Rat anti mouse cd103 m290

Single-cell suspensions (1 × 10⁶ cells) from lungs were blocked with 2% rat serum for 30 min at 4°C. Following blocking, cells were stained with the following antibodies: rat anti-mouse CD4 (GK1.5; BioLegend), rat anti-mouse CD8α (53-6.7; BioLegend), rat anti-mouse CD49d (R1-2, BioLegend), rat anti-mouse CD11a (M17/4; BD Biosciences, San Jose, CA), rat anti-mouse CD103 (M290; BD Biosciences), and rat anti-mouse CD69 (H1.2F3; eBioscience), to identify CD4 and CD8 T cell subsets. Antigen experienced T cells were identified via expression of surrogate markers as previously described (38 (link), 39 (link)). Briefly, CD11a^hiCD49d^pos expression was utilized to identify antigen-experienced CD4 T cells, while CD11a^hiCD8α^lo expression was utilized to quantify antigen-experienced CD8 T cells. To identify B cell subsets, cells were stained with rat anti-mouse CD19 (1D3; BD Biosciences), rat anti-mouse B220 (RA3-6B2; BioLegend), rat anti-mouse IgM (B7-6), and FITC-conjugated peanut agglutinin (PNA; Vector Laboratories, Burlingame, CA). Cells were then fixed with BD FACS™ Lysing Solution per manufacturer's instructions and resuspended in PBS. Data were acquired on a LSRII (BD Biosciences) and analyzed using FlowJo software (Tree Star, Ashland, OR).