Anti cd8 bs 0648 r

For histological analysis, the kidneys were fixed in 20% buffered formalin and embedded in paraffin. Tissue samples were stained with hematoxylin/eosin (HE; Merck, Darmstadt, Germany) and periodic-acid-Schiff (PAS; Roth, Karlsruhe, Germany) for light microscopy. For detection of infiltrating T cells and macrophages, tissues were incubated with CD3 (anti-CD3 #ab16669, Abcam), CD4 (anti-CD4-Aff–Purified #AP20210PU-N, Acris, Herford, Germany), CD8 (anti-CD8 #bs-0648 R, Bioss Antibodies Inc., Woburn, MA, USA), forkhead box P3 (anti-FoxP3 #ab54501, Abcam), and MAC2 (anti-MAC2 #CL8942AP, Cedarlane, Burlington, Ontario, Canada) antibodies. To verify that CD4+, CD8+, and FoxP3+ cells were T cells, we performed a double staining with CD3 (anti-CD3 #ab5690 and anti-FoxP3 #ab54502, Abcam; anti-CD4 #14-9766, eBioscience, San Diego, CA, USA; anti-CD8 #bs-0648 R, Bioss Antibodies Inc., Woburn, MA, USA). Additionally, apoptosis was detected by means of a terminal deoxynucleotidyl transferase deoxyuridine triphosphate nucleotide (dUTP) nick end labeling (TUNEL) assay (TUNEL #G7130, Promega, Mannheim, Germany). In each kidney, the number of infiltrating CD3+ T cells, CD4+ T cells, CD8+ T cells, FoxP3+ T cells, and MAC2+ macrophages as well as apoptotic cells was counted in 20 randomly selected interstitial fields of view at 40× magnification.