Background
Materials and methods
Study population
Immunohistochemistry
Quantification and scoring of TILs and TLS
Statistical analysis
Results
Characteristics | Total study cohort (n = 57) | |
---|---|---|
n | % | |
Median age at surgery (range) | 64 (33–83) | |
Median follow-up after metastasectomy in months (range) | 30 (4–137) | |
Sex | ||
Male | 33 | 57.9 |
Female | 24 | 42.1 |
Localization of primary tumor | ||
Colon | 32 | 56.1 |
Rectum | 25 | 43.9 |
UICC stage of primary tumor | ||
I | 4 | 7.4 |
II | 14 | 25.9 |
III | 27 | 50.0 |
IV | 9 | 16.7 |
Unknown | 3 | – |
Previous liver metastasis | ||
Yes | 17 | 29.8 |
No | 40 | 70.2 |
DFI | ||
<36 months | 38 | 66.7 |
36–60 months | 9 | 15.8 |
60 months | 10 | 17.5 |
No. of pulmonary metastases | ||
Singular | 37 | 64.9 |
Multiple | 20 | 35.1 |
Chemotherapy before metastasectomy | ||
Yes | 45 | 78.9 |
No | 12 | 21.1 |
Chemotherapy after metastasectomy | ||
Yes | 43 | 75.4 |
No | 14 | 24.6 |
Density and distribution of TILs and TLS in pulmonary metastases
TILs density | CD3+ TILs | CD8+ TILs | CD45RO+ TILs | FoxP3+ TILs | ||||
---|---|---|---|---|---|---|---|---|
n | % | n | % | n | % | n | % | |
Tumor center | ||||||||
0 | 0 | 0.0 | 2 | 3.5 | 4 | 7.0 | 8 | 14.0 |
1+ | 7 | 12.3 | 28 | 49.1 | 18 | 31.6 | 19 | 33.3 |
2+ | 21 | 36.8 | 15 | 26.3 | 28 | 49.1 | 24 | 42.1 |
3+ | 20 | 35.1 | 12 | 21.1 | 4 | 7.0 | 2 | 3.5 |
4+ | 7 | 12.3 | 0 | 0.0 | o | 0.0 | 0 | 0.0 |
Total | 55 | 96.5 | 57 | 100.0 | 57 | 100.0 | 57 | 100.0 |
Invasive margin | ||||||||
0 | 0 | 0.0 | 13 | 22.8 | 2 | 3.5 | 10 | 17.5 |
1+ | 3 | 5.3 | 21 | 36.8 | 13 | 22.8 | 26 | 45.6 |
2+ | 20 | 35.1 | 17 | 29.8 | 32 | 56.1 | 17 | 29.8 |
3+ | 22 | 38.6 | 6 | 10.5 | 7 | 12.3 | 0 | 0.0 |
4+ | 10 | 17.5 | 0 | 0.0 | 0 | 0.0 | 0 | 0.0 |
Total | 55 | 96.5 | 57 | 100.0 | 54 | 94.7 | 53 | 93.0 |
CD3+ TILs | CD8+ TILs | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
Tumor center | Invasive margin | Tumor center | Invasive margin | |||||||||
Low | High |
P
| Low | High |
P
| Low | High |
P
| Low | High |
P
| |
Age at surgery | ||||||||||||
<64 | 12 | 15 | 0.346 | 9 | 18 | 0.210 | 21 | 7 | 0.473 | 24 | 4 | 0.423 |
≥64 | 16 | 12 | 14 | 14 | 24 | 5 | 27 | 2 | ||||
Sex | ||||||||||||
Male | 15 | 17 | 0.480 | 12 | 20 | 0.444 | 24 | 9 | 0.177 | 29 | 4 | 1.000 |
Female | 13 | 10 | 11 | 12 | 21 | 3 | 22 | 2 | ||||
Localization of primary tumor | ||||||||||||
Colon | 12 | 19 |
0.040
| 13 | 18 | 0.984 | 23 | 9 | 0.138 | 28 | 4 | 0.686 |
Rectum | 16 | 8 | 10 | 14 | 22 | 3 | 23 | 2 | ||||
UICC stage of primary tumor | ||||||||||||
I | 1 | 3 | 0.815 | 0 | 4 | 0.165 | 3 | 1 | 0.511 | 3 | 1 | 0.292 |
II | 6 | 6 | 4 | 8 | 13 | 1 | 14 | 0 | ||||
III | 15 | 12 | 15 | 12 | 20 | 7 | 23 | 4 | ||||
IV | 4 | 5 | 3 | 6 | 7 | 2 | 8 | 1 | ||||
Unknown (n = 3) | ||||||||||||
Previous liver metastasis | ||||||||||||
No | 17 | 22 | 0.090 | 18 | 21 | 0.309 | 30 | 10 | 0.315 | 35 | 5 | 0.657 |
Yes | 11 | 5 | 5 | 11 | 15 | 2 | 16 | 1 | ||||
DFI | ||||||||||||
<36 months | 21 | 17 | 0.547 | 18 | 20 | 0.343 | 32 | 6 | 0.271 | 35 | 3 | 0.554 |
36–60 months | 4 | 4 | 3 | 5 | 7 | 2 | 8 | 1 | ||||
60 months | 3 | 6 | 2 | 7 | 6 | 4 | 8 | 2 | ||||
No. of pulmonary metastases | ||||||||||||
Singular | 17 | 18 | 0.646 | 13 | 22 | 0.352 | 29 | 8 | 1.000 | 32 | 5 | 0.410 |
Multiple | 11 | 9 | 10 | 10 | 16 | 4 | 19 | 1 | ||||
Lymphatic vessel invasion | ||||||||||||
No | 20 | 14 | 0.135 | 15 | 19 | 0.660 | 29 | 6 | 0.506 | 31 | 4 | 1.000 |
Yes | 8 | 13 | 8 | 13 | 16 | 6 | 20 | 2 | ||||
Chemotherapy before metastasectomy | ||||||||||||
No | 4 | 7 | 0.281 | 3 | 8 | 0.326 | 10 | 2 | 1.000 | 11 | 1 | 1.000 |
Yes | 24 | 20 | 20 | 24 | 35 | 10 | 40 | 5 |
CD45RO+ TILs | FoxP3+ TILs | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
Tumor center | Invasive margin | Tumor center | Invasive margin | |||||||||
Low | High |
P
| Low | High |
P
| Low | High |
P
| Low | High |
P
| |
Age at surgery | ||||||||||||
<64 | 10 | 17 | 0.580 | 6 | 21 | 0.362 | 15 | 12 | 0.494 | 19 | 8 | 0.697 |
≥64 | 12 | 15 | 9 | 18 | 12 | 14 | 17 | 9 | ||||
Sex | ||||||||||||
Male | 11 | 20 | 0.361 | 10 | 21 | 0.393 | 16 | 15 | 0.908 | 22 | 9 | 0.573 |
Female | 11 | 12 | 5 | 18 | 11 | 11 | 14 | 8 | ||||
Localization of primary tumor | ||||||||||||
Colon | 13 | 8 | 0.836 | 11 | 20 | 0.142 | 16 | 15 | 0.908 | 22 | 9 | 0.573 |
Rectum | 9 | 14 | 4 | 19 | 11 | 11 | 14 | 8 | ||||
UICC stage of primary tumor | ||||||||||||
I | 2 | 2 | 0.891 | 0 | 4 | 0.453 | 1 | 3 | 0.228 | 3 | 1 | 0.393 |
II | 4 | 9 | 2 | 11 | 4 | 8 | 6 | 6 | ||||
III | 11 | 15 | 8 | 18 | 17 | 9 | 20 | 6 | ||||
IV | 3 | 5 | 3 | 5 | 4 | 4 | 6 | 2 | ||||
Unknown (n = 3) | ||||||||||||
Previous liver metastasis | ||||||||||||
No | 14 | 24 | 0.369 | 9 | 29 | 0.333 | 16 | 21 | 0.088 | 23 | 14 | 0.172 |
Yes | 8 | 8 | 6 | 10 | 11 | 5 | 13 | 3 | ||||
DFI | ||||||||||||
<36 months | 17 | 19 | 0.298 | 12 | 24 | 0.082 | 19 | 18 | 0.641 | 24 | 13 | 0.547 |
36–60 months | 3 | 5 | 3 | 5 | 5 | 3 | 7 | 1 | ||||
60 months | 2 | 8 | 0 | 10 | 3 | 5 | 5 | 3 | ||||
No. of pulmonary metastases | ||||||||||||
Singular | 15 | 21 | 0.845 | 10 | 26 | 1.000 | 14 | 22 |
0.011
| 22 | 14 | 0.122 |
Multiple | 7 | 11 | 5 | 13 | 13 | 4 | 14 | 3 | ||||
Lymphatic vessel invasion | ||||||||||||
No | 14 | 19 | 0.752 | 11 | 22 | 0.253 | 17 | 15 | 0.695 | 25 | 7 |
0.050
|
Yes | 8 | 13 | 4 | 17 | 10 | 11 | 11 | 10 | ||||
Chemotherapy before metastasectomy | ||||||||||||
No | 6 | 6 | 0.517 | 3 | 9 | 1.000 | 4 | 7 | 0.277 | 7 | 4 | 0.730 |
Yes | 16 | 26 | 12 | 30 | 23 | 19 | 29 | 13 |
Correlation of TILs and TLS in pulmonary metastases and corresponding primary CRC
Impact of TILs and TLS in PM on outcome parameters
Recurrence-free survival | Overall survival | |||||||
---|---|---|---|---|---|---|---|---|
Univariate analysis (log-rank) | Univariate analysis (log-rank) | Multivariate Cox regression | ||||||
Months | HR (95 % CI) |
P-value | Months | HR (95 % CI) |
P-value | HR (95 % CI) |
P-value | |
Sex | ||||||||
Male | 21 | 0.65 (0.33–1.19) | 0.516 | 54 | 1.30 (0.59–2.86) | 0.516 | – | – |
Female | 13.5 | 101 | – | – | ||||
Age (years) | ||||||||
<64 | 17 | 1.01 (0.54–1.88) | 0.974 | 52 | 1.09 (0.50–2.40) | 0.831 | – | – |
≥64 | 15 | 65 | – | – | ||||
Location | ||||||||
Colon | 19 | 0.97 (0.52–1.80) | 0.917 | 65 | 0.75 (0.34–1.62) | 0.465 | – | – |
Rectum | 11 | 36 | – | – | ||||
UICC stage | ||||||||
I + II | 17 | 1.14 (0.59–2.24) | 0.683 | 36 | 1.95 (0.88–4.85) | 0.099 | – | – |
III + IV | 15 | NR | – | – | ||||
Unknown | 3 | – | – | |||||
Chemotherapy before metastasectomy | ||||||||
Yes | 15 | 1.19 (0.57–2.49) | 0.648 | 52 | 0.81 (0.30–2.09) | 0.644 | – | – |
No | 22 | 65 | – | – | ||||
Chemotherapy after metastasectomy | ||||||||
Yes | 15 | 1.34 (0.65–2.72) | 0.443 | 39 | 2.08 (0.71–4.74) | 0.214 | – | – |
No | 21 | NR | – | – | ||||
Previous liver metastasis | ||||||||
Yes | 9 | 1.99 (1.11–4.81) |
0.029
| 36 | 1.39 (0.61–3.44) | 0.414 | – | – |
No | 21 | 54 | – | – | ||||
DFI | ||||||||
<36 | 17 | 1.20 (0.62–2.31) | 0.600 | 52 | 1.15 (0.51–2.62) | 0.741 | – | – |
≥36 | 17 | 65 | – | – | ||||
No. of nodules | ||||||||
Singular | 17 | 0.93 (0.48–1.78) | 0.811 | 54 | 0.85 (0.37–1.89) | 0.681 | – | – |
Multiple | 16 | 36 | – | – | ||||
CD3+ TILs TC | ||||||||
High | 21 | 0.66 (0.35–1.23) | 0.199 | 54 | 1.10 (0.48–2.53) | 0.820 | – | – |
Low | 15 | 52 | – | – | ||||
CD3+ TILs IM | ||||||||
High | 16 | 1.46 (0.77–2.78) | 0.254 | 54 | 1.68 (0.76–3.89) | 0.213 | – | – |
Low | 19 | 101 | – | – | ||||
CD8+ TILs TC | ||||||||
High | 25 | 0.57 (0.29–1.24) | 0.181 | 65 | 0.36 (0.17–1.27) | 0.143 | – | – |
Low | 15 | 39 | – | – | ||||
CD8+ TILs IM | ||||||||
High | 43 | 0.48 (0.23–1.34) | 0.202 | NR | 0.00 (0.09–1.04) | 0.064 | – | – |
Low | 15 | 39 | – | – | ||||
CD45+ TILs TC | ||||||||
High | 17 | 0.88 (0.45–1.66) | 0.676 | 65 | 0.78 (0.34–1.69) | 0.515 | – | – |
Low | 15 | 36 | – | – | ||||
CD45+ TILs IM | ||||||||
High | 17 | 0.91 (0.44–1.84) | 0.984 | 54 | 0.79 (0.32–1.78) | 0.451 | – | – |
Low | 17 | 36 | – | – | ||||
FoxP3+ TILs TC | ||||||||
High | 17 | 0.99 (0.52–1.89) | 0.982 | 54 | 0.98 (0.43–2.27) | 0.969 | – | – |
Low | 15 | 52 | – | – | ||||
FoxP3+ TILs IM | ||||||||
High | 15 | 1.50 (0.79–3.16) | 0.211 | 35 | 2.40 (1.11–6.96) |
0.031
| 0.61 (0.23–1.61) | 0.319 |
Low | 17 | 65 | ||||||
TLS | ||||||||
Present | 17 | 1.00 (0.48–2.10) | 0.999 | 52 | 0.65 (0.26–1.71) | 0.413 | – | – |
Not present | 15 | NR | – | – | ||||
CD3+ TLS | ||||||||
High | 21 | 1.85 (0.36–7.72) | 0.525 | 65 | 0.34 (0.02–1.70) | 0.136 | – | – |
Low | 19 | 52 | – | – | ||||
CD8+ TLS | ||||||||
High | 21 | 0.58 (0.27–1.18) | 0.140 | NR | 0.30 (0.14–0.79) |
0.016
| 4.39 (1.25–15.32) |
0.020
|
Low | 11 | 35 | ||||||
CD45+ TLS | ||||||||
High | 17 | 1.05 (0.40–2.80) | 0.918 | 65 | 1.03 (0.33–3.25) | 0.949 | – | – |
Low | 17 | 30 | – | – | ||||
FoxP3+ TLS | ||||||||
High | 11 | 2.42 (0.96–5.59) | 0.069 | 39 | 1.50 (0.54–4.45) | 0.439 | – | – |
Low | 21 | 101 | – | – | ||||
CD8/FoxP3-ratio IM | ||||||||
Low | 15 | 1 | 29 | 1 | – | – | ||
Equal | 17 | 0.65 (0.30–1.26) | 0.197 | 65 | 0.39 (0.13–0.83) |
0.021
| – | – |
High | 26 | 0.32 (0.12–1.03) | 0.080 | NR | n/a |
0.042
| – | – |
CD8/FoxP3-ratio TLS | ||||||||
Low | 11 | 1 | 30 | 1 | – | – | ||
Equal | 14 | 0.89 (0.32–2.37) | 0.804 | 101 | 2.13 (0.71–7.60) | 0.191 | – | – |
High | NR | 3.65 (1.02–10.96) | 0.061 | NR | 6.99 (1.28–20.48) |
0.027
| – | – |