Accuri C6 Flow Cytometer System
Originally published online as doi:10.1189/jlb.0808501 on April 1, 2009

Published online before print April 1, 2009
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(Journal of Leukocyte Biology. 2009;85:1027-1035.)
© 2009 by Society for Leukocyte Biology

Effects of HIV-1 infection in vitro on transendothelial migration by monocytes and monocyte-derived macrophages

Clare L. V. Westhorpe*,{dagger}, Jingling Zhou*, Nicole L. Webster{ddagger}, Bill Kalionis§, Sharon R. Lewin*,{dagger},||, Anthony Jaworowski*,{dagger}, William A. Muller and Suzanne M. Crowe*,{dagger},1

* Centre for Virology, Macfarlane Burnet Institute for Medical Research and Public Health, Melbourne, Australia; Department of
{dagger} Medicine, Monash University, Melbourne, Australia;
{ddagger} Department of Microbiology and Immunology, Melbourne University, Melbourne, Australia;
§ Pregnancy Research Centre, Department of Perinatal Medicine, Royal Women’s Hospital and Department of Obstetrics and Gynaecology, The Royal Women’s Hospital and University of Melbourne, Carlton, Australia,
|| Infectious Diseases Unit, The Alfred Hospital, Melbourne, Australia; and
Department of Pathology, Northwestern University, Chicago, Illinois, USA

1 Correspondence: Centre for Virology, Macfarlane Burnet Institute for Medical Research and Public Health, GPO Box 2284, Melbourne 3001, Australia. E-mail: crowe{at}burnet.edu.au

ABSTRACT

Monocytes constitutively migrate from the bloodstream across the vascular endothelium for systemic immune surveillance and maintenance of macrophage populations. They also perform reverse transendothelial migration (TEM) across the endothelium, which is required for entry of tissue monocytes/macrophages into the lymphatics or back into the bloodstream. We have modeled these processes previously using HUVEC monolayers grown on three-dimensional collagen matrices. The aim of the present study was to determine whether HIV-1 infection of monocytes/macrophages in vitro affects TEM. Purified primary human monocytes and monocyte-derived macrophages (MDM) expressed important TEM proteins such as CD62L, CD18, PECAM-1, CCR2, and CCR8. Purified monocytes underwent efficient forward and reverse TEM across HUVEC, and this function was maintained by MDM after up to 15 days of culture. Monocytes exposed to HIV-1 for 2 days had unaltered forward or reverse TEM. However, HIV-1 infection of MDM for 7 days decreased reverse TEM by an average of 66.5% compared with mock-infected MDM (n=9 independent donors; P=0.004), without affecting forward TEM. Decreased reverse TEM by HIV-infected MDM required viral RT and was not a result of alterations in surface expression of CCR8 or p-glycoprotein or a general impairment in mobility, as assessed by migration toward fMLP. This study indicates that HIV-1 infection of macrophages reduces their capacity to emigrate from the subendothelial extracellular matrix in vitro, which could result in defective cell-mediated immune responses to infections and promote establishment of viral reservoirs of HIV in tissue macrophages in vivo.

Key Words: cells of macrophage lineage • CCR8 • HUVEC • innate immunity