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Cytokine Expression Profiling in Human Leukocytes after Exposure to Hypertonic and Isotonic Fluids

Gushchin, Vadim MD; Stegalkina, Svetlana MS; Alam, Hasan B. MD; Kirkpatrick, John R. MD; Rhee, Peter M. MD, MPH; Koustova, Elena PhD

The Journal of Trauma: Injury, Infection, and Critical Care: May 2002 - Volume 52 - Issue 5 - p 867-871
Annual Meeting Articles

Background  Resuscitation from hemorrhagic shock causes profound immunologic changes. The tonicity of fluids used for resuscitation clearly influences the immune response. Our study was designed to determine whether isotonic and hypertonic fluids exert their differential effects on immune response by altering the cytokine gene profile of human leukocytes. The cDNA array method was used to profile transcriptional responses after exposure to hypertonic and isotonic fluids.

Methods  Blood from seven healthy volunteers was incubated for 30 minutes with isotonic (10% dextran-40 and lactated Ringer’s [LR] solution) and hypertonic (7.5% hypertonic saline and hypertonic dextran [HTD]) fluids. The volumes of isotonic fluids used were equal to the volume of blood, whereas the volumes of hypertonic fluids were adjusted to keep the salt load identical to the LR group. The cDNA array technique was used to measure the gene expression of 23 common cytokines.

Results  Increased gene transcription of proinflammatory cytokines (interleukin [IL]-1α, IL-6, IL-10, and tumor necrosis factor-α) as well as others (IL-5, IL-7, and IL-16) was found after incubation with resuscitation fluids. Variances were noted depending on the type of fluid: HTD and LR solution did not induce expression of IL-5, and HTD also did not induce IL-1β expression. Genes encoding IL-1α, IL-6, IL-9, and tumor necrosis factor-α had low level baseline expression in leukocytes isolated from unstimulated blood, and their expression increased markedly after exposure to resuscitation fluids. The inducible transcripts included IL-1β, IL-7, IL-10, and IL-16. However, there was no difference in cytokine expression profile between isotonic and hypertonic fluids.

Conclusion  Exposure of human leukocytes to resuscitation fluids causes an increase in cytokine gene expressions compared with undiluted blood. This expression profile is largely independent of the type of fluid used.

From the Departments of Surgery, Uniformed Services University of the Health Sciences (V.G., S.S., H.B.A., P.M.R., E.K.), Bethesda, Maryland, and Department of Surgery, Washington Hospital Center (V.G., H.B.A., J.R.K., P.M.R.), Washington, D.C.

Submitted for publication October 5, 2001.

Accepted for publication November 29, 2001.

Supported by Office of Naval Research Grant MDA 905-99-1-0022.

The opinions and assertions contained herein are the private ones of the authors and are not to be construed as official or reflecting the views of the Department of Defense at large. This manuscript was prepared by United States Government employees and, therefore, cannot be copyrighted and may be copied with restriction.

This work was scheduled for presentation at the 61st Annual Meeting of the American Association for the Surgery of Trauma, which was canceled because of the terrorist attacks of September 11, 2001.

Address for reprints: Elena Koustova, PhD, Department of Surgery, Uniformed Services University of the Health Sciences, Bethesda, MD 20814; email:

© 2002 Lippincott Williams & Wilkins, Inc.