Morgan-Bathke Maria, Harteneck Debra, Jaeger Philippa, Sondergaard Esben, Karwoski Ron, Espinosa De Ycaza Ana, Carranza-Leon B Gisella, Faubion William A, Oliveira Andre M, Jensen Michael D
Endocrine Research Unit, Mayo Clinic, Rochester, Minnesota, USA.
Department of Endocrinology and Internal Medicine, Aarhus University Hospital, Aarhus, Denmark.
Obesity (Silver Spring). 2017 Dec;25(12):2100-2107. doi: 10.1002/oby.22012. Epub 2017 Oct 6.
The relationship between inflammation, obesity, and adverse metabolic conditions is associated with adipose tissue macrophages (ATM). This study compared the measurements of human ATM using flow cytometry, immunohistochemistry (IHC), and real-time polymerase chain reaction (RT-PCR) of ATM markers.
A new software program (AMCounter) was evaluated to help measure ATM using IHC, and this was compared to flow cytometry and RT-PCR.
IHC had good intraindividual reproducibility for total (CD68), proinflammatory (CD14), and anti-inflammatory (CD206) ATM. The AMCounter improved interreader agreement and was more time efficient. Flow cytometry had acceptable intraindividual reproducibility for the percentage of CD68 cells that were CD14 or CD206 , but not for ATMs per gram of tissue. ATMs per gram of tissue was much greater using IHC than flow cytometry. The flow cytometry and IHC measures of ATM from the same biopsies were not correlated. There were statistically significant correlations between RT-PCR CD68 and IHC CD68, CD14, and CD206 ATMs per 100 adipocytes. Also of interest were statistically significant correlations between RT-PCR CD68 and IHC CD68, CD14, and adipose flow cytometry measures of CD68 , CD68 /CD14 , and CD68 /CD206 ATMs per gram of tissue.
The AMCounter software helps provide reproducible and efficient measures of IHC ATMs. Flow cytometry, IHC, and RT-PCR measures of adipose inflammation provide somewhat different information.
炎症、肥胖与不良代谢状况之间的关系与脂肪组织巨噬细胞（ATM）有关。本研究比较了使用流式细胞术、免疫组织化学（IHC）和ATM标志物的实时聚合酶链反应（RT-PCR）对人ATM的测量结果。
评估了一种新的软件程序（AMCounter），以帮助使用IHC测量ATM，并将其与流式细胞术和RT-PCR进行比较。
IHC对总（CD68）、促炎（CD14）和抗炎（CD206）ATM具有良好的个体内重复性。AMCounter提高了阅片者间的一致性，且更节省时间。流式细胞术对CD14或CD206阳性的CD68细胞百分比具有可接受的个体内重复性，但对每克组织中的ATM则不然。使用IHC测量的每克组织中的ATM比流式细胞术多得多。来自相同活检组织的ATM的流式细胞术和IHC测量结果不相关。RT-PCR CD68与每100个脂肪细胞中的IHC CD68、CD14和CD206 ATM之间存在统计学显著相关性。同样有趣的是，RT-PCR CD68与每克组织中IHC CD68、CD14以及脂肪流式细胞术测量的CD68、CD68/CD14和CD68/CD206 ATM之间存在统计学显著相关性。
AMCounter软件有助于提供可重复且高效的IHC ATM测量方法。脂肪炎症的流式细胞术、IHC和RT-PCR测量提供了略有不同的信息。
