ABSTRACT
Background: This study aimed to assess the diagnostic value of Krebs von den Lungen-6 (KL-6), Surfactant protein-A (SP-A), SP-D and molecular matrixmetalloproteinase-7 (MMP-7) in discriminating patients with interstitial lung diseases (ILDs) from disease control subjects. Methods: Serum levels of KL-6, SP-A, SP-D and MMP-7 were measured in both the ILD and non-ILD (NILD) groups. Receiver operating characteristic (ROC) curve analysis was conducted to evaluate the diagnostic potential of these markers and laboratory indices. High-resolution computed tomography (HRCT) fibrosis scores were determined, and their correlation with the serum markers was analyzed. Results: Serum levels of KL-6 and MMP-7 were significantly elevated in the ILD group compared to the control group, while no significant differences were observed for SP-A and SP-D. ROC analysis of KL-6 demonstrated superior diagnostic accuracy, with a sensitivity of 76.36%, specificity of 91.07%, and an area under curve (AUC) of 0.902 (95%CI 0.866-0.945). These findings were consistent across an additional cohort. Correlation analysis revealed a link between KL-6 levels at initial diagnosis and HRCT fibrosis scores, indicating disease severity. Moreover, a negative correlation was found between KL-6 and pulmonary function indices, reflecting disease progression. Patients with increased 12-month HRCT fibrosis score showed higher lactate dehydrogenase (LDH) levels, with LDH exhibiting an AUC of 0.767 (95% CI: 0.520-0.927) as a predictor of progression. Conclusions: Serum KL-6 detection proves to be a valuable tool for accurately distinguishing ILDs from control subjects. While KL-6 shows a correlation with HRCT fibrosis scores and a negative association with pulmonary function indices, its predictive value for ILDs prognosis is limited. Trial registration: This study received retrospective approval from the Ethical Committee of Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (institutional review board ID: TJ-IRB20210331, date: 2021.03.30).
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BACKGROUND: The study aimed to investigate the diagnostic efficiency of human neutrophil lipocalin (HNL) in bacterial infections in children. METHODS: This study included 49 pediatric patients with bacterial infections, 37 patients with viral infections, 30 patients with autoimmune diseases (AID) and 41 healthy controls (HCs). HNL, procalcitonin (PCT), C-reactive protein (CRP), white blood cell (WBC) and neutrophil counts were detected in the initial diagnosis and the following days. RESULTS: In the patients with bacterial infections, the levels of HNL, PCT, CRP, WBC and neutrophils were significantly increased than that of disease controls and HCs. The dynamic of these markers was monitored during antibiotic treatment. The level of HNL was decreased rapidly in patients with effective treatment, but maintained at high levels in deteriorated patients according to the clinical progression. CONCLUSIONS: HNL detection is an effective biomarker to identify bacterial infections from viral infections and other AIDs, and has potential value to evaluate the effect of antibiotic treatment in pediatric patients.
Subject(s)
Bacterial Infections , Virus Diseases , Humans , Child , Lipocalins/metabolism , Neutrophils/metabolism , Biomarkers , C-Reactive Protein , Bacterial Infections/microbiology , Virus Diseases/diagnosis , ProcalcitoninABSTRACT
Background: The accurate detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the key to control Coronavirus Disease-2019 (COVID-19). The performance of different antibody detection methods for diagnosis of COVID-19 is inconclusive. Methods: Between 16 February and 28 February 2020, 384 confirmed COVID-19 patients and 142 healthy controls were recruited. 24 different serological tests, including 4 enzyme-linked immunosorbent assays (EIAs), 10 chemiluminescent immunoassays (CLIAs), and 10 lateral flow immunoassays (LFIAs), were simultaneously performed. Results: The sensitivities of anti-SARS-CoV-2 IgG and IgM antibodies with different reagents ranged from 75 to 95.83% and 46.09 to 92.45%, respectively. The specificities of both anti-SARS-CoV-2 IgG and IgM were relatively high and comparable among different reagents, ranged from 88.03 to 100%. The area under the curves (AUCs) of different tests ranged from 0.733 to 0.984, and the AUCs of EIAs or CLIAs were significantly higher than those of LFIAs. The sensitivities of both IgG and IgM gradually increased with increase of onset time. After 3-4 weeks, the sensitivities of anti-SARS-CoV-2 IgG were maintained at a certain level but the sensitivities of IgM were gradually decreased. Six COVID-19 patients who displayed negative anti-SARS-CoV-2 results were associated with the factors such as older age, having underlying diseases, and using immunosuppressant. Conclusion: Besides the purpose of assessing the impact of the SARS-CoV-2 pandemic in the population, SARS-CoV-2 antibody assays may have an adjunct role in the diagnosis and exclusion of COVID-19, especially by using high-throughput technologies (EIAs or CLIAs).
ABSTRACT
Background: Novel approaches for tuberculosis (TB) diagnosis, especially for distinguishing active TB (ATB) from latent TB infection (LTBI), are urgently warranted. The present study aims to determine whether the combination of HLA-DR on Mycobacterium tuberculosis (MTB)-specific cells and TB antigen/phytohemagglutinin (TBAg/PHA) ratio could facilitate MTB infection status discrimination. Methods: Between June 2020 and June 2021, participants with ATB and LTBI were recruited from Tongji Hospital (Qiaokou cohort) and Sino-French New City Hospital (Caidian cohort), respectively. The detection of HLA-DR on MTB-specific cells upon TB antigen stimulation and T-SPOT assay were simultaneously performed on all subjects. Results: A total of 116 (54 ATB and 62 LTBI) and another 84 (43 ATB and 41 LTBI) cases were respectively enrolled from Qiaokou cohort and Caidian cohort. Both HLA-DR on IFN-γ+TNF-α+ cells and TBAg/PHA ratio showed discriminatory value in distinguishing between ATB and LTBI. Receiver operator characteristic (ROC) curve analysis showed that HLA-DR on IFN-γ+TNF-α+ cells produced an area under the ROC curve (AUC) of 0.886. Besides, TBAg/PHA ratio yield an AUC of 0.736. Furthermore, the combination of these two indicators resulted in the accurate discrimination with an AUC of 0.937. When the threshold was set as 0.36, the diagnostic model could differentiate ATB from LTBI with a sensitivity of 92.00% and a specificity of 81.82%. The performance obtained in Qiaokou cohort was further validated in Caidian cohort. Conclusions: The combination of HLA-DR on MTB-specific cells and TBAg/PHA ratio could serve as a robust tool to determine TB disease states.
Subject(s)
Antigens, Bacterial/immunology , HLA-DR Antigens/immunology , Latent Tuberculosis/immunology , Mycobacterium tuberculosis/immunology , Phytohemagglutinins/immunology , Tuberculosis/immunology , Adult , Aged , Cohort Studies , Diagnosis, Differential , Diagnostic Tests, Routine/methods , Female , HLA-DR Antigens/metabolism , Humans , Interferon-gamma/immunology , Interferon-gamma/metabolism , Latent Tuberculosis/diagnosis , Latent Tuberculosis/microbiology , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/metabolism , Leukocytes, Mononuclear/microbiology , Male , Middle Aged , Mycobacterium tuberculosis/physiology , ROC Curve , Tuberculosis/diagnosis , Tuberculosis/microbiology , Tumor Necrosis Factor-alpha/immunology , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Background: This study aimed to assess the host immune signatures associated with EBV infection and its clinical value in indicating the severity of children with acute infectious mononucleosis (IM). Methods: Twenty-eight pediatric patients with IM aged 3-8 years were enrolled. The immune phenotypes and cytokine secretion capability of T cells were detected. Results: The percentages and absolute numbers of CD3+ and CD8+ T cells were significantly increased in IM patients compared with HCs. The percentages of Naïve CD4+ and CD8+ T cells were decreased but with increased percentages of memory CD4+ and CD8+ T subsets. Our results showed the upregulation of active marker HLA-DR, TCR-αß, and inhibitory receptors PD-1, TIGIT in CD8+ T cells from IM patients, which suggested that effective cytotoxic T cells were highly against EBV infection. However, EBV exposure impaired the cytokine (IFN-γ, IL-2, and TNF-α) secretion capability of CD4+ and CD8+ T cells after stimulation with PMA/ionomycin in vitro. Multivariate analysis revealed that the percentage of HLA-DR+ CD8+ T cells was an independent prognostic marker for IM. The percentage of HLA-DR+ CD8+ T cells was significantly correlated with high viral load and abnormal liver function results. Conclusion: Robust expansion and upregulation of HLA-DR in CD8+ T cells, accompanied with impaired cytokine secretion, were typical characteristics of children with acute IM. The percentage of HLA-DR+ CD8+ T cells might be used as a prominent marker not only for the early diagnosis but also for indicating the severity of IM.
Subject(s)
CD8-Positive T-Lymphocytes/metabolism , HLA-DR Antigens/biosynthesis , Infectious Mononucleosis/immunology , Lymphocyte Subsets/metabolism , B-Lymphocyte Subsets/immunology , Case-Control Studies , Child , Child, Preschool , Cytokines/metabolism , Early Diagnosis , Female , Gene Expression Regulation/immunology , Genes, MHC Class II , HLA-DR Antigens/genetics , Humans , Immunologic Memory , Immunophenotyping , Infectious Mononucleosis/diagnosis , Lymphocyte Activation , Lymphocyte Count , Male , Monocytes/immunology , Severity of Illness IndexABSTRACT
BACKGROUND: This study aimed to investigate the efficiency of combining tumor-associated antigens (TAAs) and autoantibodies in the diagnosis of lung cancer. METHODS: The serum levels of TAAs and seven autoantibodies (7-AABs) were detected from patients with lung cancer, benign lung disease and healthy controls. The performance of a new panel by combing TAAs and 7-AABs was evaluated for the early diagnosis of lung cancer. RESULTS: The positive rate of 7-AABs was higher than the single detection of antibody. The positive rate of the combined detection of 7-AABs in lung cancer group (30.2%) was significantly higher than that of healthy controls (16.8%), but had no statistical difference compared with that of benign lung disease group (20.8%). The positive rate of 7-AABs showed a tendency to increase in lung cancer patients with higher tumor-node-metastasis (TNM) stages. For the pathological subtype analysis, the positive rate of 7-AABs was higher in patients with squamous cell carcinoma and small cell lung cancer than that of adenocarcinoma. The levels of carcinoembryonic antigen (CEA) and cytokeratin 19 fragment 211 (CYFRA 21-1) were significantly higher than that of benign lung disease and healthy control groups. An optimal model was established (including 7-AABs, CEA and CYFRA21-1) to distinguish lung cancer from control groups. The performance of this model was superior than that of single markers, with a sensitivity of 52.26% and specificity of 77.46% in the training group. Further assessment was studied in another validation group, with a sensitivity of 44.02% and specificity of 83%. CONCLUSIONS: The diagnostic performance was enhanced by combining 7-AABs, CEA and CYFRA21-1, which has critical value for the screening and early detection of lung cancer.
Subject(s)
Autoantibodies/immunology , Epithelial Cell Adhesion Molecule/immunology , Lung Neoplasms/diagnosis , Female , Humans , Lung Neoplasms/immunology , Male , Middle Aged , Retrospective StudiesABSTRACT
OBJECTIVES: T cell immunoglobulin and mucin domain 3 (TIM-3) has been reported as an important regulatory molecule on T cells and plays a pivotal role in autoimmune diseases, but the impact on dendritic cells (DCs) is poorly explored. The formation of neutrophil extracellular traps (NETs) is considered as strongly implicated in the pathogenesis of autoimmune diseases, such as in myeloperoxidase-antineutrophil cytoplasmic autoantibody associated vasculitis (MPO-AAV). This study thus aimed to investigate the potential regulation roles of TIM-3 in the regulation of NETs-mediated DC activation in MPO-AAV. METHODS: Twenty untreated patients with MPO-AAV and 20 healthy controls were enrolled in this study. The expressions of TIM-3 and toll-like receptor 4 (TLR4) in peripheral blood dendritic cells were analysed by flow cytometry, and the release of NETs by neutrophils was evaluated by immunofluorescence. In animal experiments, we measured the DC activation markers after the stimulation of NETs. Furthermore, we detected the NETs-mediated DC activation after TIM-3 blockade. RESULTS: Here we found an increased spontaneous NET production in MPOAAV patients. We also revealed a markedly reduced expression of TIM-3 and an increased expression of TLR4 on DCs of active MPO-AAV patients. We found the NETs could induce the activation of DCs and promote Toll-like receptor 4 expression on DC surface. More interestingly, the blockade of TIM-3 could further enhance the NETs-mediated DC cytokine expression. CONCLUSIONS: Our results demonstrated DC surface TIM-3 plays an important role in maintaining the NETs mediated immune homeostasis in MPO-AAV, suggesting an important role in MPOAAV development.
Subject(s)
Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis , Extracellular Traps , Animals , Antibodies, Antineutrophil Cytoplasmic , Dendritic Cells , Hepatitis A Virus Cellular Receptor 2 , Humans , Neutrophils , PeroxidaseABSTRACT
The performance of the automated indirect immunofluorescence system was compared with the manual method for detection of antinuclear antibodies (ANA) from 354 clinical serum samples. We compared the results (negative or positive), ANA patterns, and titers for the two methods. The coincidence rates for ANA positive and negative samples were 93.4% and 98.7%, respectively. The coincidence rates for single patterns, mixed patterns, and the final titers were 85.1%, 87.6%, and 87.6%, respectively. The homogeneous, speckled, cytoplasmic, centromere, multiple nuclear dots, and nucleolar pattern coincidence rates were 79.3%, 83.0%, 87.8%, 72.7%, 50%, and 56.3%, respectively. The automated indirect immunofluorescence system had acceptable accuracy for detection of ANA.
Subject(s)
Antibodies, Antinuclear/blood , Autoantibodies/blood , Autoimmune Diseases/diagnosis , Automation, Laboratory/standards , Fluorescent Antibody Technique, Indirect/methods , Antibodies, Antinuclear/immunology , Autoantibodies/immunology , Autoimmune Diseases/blood , Autoimmune Diseases/immunology , HumansABSTRACT
Accurate monitoring of host immunity is hampered by the flaws of conventional tests. The relationship between lymphocyte number and function is unknown. The function of lymphocytes was analyzed based on IFN-γ secretion assay. Lymphocyte number and function was investigated in individuals under various states. The number of CD4+ and CD8+ T cells was gradually decreased, whereas the function of them was gradually increased with increasing age. A significantly negative correlation existed between the number and function of both CD4+ and CD8+ T cells. Differently, both the number and function of NK cells are maintained at a high level after birth. Staying up all night was found to impair the function of CD4+, CD8+ T cells, or NK cells. Lymphocyte number and function were both decreased in patients with immunosuppressive conditions or opportunistic infections, while the opposite phenomenon was observed in patients with some autoimmune diseases (except for NK cells). In kidney transplant recipients, the number and function of CD4+ and CD8+ T cells were increased or decreased when rejection or infection occurred. We demonstrated that evaluation of host immunity based on combination of lymphocyte number and function plays an important role in the diagnosis, treatment, and prognosis of diseases.
Subject(s)
Lymphocytes/physiology , Adolescent , Adult , Aged , Aged, 80 and over , Calcium Ionophores/pharmacology , Child , Child, Preschool , Gene Expression Regulation/drug effects , Humans , Immunity, Cellular , Infant , Interferon-gamma/genetics , Interferon-gamma/metabolism , Ionomycin/pharmacology , Male , Middle Aged , Phorbol Esters/pharmacology , Risk , Young AdultABSTRACT
BACKGROUND: Anti-dsDNA antibody is a specific antibody in systemic lupus erythematosus (SLE). Indirect immunofluorescence test (IIFT) is a highly specific method in detecting anti-dsDNA antibody. The application of automated system has gained better consistency than manual operation. This study detected anti-dsDNA antibodies using EUROPattern Computer-aided immunofluorescence microscopy (EPA), and evaluated the performance of the automated system. METHODS: The sera of 96 patients with suspected SLE and 102 control patients were examined using IIFT. The consistency between the EPA and manual reading was analyzed. RESULTS: Analysis of 198 samples showed that the overall consistency of the negative/positive results between the EPA and manual reading was 94.95%. Based on the manual reading results, the sensitivity and specificity of EPA were 95.70% and 94.29%, respectively. The analysis of 57 samples with non-specific fluorescence showed that the overall consistency of the negative/positive results was 96.49%. The analysis of the antibody titer of 89 positive samples showed that the consistency between the EPA and manual reading was 97.75%. CONCLUSION: EPA was consistent with the manual reading with regard to qualitative reading and antibody titer. With low-exposure function, EPA could read samples with non-specific fluorescence. EPA was superior to manual reading in automation and standardization.
Subject(s)
Antibodies, Antinuclear/blood , Fluorescent Antibody Technique, Indirect/methods , Lupus Erythematosus, Systemic/diagnosis , Microscopy, Fluorescence/methods , Automation , Case-Control Studies , Fluorescent Antibody Technique, Indirect/standards , Humans , Microscopy, Fluorescence/standards , Reference Standards , Sensitivity and SpecificityABSTRACT
Extrapulmonary tuberculosis (EPTB) has become more common in recent years; however, the diagnosis of EPTB remains a challenge. In this study, we analyzed the performance of the ratio of TB-specific antigen (TBAg) to phytohemagglutinin (PHA) (TBAg/PHA ratio) in T-SPOT.TB (T-SPOT) assay for diagnosis and treatment monitoring of EPTB. Between 2012 and 2017, 734 EPTB patients were diagnosed and recruited from Tongji hospital, and 1,137 suspected EPTB patients who had other diagnoses were recruited as non-EPTB controls. To validate the study, another small group of EPTB patients and non-EPTB controls were recruited from Sino-French New City Branch of Tongji Hospital. The positive rate of peripheral blood T-SPOT in EPTB and non-EPTB were 88.15 and 32.28%. In T-SPOT positive patients, the direct T-SPOT results had limited value in distinguishing these two conditions. A further calculation of the TBAg/PHA ratio of T-SPOT showed improved performance in each form of EPTB. If using 0.20 as the threshold value of the TBAg/PHA ratio, the pooled sensitivity and specificity were 70.79 and 91.55% in distinguishing EPTB from non-EPTB. The validation results showed a better performance of the TBAg/PHA ratio in distinguishing these two conditions, with a sensitivity and specificity of 81.82 and 97.56%, respectively. Comparing with directly using T-SPOT results, the TBAg/PHA ratio was less affected by immunosuppression. Furthermore, PHA value reflected immunosuppression and could help to judge the credibility of T-SPOT results in EPTB patients with different immune status. The TBAg/PHA ratio was significantly decreased during anti-tuberculosis (TB) treatment, which suggests that it can also be used to monitor therapeutic efficacy. These data provide new insights into the role of T-SPOT assay in TB disease, and the TBAg/PHA ratio might be a useful tool for diagnosis and treatment monitoring of EPTB.
Subject(s)
Antigens, Bacterial/analysis , Phytohemagglutinins/analysis , Tuberculosis/diagnosis , Tuberculosis/drug therapy , Adult , Female , Humans , Interferon-gamma Release Tests , Male , Middle Aged , Mycobacterium tuberculosis , Sensitivity and SpecificityABSTRACT
INTRODUCTION: Differentiation of tuberculoma from cancer in solitary pulmonary nodule or mass still remains a major challenge in diagnostic laboratories. OBJECTIVES: The objective of this study is to determine the performance of T-SPOT.TB assay in discriminating these 2 diseases. METHODS: We prospectively enrolled 331 patients with a solitary pulmonary nodule or mass on computed tomography scans. Conventional tests and T-SPOT.TB assay were simultaneously performed in all participants. RESULTS: Our results showed that the performance of directly using T-SPOT.TB results in distinguishing tuberculoma from cancer in solitary pulmonary nodule or mass was not satisfactory because of moderate sensitivity and specificity. However, a further calculation of the ratio of TB-specific antigen (TBAg) to phytohemagglutinin (PHA) (TBAg/PHA ratio) of T-SPOT.TB assay may lead to improvement in distinguishing these 2 diseases. If using the threshold value of 0.236, the sensitivity and specificity of the TBAg/PHA ratio in distinguishing tuberculoma from cancer in solitary pulmonary nodule or mass were, respectively, 80.6% and 93.3%. The area under the curve (AUC) of the receiver operating characteristic curve was 0.921 (95% confidence interval, 0.875-0.967). Furthermore, the TBAg/PHA ratio may also be used to distinguish tuberculoma from other benign diseases (AUC: 0.909, sensitivity: 85.07%, specificity: 90%). CONCLUSIONS: Calculation of the TBAg/PHA ratio might provide a useful non-invasive tool for distinguishing tuberculoma from cancer in patients with a solitary pulmonary nodule or mass in TB-endemic countries.
Subject(s)
Antigens, Bacterial/analysis , Lung Neoplasms/diagnosis , Phytohemagglutinins/analysis , Solitary Pulmonary Nodule/diagnosis , Tuberculoma/diagnosis , Tuberculosis, Pulmonary/diagnosis , Biopsy, Needle , Diagnosis, Differential , Female , Follow-Up Studies , Humans , Lung Neoplasms/metabolism , Male , Middle Aged , Mycobacterium tuberculosis/immunology , Mycobacterium tuberculosis/isolation & purification , Prospective Studies , ROC Curve , Solitary Pulmonary Nodule/metabolism , Tomography, X-Ray Computed , Tuberculoma/metabolism , Tuberculoma/microbiology , Tuberculosis, Pulmonary/metabolism , Tuberculosis, Pulmonary/microbiologyABSTRACT
Mycobacterium tuberculosis (Mtb)-specific IFN-γ secretion plays important roles in anti-tuberculosis (TB) immunity. Mtb-specific IFN-γ response can be induced in HIV/TB co-infected patients with a low CD4 lymphocyte count; this suggests that the source of Mtb-specific IFN-γ production is not limited in CD4(+) T lymphocytes. Currently, the major sources of Mtb-specific IFN-γ production and the function and phenotype of Mtb-specific IFN-γ-producing cells still remain unclear. Thirty-nine participants (24 active TB patients, 10 HIV/TB co-infected patients, and 5 healthy volunteers) were recruited according to conventional tests and Mtb-specific IFN-γ ELISPOT assay. Multicolor flow cytometry was used to investigate the production of intracellular IFN-γ in peripheral blood mononuclear cells (PBMCs) after Mtb-specific antigen stimulation. Our results showed that CD4(+), CD8(+) T cells and NK cells are all major sources of Mtb-specific IFN-γ production in PBMCs of TB patients. Moreover, CD8(+) T cells are the highest number of Mtb-specific IFN-γ-producing cells in HIV/TB co-infected patients. Although the activity of NK cells is significantly reduced in TB patients when compared with healthy controls, Mtb-specific antigen stimulation induces a significant increase in NK cell activity. We also showed that CD45RO is the characteristic marker of Mtb-specific IFN-γ-producing T cells but not that of Mtb-specific IFN-γ-producing NK cells in peripheral blood. High expression of CD11a may be the characteristic feature of Mtb-specific IFN-γ-producing NK cells. This study put forward a new insight on the source of antigen-specific IFN-γ-production in PBMCs of TB patients.
Subject(s)
CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Interferon-gamma/immunology , Killer Cells, Natural/immunology , Mycobacterium tuberculosis/immunology , Tuberculosis, Pulmonary/immunology , Adult , Aged , CD11a Antigen/immunology , Female , HIV Infections/immunology , Humans , Leukocyte Common Antigens/immunology , Male , Middle Aged , Young AdultABSTRACT
Human NK cells display extensive phenotypic and functional heterogeneity among healthy individuals, but the mechanism responsible for this variation is still largely unknown. Here, we show that a novel immune receptor, T-cell immunoglobulin and ITIM domain (TIGIT), is expressed preferentially on human NK cells but shows wide variation in its expression levels among healthy individuals. We found that the TIGIT expression level is related to the phenotypic and functional heterogeneity of NK cells, and that NK cells from healthy individuals can be divided into three categories according to TIGIT expression. NK cells with low levels of TIGIT expression show higher cytokine secretion capability, degranulation activity, and cytotoxic potential than NK cells with high levels of TIGIT expression. Blockade of the TIGIT pathway significantly increased NK-cell function, particularly in NK cells with high levels of TIGIT expression. We further observed that the TIGIT expression level was inversely correlated with the IFN-γ secretion capability of NK cells in patients with cancers and autoimmune diseases. Importantly, we propose a novel mechanism that links TIGIT expression with NK-cell functional heterogeneity, and this mechanism might partially explain why individuals have different susceptibilities to infection, autoimmune disease, and cancer.
Subject(s)
Gene Expression Regulation/immunology , Interferon-gamma/immunology , Killer Cells, Natural/immunology , Receptors, Immunologic/immunology , Signal Transduction/immunology , Female , Humans , Killer Cells, Natural/cytology , MaleABSTRACT
Active tuberculosis (TB) patients show impaired NK cell function, and the underlying mechanism remains largely unknown. In this study, we confirmed the decrease in activation, cytokine secretion, and degranulation potential of NK cells in active TB patients. We further investigated whether coinhibitory receptor Tim-3 was involved with impairment of NK cells. Our results revealed that the expression of Tim-3 on NK cells was increased in active TB patients. Tim-3 expression was inversely correlated with IL-12-stimualted IFN-γ production. Moreover, blocking the Tim-3 pathway restored IFN-γ secretion and degranulation of NK cells. Blocking this pathway also increased NK cell cytotoxicity against K562 target cells, and improved the ability of NK cells to control Mtb growth in monocyte-derived macrophages. The Tim-3 expression on NK cells was also observed to be significantly decreased in TB patients post-treatment. In this study, we have identified that Tim-3 is involved with NK cell impairment in TB patients.
Subject(s)
Killer Cells, Natural/immunology , Membrane Proteins/metabolism , Tuberculosis/metabolism , Cell Degranulation , Hepatitis A Virus Cellular Receptor 2 , Humans , Interferon-gamma/biosynthesis , Interleukin-12/physiology , K562 Cells , Lymphocyte Activation , Mycobacterium tuberculosis/growth & development , Tuberculosis/drug therapy , Tuberculosis/immunologyABSTRACT
The clinical diagnosis of hepatitis C virus (HCV) infection is important to direct an accurate course of therapy. Previous studies have reported a correlation between the signal-to-cutoff (S/CO) ratios of the anti-HCV screening test and confirm HCV infections for American anti-HCV screening kits as well as for those in China. It is currently unknown whether clinical laboratories use the same threshold S/CO ratios under routine conditions and if these values are acceptable for the analysis of Chinese samples. A total of 336 anti-HCV screening-test-positive serum samples were tested in duplicate using different lots of three most commonly used enzyme immunoassay (EIA) kits available in China. Samples were also tested using the Architect Anti-HCV chemiluminescent microparticle immunoassay (CMIA) kit and measured for HCV RNA. Recombinant immunoblot assays (RIBA) were additionally performed on samples with HCV RNA-negative results with RIBA HCV 3.0. The relationship between S/CO ratios and confirmed HCV infection rates were analyzed. The threshold S/CO ratio for each screening kit correlated with the ≥ 95% positive predictive value was InTec 12.0, KHB 4.0, Wantai 5.0, and Abbott Architect 5.0. Therefore, the same threshold S/CO ratios for manufactured domestically EIA kits was difficult to attain. A multi-center study with a large sample size is required to identify a uniform threshold S/CO ratio for use in different diagnostic laboratories. Alternatively, individual laboratories may be required to establish threshold S/CO ratios in their own laboratories to obtain consistent diagnostic results.
Subject(s)
Clinical Laboratory Techniques/methods , Hepacivirus/immunology , Hepatitis C Antibodies/blood , Hepatitis C/diagnosis , Mass Screening/methods , China , Hepatitis C/immunology , Humans , Immunoassay/methods , Predictive Value of TestsABSTRACT
BACKGROUND: ELISA and CMIA are commonly used for detection of HBsAg. However, few investigations have been performed to evaluate their value in clinical practice, especially when jointly used. A reasonable and economic HBsAg testing algorithm is in great need. METHODS: A total of 161,426 specimens in China were tested for 5 serum HBV markers with commonly used ELISA kits. 498 of these specimens were further tested for HBsAg by another ELISA kit, a CMIA kit and an HBsAg confirmatory assay. RESULTS: The sensitivities of the 2 ELISA kits were 76.21% and 88.42%, respectively. However, when using "gray-zones", the sensitivities were significantly improved to 97.43% and 96.43%. Furthermore, the combined use of the 2 ELISA kits and their "gray-zones" improved the sensitivity to 99.04%. Nevertheless, 2.91% of the samples with S/CO values below the lower "gray-zone" limits were reactive by the CMIA kit and then confirmed as HBsAg positive. However, 71.43% of the samples with HBsAg values within 0.05 and 0.10 IU/ml detected by the CMIA kit could not be confirmed. CONCLUSIONS: As a rational and economic strategy, combined use of "gray-zones" in ELISA and several different detection assays can significantly increase the efficiency of HBsAg detection.
Subject(s)
Enzyme-Linked Immunosorbent Assay/methods , Hepatitis B Surface Antigens/blood , Hepatitis B/diagnosis , Carrier State , Hepatitis B/blood , Humans , Sensitivity and SpecificityABSTRACT
The variable heavy chain region (VH) genes of 3 untreated patients with B-cell chronic lymphocytic leukemia (B-CLL) were cloned and analyzed. The VH family used was VH3-11, VH3-72 and VH3-33. More than 2% difference from the corresponding germline gene was detected in all the 3 obtained potential functional genes (average 16.7). Mutation pattern analysis indicated evidence of antigen selective pressure observed in 1 of 3 cases. Our findings suggested that the tumor cells originate from post-GC cells.