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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.
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Objective:To study the influence of different culture conditions on charcic and inhibition activity of nature killer cells ( NK) ,whether to join the modified K562 cells with IL-6 cytokine.Methods:According to the 5′end of the human IL-6 cDNA sequence,PCR primers designed to amplificate,express and transfect K562 cells cDNA library as a template for DNA.Genetic modified K562 cells as stimulating cells were prepared by expressing IL-6.To extract peripheral blood mononuclear cells( PBMC) from human peripheral blood.PBMC were explanted by genetic modified K562 stimulated.The expansion was initiated by CO-culture of PBMC and irradiate genetic modified K562 cell.The number of NK cell increased by directed induced generation of genetic modified K562 cell.Immunophenotypic analysis of NK cell surface markers was performed by flow cytometry (FCM).51Cr release assay was employed to measure the specific lysis skilling of NK cell target K562 cells.Results:We have constracted genetic modified K562 cells by genetic engineering.As stimulated cell added into the PBMC,an average of 760 ±18 fold expansion of CD56+CD16+CD3-cells was observed after 3 weeks of co-culture system.The NK cells population could proliferated more 91%±2% after expansion comparing with 6%± 0.4%in PBMC before expansion by FCM.The cytotoxical activity of NK cells which was induced by genetic modified K562 cell was the strongest than induced by IL-6 cytokine alone.The expanded NK cells lysed 92%±2% of K562 targets in a 5∶1 effector to target ratio.In this case,the NK cells induced by genetic modified K562 cells against tumor cells was more lethal.Conclusion:PBMC based in vitro expansion of natural killer cells was set up by genetic modified K562 cells.The cytotoxicity of NK cells was the strongest induced by genetic modified K562 cell treated.These results had important guiding significance for the the NK large number of amplification and used in clinical.