Introduction: The objective of this study was the comparison of the results of T-SPOT.TB using pleural effusion (PE) with those of IGRAs using peripheral blood (PB) or other diagnostic methods for the diagnosis of tub...Introduction: The objective of this study was the comparison of the results of T-SPOT.TB using pleural effusion (PE) with those of IGRAs using peripheral blood (PB) or other diagnostic methods for the diagnosis of tuberculous (TB) pleurisy. Methods: We measured adenosine deaminase (ADA) in PE, QuantiFERON TB-Gold In-Tube (QFT), and T-SPOT.TB using PB, and T-SPOT.TB using PE. The definite group of TB pleurisy included 12 patients and other disease group 33 patients. Main find-ings: Sensitivity for QFT using PB was 83% and specificity was 85%, sensitivity for T-SPOT.TB using PB was 92% and specificity was 82%, while sensitivity for ADA in PE was 83% and specificity was 76%. When we adopted the same cut-off level of a positive response for T-SPOT.TB as PB using PE, sensitivity for T-SPOT.TB using PE was 100% and specificity was 82%, respectively. Although there were no significant differences among the four diagnostic methods, sensitivity for T-SPOT.TB using PE gave the most accurate diagnosis of TB-definite patients compared to ADA in PE or QFT using PB. Conclusions: If we performed T-SPOT.TB using a local specimen from the infection site, we could obtain a higher sensitivity than IGRAs using PB or ADA in PE and the numbers of ESAT-6 and CFP-10-positive SFCs were 3 to 5 fold higher in PEMCs than in PBMCs. T-SPOT.TB using PE may become a useful diagnostic method for TB pleurisy.展开更多
Objective： Low-density granulocytes （LDGs） can form neutrophil extracellular traps （NETs） spontaneously and excessively. When peripheral blood mononuclear calls （PBMCs） are used for studying T lymphocytes, LDGs...Objective： Low-density granulocytes （LDGs） can form neutrophil extracellular traps （NETs） spontaneously and excessively. When peripheral blood mononuclear calls （PBMCs） are used for studying T lymphocytes, LDGs contained in the PBMCs may decrease the threshold of activating T lymphocytes by forming NETs. This study focused on the profiles of LDGs in common autoimmune diseases and methods for removing LDGs from PBMCs. Methods： The percentages of LDGs in PBMCs from 55 patients with dermatomyositis （DM）, 15 with polymyositis （PM）, 42 with rheumatoid arthritis （RA）, 25 with systemic lupus erythematosus （SLE）, and 19 healthy controls were determined by flow cytometry. Three methods of removing LDGs were explored and compared. After removal, PBMCs from six patients with positive T-SPOT.TB were tested again to find out if LDGs contained in the PBMCs could influence T lymphocyte reactions. Results： Significantly higher LDG percentages were found in PBMCs from patients with DM （（8.41±10.87）%, P〈0.0001 ）, PM （（8.41±10.39）%, P〈0.0001 ）, RA （（4.05±6.97）%, P=0.0249）, and SLE （（7.53±11.52）%, P=0.0006）, compared with the controls （（1.28±0.73）%）. The T-SPOT.TB values significantly decreased after LDGs were removed. Increasing relative centrifugal force （RCF） within a limited range can decrease the LDG percentage from an initial high level, but not markedly increase the LDG clearance rate. Compared with the whole blood sediment method, the PBMC adherence method can significantly remove LDGs yet scarcely influence the T lymphocyte percentage in PBMCs. Conclusions： The LDG percentage in PBMCs is significantly increased in patients with S/E, DM, PM, and RA. The influence of LDGs on T lymphocytes cannot be ignored in PBMC cultures. The adherence method is a simple and easy-to-use method for removing LDGs and purifying T lymphocytes from PBMCs.展开更多
文摘Introduction: The objective of this study was the comparison of the results of T-SPOT.TB using pleural effusion (PE) with those of IGRAs using peripheral blood (PB) or other diagnostic methods for the diagnosis of tuberculous (TB) pleurisy. Methods: We measured adenosine deaminase (ADA) in PE, QuantiFERON TB-Gold In-Tube (QFT), and T-SPOT.TB using PB, and T-SPOT.TB using PE. The definite group of TB pleurisy included 12 patients and other disease group 33 patients. Main find-ings: Sensitivity for QFT using PB was 83% and specificity was 85%, sensitivity for T-SPOT.TB using PB was 92% and specificity was 82%, while sensitivity for ADA in PE was 83% and specificity was 76%. When we adopted the same cut-off level of a positive response for T-SPOT.TB as PB using PE, sensitivity for T-SPOT.TB using PE was 100% and specificity was 82%, respectively. Although there were no significant differences among the four diagnostic methods, sensitivity for T-SPOT.TB using PE gave the most accurate diagnosis of TB-definite patients compared to ADA in PE or QFT using PB. Conclusions: If we performed T-SPOT.TB using a local specimen from the infection site, we could obtain a higher sensitivity than IGRAs using PB or ADA in PE and the numbers of ESAT-6 and CFP-10-positive SFCs were 3 to 5 fold higher in PEMCs than in PBMCs. T-SPOT.TB using PE may become a useful diagnostic method for TB pleurisy.
基金Project supported by the National Natural Science Foundation of China(Nos.81560267 and 81401363)the Natural Science Foundation of Gansu Province(No.1606RJZA213)the Asia Pacific League of Associations for Rheumatology(APLAR) Research Grant 2015
文摘Objective： Low-density granulocytes （LDGs） can form neutrophil extracellular traps （NETs） spontaneously and excessively. When peripheral blood mononuclear calls （PBMCs） are used for studying T lymphocytes, LDGs contained in the PBMCs may decrease the threshold of activating T lymphocytes by forming NETs. This study focused on the profiles of LDGs in common autoimmune diseases and methods for removing LDGs from PBMCs. Methods： The percentages of LDGs in PBMCs from 55 patients with dermatomyositis （DM）, 15 with polymyositis （PM）, 42 with rheumatoid arthritis （RA）, 25 with systemic lupus erythematosus （SLE）, and 19 healthy controls were determined by flow cytometry. Three methods of removing LDGs were explored and compared. After removal, PBMCs from six patients with positive T-SPOT.TB were tested again to find out if LDGs contained in the PBMCs could influence T lymphocyte reactions. Results： Significantly higher LDG percentages were found in PBMCs from patients with DM （（8.41±10.87）%, P〈0.0001 ）, PM （（8.41±10.39）%, P〈0.0001 ）, RA （（4.05±6.97）%, P=0.0249）, and SLE （（7.53±11.52）%, P=0.0006）, compared with the controls （（1.28±0.73）%）. The T-SPOT.TB values significantly decreased after LDGs were removed. Increasing relative centrifugal force （RCF） within a limited range can decrease the LDG percentage from an initial high level, but not markedly increase the LDG clearance rate. Compared with the whole blood sediment method, the PBMC adherence method can significantly remove LDGs yet scarcely influence the T lymphocyte percentage in PBMCs. Conclusions： The LDG percentage in PBMCs is significantly increased in patients with S/E, DM, PM, and RA. The influence of LDGs on T lymphocytes cannot be ignored in PBMC cultures. The adherence method is a simple and easy-to-use method for removing LDGs and purifying T lymphocytes from PBMCs.
