Figure 5

Concurrent immunofluorescence staining of lymphocytes and LIGHT: Representative images of CD4+ staining (A,D red), LIGHT positive staining (B,E green) and co-expression (C,F yellow). DAPI was utilized as a nuclear counterstain (blue). Upper panels at low power identify an area of CD4+ TIL infiltration in a resected colorectal liver metastasis (200x). Lower panels demonstrate a single high power field in a peritumoral specimen (400x). Intratumoral CD3+ (14 ± 2.08 vs. 2.33 ± .67, p = .00006), CD4+ (8 ± 1.86 vs. 1.67 ± 0.33, p = .0009) and CD8+ (7 ± 2.33 vs. 2.0 ± 0.58, p = .029) lymphocytes were decreased in CRLM compared to lymphocytes from corresponding and equal areas of healthy control liver. Total CD3 + LIGHT + (0.33 ± 0.33 vs. 9 ± 2.65, p = .00006), CD4 + LIGHT + (0. 33 ± 0.33 vs. 5.33 ± .88, p = .0009) and CD8 + LIGHT + (1.33 ± .67 vs. 3.67 ± 1.33, p = .029) cells were decreased in tumor bearing liver compared to control. LIGHT-expressing CD3+ (6.33 ± 2.40 vs. 0.33 ± .33, p = .00006), CD4+ (5.33 ± 1.20 vs. 0.33 ± .33, p = .0009), and CD8+ (5.67 ± 2.40 vs. 1.67 ± 0.33, p = .029) lymphocytes were significantly higher in the peritumor region compared to the intratumor region (panels G-I)(n = 6).