Improved perforin expression on HTLV-1 specific CD8+ T cells on ligand blockade in ATLL and AC subjects.

<p>PBMC from 6 ATLL and 5 AC subjects were stimulated for 6 hours with and without HTLV-1 peptide in the presence or absence of CD48 in 7 independent experiments. In a) Representative dot plots showing perforin release in peptide only stimulated and peptide stimulated samples to which anti-CD48 was added. In b) Summary of data for all 6 ATLL subjects in 7 independent experiments. In c) summary of data for all 5 asymptomatic carriers in 7 independent experiments (p-value based on Wilcoxon matched-pairs signed rank test).</p

Expression patterns of 2B4 on virus-specific CD8+T lymphocytes.

<p>A), Representative dot plots showing gating strategy for 2B4 and percentage of 2B4 expression on virus-specific CD8+ T lymphocytes, HTLV-1, CMV and EBV. B) Summary data of expression of 2B4 on HTLV-1 infected (ATLL, AC); CMV− and EBV-specific CD8+ T cells in ATLL, AC and HD. Horizontal line represents the mean. (p-value based on by Mann-Whitney test).</p

SAP expression on CD8+ T cells and HTLV-1 specific CD8+T cells in HTLV-1 infection.

<p>In A) Summary of data for percentage SAP expression on total CD8+ T cells in HTLV-1 infected and healthy individuals. In B) Representative dot plots showing gating strategy for HTLV-1 tetramer positive CD8+T cells, isotype control, and SAP expression on HTLV-1 specific CD8+ T cells. In C) Summary data of SAP expression on subjects for whom HTLV-1 specific cluster could be demonstrated matched with SAP expression on CD8+ T cells in HTLV-1 infected subjects (ATLL (9), AC (8)) (p-values based on Mann-Whitney and Wilcoxon matched-pairs signed rank test).</p

2B4 expression on total CD8+Tcells in HTLV-1 infection.

<p>Appropriate isotype controls were used to set quadrant gates, A) Dot plot of isotype and positive 2B4 staining in a representative sample. Numbers indicate frequency of 2B4 expression on total CD8+ T cells of an HTLV-1 infected subject B),Summary data showing 2B4+ expression on total CD8+ lymphocytes in 52 HTLV-1 infected subjects (21 ATL and 31 AC) and 12 uninfected healthy donors (HD). Each dot represents a single individual. Horizontal bars represent mean values (p-value based on Mann-Whitney test).</p

Increased CD107a degranulation of HTLV-1 specific CD8+ T cells on ligand blockade in both ATLL and AC.

<p>In A), Representative dot plots show CD107a surface degranulation on HTLV-1 specific CD8+ T cells in varying conditions. In B) Summary plot of CD107a surface degranulation in tax-stimulated samples in the presence and absence of CD48 antibody addition in ATLL (11 independent experiments in 7 patients) and AC (12 independent experiments in 8 subjects). Experiments were done in duplicates (p-value based on Wilcoxon matched-pairs signed rank test).</p

Characteristics of the Study Groups.

a<p>M/F = male/female,</p>b<p>ATLL = adult T-cell leukemia/lymphoma,</p>c<p>AC = asymptomatic carrier.</p>d<p>HD = healthy donor.</p

CD48 MFI (mean fluorescence intensity) on CD4 T cells.

<p>In A) Representative histograms of CD48 MFI on CD4+ T cell; ATLL (opened histogram), AC (grayed histogram), HD (black histogram) B) summary data comparing CD48 MFI (mean fluorescence intensity) on total CD4+ T cells in HTLV-1 infected and uninfected subjects (13 ATLL, 13 AC and 12HD). CD48 MFI was visibly lower in ATLL and AC relative to HD. The horizontal line represents the mean ± S.D (p-value based on Mann-Whitney test).</p

Characterization of memory population of 2B4+CD8+ T cells by CCR7, CD45RA, CD62L and CD57 expression.

<p>2B4+CD8+ T cells tended towards effector and terminally differentiated memory phenotype relative to 2B4-CD8+ T cells. A, B) Summary of frequency of CD45RA and CCR7 coexpression on 2B4+CD8+ and 2B4-CD8+ T cells in HTLV-1 infected (6 ATL and 5 AC) and 5 healthy donors (HD). 2B4+CD8+ T cells expressing CD45RA and CCR7 showed a mature effector phenotype in the effector memory and terminally differentiated compartments compared to 2B4-CD8+ T lymphocytes. C) Summary of frequency of CD57 on CD8+ T cells were significantly higher in 2B4+ compared to 2B4− populations in both ATLL and AC D) Lower CD62L expression on 2B4+CD8+ compared to 2B4-CD8+ T cells. Data for CD57 and CD62L was obtained from 6 ATL and 5 AC. Phenotypic patterns are indicated by legends. Horizontal lines represent mean ± S.D; statistical comparisons, p- value based on Mann-Whitney and Wilcoxon matched-pairs signed rank test.</p

Detectable Tax-specific CD8+ T cells in all samples used in this study.

a<p>human leucocyte antigen.</p>b<p>cytomegalovirus.</p>c<p>epstein-barr virus.</p