COMPREHENSIVE ANALYSIS OF CHEMOKINES AND CYTOKINES SECRETED IN THE PERITONEAL CAVITY DURING LAPAROTOMY

Figures & data

TABLE 1 Patient Demographics

Case	Age	Sex	Operation	Stage	Operation Time (h)
1	51	Male	Intersphincteric resection with loop ileostomy	I	5.00
2	74	Female	Sigmoid colectomy	II	2.25
3	57	Male	Low anterior resection with loop ileostomy	II	3.30
4	76	Male	Low anterior resection	IV	5.00
5	65	Male	Left hemicolectomy with extended left lobectomy of the liver	IV	8.75
6	62	Male	Anterior resection	III	3.17
7	40	Female	Low anterior resection	III	4.08

FIGURE 1 Relative amount of measured cytokines (A) and chemokines (B) listed in Table 1. Percentage was calculated by dividing the concentration of cytokines or chemokines (pg/mL) with sum of the concentrations (total concentration) of measured cytokines or chemokines (pg/mL). (C) Data from an individual case for CCL5 and IL-6 are shown. Blank bars, preoperative concentration; solid bars, postoperative concentration. The case number indicates the patient ID in Table 1 (color figure available online).

TABLE 2 Concentration of Cytokines and Chemokines (pg/mL) in Peritoneal Lavage Fluid

	Pre-operation ^a	Post-operation ^a	P value	Fold Increase
Cytokines
IL-6	1776 ± 2624	20074 ± 7240	0.001*	11.3
IL-1β	333 ± 155	2857 ± 4010	0.153	8.6
PDGF	185 ± 67	1084 ± 776	0.025*	5.8
G-CSF	435 ± 410	2098 ± 2119	0.100	4.8
IL-1Ra	220 ± 119	874 ± 709	0.058	4.0
IFN-γ	176 ± 30	567 ± 275	0.013*	3.2
IL-13	333 ± 158	763 ± 541	0.046*	2.3
IL-4	163 ± 69	328 ± 192	0.021*	2.0
IL-10	489 ± 149	982 ± 482	0.071	2.0
VEGF	822 ± 246	1609 ± 884	0.101	2.0
IL-12	217 ± 75	372 ± 221	0.121	1.7
TNF-α	742 ± 132	1222 ± 576	0.104	1.6
IL-15	874 ± 567	1352 ± 857	0.014*	1.5
IL-9	323 ± 61	495 ± 123	0.001*	1.5
IL-7	201 ± 75	273 ± 141	0.368	1.4
IL-2	223 ± 133	280 ± 105	0.157	1.3
GM-CSF	1218 ± 859	1012 ± 628	0.438	0.8
IL-5	90 ± 80	63 ± 10	0.405	0.7
FGFbasic	3882 ± 1886	2707 ± 3004	0.363	0.7
IL-17	669 ± 446	418 ± 333	0.122	0.6
Chemokines
CCL5 (RANTES)	2241 ± 1921	24066 ± 2979	P < 0.0001*	10.7
CCL3 (MIP1α)	160 ± 69	1431 ± 2865	0.284	8.9
CCL2 (MCP-1)	2250 ± 1534	14511 ± 8692	0.008*	6.4
CXCL8 (IL-8)	1881 ± 2446	11647 ± 7836	0.024*	6.2
CCL4 (MIP1β)	1409 ± 1287	7216 ± 5660	0.033*	5.1
CCL11 (eotaxin)	318 ± 185	584 ± 253	0.031*	1.8
CXCL10 (IP-10)	980 ± 1267	827 ± 1448	0.109	0.8
Total protein mg/mL	1.593 ± 1.721	3.39 ± 2.99	0.269	2.1

FIGURE 2 TNF-α, IL-6, and CCL5 production by peritoneal exudate cells. Peritoneal exudate cells harvested from preoperative lavage fluid were cultured with (LPS) or without LPS (none). Culture supernatant was subjected to the assay for TNF-α (A), IL-6 (B), and CCL5 (C).

FIGURE 3 CCL5-driven chemotaxis of human PBMCs. (A) Chemotaxis of PBMCs induced by recombinant CCL5 was measured in the presence of indicated concentrations of anti-CCL5 antibody (mg/mL). (B) Pre- (blank) and postoperative (solid) peritoneal fluid from case 1 was tested for the chemotactic potential of human PBMCs in the presence of 0.1 mg/mL of anti-CCL5 antibody or anti-CCL2 antibody. Data are shown as an average of duplicated assay.