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.

Significance of T-cell dot test and combined detection of adenosine deaminase in tuberculous pleurisy at different ages / 中国综合临床

Objective To analyze the clinical value of combined detection of tuberculous T cell enzyme-linked immuno spot assay (T-SPOT.TB) and adenosine deaminase (ADA) in tuberculous pleurisy patients of different ages.Methods From February 2014 to February 2018,three hundred and thirty-six patients with pleural effusion were admitted to Hebei Thoracic Hospital.Among them,two hundred and fifty five cases were diagnosed as tuberculous pleurisy and 81 cases were diagnosed as non-tuberculous pleurisy.The patients were divided into two groups according to their age.The younger group (214 cases) was 16-59 years old and the older group (122 cases) was over 60 years old.The sensitivity and specificity of T-SPOT.TB combined with ADA in the diagnosis of tuberculous pleurisy were compared between the two groups.Results The sensitivity and specificity of T-SPOT.TB were 85.5％ (153/179) and 71.4％ (25/35) in the young and middle-aged group,73.7％ (56/76) and 58.7％ (27/46) in the old group,respectively.The sensitivity of the young and middle-aged group was significantly higher than that of the old group (x2 =4.990,P =0.045).The sensitivity and specificity of T-SPOT.TB combined with ADA were 98.9％ (177/179) and 94.3％ (33/35) in the young and middle-aged group,96.1％ (73/76) and 89.1％ (41/46) in the elderly group,respectively.There was no significant difference in sensitivity and specificity between the two groups (x2 =0.256,P=0.393、x2=0.655,P=0.218).Conclusion The diagnostic efficacy of T-SPOT.TB combined with ADA in patients with tuberculous pleurisy at different ages has been improved,especially for those who can not tolerate pleural biopsy and elderly patients.

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.

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

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 / 감염과화학요법

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

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.

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 effects between IL-2 and IL-7 on glutamic acid decarboxylase 65 reactive T cell responses in patients with Type Ⅰ diabetes / 中南大学学报(医学版)

Objective:To explore the type of cytokine (IL-2 or IL-7) and its most optimal concentration regarding the improvement of the signal-to-noise ratio of glutamic acid decarboxylase 65 (GAD65) in enzyme-linked immunospot (ELISPOT) assay in Type 1 diabetic (T1DM) patients.Methods:Twenty T1DM patients (Group A) and sixteen healthy controls matched with age and sex (Group B) were enrolled in our study,and their peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll method.GAD65,internal control and Pediacel served as "five-for-one" vaccine were selected as the stimulating antigen.Different concentrations of IL-2 [0 U/mL (Group 1),0.5 U/mL (Group 2),2.5 U/mL (Group 3) and 12.5 U/mL (Group 4)] were added to the culture system.The CD4+ T cells of secreting interferon-gamma (IFN-γ) in the above groups were determined by ELISPOT.The spots number,net values and stimulating index (SI) were compared in GAD65 (signal) and internal control (background).Next,another 21 T1DM patients (Group C) and 12 healthy controls matched with age and sex (Group D) were enrolled,and the specific T cell response to the GAD65 antigen was detected.The net values and SI were compared between the best optimal concentration of IL-2 (2.5 U/mL,Group 5) and IL-7 (0.5 ng/mL,Group 6).Results:1) After adding IL-2 into the Group A,the amount of GAD65 reactive T cells in different groups increased compared with Group A1,while the background in the internal control also increased gradually with the increased concentration of IL-2.There was no significant difference in net value (signal-noise) in the different concentration between the Group A3 and the Group A4 (P＞0.05).The SI in the Group A3 (2.8),the highest one,was significantly higher than that in the Group B3 (1.3) (P＜0.05).2) Although the number of GAD65 spots in the Group C6 and the Group D6 were slightly higher than that in the Group C5 and the Group D5,respectively,the background in the Group C6 and the Group D6 also increased,without statistical significance (P＞0.05).The mean net value spot and SI in the Group C5 (net value:5.5;SI:2.8) were both significantly higher than those in the Group C6 (net value:4.3;SI:1.8) (bothP9＜0.05).Conclusion:The concentration of 2.5 U/mL for IL-2 is proved to be the best optimal concentration for GAD65 specific T-cell responses in ELISPOT in patients with T1DM.IL-2 is much better than IL-7 in improvement of the SI in the ELISPOT.

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

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.

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.

Percentage and functions of natural killer T cells in peripheral blood and bone marrow of children with severe aplas-tic anemia / 临床儿科杂志

Objective To analyze the percentage and functional changes of natural killer T (NKT) cells in peripheral blood and bone marrow of severe aplastic anemia (SAA) children before immunosuppressive therapy (IST) comparing to that of healthy children.Methods Ten children with severe aplastic anemia were included in the study and ten healthy children at the same age were selected as the control group. By lfow cytometry, the percentage of CD3+CD1d tetramer+ NKT cell in peripheral blood and bone marrow were detected from March 2014 to December 2014 in our hospital. Immune magnetic bead separation was used to isolate and purify iNKT cells .The puriifed iNKT cells were cultured in the OCH(50 ng/ml,100 ng/ml or 200 ng/ml)+rhIL-2+rhG-CSF culture systems. The ampliifcation of iNKT cells after cultured in different systems were calculated. Elispot method was used to analyze the spotting form cells (SFCs) of IFN-γ or IL-4 expressed by activated iNKT cells.Results The percentage of CD3+CD1d tetramer+ NKT cells in peripheral blood of SAA group(0.72±0.03)% was signiifcantly lower than that of the control group(0.92±0.02)%(P=0.000). The percentage of CD3+CD1d tetramer+ NKT cells in bone marrow of SAA group(0.82±0.02)% was signiifcantly lower than that of the control group(1.05±0.05)%(P=0.000).In vitro iNKT cell ampliif-cation ability of bone marrow in SAA group was signiifcantly lower than the control group, and in medium concentration(50±6) and high concentration OCH group(52±6), the ampliifcation ability was higher than that in low concentration OCH group(30±5) (P<0.05). The secretion of IFN-γ in the iNKT cells of SAA bone marrow was signiifcantly lower in medium concentration(33±3) and high concentration(35±3)OCH group than that of the low concentration(50±3)OCH group(P<0.01). The secretion of IL-4 in the iNKT cells of SAA bone marrow was signiifcantly higher in medium concentration(50±3)and high concentration(75±3) OCH group than that of the low concentration(33±3) OCH group(P<0.01).Conclusions The quantity and function of NKT cells from children with SAA are lower than that of the healthy children.In vitro, they had better ampliifcation ability and could improve IL-4/IFN-γ imbalance in medium concentration and high concentration OCH group than in low concentration OCH group.

Potential Role of Immunodiagnosis for Pulmonary Tuberculosis Using Induced Sputum Cells

PURPOSE: To evaluate the diagnostic utility and predictors for determinate results of an enzyme-linked immunospot assay using induced sputum cells (IS ELISPOT) for a rapid diagnosis of pulmonary tuberculosis (TB). MATERIALS AND METHODS: Subjects suspected of pulmonary TB who had either sputum acid fast bacilli smear-negative or not producing sputum spontaneously were prospectively enrolled. ELISPOT assay was performed using cells from induced sputum. RESULTS: A total of 43 subjects, including 25 with TB (TB group) and 18 with non-TB disease (non-TB group) were enrolled. Results of IS ELISPOT were determinate in only 17/43 (39%) subjects, but all of determinate results were consistent with the final diagnosis. Of the 43 sputum samples, 11 (26%) were inadequate to perform IS ELISPOT. Of 32 adequate sputum samples, the proportion of determinate results was significantly higher in the TB group (75%, 15/20) than in the non-TB group (17%, 2/12) (p=0.002). The status of active TB was a unique predictor but smear positivity was not a significant predictor for determinate results. In addition, sensitivity of IS ELISPOT (75%, 9/12) in smear negative TB was higher than that of TB-polymerase chain reaction (25%, 3/12). CONCLUSION: IS ELISPOT showed relatively high diagnostic value and accuracy in the TB group, independent of smear positivity. IS ELISPOT may provide additional diagnostic yield for microbiological tools in the rapid diagnosis of smear-negative TB.

The diagnostic value of enzyme-linked immunospot assay in AIDS patients with Mycobacterium tuberculosis infection / 国际检验医学杂志

Objective To explore the value of enzyme‐linked immunospot(ELISPOT ) assay in the diagnosis of acquired immu‐nodeficiency syndrome(AIDS) patients with Mycobacterium tuberculosis infection .Methods 42 hospitalized AIDS patients with Mycobacterium tuberculosis infection and 60 non‐tuberculosis hospitalized patients with diseases of respiratory system in the same period were recruited as observation group and control group ,respectively .ELISPOT assay ,tuberculosis antibody(TBAb) test and tuberculin skin test(TST) were performed for all the patients ,respectively .The sensitivity ,specificity and accuracy of the 3 meth‐ods were comparatively analyzed .Results The sensitivity ,specificity and accuracy of ELISPOT were 92 .9% ,93 .3% and 93 .1% . These of TBAb were 54 .8% ,71 .7% and 64 .7% ,and of TST were 47 .6% ,53 .3% and 51 .0% ,respectively .There were significant differences among three methods in sensitivity ,specificity and accuracy(P0 .05) .Conclusion ELISPOT assay has relatively high diagnostic value in AIDS patients with My‐cobacterium tuberculosis infection .

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.

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.

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.