Journal of Leukocyte Biology Myeloid cells, immune suppression, tumor immunology
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Originally published online as doi:10.1189/jlb.0904494 on June 16, 2005 Originally published online as doi:10.1189/jlb.0904494 on April 4, 2005

Published online before print April 4, 2005
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(Journal of Leukocyte Biology. 2005;78:51-61.)
© 2005 by Society for Leukocyte Biology

Human neutrophil gene expression profiling following xenogeneic encounter with porcine aortic endothelial cells: the occult role of neutrophils in xenograft rejection revealed

F. Al-Mohanna1, S. Saleh, R. S. Parhar, K. Khabar and K. Collison

Biological & Medical Research Department, King Faisal Specialist Hospital and Research Centre, Riyadh, Saudi Arabia

1 Correspondence: Biological & Medical Research, MBC 03, King Faisal Specialist Hospital & Research Centre, P.O. Box 3354, Riyadh 11211, Saudi Arabia. E-mail: futwan{at}kfshrc.edu.sa

The role of innate immune cells in the recognition and activation of xenogeneic endothelium has always been considered secondary to the initial insult of xenoreactive natural antibodies (XNA) and complement. It was argued, however, that innate immune cells are capable of recognizing and activating xenogeneic endothelium in the absence XNA and complement. Here, we show that porcine aortic endothelial cells (PAECs) activate human neutrophils directly. This contact-dependent activation causes a transient calcium rise leading to increased reactive oxygen metabolite (ROM) production. Neutrophil gene-expression profiling using an adenylate uridylate-rich element-based microarray revealed a dramatic change in the neutrophil gene profiles upon exposure to PAECs. The PAEC-dependent neutrophil transcriptional activity was further confirmed by real-time polymerase chain reaction, which revealed a rapid increase in the mRNA message of a number of inflammatory cytokines. The activation of human neutrophils by PAECs was independent of galactose {alpha}1,3-galactose (Gal{alpha}1,3-gal) structures, as inclusion of saturating concentrations of anti-Gal{alpha}1,3-gal l antibodies had no significant effect. Furthermore, this activation was inhibited in the presence of the calcium chelator 1,2-bis(O-aminophenyl-ethane-ethane)-N,N,N',N'-tetraacetic acid-acetoxymethyl ester and the ROM inhibitor diphelylene iodonium. Our data illustrate the direct activation of innate immune cells by PAECs in the absence of XNA and complement and suggest alternative recognition sites between PAECs and human innate immune cells.

Key Words: Gal{alpha}1 • 3-gal • calcium • ROM • PAECs




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