Background
Methods
Patient Selection
-
squamous cell carcinoma
-
primary treatment by radical surgery
-
T category: pT4 or pT3 R1 or
-
N category:○ ≥ 3 pathologically involved lymph nodes or○ extracapsular growth of metastasis in N+ nodes or○ lymphangiosis carcinomatosa
n | (%) | ||
---|---|---|---|
All patients | 33 | 100 | |
Gender
| Male | 32 | (97) |
Female | 1 | (3) | |
Age (median, 53 years)
| |||
36–40 | 1 | (3) | |
41–50 | 10 | (30) | |
51–60 | 15 | (46) | |
61–70 | 7 | (21) | |
Tumor site
| Tonsils | 8 | (24) |
Oropharynx | 12 | (36) | |
Hypopharynx | 5 | (16) | |
Mouth | 8 | (24) | |
pTN category
| T1 N2 | 7 | (21) |
T1 N3 | 1 | (3) | |
T2 N0 | 1 | (3) | |
T2 N1 | 2 | (6) | |
T2 N2 | 5 | (16) | |
T3 N1 | 1 | (3) | |
T3 N2 | 5 | (16) | |
T4 N0 | 1 | (3) | |
T4 N1 | 1 | (3) | |
T4 N2 | 6 | (19) | |
TX N1 | 1 | (3) | |
TX N2 | 2 | (6) | |
AJCC Stage
| I | 0 | (0) |
II | 1 | (3) | |
III | 4 | (12) | |
IV | 28 | (84) | |
p16 Status
| positive | 10 | (31) |
negative | 22 | (69) |
Tissue miroarrays and evaluation of tumour infiltrating lymphocytes
TIL | Primary- tumor | Intraepithelial N+ node | Peritumoral N+ node | Non-metastatic node | |
---|---|---|---|---|---|
cells per 100 tumor cells | cells per image (0.087 mm2) | ||||
CD3
| |||||
Median (range) |
3.04
(0–19) |
3.52
(0.16–13.52) |
204.5
(60–341) |
183
(54–348) | |
Mean ± SE |
4.36 ± 4.92
|
4.34 ± 4.64
|
209.1 ± 68.5
|
183.4 ± 69.3
| |
CD8
| |||||
Median (range) |
2.49
(0.19–17.79) |
1.29
(0.16–16.23) |
37.5
(10–149) |
88
(25–308) | |
Mean ± SE |
3.54 ± 4.48
|
3.31 ± 4.71
|
45.8 ± 37.5
|
108.5 ± 69.8
| |
CD20
| |||||
Median (range) |
0.23
(0.11–5.26) |
1.76
(0.15–5.22) |
192
(19–631) |
148.5
(14–321) | |
Mean ± SE |
0.91 ± 1.48
|
2.53 ± 2.93
|
200.6 ± 124.3
|
148.9 ± 83.9
| |
CD68
| |||||
Median (range) |
1.02
(0.15–5.22) |
1.05
(0–4.93) |
5.5
(1–17) |
4
(1–23) | |
Mean ± SE |
1.17 ± 1.26
|
1.37 ± 1.26
|
6.5 ± 4.7
|
7 ± 6.2
| |
FoxP3
| |||||
Median (range) |
1.81
(0.15–13.0) |
2.26
(0.32–18.4) |
44
(4–134) |
49.5
(10–139) | |
Mean ± SE |
2.88 ± 2.90
|
2.96 ± 3.29
|
49.0 ± 33.2
|
53.6 ± 26.4
| |
Granzyme B
| |||||
Median (range) |
1
(0–21) |
0
(0–10) |
0
(0–54) |
3
(0–39) | |
Mean ± SE |
2.03 ± 3.93
|
1.39 ± 2.70
|
4.79 ± 11.52
|
5.27 ± 7.96
|
Statistical analysis
Results
Patterns of TIL-infiltration in different compartments
Prognostic impact of TIL in different tumour compartments
Cut-off value* | DFS | p | |
---|---|---|---|
Immune cells in primary tumor | |||
CD3 | >3.04 (n = 16) vs. ≤ 3.04 (n = 16) | 85% vs. 67% | 0.4 |
CD8 | >2.49(n = 16) vs. ≤ 2.49(n = 16) | 79% vs. 69% | 0.29 |
FoxP3 | >1.81 (n = 17) vs. ≤ 1.81(n = 15) | 86% vs. 63% | 0.27 |
CD20 | >0.23 (n = 18) vs. ≤ 0.23(n = 13) | 83% vs. 81% | 0.8 |
CD68 | >1.02 (n = 16) vs. ≤ 1.02(n = 16) | 73% vs. 75% | 0.8 |
Granzyme B | >0 (n = 18) vs. = 0(n = 15) | 82% vs. 68% | 0.53 |
Intraepithelial immune cells in metastatic lymph nodes
| |||
CD3 | >3.52 (n = 13) vs. ≤ 3.52 (n = 14) | 77% vs. 62% | 0.26 |
CD8 | >1.29 (n = 14) vs. ≤ 1.29 (n = 14) | 91% vs. 53% |
0.07
|
FoxP3 | >2.26 (n = 14) vs. ≤ 2.26 (n = 13) | 84% vs. 53% | 0.17 |
CD20 | >1.76 (n = 12) vs. ≤ 1.76 (n = 11) | 82% vs. 60% | 0.29 |
CD68 | >1.05 (n = 14) vs. ≤ 1.05 (n = 13) | 67% vs.73% | 0.9 |
Granzyme B | >0 (n = 11) vs. = 0(n = 19) | 80% vs. 68% | 0.46 |
Peritumoral immune cells in lymphatic tissue of metastatic nodes
| |||
CD3 | >204.5 (n = 13) vs. ≤ 204.5 (n = 13) | 66.7% vs. 80.8% | 0.25 |
CD8 | > 37.5 (n = 12) vs. ≤ 37.5 (n = 12) | 72.7% vs. 76.4% | 0.53 |
FoxP3 | > 44 (n = 11) vs. ≤ 44 (n = 12) | 90.9% vs. 62.3% | 0.18 |
CD20 | > 192 (n = 11) vs. ≤ 192 (n = 11) | 100% vs. 63.6% |
0.039
|
CD68 | > 5.5 (n = 13) vs. ≤ 5.5 (n = 13) | 76.9% vs. 81.5% | 0.5 |
Granzyme B | >0 (n = 15) vs. = 0 (n = 15) | 78.6% vs.67.3% | 0.65 |
Immune cells in uninvolved regional lymph nodes
| |||
CD3 | >183 (n = 15) vs. ≤ 183 (n = 16) | 75.0% vs. 73.3% | 0.675 |
CD8 | >88 (n = 15) vs. ≤ 88 (n = 16) | 84.6% vs. 65.6% | 0.337 |
FoxP3 | >49 (n = 15) vs. ≤ 49 (n = 16) | 78.6% vs. 71.1% | 0.905 |
CD20 | >147 (n = 15) vs. ≤ 147 (n = 16) | 79% vs. 68.6% | 0.779 |
CD68 | >4 (n = 12) vs. ≤ 4 (n = 17) | 75.8% vs. 68.8% | 0.424 |
Granzyme B | >0 (n = 13) vs. = 0 (n = 15) | 73.3% vs.68.8% | 0.939 |