Attempt at simultaneous quantitative determination of multiple cytokines and chemokines in peripheral blood with a suspension array system

Purpose

Blood cytokine and chemokine values have been examined in pathological analyses of surgical invasion, but making multiple simultaneous measurements has been difficult from the standpoint of cost, measurement time, specimen size, etc. Microbead suspension arrays have been developed in recent years, and they have made quantitative determination of multiple substances possible.

Subjects and methods

The subjects were 16 surgical patients with severe septicemia secondary to peritonitis. The patients were divided into groups according to whether their preoperative leukocyte count was below 4000/μl (LW group) or 4000/μl or greater (HW group) and changes between before and after surgery, and the changes between before and after performance of postoperative endotoxin adsorption therapy (PMX) were investigated.

The specimens consisted of frozen plasma, and the measurements were made in 25 μl samples by Bio-Plex assays (Bio-Rad Laboratories). The Wilcoxon test was used to perform the statistical analysis.

Results

There were five cases in the LW group, and 11 in the HW group. There were no significant differences in age, interval between onset of symptoms and arrival at the hospital, or APACHE II scores, and the differences between groups in the operation time and blood loss were not significant either. The difference between groups in TNF-α was not significant, but the IL-1β, IL-4, IL-5, IL-6, IL-8, IL-10, and IL-13, G-CSF, GM-CSF, INF, MCP-1, and MIP-1β values were significantly higher in the LW group both preoperatively and postoperatively (P < 0.05). In the HW group IL-12 and IL-17 tended to be higher before surgery. Before surgery the Th1/Th2 ratio (INF/IL-4) was higher in the LW group, but after surgery it was higher in the HW group. Comparisons between before and after PMX therapy showed a significant decrease in endotoxin values (P = 0.03), but there were no significant changes in cytokine or chemokine values.

Conclusion

A decreased leukocyte count in peritonitis appeared to reflect a state of increased cytokine response, especially an increased response of the Th1 system. Postoperatively, however, Th2 tended to predominate. PMX therapy had no effect on cytokines. It appeared that this type of analysis will elucidate the indications and optimal timing for PMX therapy and G-CSF and INF administration in severe pathological states.