Analysis of TNF-receptor and ligand superfamily molecules in patients with lymphoproliferative disease of granular lymphocytes by Renato Zambello, Livio Trentin, Monica Facco, Marta Siviero, Silvia Galvan, Francesco Piazza, Alessandra Perin, Carlo Agostini, and Gianpietro Semenzato Blood Volume 96(2):647-654 July 15, 2000 ©2000 by American Society of Hematology

Phenotypic expression, redirected cytoxicity, and proliferative activity.Phenotypic expression (A), redirected cytotoxicity against P815 target cells (B), and proliferative activity (C) in the presence of mAbs to CD30, CD30L, CD40, CD40L, CD70, CD95, and CD... Phenotypic expression, redirected cytoxicity, and proliferative activity.Phenotypic expression (A), redirected cytotoxicity against P815 target cells (B), and proliferative activity (C) in the presence of mAbs to CD30, CD30L, CD40, CD40L, CD70, CD95, and CD95L in a representative CD3+TCRαβ+ patient (case 11). Because the expression of TNF-R and TNF-L antigens was different among the patients, the relationship among phenotype, cytotoxicity, and proliferation are shown for a correct interpretation of data. The differences between the peaks of cell fluorescence (continuous lines) with respect to controls (dotted lines) were analyzed using the Kolmogorov-Smirnov test for analysis of histograms and a D/s value ≥ 15 was accepted as significant, as reported in “Patients, materials, and methods.” Cytotoxicity and proliferation experiments were performed in triplicate and mean ± SEM are shown. Renato Zambello et al. Blood 2000;96:647-654 ©2000 by American Society of Hematology

Case 13.Phenotypic expression (A), redirected cytotoxicity against P-815 target cells (B), and proliferative activity (C) in the presence of mAbs to CD30, CD30L, CD40, CD40L, CD70, CD95, and CD95L in a representative CD3+TCRγδ+ patient (case 13). Case 13.Phenotypic expression (A), redirected cytotoxicity against P-815 target cells (B), and proliferative activity (C) in the presence of mAbs to CD30, CD30L, CD40, CD40L, CD70, CD95, and CD95L in a representative CD3+TCRγδ+ patient (case 13). The experimental conditions were the same as in Figure 1. Renato Zambello et al. Blood 2000;96:647-654 ©2000 by American Society of Hematology

Case 17.Phenotypic expression (A), redirected cytotoxicity against P-815 target cells (B), and proliferative activity (C) in the presence of mAbs to CD30, CD30L, CD40, CD40L, CD70, CD95, and CD95L in a representative CD3− patient (case 17). Case 17.Phenotypic expression (A), redirected cytotoxicity against P-815 target cells (B), and proliferative activity (C) in the presence of mAbs to CD30, CD30L, CD40, CD40L, CD70, CD95, and CD95L in a representative CD3− patient (case 17). The experimental conditions were the same as in Figure 1. Renato Zambello et al. Blood 2000;96:647-654 ©2000 by American Society of Hematology

RT-PCR analysis of the expression of CD30, CD30L, CD40, CD40L, Fas, FasL, and control β-actin mRNA in GLs from 6 LDGL patients.Lane 1 indicates molecular weight marker (100 base pairs [bp] DNA ladder); lane 2: control peripheral blood mononuclear cells stim... RT-PCR analysis of the expression of CD30, CD30L, CD40, CD40L, Fas, FasL, and control β-actin mRNA in GLs from 6 LDGL patients.Lane 1 indicates molecular weight marker (100 base pairs [bp] DNA ladder); lane 2: control peripheral blood mononuclear cells stimulated for 12 hours with PHA (5 μg/mL) (positive control) for CD30, CD30L, CD40L, and FasL; positive control for CD40 was represented by purified B lymphocytes; lane 3: the negative control (sample without cDNA), lanes 4 to 6: 3 CD3+ LDGL cases (cases 3, 8, and 13 in Table 1), lane 7 to 9: 3 CD3− LDGL (cases 15, 17, and 20 in Table 1). Renato Zambello et al. Blood 2000;96:647-654 ©2000 by American Society of Hematology

Expression of Annexin V on purified GLs in a representative LDGL case (no 4) of 5 different patients tested.Purified GLs were stimulated with rIL-2 (“Patients, materials, and methods”) for 72 hours, then cells were stained with FITC Annexin V and propidium ... Expression of Annexin V on purified GLs in a representative LDGL case (no 4) of 5 different patients tested.Purified GLs were stimulated with rIL-2 (“Patients, materials, and methods”) for 72 hours, then cells were stained with FITC Annexin V and propidium iodide. The percentage of double positive cells ranged between 22% and 29%. Renato Zambello et al. Blood 2000;96:647-654 ©2000 by American Society of Hematology