Inverse correlation between CD8+ inflammatory cells and E-cadherin expression in gallbladder cancer: Tissue microarray and imaging analysis

Table 1 The primary antibodies used for immunohistochemistry

Antibody	Dilution	Clone	Manufacturer
LCA	Prediluted	PD7/26, 2B11	Nichirei Biosciences, Tokyo
MPO	1:200	Polyclonal	DakoCytomation, Glostrup, Denmark
CD3	Prediluted	PS1	Nichirei Biosciences
CD4	Prediluted	4B12	Nichirei Biosciences
CD8	1:50	C8/144B	DakoCytomation
CD20	1:50	L26	DakoCytomation

LCA: Leukocyte common antigen; MPO: Myeloperoxidase.

Table 2 The numbers of inflammatory cells per tissue microarray core

	Maximun positive cell counts/core	Minimun positive cell counts/core	Average/core	Standard deviation/core	No. of analyzable cores
LCA	13962	73	3108.27	2734.79	136
MPO	10911	388	2692.39	1830.47	131
CD3	14702	74	1967.06	2058.45	130
CD4	6583	0	583.01	934.52	134
CD8	5572	10	919.44	1034.35	131
CD20	7097	4	467.95	834.74	133
CD68	11355	97	2452.99	2043.92	135

LCA: Leukocyte common antigen; MPO: Myeloperoxidase.

Table 3 Single linear regression analysis of the relationship between E-cadherin expression and the type of inflammatory cells

Cells	Regression equation	Adjusted R2	R	Standard error	P-value
LCA	EHS = 117.67879 − 0.0029223 × LCA	0.0216	0.15	46.5793	0.0482
CD20	EHS = 112.37344 − 0.0083497 × CD20	0.0153	0.12	45.8466	0.083
CD3	EHS = 119.86794 − 0.0050479 × CD3	0.0421	0.21	45.7107	0.011
CD4	EHS = 109.63264 − 0.0003939 × CD4	< 0.0001	n/a	47.1913	0.9285
CD8	EHS = 121.37735 − 0.0146326 × CD8	0.1011	0.32	43.6707	0.0001
MPO	EHS = 104.23773 + 0.0012811 × MPO	−0.00523	n/a	46.9647	0.5701
CD68	EHS = 113.83072 − 0.0023317 × CD68	0.0029	0.05	46.8666	0.2412

EHS: E-cadherin histological score; n/a: Not available; LCA: Leukocyte common antigen; MPO: Myeloperoxidase.

Table 4 Clinicopathologic features of the patients with gallbladder cancer (n = 50) n (%)

Age	Mean ± SD	68.7 ± 8.4
Gender	Male	15 (30.0)
	Female	35 (70.0)
Differentiation of invasive front	G1	16 (32.0)
	G2	19 (38.0)
	G3	15 (30.0)
T-stage	T2	18 (36.0)
	T3	28 (56.0)
	T4	4 (8.0)
N	N0	17 (34.0)
	N1	33 (66.0)
No. of CD8-positive cells (means of analyzable cores in each case)	Median ± SD	580.0 ± 1154.5
E-cadherin histological score	Median ± SD	114.3 ± 37.8