Figure 5

Anti-MCAM treatment affects T cell migration to the lungs but not brain. Donor CD4+ cells were isolated from spleens and lymph nodes of C57BL/6 male mice. T cells were activated and polarized by culture for 5 days in the presence of anti-CD3 and a Th17-polarizing cocktail (3 ng/ml TGF-β, 10 ng/ml IL-6, 5 ng/ml IL-1β, 10 ng/ml IL-23, 3 μg/ml anti IL-4, 3 μg/ml anti IL-12, 3 μg/ml anti IFN-ɣ, and 3 μg/ml anti IL-2). Th17 Teffs were harvested and labeled with ¹¹¹In-oxyquinoline and 20 × 10⁶¹¹¹In-labeled Th17 Teffs were adoptively transferred to recipients treated with MPTP at dosages of 18 mg/kg every 2 hours for 4 doses. One hour prior to adoptive transfer and every 24 hours thereafter, recipients were treated ip with 10 mg/kg of either anti-MCAM or rat isotype control antibody. CT/SPECT images of each animal were acquired at 24, 48, and 72 hours post-transfer. Within tomographic images, electronic bit maps were drawn to circumscribe regions of interest that encompassed (A) brain, (B) lungs, (C) kidneys, (D) spleen, (E) cervical lymph nodes, (F) remaining lymph nodes, and included the entire body. Counts of radiolabeled T cells for each organ and entire body were determined by digital image analysis software (VIVID, GE Healthcare) and corrected for decay from the time of labeling. Counts for each organ were normalized as the percentage of total body counts at each time (A-F). Means ± SEMs of radiolabel percentages were determined for 3–4 mice/treatment group and differences of percentages between isotype antibody and anti-MCAM treatment groups were determined by Student’s t-test where p ≤0.05 was considered significant.