Role of CCL5 (RANTES) in Viral Lung Disease

Fiona Culley

Alasdair Pennycook

John Tregoning

Jonathan Dodd

Department of Respiratory Medicine, National Heart and Lung Institute, Imperial College London, St Mary's Campus, Norfolk Place, London, United Kingdom,^{Serono Pharmaceutical Research Institute, Geneva, Switzerland}²

^{Corresponding author. Mailing address: Department of Respiratory Medicine, Imperial College London, St Mary's Campus, Norfolk Place, London W2 1PG, United Kingdom. Phone: 44 20 7594 3854. Fax: 44 20 7262 8913. E-mail:}ku.ca.lairepmi@wahsnepo.p.

^{Present address: Division of Cell and Molecular Biology, Sir Alexander Fleming Building, Faculty of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, United Kingdom.}

^{These authors contributed equally to this work.}

^{Present address: Institute of Internal Medicine, University of Stellenbosch Medical School, Cape Town, South Africa.}

^{Present address: The Kennedy Institute, Aspenlea Road, London W6 8LH, United Kingdom.}

Received 2005 Mar 9; Accepted 2006 Jun 2.

Abstract

CCL5/RANTES is a key proinflammatory chemokine produced by virus-infected epithelial cells and present in respiratory secretions of asthmatics. To examine the role of CCL5 in viral lung disease, we measured its production during primary respiratory syncytial virus (RSV) infection and during secondary infection after sensitizing vaccination that induces Th2-mediated eosinophilia. A first peak of CCL5 mRNA and protein production was seen at 18 to 24 h of RSV infection, before significant lymphocyte recruitment occurred. Treatment in vivo with Met-RANTES (a competitive chemokine receptor blocker) throughout primary infection decreased CD4^{and CD8}^{cell recruitment and increased viral replication. In RSV-infected, sensitized mice with eosinophilic disease, CCL5 production was further augmented; Met-RANTES treatment again reduced inflammatory cell recruitment and local cytokine production. A second wave of CCL5 production occurred on day 7, attributable to newly recruited T cells. Paradoxically, mice treated with Met-RANTES during primary infection demonstrated increased cellular infiltration during reinfection. We therefore show that RSV induces CCL5 production in the lung and this causes the recruitment of RSV-specific cells, including those making additional CCL5. If this action is blocked with Met-RANTES, inflammation decreases and viral clearance is delayed. However, the exact effects of chemokine modulation depend critically on time of administration, a factor that may potentially complicate the use of chemokine blockers in inflammatory diseases.}

Abstract

Bronchiolitis resulting from respiratory syncytial virus (RSV) infection is the single major cause of infant hospitalization in the developed world (25). It is characterized by excessive cell recruitment to the lung, leading to bronchiolar obstruction and sometimes ventilatory failure (24). RSV bronchiolitis is associated with the recurrent wheezing and asthma diagnosis in later childhood (33).

CCL5 (RANTES) is a potent chemoattractant cytokine that recruits monocytes, T cells, and eosinophils, acting via the receptors CCR1, CCR3, and CCR5 (30). Infection of respiratory epithelial cells with RSV causes upregulation of CCL5 secretion (21) by NF-κB translocation (39) and by increasing the stability of CCL5 mRNA (16), as does stimulation of epithelial cells with the Th1 cytokine gamma interferon (IFN-γ) (37). Children with RSV infections have increased CCL5 protein levels in both the upper and lower airway secretions, and levels of CCL5 in upper airway secretions correlate positively with disease severity (2, 9, 11, 36). In mice, CCL5 induction by RSV infection contributes to subsequent allergic pulmonary inflammation (14). CCL5 is a key chemokine in recruitment of CD8 T cells to the lung (6) and has been implicated in classical IFN-γ dominant Th1 responses, and yet it is also involved in eosinophilic disease driven by Th2 cells (7, 8, 17, 27).

In mice, RSV infection can prime for Th1- or Th2-biased T-cell populations that control infection but also enhance inflammation upon subsequent exposure to RSV, allowing us to examine situations in which polarized cytokine responses can be achieved in the context of identical viral challenge (24). In this system, we have recently shown that the pattern of chemokine release is directly affected by priming with individual RSV proteins (4).

To further explore the timing of CCL5 production, its cellular source, and the associated pattern of pathology, we used the inhibitory CCL5 analogue Met-RANTES (28) to block CCL5 activity in a mouse model of acute viral lung disease. To further investigate the role of CCL5 in Th2-biased immunopathology, we primed mice with RSV-G prior to RSV challenge. In the first 2 days of RSV infection, CCL5 protein was abundant in bronchoalveolar lavage fluid but not in the eluted cells by intracellular staining or by enzyme-linked immunospot (ELISPOT) assay; at 7 days this situation was reversed. Therefore, CCL5 and its coligands have potent time-dependent actions in viral lung disease, both in recruitment of inflammatory cells and in controlling virus infection.

Acknowledgments

This work was supported by Wellcome Trust program grants to P.J.M.O. (54797/Z/98/Z and 071381/Z/03/Z) and by a Wellcome Trust prize studentship (055303/Z/98/Z).

Acknowledgments

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