Distinct Spatial Distribution of Immune Infiltration
The HALO can not only quantify the number of immune cells on a panel, but also locate their position, measure their spatial distance [20,38,39]. By this way, we can further find the relative number and location of immune cells in situ tissue of CC after pLN metastasis more visually and thereby draw the relationship of immune cells which would promote LN metastasis.
The average distance (um) of CD8 to CD56 was 8.33 ± 1.26% in the pLN-negative group, 3.86 ± 1.08% in the pLN-positive group (pLN-negative versus pLN-positive, P < 0.05) (Figure 5A). The average distance (um) of CD8 to CD68 was 75.08 ± 21.91% in the pLN-negative group, 5.56 ± 1.62% in the pLN-positive group (pLN-negative versus pLN-positive, P < 0.01) (Figure 5B). The average distance (um) of CD8 to PD-1 was 4.12 ± 0.76% in the pLN-negative group, 1.30 ± 0.08% in the pLN-positive group (pLN-negative versus pLN-positive, P < 0.001) (Figure 5C). The average distance (um) of CD8 to PD-L1 was 5.58 ± 0.81% in the pLN-negative group, 1.50 ± 0.16% in the pLN-positive group (pLN-negative versus pLN-positive, P< 0.0001) (Figure 5D). In these data, CD8+T cells were significantly close to NK cells, macrophages and tumor cells with pLN metastasis.
The average distance (um) of CD56 to PD-1 was 2.11 ± 0.24% in the pLN-negative group, 0.91 ± 0.09% in the pLN-positive group (pLN-negative versus pLN-positive, P< 0.0001) (Figure 5E). The average distance (um) of CD56 to PD-L1 was 2.57 ± 0.32% in the pLN-negative group, 1.26 ± 0.18% in the pLN-positive group (pLN-negative versus pLN-positive, P< 0.0001) (Figure 5F). Thus, NK cells were significantly close to tumor cells with pLN metastasis.
The average distance (um) of PD-1 to PD-L1 was 2.09 ± 0.10% in the pLN-negative group, 1.74 ± 0.09% in the pLN-positive group (pLN-negative versus pLN-positive, P< 0.05) (Figure 5G).