Hemoperfusion with polymyxin B-immobilized fibers reduced the number of CD16+CD14 + monocytes in patients with septic shock

Abstract

Background: CD16⁺CD14⁺ monocytes dramatically increase in number in patients with severe infection. Hemoperfusion with PMX-F (direct hemoperfusion with polymyxin B immobilized fibers) has been reported to be a safe and effective treatment for patients with septic shock, although the molecular mechanism that accounts for its effectiveness is still unclear. The purpose of this study was to quantify the number of CD16⁺CD14⁺ monocytes in patients with an intra-abdominal infection and to evaluate the effects of PMX-F treatment on clinical parameters and leukocyte surface antigen expression in these patients.

Materials and Methods : Seventeen septic patients who had an intra-abdominal infection were enrolled in this study; 7 of these patients received PMX-F treatment. Peripheral blood samples were obtained immediately after admission, and were also collected from the above 7 patients before, during, and immediately after their PMX-F treatment. The expression of CD14, CD16, and Toll-like receptor (TLR)-4 on these patients' monocytes was evaluated using flow cytometry. In addition, lipopolysaccharide (LPS)-induced production of TNF-α and IL-1β by these cells was measured by ELISA.

Results: Monocytic expression of CD16 and TLR-4 was significantly greater in septic patients than in healthy controls, and their proportion of CD16⁺CD14⁺ monocytes was similarly elevated. LPSinduced production of TNF-α and IL-1β by peripheral blood mononuclear cells (PBMCs) of septic patients was significantly reduced compared to controls. Furthermore, there was a reduction in the proportion of CD 16⁺CD 14⁺ monocytes during PMX-F treatment, and in the expression of TLR-4 on monocytes after PMX-F treatment.

Conclusions: These results showed that the number of peripheral blood CD16⁺CD14⁺ monocytes and monocytic TLR-4 expression were markedly increased, and the production of pro-inflammatory cytokines in response to LPS significantly reduced in patients with sepsis. PMX-F treatment was found to be effective in reducing the number of CD 16⁺CD 14⁺ monocytes and in decreasing the monocytic expression of TLR-4 in patients with septic shock.

Keywords

Polymyxin B-immobilized fiber (PMX)sepsis CD16+CD14+ monocytes

References

Kostyal DA , Butler GH , Beezhold DH Mycoplasma hyorhinis molecules that induce tumor necrosis factor alpha secretion by human monocytes. Infect Immun 1995; 63: 3858—3863.

Wang SY , Mak KL , Chen LY , Chou MP , Ho CK Heterogeneity of human blood monocyte: two subpopulations with different sizes, phenotypes and functions. Immunology 1992; 77: 298—303.

Fingerle-Rowson G. , Angstwurm M. , Andreesen R. , Ziegler-Heitbrock HW. Selective depletion of CD14+ CD16+ monocytes by glucocorticoid therapy. Clin Exp Immunol 1998; 112: 501—506.

Haziot A. , Hijiya N. , Schultz K. , Zhang F. , Gangloff SC , Goyert SM CD14 plays no major role in shock induced by Staphylococcus aureus but down-regulates TNF-1 production. J Immunol 1999; 162: 4801—4805.

Locher C. , Vanham G. , Kestens L. et al. Expression patterns of Fc gamma receptors, HLA-DR and selected adhesion molecules on monocytes from normal and HIV-infected individuals. Clin Exp Immunol 1994; 98: 115—122.

Blumenstein M. , Boekstegers P. , Fraunberger P. , Andreesen R. , Ziegler-Heitbrock HW , Fingerle-Rowson G. Cytokine production precedes the expansion of CD14+CD16+ monocytes in human sepsis: a case report of a patients with self-induced septicemia. Shock 1997; 8: 73—75.

Belge K. , Dayyani F. , Horelt A. et al. The proinflammatory CD14+CD16+Dr++ monocytes are a major source of TNF. J Immunol 2002; 168: 3536—3542.

Horelt A. , Belge KU , Steppich B. , Prinz J. , Ziegler-Heitbrock L. The CD14+CD16+ monocytes in erysipelas are expanded and show reduced cytokine production. Eur J Immunol 2002; 32: 1319—1327.

Hanasawa K. , Tani T. , Kodama M. New approach to endotoxic and septic shock by means of polymyxin B immobilized fiber. Surg Gynecol Obstet 1989; 168: 323—331.

10.

Aoki H. , Kodama M. , Tani T. , Hanasawa K. Treatment of sepsis by extracorporeal elimination of endotoxin using polymyxin B-immobilized fiber. Am J Surg 1994; 167: 412—417.

11.

Tsuzuki H. , Tani T. , Ueyama H. , Kodama M. Lipopolysaccharide: neutralization by polymyxin B shuts down the signaling pathway of nuclear factor kappaB in peripheral blood mononuclear cells, even during activation. J Surg Res 2001; 100: 127—134.

12.

Wang Y. , Liu Y. , Sarker KP et al. Polymyxin B binds to anandamide and inhibits its cytotoxic effect . FEBS Lett 2000; 470: 151—155.

13.

Tani T. , Hanasawa K. , Kodama M. et al. Correlation between plasma endotoxin, plasma cytokines, and plasminogen activator inhibitor-1 activities in septic patients. World J Surg 2001; 25: 660—668.

14.

Nakamura T. , Ebihara I. , Shimada N. , Shoji H. , Koide H. Modulation of plasma metalloproteinase-9 concentrations and peripheral blood monocyte mRNA levels in patients with septic shock: effect of fiber-immobilized polymyxin B treatment. Am J Med Sci 1998; 316: 355—360.

15.

Bone RC , Sprung CL , Sibbald WJ Definitions for sepsis and organ failure. Crit Care Med 1992; 20: 724—726.

16.

Ziegler-Heitbrock H. , Fingerle G. , Strobel M. et al. The novel subset of CD14+CD16+ blood monocyte exhibits features of tissue macrophages. Eur J Immunol 1993; 23: 2053—2058.

17.

Fingerle G. , Pforte A. , Passlick B. , Blumenstein M. , Strobel M. , Ziegler-Heitbrock HW. The novel subset of CD14+CD16+ blood monocytes is expanded in sepsis patients. Blood 1993; 82: 3170—3176.

18.

Mackensen A. , Galanos C. , Wehr U. , Engelhardt R. Endotoxin tolerance: regulation of cytokine production and cellular changes in response to endotoxin application. Eur Cytokine Netw 1992; 3: 571.

19.

Frankenberger M. , Sternsdorf T. , Pechumer H. , Pforte A. , Ziegler-Heitbrock HW Differential cytokine expression in human blood monocyte subpopulations: a PCR-analysis. Blood 1996; 87: 373—377.

20.

Kodama M. , Tani T. , Hanasawa K. et al. Treatment of sepsis by plasma endotoxin removal: hemoperfusion using a polymyxin-B immobilized column. J Endotoxin Res 1997; 4: 293—300.

21.

Nakamura T. , Ebihara I. , Shimada N. , Koide H. Changes in plasma erythropoietin and interleukin-6 concentrations in patients with septic shock after hemoperfusion with polymyxin B immobilized fiber. Intensive Care Med 1998 ; 24: 1272—1276.

22.

Nemoto H. , Nakamoto H. , Okada H. et al. Newly developed immobilized polymyxin B fiber improve the survival of patients with sepsis. Blood Purif 2001; 19: 361—369.