Increased chemokine receptor CCR7/EBI1 expression enhances the infiltration of lymphoid organs by adult T-cell leukemia cells by Hitoshi Hasegawa, Tetsuhiko.

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Increased chemokine receptor CCR7/EBI1 expression enhances the infiltration of lymphoid organs by adult T-cell leukemia cells by Hitoshi Hasegawa, Tetsuhiko Nomura, Masashi Kohno, Norihiko Tateishi, Yoji Suzuki, Nobuji Maeda, Ryuichi Fujisawa, Osamu Yoshie, and Shigeru Fujita Blood Volume 95(1):30-38 January 1, 2000 ©2000 by American Society of Hematology

Hitoshi Hasegawa et al. Blood 2000;95:30-38 ©2000 by American Society of Hematology

Hitoshi Hasegawa et al. Blood 2000;95:30-38 ©2000 by American Society of Hematology

Hitoshi Hasegawa et al. Blood 2000;95:30-38 ©2000 by American Society of Hematology

Comparison of secondary lymphoid-tissue chemokine (SLC)- and EBI1-ligand chemokine (ELC)-induced adhesion of adult T-cell leukemia (ATL) cells under static conditions.(A) Comparison of SLC- and ELC-induced adhesion of ATL cells from patients with and withou... Comparison of secondary lymphoid-tissue chemokine (SLC)- and EBI1-ligand chemokine (ELC)-induced adhesion of adult T-cell leukemia (ATL) cells under static conditions.(A) Comparison of SLC- and ELC-induced adhesion of ATL cells from patients with and without lymphoid organ involvement to human umbilical vein-derived endothelial cells (HUVECs) under static conditions. Cells (2 × 105/well), which had been labeled with 51Cr, were incubated with SLC or ELC at a concentration of 30 ng/ml at 37°C for 30 minutes and were added to the well containing IL-1β–activated HUVECs. The plates were incubated at 37°C for 30 minutes and then gently washed, and radioactivity of the adherent cells in each well was counted. The cells are as follows: control CD4+CD45RO+ T cells (A, closed squares) and ATL cells from patients without (B, patients 1 through 12) and with lymphoid organ involvement (C, patients 13 through 28). ATL cells from patients with lymphoid organ involvement were blocked with 100 ng/ml pertussis toxin (PTX) for 1.5 hours at 37°C (C+PT) or with anti-CD18 mAb, 7E4 (20 μg/ml) for 20 minutes at room temperature (C+CD18 Ab) before adding of chemokines. The data are expressed as the mean percentage of the binding of indicated cells from triplicate experiments. The statistical analysis was performed using Student t test. (B) Comparison of SLC- and ELC-induced adhesion of ATL cells from patients with and without lymphoid organ involvement to ICAM-1 under static conditions. The 96-well culture plates were coated with ICAM-1 protein (25 ng/well). Hitoshi Hasegawa et al. Blood 2000;95:30-38 ©2000 by American Society of Hematology

Comparison of secondary lymphoid-tissue chemokines (SLC)- and EBI1-ligand chemokine (ELC)-induced adhesion of adult T-cell leukemia (ATL) cells from patients with and without lymphoid organ involvement to ICAM-1 under flow conditions.Cells (106/ml) were per... Comparison of secondary lymphoid-tissue chemokines (SLC)- and EBI1-ligand chemokine (ELC)-induced adhesion of adult T-cell leukemia (ATL) cells from patients with and without lymphoid organ involvement to ICAM-1 under flow conditions.Cells (106/ml) were perfused onto the coated dish with substrates consisting of ICAM-1 (10 μg/ml) alone or also with SLC (10 μg/ml) or ELC (10 μg/ml) at 1 dyne/cm2 for the first 4 minutes. The number of arrested cells is shown by the total number accumulated as arrested cells for the first 4 minutes in a single field of view (4X objective). The cells are as follows: control CD4+CD45RO+ T cells (A, closed squares) and ATL cells from patients without (B, patients 1 through 12) and with lymphoid organ involvement (C, patients 13 through 28). ATL cells from patients with lymphoid organ involvement were blocked with 100 ng/ml pertussis toxin (PTX) for 1.5 hours at 37°C (C+PT) or with anti-CD18 mAb, 7E4 (20 μg/ml) for 20 minutes at room temperature (C+CD18 Ab) before binding to ICAM-1 with SLC or ELC. Experiments were performed in triplicate. The bars indicate the mean value of each group; the statistical analysis was performed using Student ttest. Hitoshi Hasegawa et al. Blood 2000;95:30-38 ©2000 by American Society of Hematology

Comparison of secondary lymphoid-tissue chemokine (SLC)- and EBI1-ligand chemokine (ELC)-evoked chemotaxis of adult T-cell leukemia (ATL) cells from patients with and without lymphoid organ involvement.Chemotactic assays were performed in the presence of SL... Comparison of secondary lymphoid-tissue chemokine (SLC)- and EBI1-ligand chemokine (ELC)-evoked chemotaxis of adult T-cell leukemia (ATL) cells from patients with and without lymphoid organ involvement.Chemotactic assays were performed in the presence of SLC or ELC at a low concentration (30 ng/ml) as described in Materials and Methods. The cells are as follows: control CD4+CD45RO+ T cells (A, closed squares) and ATL cells from patients without (B, patients 1 through 12) and with lymphoid organ involvement (C, patients 13 through 28). ATL cells from patients with lymphoid organ involvement were blocked with 100 ng/ml pertussis toxin (PTX) for 1.5 hours at 37°C (C+PT) before chemotactic assays. The data are expressed as the mean percentage of migrated cells toward SLC or ELC from triplicate experiments. The bars indicate the mean value of each group; the statistical analysis was performed using Student t test. Hitoshi Hasegawa et al. Blood 2000;95:30-38 ©2000 by American Society of Hematology

Flow cytometric analysis of the infiltrating adult T-cell leukemia (ATL) cells in lymph nodes.CCR7/EBI1 expression levels of infiltrating ATL cells prepared from lymph nodes of 2 representative patients, patients 26 and 27, and control CD4+CD45RO+ T cells f... Flow cytometric analysis of the infiltrating adult T-cell leukemia (ATL) cells in lymph nodes.CCR7/EBI1 expression levels of infiltrating ATL cells prepared from lymph nodes of 2 representative patients, patients 26 and 27, and control CD4+CD45RO+ T cells from reactive lymph nodes of HTLV-1-seronegative individuals were compared by flow cytometric analysis using an anti-CCR7 mAb, CCR7.6B3. The infiltrating ATL cells and control CD4+CD45RO+ T cells were purified by the negative immunoselection technique as described in Materials and Methods. Hitoshi Hasegawa et al. Blood 2000;95:30-38 ©2000 by American Society of Hematology