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Mouse model to study pulmonary intravascular macrophage recruitment and lung inflammation in acute necrotizing pancreatitis

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Abstract

Patients suffering from severe acute pancreatitis (AP) can develop acute lung injury (ALI) with poor outcomes and the mechanisms involved remain incompletely understood. Pulmonary intravascular macrophages (PIMs), which are credited as promoters of ALI, are not constitutively present in humans and rodents; however, there is evidence of PIM recruitment in rodents during some pathological conditions, such as hepatic diseases. Therefore, this study assesses PIM recruitment in the lungs of a mouse model of acute necrotizing pancreatitis (ANP) induced with l-arginine monohydrochloride. Mice were euthanized after 24 h, 72 h and 120 h. Control mice received sham injections of saline. Pancreatic histopathological grading and plasma amylase were used to confirm the development of ANP in l-arginine-treated mice. Histopathological grading of lungs from the ANP mice at 72 h showed increased mononuclear phagocytes in alveolar septa, compared to that from the controls. Lungs from the ANP mice also showed increased numbers of CD68-immunopositive alveolar septal macrophages, suggestive of PIM recruitment, compared to those from the controls. Lungs from the ANP mice showed increased expression of IL-6, IL-10, monocyte chemoattractant protein 1 (MCP-1) and von Willebrand factor compared to those from the controls. The recruitment of CD68-positive septal macrophages was not observed in MCP-1 knockout mice with ANP at 72 h when compared to C57BL/6 wild-type mice. Taken together, we developed a mouse model of PIM recruitment dependent on MCP-1 that allows us to explore their roles in ANP-associated ALI.

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Funding

The research project was supported through a Discovery Grant from the Natural Sciences and Engineering Research Council of Canada. Dr. Vrolyk was supported by an Interprovincial Graduate Scholarship from the Western College of Veterinary Medicine, University of Saskatchewan.

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Correspondence to Baljit Singh.

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Figure S1

Electron microscopy showing pulmonary intravascular macrophages (PIM) located within the lumina of an alveolar septa capillary, which is lined by endothelial cells (En). Ep: epithelium; AS: alveolar space. The scale bar is 5 μm. (PNG 2638 kb)

High Resolution Image (TIF 2227 kb)

Figure S2

Bronchoalveolar lavage white blood cell count in control mice compared to l-arginine-treated mice showing no statistical difference between groups (p = 0.08). (PNG 805 kb)

High Resolution Image (TIF 2282 kb)

Figure S3

Immunohistochemistry against vWF. MCP-1 KO control mice (a) and MCP-1 KO mice treated with l-arginine (b) only present labeling for vWF in endothelial cells of medium-large blood vessels (arrows). All scale bars are 100 μm. (PNG 67 kb)

High Resolution Image (TIF 4866 kb)

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Vrolyk, V., Schneberger, D., Le, K. et al. Mouse model to study pulmonary intravascular macrophage recruitment and lung inflammation in acute necrotizing pancreatitis. Cell Tissue Res 378, 97–111 (2019). https://doi.org/10.1007/s00441-019-03023-9

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  • DOI: https://doi.org/10.1007/s00441-019-03023-9

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