P0 protein promotes the expression of INF-γ and IL-10 in peripheral blood of n-hexane neuropathy patients / 中华劳动卫生职业病杂志

Objective@#To study the changes of monocyte cytokines in peripheral blood of n-hexane neuropathy patients induced by P0 protein, and to explore the role of autoimmunity in n-hexane neuropathy patients.@*Methods@#In May 2018, 5 patients with peripheral neuropathy diagnosed as n-hexane poisoning were selected as case group in Shenzhen Prevention and Treatment Center for Occupational Disease in 2017. 6 workers exposure to n-hexane and 6 workers without n-hexane exposure were selected as contact group and control group. Peripheral blood mononuclear cells(PBMC) were isolated from venous blood.@*Results@#The number of spots produced by INF-γ and IL-10 increased after stimulation with P0 protein in case group, and the positive rate was significantly higher than control group and the contact group.@*Conclusion@#Autoimmunity induced by P0 protein may be involved in the occurrence of myelin sheath damage in n-hexane neuropathy patients.

Application of enzyme-linked immunospot assay in the identification of culprit drugs in severe drug eruptions / 中华皮肤科杂志

Early identification of culprit drugs is crucial for the treatment and prevention of severe drug eruptions.At present,no accurate and effective methods are available for identifying the culprit drugs in severe drug eruptions.Commonly used tests include patch test,lymphocyte transformation test and so on.However,low sensitivity and specificity limit their clinical application.Enzyme-linked immunospot assay,an in vitro technique,can identify culprit drugs in cutaneous adverse drug reactions by detecting cytokines secreted by drug-specific T lymphocytes.It has high sensitivity and specificity in patients with severe drug eruptions,and can be carried out during the acute stage of disease or among immunocompromised patients.Therefore,enzyme-linked immunospot assay may be an effective method for identifying culprit drugs in severe drug eruptions.

Optimal Timing of Zoster Vaccination After Shingles: A Prospective Study of the Immunogenicity and Safety of Live Zoster Vaccine

BACKGROUND: Zoster vaccination is recommended for people with a history of herpes zoster (HZ), but the most effective timing of vaccine administration after zoster illness is unresolved. This prospective observational study compared the immunogenicity and safety of administering HZ vaccine at 6-12 months and 1-5 years after zoster illness. MATERIALS AND METHODS: Blood samples were collected before the administration of live zoster vaccine and 6 weeks after vaccination. Varicella-zoster virus (VZV) IgG concentrations and T-cell responses were assessed by glycoprotein enzyme-linked immunosorbent assay and interferon-γ enzyme-linked immunospot assay (ELISPOT), respectively. RESULTS: The baseline geometric mean value (GMV) of VZV IgG was higher in the 6-12 months group than in the 1-5 years group (245.5 IU/mL vs. 125.9 IU/mL; P = 0.021). However, the GMV increased significantly in both groups (P = 0.002 in the 6-12 months group; P 1 year after zoster illness. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT02704572

Optimal Timing of Zoster Vaccination After Shingles: A Prospective Study of the Immunogenicity and Safety of Live Zoster Vaccine

BACKGROUND: Zoster vaccination is recommended for people with a history of herpes zoster (HZ), but the most effective timing of vaccine administration after zoster illness is unresolved. This prospective observational study compared the immunogenicity and safety of administering HZ vaccine at 6-12 months and 1-5 years after zoster illness. MATERIALS AND METHODS: Blood samples were collected before the administration of live zoster vaccine and 6 weeks after vaccination. Varicella-zoster virus (VZV) IgG concentrations and T-cell responses were assessed by glycoprotein enzyme-linked immunosorbent assay and interferon-γ enzyme-linked immunospot assay (ELISPOT), respectively. RESULTS: The baseline geometric mean value (GMV) of VZV IgG was higher in the 6-12 months group than in the 1-5 years group (245.5 IU/mL vs. 125.9 IU/mL; P = 0.021). However, the GMV increased significantly in both groups (P = 0.002 in the 6-12 months group; P 1 year after zoster illness. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT02704572

Clinical application of T-spot test of Mycobacterium tuberculosis infection for diagnosis of suspected pulmonary tuberculosis patients / 解放军医学杂志

Objective To explore the application value of T-spot test of Mycobacterium tuberculosis infection (T-SPOT.TB) on diagnosis and differential diagnosis of pulmonary tuberculosis.Methods From Apr.2014 to Dec.2016,700 patients with suspected pulmonary tuberculosis were collected,venous blood (5ml) was drawn off and sputum was collected from each patient separately for T-SPOT.TB and pathogens identification (including TB).Chest CT,bronchoscopy brush or biopsy histopathological examination were followed up,cultivation of My.tuberculosis and of common bacteria with sputum or lavage fluid when needed.T-SPOT.TB test was performed according to the kit instruction operation.2.5 × 105 peripheral blood mononuclear cells (PBMCs) were added into the pre-coated anti-human γ-interferon antibody,and co-incubated separately with two specific My.tuberculosis antigens,namely early secretory targeting 6 (ESAT-6) and culture filtration protein 10 (CFP-10),and then the spot forming cells (SFCs) were counted.The gold standard for present study were set as follows:1) My.tuberculosis smear positive or culture positive;2) Clinical diagnosis (meet any one is positive).The efficacy of T-SPOT.TB on diagnosing active TB was observed,and then the optimal critical value for diagnosing active TB was determined.Patients diagnosed as active TB were divided into 4 subgroups:initial treatment group,retreatment group,smear or culture positive group,and smear or culture negative group.T-SPOT.TB was carried out to detect A and B antigen,and the difference of formed SFCs was then compared.The present study was approved by the Ethics Committee of Xinjiang Uygur Autonomous Region Chest Hospital.Results Of 700 cases suspected of pulmonary tuberculosis enrolled in present study,528 out of 624 definite cases (84.6％) were finally diagnosed as active tuberculosis (active TB group) and 96 cases (15.4％) were as without TB infection (non-TB group).Positive results of T-SPOT.TB test were found in 414 cases in active TB group,and 47 cases in non-TB group were reported with T-SPOT.TB negative.The sensitivity and specificity of T-SPOT.TB test for diagnosing active TB were 78.4％ and 49％,respectively.The positive predictive value,negative predictive value,positive likelihood ratio and negative likelihood ratio were 89.4％,29.2％,1.537 and 0.441,respectively.ROC curve showed that the specificity increased significantly (from 49％ to 62.5％) while the sensitivity decreased (from 78.4％ to 72.7％) when antigen A (cut-off:16.0 SFCs/2.5 × 105 PBMC) was combined with antigen B (cut-off:7.0 SFCs/2.5 × 105 PBMC) for analysis.In addition,the number of A and B antigen spots in active TB group was significantly higher than that in non-TB group (P＜0.01).The number of B antigen spots in positive TB group was significantly higher than that in negative TB group (P＜0.05).There was no significant difference among the other groups.Conclusions Since the high sensitivity and low specificity of T-SPOT.TB in the diagnosis of active TB,the final diagnosis should be combined with clinical manifestations.When the A antigen is 16.0 SFCs/2.5 × 105 PBMC and the B antigen is 7.0 SFCs/2.5 × 105 PBMC,the specificity of T-SPOT.TB will be improved.Higher number of spots has a certain reference diagnostic value for active TB.

Comparison of the Commercial QuantiFERON-CMV and Overlapping Peptide-based ELISPOT Assays for Predicting CMV Infection in Kidney Transplant Recipients

Cytomegalovirus (CMV) is one of the most important opportunistic infections in transplant recipients. Tests for CMV-specific T cell responses have been proposed to change the current risk stratification strategy using CMV assays. We evaluated the usefulness of pre-transplant CMV-specific T cell assays in kidney transplant (KT) candidates for predicting the development of CMV infection after transplantation comparing the results of the overlapping peptides (OLPs)-based enzyme-linked immunospot (ELISPOT) assay and the commercial QuantiFERON-CMV assay. We prospectively enrolled all cases of KT over a 5-month period, except donor CMV-seropositive and recipient seronegative transplants that are at highest risk of CMV infection. All the patients underwent QuantiFERON-CMV, CMV OLPs-based pp65, and immediate-early 1 (IE-1)-specific ELISPOT assays before transplantation. The primary outcome was the incidence of CMV infection at 6 months after transplant. The total of 47 KT recipients consisted of 45 living-donor KTs and 2 deceased-donor KTs. There was no association between positive QuantiFERON-CMV results and CMV infection. However, 10 of 34 patients with phosphoprotein 65 (pp65)- or IE-1-specific ELISPOT results higher than cut-off value developed CMV infections compared with none of 13 patients with results lower than cut-off value developed CMV. The OLPs-based ELISPOT assays are more useful than the QuantiFERON-CMV assay for predicting CMV infection. Patients with higher CMV-specific T cell immunity at baseline appear to be more likely to develop CMV infections after KT, suggesting that the abrupt decline in CMV-specific T cell responses after immunosuppression, or high CMV-specific T cell responses due to frequent CMV activation before KT, may promote CMV infection.

Clinical applications of interferon-γ releasing assays for cytomegalovirus to differentiate cytomegalovirus disease from bystander activation: a pilot proof-of-concept study

BACKGROUND/AIMS: We evaluated the proposed clinical application of the combined interpretation of host factors and viral factors in two different cytomegalovirus (CMV) co-infection models. METHODS: We prospectively enrolled all human immunodeficiency virus non-infected patients with confirmed Pneumocystitis jirovecii pneumonia (PCP) and those with suspected gastrointestinal CMV disease in a tertiary hospital. All patients underwent CMV interferon-γ releasing assay (IGRA) for CMV (T-track CMV, Lophius Biosciences). We created the 2-axis model with the CMV IGRA results as the x-axis and the results for CMV virus replication as the y-axis, and hypothesized that cases falling in the left upper quadrant (high viral load and low CMV-specific immunity) of the model would be true CMV infections. The CMV IGRA results were concealed from the attending physicians. RESULTS: Of 39 patients with PCP, four (10%) were classified as combined CMV pneumonia, 13 (33%) as bystander activation, and the remaining 22 (56%) as no CMV infection. The data for all four patients with PCP and CMV pneumonia fell in the left upper quadrant of the 2-axis model. Of 24 patients with suspected gastrointestinal CMV disease, 12 (50%) were classified as gastrointestinal CMV disease and the remaining 12 (50%) as bystander activation with no gastrointestinal CMV disease. The data for 11 of the 12 patients (92%) with gastrointestinal CMV disease were located in the left upper quadrant of the 2-axis model. CONCLUSIONS: Cases yielding low CMV IGRA results and high CMV viral replication appear to be true CMV infections. Further studies with large number of cases in different types of CMV disease should be proposed.

Comparison of effects of ELISPOTs using pleural fluid and peripheral blood in the diagnosis of tuberculous pleurisy / 天津医药

Objective To evaluate the effects of ELISPOT (enzyme-link immunospot) test using different samples in diagnosing tuberculous pleurisy. Methods Using T-Spot-TB kit to detect interferon-γlevel in pleural effusion and periph?eral blood from 164 patients with tuberculous pleural effusion and 102 patients without tuberculous pleural effusion. Number of spot forming cells (SFCs) as well as the specificity and sensitivity of the tests were compared between these two methods (ELISPOT using leural effusion or peripheral blood). Results The area under the ROC curve was 0.947 in pleural effusion Elispot test while it was 0.905 in peripheral blood Elispot test. The sensitivity of pleural effusion ELISPOT test in diagnosis of tuberculous pleurisy (95.1%) was significantly higher than that of peripheral blood ELISPOT test (89.0%). What’s more, the specificity of pleural effusion ELISPOT test in diagnosis of tuberculous pleurisy (90.2%) was higher than that in diagno?sis of peripheral blood ELISPOT test (88.2%). Conclusion The pleural effusion ELISPOT test is more valuable than periph?eral blood ELISPOT in the diagnosis of tuberculous pleuritis.

Diagnostic Usefulness of Cytomegalovirus (CMV)-Specific T Cell Immunity in Predicting CMV Infection after Kidney Transplantation: A Pilot Proof-of-Concept Study

BACKGROUND: Cytomegalovirus (CMV) is one of the most important opportunistic infections in transplant recipients. Currently sero-positivity for CMV IgG before solid organ transplantation is the laboratory test of choice for stratifying the risk of CMV reactivation after solid organ transplantation. Theoretically, CMV-specific cell-mediated immune responses before solid organ transplantation should further categorize patients as high or low risk of CMV development. We therefore evaluated the usefulness of the CMV-specific enzyme-linked immunospot (ELISPOT) assay in kidney transplant (KT) candidates for predicting the development of CMV infections after transplantation. MATERIALS AND METHODS: All adult CMV IgG (+) recipients admitted to the KT institute between March 2014 and June 2014 were enrolled, and CMV infections after KT were observed between March 2014 and December 2014. All patients underwent CMV pp65 and IE1-specific ELISPOT assays before transplantation. CMV infection was defined in the presence of CMV antigenemia, CMV syndrome, or tissue-invasive CMV disease. We used the data to select optimal cut-off values for pp65 and IE1, respectively, on ROC curves. RESULTS: A total of 69 transplant recipients involving 54 (78%) living-donor KT, 9 (13%) deceased-donor KT, 3 (4%) kidney-pancreas transplants, and 3 (4%) pancreas transplants were enrolled. Of the 69 patients, 27 (39%) developed CMV infections. There was no association between the IE1-specific ELISPOT assay and CMV infection. However, only 15 (31%) of the 48 patients with positive pp65-specific ELISPOT results (>10 spots/2.0 x 105 cells) developed CMV infections, whereas 12 (57%) of the 21 patients with negative pp65-specific ELISPOT results developed CMV infection (P = 0.04). CONCLUSION: Negative pp65-specific ELISPOT assay results before transplantation appear to predict the subsequent development of CMV infections after transplantation in CMV IgG (+) KT recipients. Therefore, risk stratification of CMV IgG (+) recipients using the CMV-specific ELISPOT, together with preventive strategies, may further reduce CMV development.

Interferon-gamma Enzyme-Linked Immunospot Assay in Patients with Tuberculosis and Healthy Adults / 결핵및호흡기질환

BACKGROUND: Interferon-gamma assays based on tuberculosis (TB)-specific antigens have been utilized for diagnosing and ruling out latent TB and active TB, but their utility is still limited for TB incidence countries. The aim of this study is to understand the clinical utility of enzyme-linked immunospot (ELISpot) assays among patients with clinically suspected TB and healthy adults in clinical practices and community-based settings. METHODS: The ELISpot assays (T SPOT.TB, Oxford Immunotec, UK) were prospectively performed in 202 patients. After excluding those with indeterminate results, 196 were included for analysis: 41 were TB patients, 93 were non-TB patients, and 62 were healthy adults. RESULTS: The sensitivity and negative predictive values of the T SPOT.TB assays for the diagnosis of TB were 87.8% and 89.1%, respectively, among patients with suspected TB. The agreement between the tuberculin skin test (10-mm cutoff) and the T SPOT.TB assay was 66.1% (kappa=0.335) in all participants and 80.0% (kappa=0.412) in TB patients. Among those without TB (n=155), a past history of TB and fibrotic TB scar on chest X-rays were significant factors that yielded positive T SPOT.TB results. There was a significant difference in the magnitude of T SPOT.TB spot counts between TB patients and non-TB patients or healthy adults. CONCLUSION: The T SPOT.TB assay appeared to be a useful test for the diagnostic exclusion of TB. A positive result, however, should be cautiously interpreted for potential positives among those without active TB in intermediate TB incidence areas.

Clinical application of T cell enzyme-linked immunospot assay in extrapulmonary on rapidly diagnosis of patients with tuberculosis / 中国综合临床

Objective To investigate the diagnostic value of T cell enzyme-linked immunospot (T-SPOT.TB) assay on extrapulmonary tuberculosis patients.Methods Thirty patients suffered from extrapulmonary mycobacterium tuberculosis(MTB) infection and 30 with non-MTB infection were recruited this study.T-SPOT.TB assay was used to detect early secreting antigen target-6 (ESAT-6) and culture filtrate protein-10(CFP-10) specific T cells in blood samples.PPD skin test was also used.Results (1)The positive rate of MTB detected by T-SPOT.TB assay was 91.89％ (34/37),higher than that of un-tuberculosis group (6.67 ％ (2/30)),and the difference was significant (x2 =48.403,P ＜ 0.001).(2) The sensitivity,specificity,positive prospective value and negative prospective value of T-SPOT.TB assay were 91.89％,93.33％,94.44％ and 90.32％ respectively,better than those of PPD skin test (67.57％,56.67％,65.79％,58.62％),and the differences were markedly (x2 =6.773,10.756,9.392,8.031 respectively ; P =0.009,0.001,0.002,0.005 respectively).Meanwhile T-SPOT.TB assay has low agreement with means of PPD skin test(Kappa =0.311,x2 =6.801,P =0.009).Conclusion T-SPOT.TB assay has a higher sensitivity and specificity in the rapid diagnosis of extrapulmonary tuberculosis.Therefore,it is with great value and applicability as a screening test.

Evaluation of an enzyme-linked immunospot assay for the immunodiagnosis of atypical spinal tuberculosis (atypical clinical presentation/atypical radiographic presentation) in China

BACKGROUND: Atypical spinal tuberculosis (TB) usually presents in a slowly indolent manner with nonspecific clinical presentations making the diagnosis a great challenge for physicians. New technologies for the detection of atypical spinal TB are urgently needed. The aim of this study was to assess the diagnostic value of an enzyme-linked immunospot (ELISPOT) assay in clinically suspected cases of atypical spinal TB in China. METHODS: From March 2011 to September 2012, a total of 65 patients with suspected atypical spinal TB were enrolled. In addition to conventional tests for TB, we used ELISPOT assays to measure the IFN-I response to ESAT-γ and CFP-10 in T-cells in samples of peripheral blood mononuclear cells. Patients with suspected atypical spinal TB were classified by diagnostic category. Data on clinical characteristics of the patients and conventional laboratory results were collected. RESULTS: Out of 65 patients, 4 were excluded from the study. 18 (29.5%) subjects had cultureconfirmed TB, 11 (18.0%) subjects had probable TB, and the remaining 32 (52.5%) subjects did not have TB. Generally, the features of atypical spinal TB include the following aspects: (1) worm-eaten destruction of vertebral endplate; (2) destruction of centricity of the vertebral body or concentric collapse of vertebral body; (3) tuberculous abscess with no identifiable osseous lesion; (4) contiguous or skipped vertebral body destruction. 26 patients with atypical spinal TB had available biopsy or surgical specimens for histopathologic examination and 23 (88.5%) specimens had pathologic features consistent with TB infection. The sensitivities of the PPD skin test and ELISPOT assay for atypical spinal TB were 58.6% and 82.8%, and their specificities were 59.4% and 81.3%, respectively. Malnutrition and age were associated with ELISPOT positivity in atypical spinal TB patients. CONCLUSIONS: The ELISPOT assay is a useful adjunct to current tests for diagnosis of atypical spinal TB.

Immune response of human immunodeficiency virus-1 specific T lymphocytes in human immunodeficiency virus-1 monoinfected or human immunodeficiency virus-1/hepatitis C virus coinfected individuals / 中华传染病杂志

Objective To investigate the features of immune response of human immunodeficiency virus type-1 (HIV-1) antigen specific T lymphocytes in HIV-1 monoinfected or HIV 1/hepatitis C virus (HCV) coinfected individuals.Methods Twenty-six HIV-1 monoinfected and 23 HIV-1/HCV coinfected individuals were enrolled.Immunomagnetic microbeads were used to isolate T lymphocyte subpopulation CD4+ T cells and CD8+ T cells from human peripheral blood mononuclear cells (PBMC).Frequencies of interferon-γ (IFN-γ) secreting cells of CD4+,CD8+ T lymphocytes and PBMC stimulated by a peptide pool containing 12 overlapping peptides in HIV-1 P24 from 49 patients were assessed by enzyme-linked immunospot (ELISPOT) assay.HIV-1 RNA levels of these patients were also detected by real-time fluorescence quantitative polymerase chain reaction.The data were compared by one-way ANOVA and Mann-Whitney U test,and Spearman test was used for correlation analysis.Results Frequencies of HIV-1 antigen specific CD4+ T lymphocytes [median =25 spot-forming cells (SFC)/106 cells] were significantly lower than those of CD8+T lymphocytes (median=38SFC/106 cells,F=4.592,P=0.037) and PBMC (median=53 SFC/106 cells,F=5.436,P=0.025) in HIV-1 monoinfected group.Frequencies of HIV-1 antigen specific CD4+ T lymphocytes (median=5 SFC/106 cells,Z=-2.432,P=0.015),CD8+T lymphocytes (median=5 SFC/106 cells,Z=-1.996,P=0.046) and PBMC (median=10 SFC/106 cells,Z=-2.306,P=0.021) in HIV-1/HCV coinfected group were significantly lower than those in HIV-1 monoinfected group.Conclusions In HIV-1 infection,antigen specific immune response of CD4+ T cells can be activated,but weaker than that of CD8+ T cells.Co-infection with HCV might down-regulate the responses of HIV-1 antigen specific T lymphocytes in HIV-1 infected individuals.

A Case of Sulfasalazine-Induced Hypersensitivity Syndrome Confirmed by Enzyme-Linked Immunospot Assay

A 24-year-old male with a history of spondyloarthropathy presented with high fever, cervical lymphadenopathy and generalized maculopapular rash. He was treated with prednisolone for chronic uveitis before being switched to sulfasalazine 3 weeks prior to admission. Laboratory findings revealed marked leukocytosis with frequent atypical lymphocytes. Sulfasalazine was discontinued and the etiology of mononucleosis syndrome explored. During admission, he developed acalculous cholecystitis and hypotension. All symptoms quickly improved following administration of systemic corticosteroids. The investigation for infectious mononucleosis yielded negative results and a diagnosis of sulfasalazine-induced hypersensitivity syndrome was confirmed using enzyme-linked immunospot assays.

A Case of Sulfasalazine-Induced Hypersensitivity Syndrome Confirmed by Enzyme-Linked Immunospot Assay

A 24-year-old male with a history of spondyloarthropathy presented with high fever, cervical lymphadenopathy and generalized maculopapular rash. He was treated with prednisolone for chronic uveitis before being switched to sulfasalazine 3 weeks prior to admission. Laboratory findings revealed marked leukocytosis with frequent atypical lymphocytes. Sulfasalazine was discontinued and the etiology of mononucleosis syndrome explored. During admission, he developed acalculous cholecystitis and hypotension. All symptoms quickly improved following administration of systemic corticosteroids. The investigation for infectious mononucleosis yielded negative results and a diagnosis of sulfasalazine-induced hypersensitivity syndrome was confirmed using enzyme-linked immunospot assays.

T-SPOT.TB assay in diagnosis and efficacy assessment of pulmonary tuberculosis / 中华临床感染病杂志

Objective To evaluate the application of interferon-γ release assay T-SPOT.TB in diagnosis and efficacy assessment of pulmonary tuberculosis. Methods T-SPOT.TB assay was used to determine spot-forming cells (SFCs) formed by T-cells when stimulated by Mycobacterium tuberculosisspecific antigens in 55 patients with active tuberculosis,14 patients with non-tuberculosis lung diseases and 12 healthy controls. Meanwhile 20 sputum culture-positive and qualitative assay-positive pulmonary tuberculosis patients were tested with T-SPOT.TB before and at 2-month and 6-month after treatment.Kruskal-Wallis H and Mann-Whitney U test were used in group comparison.Wilcoxon test was used in comparison between pre- and post-treatment.Results The positive rate of T-SPOT.TB was significantly higher in patients with tuberculosis (85.5％,47/55 ) than that in patients with non-tuberculosis lung diseases (2/14) and the healthy controls (1/12) (x2 =40.926,P ＜0.05).The SFCs of hole A in response to ESAT-6 and hole B in response to CFP-10 in pulmonary tuberculosis group were 70.00 (27.00 -125.00) and 80.00 ( 17.00 - 180.00),respectively,which were all significantly higher than those in nontuberculosis lung diseases group and the healthy controls (x2 =35.376 and 30.485,P ＜ 0.05 ).The sensitivity,specificity,positive predictive value and negative predictive value of T-SPOT.TB in diagnosis of smear-positive tuberculosis were 88.6％,88.5％,91.2％ and 85％,while in diagnosis of sputum smearnegative tuberculosis,the sensitivity was 80％,specificity was 88.5％,positive predictive value was 84.2％ and negative predictive value was 85.2％ ( P ＞ 0.05 ).SFCs of hole A and hole B in 20 patients with sputum culture-positive and qualitative assay-positive pulmonary tuberculosis were 75.50 (41.25 -116.25 ) and 56.25 ( 105.00 -225.00) before the treatment.After 2-month antituberculosis treatment,the SFCsofhole A and hole B were 41.0 (18.0-68.75) and 72.50 (42.25- 158.75),which were significantly lower than those before treatment (Z =- 3.213 and - 3.622,P ＜ 0.05 ).Ater 6-month antituberculosis treatment,the SFCs of hole A and hole B were 25.00 (5.75 - 52.25) and 55.00 (6.25 -122.50),which were significantly lower than those before and 2-month after antituberculosis treatment (vs.before treatment:Z =- 3.921 and - 3.923,P ＜ 0.05 ; vs.2-month antituberculosis treatment:Z =- 3.926 and - 3.884,P ＜ 0.05 ).Conclusions T-SPOT.TB assay possess satisfactory sensitivity and specificity in diagnosis of tuberculosis infection,especially for sputum-negative pulmonary tuberculosis.It is also of value in monitoring antituberculosis treatment.

The relationship between CD4+ T lymphocyte count and Mycobacterium tuberculosis coinfection in human immunodeficiency virus-infected patients / 中华传染病杂志

Objective To evaluate the relationship between CD4+ T lymphocyte count and results of enzyme-linked immunospot (ELISPOT) assay in human immunodeficiency virus (HIV)-Mycobacterium tuberculosis (M.tb) coinfected patients.Methods A total of 193 HIV-infected individuals in Yunnan Province and Shanghai were enrolled.T-SPOT.TB assay was employed to detect M.tb specific T lymphocyte in the peripheral blood mononuclear cells (PBMC).CD4+ T lymphocyte in PBMC from the enrolled subjects was detected by flow cytometry.Data were analyzed using t test.ResultsThe incidence of latent tuberculosis in HIV-infected individuals was 30.6％.The CD4+ T lymphocyte counts in HIV-infected individuals with active tuberculosis were 190×106/L,which were significantly lower than those in HIV-infected individuals with latent tuberculosis (484×106/L; t=6.665,P＜0.01).The HIV-infected individuals were stratified according to CD4+ T lymphocyte counts of ＞500×106/L,200×106-500×106/L,and ＜200×106/L and the constituent ratios of active tuberculosis/latent tuberculosis were 1∶16.2,1∶1.3 and 5.6∶1,respectively.Among 79 subjects with positive T-SPOT.TB results,20 were coinfected with active tuberculosis,in which 14 had CD4+ T lymphocyte counts of ＜200 ×106/L,5 had 200×105-500×106/L and 1 had ＞500×106/L.Fifty-two in 59 HIV/latent tuberculosis patients individuals had CD4+ T lymphocyte counts of ＞200×106/L.ConclusionsThe prevalence of latent tuberculosis in HIV-infected individuals is high in China.Cellular immunity in HIV-infected individuals with active tuberculosis is severely impaired.With the decrease of CD4 ′ T lymphocyte counts,patients with latent tuberculosis are prone to develop active tuberculosis in HIV-infected individuals.The negative predictive value of T-SPOT.TB is significantly diminished in patient with low CD4+ T lymphocyte counts,especially less than 200×106/L.

Expression of Tim-3 in peripheral CD8 + T cells and its significance in patients with chronic hepatitis B / 中华临床感染病杂志

Objective To investigate the expression of T cell immunoglobulin-and mucin-domaincontaining molecule-3 (Tim-3) in peripheral CD8 +T cells and its significance in patients with chronic hepatitis B (CHB).Methods Fifty-eight CHB patients and 16 healthy controls were enrolled.Tim-3 expression in CDs + T cells was detected by flow cytometry,and quantities of IFNγ-producing HBV-specific cytotoxic T lymphocytes (CTLs) in HLA-A2 positive subjects were detected by enzyme-linked immunosorbent spot (ELISPOT) test before and after the blockade of Tim-3/Tim-3L pathway.Paired t test was performed to compare the quantities of CTLs before and after the blockade,and nonparametric Spearman correlation analysis was performed to explore the correlation in quantitive data.Results Tim-3 expression in CHB patients was (14.2 ± 8.98 )％,which was higher than that of healthy controls (4.80 ± 2.92)％,and the difference was of statistical significance (x2 =92.48,P ＜ 0.05 ) Tim-3 expressions in 16 severe CHB patients and 42 mild CHB patients were ( 19.54 ± 10.95) ％ and (9.58 ± 7.30) ％,respectively,and the difference was statistically significant (x2 =77.24,P ＜ 0.05 ). Before the blockade of Tim-3/Tim-3L pathway,IFNγ-producing HBV-specific CTLs were 7.27 ± 3.14,and it increased to 19.62 ± 4.97 after the blockade ( t =2.95,P ＜ 0.05 ).Conclusion The upregulation of Tim-3 on peripheral CD8 + T cells may inhibit HBV-specific CTLs,and the blockade of Tim-3 pathway can enhance the proliferation of IFNγ-producing HBV-specific CTLs,thus can enhance antiviral effect.

Prevalence of latent tuberculosis infection and its risk factors among senior students from four primary schools in Shanghai / 中华传染病杂志

Objective To investigate the prevalence of latent tuberculosis infection(LTBI),and to identify the risk factors in primary schoolchildren from Shanghai through the population-based field investigation combined with the tuberculosis infection enzyme-linked immunospot assay(T-SPOT.TB)assay.Methods The children in grade 4 and 5 were enrolled from four primary schools in Pudong new district and Yangpu district of Shanghai.Questionnaire interview was applied to investigate the soeiodemographic and clinical information related to LTBI.The T-SPOT.TB assay was used to detect LTBI in the enrolled subjects.Univaitate and multivariate analyses were used to identify the risk factors associated with LTBI among the primary schoolchildren.Results Totally 472 schoolchildren were enrolled in the present study,with 439(93.0%)being vaccinated with bacillus calmette-guerin (BCG) and ten (2.1%) having contact history with tuberculosis (TB) patients.Among the 472 eligible subjects,16(3.4%) children were T-SPOT.TB positive,who had no clinical symptoms andsigns relevant to TB and were defined as LTBI.The LTBI prevalence in BCG vaccinated and unvaccinated children were 2.7% and 12.1%,respectively (OR:6.972;95%CI:1.834-26.500);those in TB contacts and children without TB contact history were 30.0% and 2.8%, respectively (OR: 16. 38; 95% CI: 3. 692-72. 700). Conclusions The prevalence of LTBI among senior schoolchildren in Shanghai is 3.4%. BCG vaccination is protective for children from LTBI, while daily contacts with TB patients increases the risk of LTBI in schoolchildren.

The clinical significance of immune-related marker detection in idiopathic thrombocytopenic purpura / 中华内科杂志

Objective To assess the clinical significance of detecting the immune markers in idiopathic thrombocytopenic purpura (ITP). Methods The frequencies of circulating B cells secreting platelet-specific antibody, platelet-specific antibody, the percentage of T lymphocyte subsets, the percentage of reticulated platelet and the level of thrombopoietin in 64 ITP patients and 31 healthy controls were measured with enzyme-linked immunospot assay (ELISPOT),modified monoclonal antibody immunobilization of platelet antigens assay (MAIPA), flow cytometry and sandwich enzyme-linked innnunosorbent assay respectively. Results Compared with the controls[1.3 ± 0. 5/105 peripheral blood mononuclear cell (PBMC), (0.33±0.06,0.41±0.03), (22.08±4.54)% and (8.19±2.46)%], the frequencies of circulating B cells secreting platelet-specific antibody (7.6±4.6/105 PBMC in acute ITP group, 5.3±3.0/105 PBMC in chronic ITP group), platelet-specific antibody (including the anti-GP Ⅱ b/Ⅲa antibody, anti-GP Ⅰ b/X antibody) (0.51 ±0.11, 0.48±0.06 in acute ITP group; 0.49±0.10,0.46±0.09 in chronic ITP group), the percentage of CD8+ T Lymphocyte (27.09±9.86 ) %, the percentage of reticulated platelet in ITP patients[the megakaryocyte cytosis group (24. 85 ± 19. 18)%, the normal megakaryocyte group (23.89±18.90)%]were significantly increased ( all P＜0.05).The frequencies of circulating B cells secreting platelet-specific antibody in acute ITP patients were notably increased (P＜0.05) compared to the chronic ITP patients. In T lymphocyte subsets, the percentage of CD3+T lymphocyte and CD4+ T lymphocyte and the ratio of CD4+/CD8+ in the patients with ITP[(60.88±14.59)%, (28.41±10.55)%, 1.18±0.59]were notably decreased than those in the healthy controls [(69.89±6.43)%, (35.38±5.05) %, 1.64±0.29, P＜0.05]. There was no apparent difference of the level of thrombopoietin between ITP patients with megakaryocyte cytosis (72. 09 ± 41.64 ) and health controls (75.37± 26. 32, P ＞ 0. 05 ), however, the level of thrombopoietin of ITP patients with normal megakaryocyte apparently increased (118.60±70.72, P＜0.05). Conclusion Detecting the frequencies of circulating B cells secreting platelet-specific antibody, platelet-specific antibody, the percentage of T lymphocyte subsets, the percentage of reticulated platelet and the level of thrombepoietin in the patients with ITP may improve the diagnosis and guide clinical therapy.