Collagen inhibits phagocytosis of amyloid in vitro and in vivo and may act as a 'don't eat me' signal.

BACKGROUND: Systemic amyloidosis refers to a group of protein misfolding disorders characterized by the extracellular deposition of amyloid fibrils in organs and tissues. For reasons heretofore unknown, amyloid deposits are not recognized by the immune system, and progressive deposition leads to organ dysfunction.

METHODS: In vitro and in vivo phagocytosis assays were performed to elucidate the impact of collagen and other amyloid associated proteins (eg serum amyloid p component and apolipoprotein E) had on amyloid phagocytosis. Immunohistochemical and histopathological staining regimens were employed to analyze collagen-amyloid interactions and immune responses.

RESULTS: Histological analysis of amyloid-laden tissue indicated that collagen is intimately associated with amyloid deposits. We report that collagen inhibits phagocytosis of amyloid fibrils by macrophages. Treatment of 15 patient-derived amyloid extracts with collagenase significantly enhanced amyloid phagocytosis. Preclinical mouse studies indicated that collagenase treatment of amyloid extracts significantly enhanced clearance as compared to controls, coincident with increased immune cell infiltration of the subcutaneous amyloid lesion.

CONCLUSIONS: These data suggest that amyloid-associated collagen serves as a 'don't eat me' signal, thereby hindering clearance of amyloid. Targeted degradation of amyloid-associated collagen could result in innate immune cell recognition and clearance of pathologic amyloid deposits.Abbreviationsr: AL: Immunoglobulin light chain amyloidosis; ANOVA: Analysis of variance; ApoE: Apolipoprotein E; ATTR: Transthyretin amyloidosis; ATTRv: Hereditary transthyretin amyloidosis; ATTRwt: Wildtype transthyretin amyloidosis; BSA: Bovine serum albumin; DL800: Dylight 800; H&E: Haematoxylin and eosin; IHC: Immunohistochemistry; LPS: Lipopolysaccharide; MRD: Mean raw density; NU/NU mice: Athymic nude mouse; PMA: Phorbolmyristate acetate; ROI: Region of interest.