Detailed Analysis of Phenotypes of Macrophages Infiltrating Glomeruli in Rat Anti-Thy1 Nephritis

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Abstract
A phenotypic analysis of infiltrating macrophages in rat anti-Thy1 glomerulonephntis induced by monoclonal antibody (mAb) 1-22-3 was carried out using recently reported macrophage-specific mAbs. This was combined with a more detailed quantitative analysis, counting positive cells in isolated glomeruli, to obtain more information on the roles played by macrophages in glomerulonephritis. In normal glomeruli a small number of ED1- or OX-3(anti-Ia)-positive cells but almost no ED2-, TRPM-3- or Mar-3-positive cells were observed. ED1-positive cells increased from 2 h and peaked between days 3 and 7 after mAb injection. TRPM-3-positive cells increased from day 3 and peaked on day 7, later than ED1. The numbers of OX-3-positive cells changed in parallel with those of ED1-positive cells. Mar-3, which stained blood monocytes and ED2, which is an indicator of tissue-fixed resident macrophages, did not react with glomerular infiltrating macrophages. In a double staining study, about 40% of ED1- or OX-3-positive cells costained with TRPM-3 on day 3 and the percentage increased on day 7, but hardly any cells were positive for TRPM-3 alone. This results in two different phenotypes (ED1+, ED2-, OX-3+, Mar-3-, TRPM-3- and ED1+, ED2-, OX-3+, Mar-3-, TRPM-3+) of infiltrating macrophages. We conclude that in rat anti-Thyl glomerulonephntis, monocytes/ macrophages may infiltrate the mesangium, rapidly changing their phenotype (Mar-3+ to Mar-3-) and resulting in a gradual shift to TRPM-3-positive, activated macrophages.

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