Molecular Imaging of Folate Receptor Beta Positive Macrophages During Acute Lung Inflammation.

Characterization of markers that identify activated macrophages could advance understanding of inflammatory lung diseases and facilitate development of novel methodologies for monitoring disease activity. We investigated whether folate receptor beta (FRβ) expression could be used to identify and quantify activated macrophages in the lungs during acute inflammation induced by E. coli lipopolysaccharide (LPS). We found FRβ expression was markedly increased in lung macrophages at 48 hours after intratracheal (IT) LPS. In vivo molecular imaging with a fluorescent probe (Cy5-PEG-folate) showed that the fluorescence signal over the chest peaked at 48 hours after IT LPS and was markedly attenuated after depletion of macrophages. Using flow cytometry, we identified the cells responsible for uptake of Cy5-conjugated folate as FRβ+ interstitial macrophages and pulmonary monocytes, which co-expressed markers associated with an M1 pro-inflammatory macrophage phenotype. These findings were confirmed using a second model of acute lung inflammation generated by inducible transgenic expression of an NF-κB activator in airway epithelium. Using CCR2 deficient mice, we found that FRβ+ macrophage/monocyte recruitment was dependent on the MCP-1/CCR2 pathway. Together, our results demonstrate that folate-based molecular imaging can be used as a noninvasive approach to detect classically activated monocytes/macrophages recruited to the lungs during acute inflammation.