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Blood, Vol. 107, Issue 2, 602-609, January 15, 2006
Skewed T-cell differentiation in patients with indolent non-Hodgkin lymphoma reversed by ex vivo T-cell culture with  c cytokines
Blood Anichini et al.
107: 602
Supplemental Materials for: Anichini et al
Files in this Data Supplement:
- Table S1. Clinical characteristics of B NHL patients (PDF, 74.6 KB)
- Table S2. Activation profile of T cells from peripheral blood of healthy donors (PDF, 54.5 KB)
- Table S3. TCRBV repertoire of T cells from lymph nodes of B NHL patients (PDF, 62.6 KB)
- Figure S1. IFN-γ and IL-4 production by T cells from tumor site in response to autologous EBV-transformed B cells (PDF, 78.5 KB) -
(A) Peripheral blood lymphocytes from a healthy donor were activated for two weeks by culture with irradiated autologous B-LCL. The resulting T cell line was then tested for intra-cellular IFN-
 and IL-4 production without any stimulus (i), or in response to autologous empty DCs (ii), or live B-LCL (iii), or DCs loaded with killed B-LCL (iv). (B) Lymphocytes from FL patient A, isolated from the same tumor containing lymph node as in Figure 2, were cultured for two weeks with autologous LCL (obtained by EBV infection of peripheral B cells), and then tested against autologous DCs either empty (i) or loaded with killed autologous B LCL (ii), or against live autologous LCL (iii). Lymphocytes were gated on CD3+ CD4+ or on CD3+ CD8+ cells. Numbers in the dot plots indicate the fraction of positive cells in each quadrant.
- Figure S2. Correspondence analysis of T cell differentiation profiles in B-NHL patients and healthy donors (PDF, 52.7 KB) -
Correspondence analysis (CA) was applied to the data set containing values for the % positive cells in each of the four possible CCR7/CD45RA maturation stages for both CD4+ and CD8+ T cells from all healthy donors (PBL) and all tissue samples (PBL, LN and bone marrow) from patients. CA results were plotted in a bi-dimensional (MDS dim.1 vs. MDS dim.2) scatterplot and a colour-coded density plot was superimposed to highlight the distribution of the data points. Green dots: T cells from PBL of healthy donors; red dots: T cells from PBL of B cell tumor patients; black dots: T cells from patients’ lymph nodes; white dots: T cells from patients’ bone marrow. The four possible maturation stages of CD4+ and CD8+ T cells are marked by black squares and indicated as 4N, 8N, 4CM, 8CM, 4EM, 8EM, 4TD, 8TD.
- Figure S3. Comparison of T cell maturation profiles in healthy donors and B NHL patients (PDF, 89.5 KB) -
The % of T cells at the TN, TCM, TEM or TTD stages in the CD8+ (A) and CD4+ (B) subsets of lymphocytes isolated from healthy donors or patients was compared by ANOVA followed by SNK test. (1), PBL from donors; (2), PBL from patients; (3), lymphocytes from involved LN or bone marrow of patients. Significance of the differences was annotated as follows: **, P<0.01; ***; P<0.001.
- Figure S4. Maturation profiles of T lymphocytes from tumor-free lymph nodes of breast cancer patients (PDF, 52.4 KB) -
Four-color flow cytometry analysis for the expression of CCR7 and CD45RA in CD3+ CD4+ or CD3+ CD8+ T cells from tumor-free lymph nodes of breast cancer patients. Numbers in the dot plots indicate the fraction of positive cells in each quadrant.
- Figure S5. Differentiation of T lymphocytes from a healthy donor to cytolytic factor+ stage by culture with IL-2 (PDF, 35.4 KB) -
T lymphocytes from PBL of a healthy donor were characterized for the CCR7 vs. CD45RA and CCR7 vs. perforin phenotype before (A) and after (B) culture for 14 days with 50 ng/mL of IL-2.
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