cell counts who remained untreated had values similar to those of the HN individuals. Our results indicate a marked perturbation of the NK cell compartment during HIV-1 infection that is multifaceted, starts early and is progressive, primarily involves the CD56(bright) subset, and is partially corrected by effective HAART.
Asunto(s)
Terapia Antirretroviral Altamente Activa , Antígeno CD56/biosíntesis , Infecciones por VIH/inmunología , VIH-1/inmunología , Células Asesinas Naturales/metabolismo , Adulto , Linfocitos T CD4-Positivos/efectos de los fármacos , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/patología , Linfocitos T CD4-Positivos/virología , Antígeno CD56/genética , Recuento de Células , Separación Celular , Citotoxicidad Inmunológica/efectos de los fármacos , Citometría de Flujo , Estudios de Seguimiento , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/patología , Infecciones por VIH/fisiopatología , Seronegatividad para VIH , VIH-1/patogenicidad , Humanos , Células Asesinas Naturales/efectos de los fármacos , Células Asesinas Naturales/inmunología , Células Asesinas Naturales/patología , Células Asesinas Naturales/virología , Persona de Mediana Edad , Subfamília C de Receptores Similares a Lectina de Células NK/genética , Subfamília C de Receptores Similares a Lectina de Células NK/metabolismo , Receptores KIR/genética , Receptores KIR/metabolismo , Receptores Gatillantes de la Citotoxidad Natural/genética , Receptores Gatillantes de la Citotoxidad Natural/metabolismoRESUMEN
Human natural killer (NK) (CD3- CD56+) cells can be divided into two functionally distinct subsets, CD3- CD56(dim) and CD3- CD56(bright). We analysed the distribution of NK cell subsets in primary and chronic human immunodeficiency virus-1 (HIV-1) infection, to determine if HIV infection stage may influence the subset distribution. In primary infection, contrary to chronic infection, the CD3- CD56(dim) subset was expanded compared to healthy controls. We also studied the effect of antiretroviral therapy administered early in infection and found that NK cell subset distribution was partially restored after 6 months of antiretroviral therapy in primary infection, but not normalized. Recently, NK cells have been divided into CD27- and CD27+ subsets with different migratory and functional capacity and CD27-mediated NK cell activation has been described in mice. We therefore investigated whether CD27 and/or CD70 (CD27 ligand) expression on NK cells, and thus the distribution of these novel NK subsets, was altered in HIV-1-infected patients. We found up-regulated expression of both CD27 and CD70 on NK cells of patients, resulting in higher proportions of CD27(high) and CD70(high) NK cells, and this phenomenon was more pronounced in chronic infection. Experiments conducted in vitro suggest that the high interleukin-7 levels found during HIV-1 infection may participate in up-regulation of CD70 on NK cell subsets. Imbalance of NK cell subsets and up-regulated expression of CD27 and CD70 initiated early in HIV-1 infection may indicate NK cell activation and intrinsic defects initiated by HIV-1 to disarm the innate immune response to the virus.
Asunto(s)
Infecciones por VIH/inmunología , VIH-1 , Células Asesinas Naturales/inmunología , Subgrupos Linfocitarios/inmunología , Ligando CD27/sangre , Recuento de Linfocito CD4 , Antígeno CD56/sangre , Células Cultivadas , Enfermedad Crónica , Humanos , Interleucina-7/inmunología , Miembro 7 de la Superfamilia de Receptores de Factores de Necrosis Tumoral/sangre , Regulación hacia Arriba/inmunologíaRESUMEN
Human and bovine colostrum (BC) contain a remarkable amount of bioactive substances, including antibodies towards many common pathogens of the intestinal and respiratory tract as well as growth factors, vitamins, cytokines and other proteic, lipidic and glucidic factors. In this study we investigated whether BC had any immunomodulatory effect on human peripheral blood mononuclear cells (PBMC) from healthy donors. To this aim we focused on the production of IL-12 and IFN-gamma, cytokines involved in the Th1 polarization required for a successful immune response towards intracellular pathogens, such as bacteria and viruses. BC induced a dose-dependent production of IL-12 by CD14+ monocytes, but was unable to induce IFN-gamma production. However, BC differentially affected stimuli-induced IFN-gamma production: it enhanced IFN-gamma in response to weak antigenic stimulation and it inhibited IFN-gamma in response to strong antigenic stimulation. These effects were not dose-dependent. We also measured PBMC proliferation, which was substantially unaffected by BC. Our data suggest that the Th1-promoting activity of BC could contribute, together with the antibodies, to the protective effect of BC on the offspring. BC could also represent an inexpensive therapeutic tool in prevention and treatment of several human microbial infections, including influenza.
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Calostro/inmunología , Factores Inmunológicos/inmunología , Leucocitos Mononucleares/inmunología , Animales , Antígenos Bacterianos/inmunología , Antígenos Virales/inmunología , Bovinos , Células Cultivadas , Relación Dosis-Respuesta Inmunológica , Femenino , Humanos , Interferón gamma/biosíntesis , Interleucina-12/biosíntesis , Leucocitos Mononucleares/metabolismo , Activación de Linfocitos , Masculino , EmbarazoRESUMEN
Identification of individuals with a tuberculosis infection is a very important element for the control of tuberculosis. The currently used tuberculin skin test has poor sensitivity and specificity. Recently, an important advance in tuberculosis diagnosis occurred with the development of in vitro T cell-based IFN-gamma release assays. The aim of this study was to compare a RD1-based in-house ELISPOT-IFN-gamma assay with a commercial (T-SPOT.TB) assay for the diagnosis of tuberculosis infection. The results showed an almost complete concordance between the two assays, confirming that our restricted but highly selected pool of peptides is sufficient to detect tuberculosis infection.
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Técnicas para Inmunoenzimas , Mycobacterium tuberculosis/metabolismo , Juego de Reactivos para Diagnóstico , Tuberculosis/diagnóstico , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Proteínas Bacterianas/inmunología , Niño , Preescolar , Femenino , Humanos , Técnicas para Inmunoenzimas/métodos , Lactante , Interferón gamma/biosíntesis , Masculino , Persona de Mediana Edad , Péptidos/síntesis química , Péptidos/inmunología , Sensibilidad y Especificidad , Linfocitos T/inmunología , Tuberculosis/sangreRESUMEN
Identification of individuals infected with Mycobacterium tuberculosis is essential for the control of tuberculosis (TB). The specificity of the currently used tuberculin skin test (TST) is poor because of the broad antigenic cross-reactivity of purified protein derivative (PPD) with BCG vaccine strains and environmental mycobacteria. Both ESAT-6 and CFP-10, two secretory proteins that are highly specific for M. tuberculosis complex, elicit strong T-cell responses in subjects with TB. Using an enzyme-linked immunospot (ELISPOT)-IFN-gamma assay and a restricted pool of peptides derived from ESAT-6 and CFP-10, we have previously demonstrated a high degree of specificity and sensitivity of the test for the diagnosis of TB. Here, 119 contacts of individuals with contagious TB who underwent TST and the ELISPOT-IFN-gamma assay were consecutively recruited. We compared the efficacy of the two tests in detecting latent TB infection and defined a more appropriate TST cutoff point. There was little agreement between the tests (k = 0.33, P < 0.0001): 53% of the contacts with a positive TST were ELISPOT negative, and 7% with a negative TST were ELISPOT positive. Furthermore, respectively 76 and 59% of the ELISPOT-negative contacts responded in vitro to BCG and PPD, suggesting that most of them were BCG vaccinated or infected with nontuberculous mycobacteria. The number of spot-forming cells significantly correlated with TST induration (P < 0.0001). Our in-house ELISPOT assay based on a restricted pool of highly selected peptides is more accurate than TST for identifying individuals with latent TB infection and could improve chemoprophylaxis for the control of TB.
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Antígenos Bacterianos/inmunología , Proteínas Bacterianas/inmunología , Interferón gamma/biosíntesis , Mycobacterium tuberculosis/inmunología , Péptidos/inmunología , Tuberculosis Pulmonar/diagnóstico , Antígenos Bacterianos/química , Proteínas Bacterianas/química , Ensayo de Inmunoadsorción Enzimática/métodos , Humanos , Leucocitos Mononucleares/inmunología , Activación de Linfocitos , Péptidos/síntesis química , Péptidos/química , Prueba de Tuberculina/normas , Tuberculosis Pulmonar/inmunología , Tuberculosis Pulmonar/prevención & controlRESUMEN
OBJECTIVE: To compare a RD1-based in-house ELISPOT-interferon-gamma (IFN-gamma) assay with a commercial (T-SPOT.TB) assay for the diagnosis of Mycobacterium tuberculosis (TB) infection and the efficacy of the tuberculin skin test (TST) and ELISPOT assay in detecting latent TB infection (LTBI). DESIGN: Eighty-six subjects (65 household contacts of contagious TB-infected patients, 13 subjects with active or previous TB infection, and 8 with suspected TB infection) were consecutively recruited in the context of a surveillance program. METHODS: Enrolled subjects underwent the Mantoux TST and two different ELISPOT-IFN-gamma assays: an in-house assay using a pool of selected M. tuberculosis peptides (MTP) and the commercial T-SPOT.TB assay. RESULTS: The in-house and commercial ELISPOT-IFN-gamma assays showed almost complete concordance (99%) in diagnosing acute or LTBI. When comparing the efficacy of the TST with the in-house ELISPOT assay in detecting TB infection, a small agreement was observed (k=0.344, P<0.0001): 36% of the subjects with a positive TST were ELISPOT-MTP negative and 12% with a negative TST were ELISPOT-MTP positive. Furthermore, 78% of the ELISPOT-MTP negative individuals were ELISPOT- Bacillus Calmette-Guérin (BCG) positive, most of whom had received BCG vaccination. CONCLUSION: Our in-house ELISPOT assay based on a restricted pool of highly selected peptides is equivalent to the commercial T-SPOT.TB assay, is cheaper and is probably not confounded, unlike the TST, by BCG vaccination in our setting.