Expression of Non-T Cell Activation Linker (NTAL) in Jurkat Cells Negatively Regulates TCR Signaling: Potential Role in Rheumatoid Arthritis.

T lymphocytes are key players in adaptive immune responses through the recognition of peptide antigens through the T Cell Receptor (TCR). After TCR engagement, a signaling cascade is activated, leading to T cell activation, proliferation, and differentiation into effector cells. Delicate control of activation signals coupled to the TCR is needed to avoid uncontrolled immune responses involving T cells. It has been previously shown that mice deficient in the expression of the adaptor NTAL (Non-T cell activation linker), a molecule structurally and evolutionarily related to the transmembrane adaptor LAT (Linker for the Activation of T cells), develop an autoimmune syndrome characterized by the presence of autoantibodies and enlarged spleens. In the present work we intended to deepen investigation into the negative regulatory functions of the NTAL adaptor in T cells and its potential relationship with autoimmune disorders. For this purpose, in this work we used Jurkat cells as a T cell model, and we lentivirally transfected them to express the NTAL adaptor in order to analyze the effect on intracellular signals associated with the TCR. In addition, we analyzed the expression of NTAL in primary CD4+ T cells from healthy donors and Rheumatoid Arthritis (RA) patients. Our results showed that NTAL expression in Jurkat cells decreased calcium fluxes and PLC-γ1 activation upon stimulation through the TCR complex. Moreover, we showed that NTAL was also expressed in activated human CD4+ T cells, and that the increase of its expression was reduced in CD4+ T cells from RA patients. Our results, together with previous reports, suggest a relevant role for the NTAL adaptor as a negative regulator of early intracellular TCR signaling, with a potential implication in RA.

Homocysteine: new tumor marker in pleural fluid.

There are no published studies examining the utility of total homocysteine (HCY) in pleural fluid. The aim was to measure the accuracy of pleural fluid HCY concentration for diagnosis of malignant pleural effusion (MPE). We studied pleural fluids obtained by thoracocentesis in patients with pleural effusion. Pleural fluid HCY concentration was measured by immunonephelometry using N Latex HCY reagent with monoclonal antibody in automated analyzers BNII (Siemens Diagnostics®). Patients were classified into two groups according to the etiology of pleural effusion: benign pleural effusions (BPE) and MPE. Pleural effusion was categorized as MPE if malignant cells were demonstrated in pleural fluid or pleural biopsy. The accuracy of pleural fluid HCY concentration for diagnosis of MPE was determined using receiver operating characteristic (ROC) techniques by analyzing the area under the ROC curve (AUC). We studied 89 patients with ages between 1 and 96 years old (median = 66). Forty-eight patients were BPE and 41 were MPE. Pleural fluid HCY concentration was significantly higher in patients with MPE (median = 13.70 µmol/L) than in those with BPE (median = 8.05 µmol/L). The AUC value was 0.833 (95 % confidence interval (CI) 0.739-0.903). The optimal cutoff value was 13.1 µmol/L exhibiting 56.1 % (95 % CI 39.8-71.5) sensitivity and 85.4 % (95 % CI 72.2-93.9) specificity. Pleural fluid HCY concentration showed high diagnostic accuracy to predict whether a pleural effusion is benign or malignant. Pleural fluid HCY concentration may be measured easily and quickly in automated analyzers and could be a tumor marker commonly used for diagnosis of MPE.

The diagnostic value of pleural fluid homocysteine in malignant pleural effusion.

BACKGROUND: Pleural fluid homocysteine (HCY) can be useful for diagnosis of malignant pleural effusion (MPE). There are no published studies comparing the diagnostic accuracy of HCY with other tumour markers in pleural fluid for diagnosis of MPE. The aim was to compare the accuracy of HCY with that of carcinoembryonic antigen (CEA), cancer antigen (CA) 15.3, CA19.9 and CA125 in pleural fluid and to develop a probabilistic model using these biomarkers to differentiate benign (BPE) from MPE. METHODS: Patients with pleural effusion were randomly included. HCY, CEA, CA15.3, CEA19.9 and CA125 were quantified in pleural fluid. Patients were classified into two groups: MPE or BPE. By applying logistic regression analysis, a multivariate probabilistic model was developed using pleural fluid biomarkers. The diagnostic accuracy was determined by receiver operating characteristic (ROC) curves and calculating the area under the curve (AUC). RESULTS: Population of study comprised 133 patients (72 males and 61 females) aged between 1 and 96 years (median = 70 years), 81 BPE and 52 MPE. The logistic regression analysis included HCY (p<0.0001) and CEA (p = 0.0022) in the probabilistic model and excluded the other tumour markers. The probabilistic model was: HCY+CEA = Probability(%) = 100×(1+e-z)-1, where Z = 0.5471×[HCY]+0.3846×[CEA]-8.2671. The AUCs were 0.606, 0.703, 0.778, 0.800, 0.846 and 0.948 for CA125, CA19.9, CEA, CA15.3, HCY and HCY+CEA, respectively. CONCLUSIONS: Pleural fluid HCY has higher accuracy for diagnosis of MPE than CEA, CA15.3, CA19.9 and CA125. The combination of HCY and CEA concentrations in pleural fluid significantly improves the diagnostic accuracy of the test.