. Author manuscript; available in PMC: 2010 Sep 1.

Published in final edited form as: J Immunol. 2009 Aug 26;183(6):3955–3962. doi: 10.4049/jimmunol.0804103

Table I.

Percentage of T cells, macrophages, and dendritic cells in MLN after EtOH intoxication and burn injury^a

Experimental Groups	T Cell (%)			Macrophage (%)(ED1⁺MHCII⁺)	Dendritic Cell (%)(OX62⁺MHCII⁺)
Experimental Groups	CD3⁺	CD3⁺CD4⁺	CD3⁺CD8⁺	Macrophage (%)(ED1⁺MHCII⁺)	Dendritic Cell (%)(OX62⁺MHCII⁺)
Saline + sham	85.21 ± 1.76	50.23 ± 3.53	29.23 ± 3.85	2.05 ± 0.21	2.27 ± 0.34
EtOH + sham	87.28 ± 2.43	56.49 ± 2.89	31.31 ± 1.35	2.07 ± 0.25	2.33 ± 0.06
Saline + burn	81.76 ± 2.23	54.53 ± 1.49	29.06 ± 1.72	1.76 ± 0.23	2.28 ± 0.16
EtOH + burn	81.97 ± 3.83	56.40 ± 3.51	28.95 ± 2.07	1.72 ± 0.18	2.00 ± 0.07

MLN harvested from rats on day 1 after injury were processed for single-cell suspension. For T cells, MLN mixed cells (1 × 10⁶/100 μl of PBS) were coincubated with Alexa Fluor 647-labeled anti-rat CD3, PE/Cy7-labeled anti-rat CD4, and PerCP-labeled anti-rat CD8. For macrophage and dendritic cells, cells were coincubated with R-PE-labeled anti-rat OX-62, Alexa Fluor 647-labeled ED1, and FITC-labeled anti-rat MHC class II. The cells were washed and analyzed using a six-color flow cytometer and FlowJo software. Cells found positive for ED1 and MHC class II were considered macrophages, whereas cells positive for OX62 and MHC class II were selected as dendritic cells. Similarly, the CD4⁺ and CD8⁺ population represents T cells that appeared positive, respectively, for CD3 and CD4 or CD3 and CD8.