Table 2 TEM of CLL cells and normal B... | American Society of Hematology

Table 2

TEM of CLL cells and normal B cells through HUVEC monolayers and the effect of EphA2-Fc or EFNA4-Fc addition

Sample/fraction no.	Treatment, %
Sample/fraction no.	hFc control	EFNA4-Fc	EphA2-Fc
CLL-1
F1	96.49 ± 0.11	76.91 ± 0.10*	99.21 ± 0.03*
F2	3.35 ± 0.05	22.94 ± 0.09*	0.78 ± 0.04*
F3	0.16 ± 0.05	0.16 ± 0.01	0.02 ± 0.01
CLL-3
F1	94.66 ± 0.91	80.73 ± 1.46†	96.58 ± 0.31
F2	3.72 ± 0.38	17.10 ± 1.20‡	2.60 ± 0.07
F3	1.63 ± 0.53	2.17 ± 0.25	0.82 ± 0.24
CLL-4
F1	96.47 ± 0.25	73.35 ± 0.15*	98.93 ± 0.08†
F2	3.20 ± 0.26	26.21 ± 0.15*	1.06 ± 0.08†
F3	0.32 ± 0.01	0.45 ± 0.00‡	0.00 ± 0.00*
CLL-5
F1	94.47 ± 0.08	65.80 ± 0.26*	98.94 ± 0.22‡
F2	5.30 ± 0.06	33.81 ± 0.26*	1.06 ± 0.23†
F3	0.24 ± 0.02	0.40 ± 0.01†	0.00 ± 0.00†
CLL-8
F1	82.62 ± 2.13	62.32 ± 1.54†	95.12 ± 0.95†
F2	11.29 ± 1.47	22.68 ± 0.45†	3.11 ± 0.64†
F3	6.10 ± 0.66	15.00 ± 1.09†	1.78 ± 0.30†
CLL-10
F1	75.49 ± 0.42	48.35 ± 0.85*	89.24 ± 0.16*
F2	16.10 ± 0.21	35.83 ± 0.58*	4.32 ± 0.02*
F3	8.41 ± 0.21	15.82 ± 0.27†	6.45 ± 0.18†
CLL-13
F1	90.83 ± 0.12	63.23 ± 0.16*	98.98 ± 0.00*
F2	8.26 ± 0.09	35.90 ± 0.14*	1.01 ± 0.01*
F3	0.91 ± 0.03	0.88 ± 0.02	0.02 ± 0.01*
NBC-1
F1	50.94 ± 0.08	44.78 ± 1.08†	51.73 ± 0.37
F2	27.25 ± 0.35	28.51 ± 0.70	22.43 ± 0.61†
F3	21.80 ± 0.43	26.72 ± 0.38†	25.85 ± 0.23†
NBC-2
F1	60.59 ± 0.56	43.17 ± 0.25*	55.62 ± 0.54†
F2	26.45 ± 0.61	40.90 ± 0.15*	18.89 ± 0.16‡
F3	12.95 ± 0.05	15.94 ± 0.10*	25.50 ± 0.70‡

Sample/fraction no.	Treatment, %
Sample/fraction no.	hFc control	EFNA4-Fc	EphA2-Fc
CLL-1
F1	96.49 ± 0.11	76.91 ± 0.10*	99.21 ± 0.03*
F2	3.35 ± 0.05	22.94 ± 0.09*	0.78 ± 0.04*
F3	0.16 ± 0.05	0.16 ± 0.01	0.02 ± 0.01
CLL-3
F1	94.66 ± 0.91	80.73 ± 1.46†	96.58 ± 0.31
F2	3.72 ± 0.38	17.10 ± 1.20‡	2.60 ± 0.07
F3	1.63 ± 0.53	2.17 ± 0.25	0.82 ± 0.24
CLL-4
F1	96.47 ± 0.25	73.35 ± 0.15*	98.93 ± 0.08†
F2	3.20 ± 0.26	26.21 ± 0.15*	1.06 ± 0.08†
F3	0.32 ± 0.01	0.45 ± 0.00‡	0.00 ± 0.00*
CLL-5
F1	94.47 ± 0.08	65.80 ± 0.26*	98.94 ± 0.22‡
F2	5.30 ± 0.06	33.81 ± 0.26*	1.06 ± 0.23†
F3	0.24 ± 0.02	0.40 ± 0.01†	0.00 ± 0.00†
CLL-8
F1	82.62 ± 2.13	62.32 ± 1.54†	95.12 ± 0.95†
F2	11.29 ± 1.47	22.68 ± 0.45†	3.11 ± 0.64†
F3	6.10 ± 0.66	15.00 ± 1.09†	1.78 ± 0.30†
CLL-10
F1	75.49 ± 0.42	48.35 ± 0.85*	89.24 ± 0.16*
F2	16.10 ± 0.21	35.83 ± 0.58*	4.32 ± 0.02*
F3	8.41 ± 0.21	15.82 ± 0.27†	6.45 ± 0.18†
CLL-13
F1	90.83 ± 0.12	63.23 ± 0.16*	98.98 ± 0.00*
F2	8.26 ± 0.09	35.90 ± 0.14*	1.01 ± 0.01*
F3	0.91 ± 0.03	0.88 ± 0.02	0.02 ± 0.01*
NBC-1
F1	50.94 ± 0.08	44.78 ± 1.08†	51.73 ± 0.37
F2	27.25 ± 0.35	28.51 ± 0.70	22.43 ± 0.61†
F3	21.80 ± 0.43	26.72 ± 0.38†	25.85 ± 0.23†
NBC-2
F1	60.59 ± 0.56	43.17 ± 0.25*	55.62 ± 0.54†
F2	26.45 ± 0.61	40.90 ± 0.15*	18.89 ± 0.16‡
F3	12.95 ± 0.05	15.94 ± 0.10*	25.50 ± 0.70‡

TEM assays were carried out by coculturing normal B cells or CLL cells with HUVEC monolayers during 2 hours, and nonadhered (F1), adhered (F2), or transmigrating cells (F3) were counted by flow cytometry. The absolute number of cells recovered within each fraction (Fn) was normalized to the total number of them recovered (Fn/F1 + F2 + F3) and expressed as percentages (%). CLL/normal B cells or HUVECs were pretreated with saturating amounts of EphA2-Fc or EFNA4-Fc, respectively. Values are mean ± SD from triplicates. Significant differences between treated and control hFc cultures were calculated by 1-sample, 2-tailed Student t test.

CLL indicates chronic lymphocytic leukemia; F, fraction; hFc, Fc fragment of human IgG immunoglobulin; HUVEC, human umbilical vein endothelial cell; NBC, normal B cell; and TEM, transendothelial migration.

Value of P < .001.

†

Value of P < .005.

‡

Value of P < .05.

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