Possible involvement of heparin-binding protein in transfusion-related acute lung injury

Yasui, Kazuta
Furuta, Rika A.
Matsuyama, Nobuki
Fukumori, Yasuo
Kimura, Takafumi
Tani, Yoshihiko
Shibata, Hirotoshi
Hirayama, Fumiya

From the Japanese Red Cross Osaka Blood Center, Osaka, Japan.

Address reprint requests to: Fumiya Hirayama, MD, Japanese Red Cross Osaka Blood, Center, 2-4-43, Morinomiya, Joto-ku, Osaka, 536-8505 Japan; e-mail: [email protected].

ABBREVIATIONS FCM = flow cytometry; fMLP = N-formyl-L-methionyl-L-leucyl-L-phenylalanine; HBP = heparin-binding protein; T-PBS = phosphate-buffered saline containing 0.5 percent Tween 20.

Received for publication July 22, 2007; revision received November 6, 2007, and accepted November 12, 2007.

Transfusion 48(5):p 978-987, May 2008.

BACKGROUND

In antibody-mediated nonhemolytic transfusion reactions, transfusion-related acute lung injury (TRALI) tends to occur typically within 2 hours after a blood transfusion. White cell antibodies or immune complexes have been frequently shown to be associated with the syndrome, although the mechanisms by which they induce TRALI are poorly understood. The aim of this study was to characterize soluble mediators that are released from cells at an early stage after immune stimulation.

STUDY DESIGN AND METHODS

To explore the mechanism of TRALI, an in vitro whole-blood cell culture assay was established in which cells were stimulated by human antibodies and the activation of neutrophils was monitored by a cell surface marker (Mac-1) with flow cytometry and by measurement of the release of soluble factors, including perforin, interleukin-6, tumor necrosis factor-α, and heparin-binding protein (HBP) with enzyme-linked immunosorbent assays. In addition, the involvement of two neutrophil FcγRs (FcγRIIIb and FcγRIIa, also known as CD16 and CD32, respectively) was examined during antibody-induced cell activation with anti-FcγR blocking antibodies.

RESULTS

Substantial amounts of HBP were released within 30 minutes of stimulation by human antibodies, although other soluble mediators were not released within the same period. Furthermore, the release of HBP was mediated via signals through both FcγRIIIb and FcγRIIa.

CONCLUSION

HBP appears to be one of the primary effector molecules of antibody-mediated nonhemolytic transfusion reactions including TRALI.