Evaluation of intrahepatic macrophages in patients with non-alcoholic steatohepatitis using multispectral imaging

• Published in: The Journal of immunology (1950) 2020-05, Vol.204 (1_Supplement), p.159-159.45
• Main Authors: Saldarriaga, Omar A, Jiao, Jingjing, Beretta, Laura, Freiberg, Benjamin, Krishnan, Santhoshi, Rao, Arvind, Booth, Adam L., Stevenson, Heather L.
• Format: Article
• Language: English
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.204.Supp.159.45

Patients with non-alcoholic steatohepatitis (NASH) have different rates of fibrosis progression. We hypothesized that this is due to intrahepatic macrophages. Kupffer cells (KCs) can affect the development of fibrosis by recruiting systemic macrophages that activate stellate cells. Liver biopsies were collected from patients with NASH and either minimal (n=6) or advanced (n=5) fibrosis. We used spectral imaging to evaluate KCs (CD68+), monocyte-derived (Mac387+), pro-fibrogenic (CD163+), pro- (CD14+), and anti- inflammatory (CD16+) macrophages. NanoString evaluated gene expression profiles in the same biopsy. t-distributed stochastic neighbor embedding (t-SNE) plots and phenotype matrices were used to visualize differences between groups. Mean normalized counts from ‘cell seg’ data were used to determine the percentage of phenotypes present. Results showed that patients have variable macrophage phenotypes in the liver depending on their propensity to develop hepatic fibrosis. Patients with minimal fibrosis showed increased CD14+ macrophages and more protective gene expression profiles. Patients with advanced fibrosis had significantly increased numbers of pathogenic pro-inflammatory/pro-fibrotic macrophages (MAC387+, CD163+) and increased expression of genes associated with mixed macrophage activation (IDO1, CD86, HLADR, IL1β, TNFα, COX-2, Dectin-1, CD209, PI3K/AKT3, IL10R, TGFB1, IRF4, CLEC7A/DECTIN-1). However, when we analyzed each patient’s liver biopsy on an individual basis, subtle variations were observed between the patients in each group. Thus, the various phenotypes of macrophages present in patients with NASH is complex and is only partially associated with fibrosis stage.