In vitro migration of cytotoxic T lymphocyte derived from a colon carcinoma patient is dependent on CCL2 and CCR2

Table 5 Induction of tumor apoptosis by CTL007 is inhibited by anti-CCR2 and CCL2 Abs and excess CCL2a

Blocking agent	Number of tumor cells/field (10 fields)	Number of apoptotic cells/field (10 fields)	% of apoptotic cells	% of tumor cell apoptosis inhibition
None	21.8 ± 9.7	8.2 ± 9	40.5 ± 9.7^b	-
Control IgG	21 ± 2.2	8.4 ± 1.1	40.1 ± 4.9^c	-
Anti-CCR1 Ab	22 ± 3.4	10.0 ± 1.6	45.6 ± 5.0	-13.7
Anti-CCR2 Ab	19.6 ± 1.1	2.6 ± 0.5	13.2 ± 2.3^c	67.1
Anti-CCR3 Ab	22.4 ± 2.8	9.2 ± 1.9	40.7 ± 4.2	1.5
Anti-CCR5 Ab	21 ± 2.6	9.6 ± 1.8	45.5 ± 3.8	-13.4
Anti-CCR7 Ab	18.8 ± 2.4	7.2 ± 1.6	38.0 ± 4.1	5.2
Anti-CXCR3 Ab	21.4 ± 1.5	9.2 ± 0.8	43.1 ± 1.8	-7.5
CCL2	21.4 ± 4.4	3.6 ± 1.1	17.2 ± 6.1^b	57.5
Anti-CCL2 Ab	17.8 ± 2.6	2.2 ± 0.4	12.7 ± 3.8^c	68.6

^a Reconstructs consisted of a bottom layer of collagen and fibroblasts, followed by a tumor cell layer and a separating layer of collagen and fibroblasts. Anti-chemokine receptor or control Abs were added, followed by a top layer containing CTL mixed with fibroblasts and collagen (E: T = 3:1). Excess CCL2 (50 ng/ml) was added to the T-cell layer. Percentage of apoptotic tumor cells in 6 day cultures was determined. ^{(b, c)}Values with same letter differ significantly from each other (p < 0.05). Data represented are from a single experiment and the Results were confirmed in at least two independent experiments.

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