Transfer of cytokine-inducing bacterial products across hemodialyzer membranes in the presence of plasma or whole blood

B. J.G. Pereira, S. Sundaram, T. W. Barrett, N. K. Butt, R. Porat, A. J. King, C. A. Dinarello

Research output: Contribution to journalArticlepeer-review

Abstract

Cytokine production by peripheral blood mononuclear cells (PBMC) is a sensitive indicator of cytokine-inducing substances which may cross from contaminated dialysate into the blood compartment. The objective of this study was to compare the transfer of cytokine-inducing substances from dialysate contaminated with a culture filtrate from Pseudomonas aeruginosa across dialyzers with low (hemophan) or intermediate ultrafiltration coefficients (modified cellulose triacetate, CTA), under conditions where either 10% plasma or whole blood was circulated in the blood compartment. Eight paired experiments of in vitro dialysis were carried out at 37°C using a countercurrent recirculating loop dialysis circuit with either a new CTA or hemophan dialyzer. 10% plasma in standard tissue culture medium was circulated through the blood compartment and bicarbonate dialysate was circulated in the dialysate compartment. The dialysate was challenged sequentially by log-fold dilutions (102, 103 or 104) of a Ps. aeruginosa culture filtrate. Samples were drawn from the blood compartment 5 and 15 minutes after each challenge and incubated with suspensions of PBMC in the absence or presence of polymyxin B, in order to block endotoxin. After 24 h at 37°C, total interleukin-1α (IL-1α) was measured by RIA. Although the dialysate contained potent cytokine-inducing substances, there was no significant IL-1α, production by PBMC incubated with the plasma mixture from the blood compartment in the majority of experiments with both dialyzers and with each of the three dilutions of the bacterial challenge. Eight experiments were also performed with CTA dialzyers using heparinized whole blood in the blood compartment. Samples of whole blood and dialysate were drawn at baseline, after one hour of dialysis with uncontaminated dialysate and 15 minutes and three hours after dialysis with dialysate contaminated with Ps. aeruginosa filtrate. There was no significant IL-1α production by PBMC isolated from the whole blood 1 h after dialysis with uncontaminated dialysate, and 15 min and 2 h after adding the Ps. aeruginosa filtrate to the dialysate side. In contrast, production of IL-1α by PBMC from the same donors incubated with samples from the dialysate were 263 ± 50, 1074 ± 306, 2333 ± 774 and 2602 ± 702 pg/2.5 x 106 PBMC, respectively at the same four time points. These data suggest that although the Ps. aeruginosa culture filtrate present in the dialysate was a potent inducer of IL-1α, neither dialyzer permitted transfer of cytokine inducing substances from the dialysate into the blood compartment.

Original languageEnglish
Pages (from-to)394-401
Number of pages8
JournalClinical Nephrology
Volume46
Issue number6
StatePublished - 1996
Externally publishedYes

Keywords

  • Cytokine-inducing bacterial products
  • Hemodialyzer membranes
  • Pseudomonas aeruginosa

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