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CCL2 (MCP-1), typically regarded as a leukocyte-derived pro-inflammatory chemokine, can also be directly upregulated by hypoxia-induced transcription. We hypothesized that such a hypoxia-induced upregulation of CCL2 is required for HPC-induced ischemic tolerance.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Methods<\/jats:title>\n            <jats:p>Adult male SW\/ND4, CCL2-null, and wild-type mice were used in these studies. Cortical CCL2\/CCR2 message, protein, and cell-type specific immunoreactivity were determined following HPC (4 h, 8% O<jats:sub>2<\/jats:sub>) or room air control (21% O<jats:sub>2<\/jats:sub>) from 6 h through 2 weeks following HPC. Circulating leukocyte subsets were determined by multi-parameter flow cytometry in na\u00efve mice and 12 h after HPC. CCL2-null and wild-type mice were exposed to HPC 2 days prior to tMCAo, with immunoneutralization of CCL2 during HPC achieved by a monoclonal CCL2 antibody.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>Cortical CCL2 mRNA and protein expression peaked at 12 h after HPC (both <jats:italic>p<\/jats:italic> &lt; 0.01), predominantly in cortical neurons, and returned to baseline by 2 days. A delayed cerebral endothelial CCL2 message expression (<jats:italic>p<\/jats:italic> &lt; 0.05) occurred 2 days after HPC. The levels of circulating monocytes (<jats:italic>p<\/jats:italic> &lt; 0.0001), T lymphocytes (<jats:italic>p<\/jats:italic> &lt; 0.0001), and granulocytes were decreased 12 h after HPC, and those of B lymphocytes were increased (<jats:italic>p<\/jats:italic> &lt; 0.0001), but the magnitude of these respective changes did not differ between wild-type and CCL2-null mice. HPC did decrease the number of circulating CCR2<jats:sup>+<\/jats:sup> monocytes (<jats:italic>p<\/jats:italic> &lt; 0.0001) in a CCL2-dependent manner, but immunohistochemical analyses at this 12 h timepoint indicated that this leukocyte subpopulation did not move into the CNS. While HPC reduced infarct volumes by 27% (<jats:italic>p<\/jats:italic> &lt; 0.01) in wild-type mice, CCL2-null mice subjected to tMCAo were not protected by HPC. Moreover, administration of a CCL2 immunoneutralizing antibody prior to HPC completely blocked (<jats:italic>p<\/jats:italic> &lt; 0.0001 vs. HPC-treated mice) the development of ischemic tolerance.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusions<\/jats:title>\n            <jats:p>The early expression of CCL2 in neurons, the delayed expression of CCL2 in cerebral endothelial cells, and CCL2-mediated actions on circulating CCR2<jats:sup>+<\/jats:sup> monocytes, appear to be required to establish ischemic tolerance to focal stroke in response to HPC, and thus represent a novel role for this chemokine in endogenous neurovascular protection.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/1742-2094-9-33","type":"journal-article","created":{"date-parts":[[2012,2,16]],"date-time":"2012-02-16T19:41:47Z","timestamp":1329421307000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":77,"title":["CCL2 upregulation triggers hypoxic preconditioning-induced protection from stroke"],"prefix":"10.1186","volume":"9","author":[{"given":"Ann M","family":"Stowe","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bradley K","family":"Wacker","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Petra D","family":"Cravens","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer L","family":"Perfater","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Min 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