Background
Material and methods
Study design, patient and tumor characteristics
Total | 87 |
Age | 53 years (range: 28–83) |
Menopausal status | |
pre-menopausal | 43 (50%) |
peri-menopausal | 6 (7%) |
post-menopausal | 38 (44%) |
T stadium pre-NACT | |
cT1a-cT1c | 20 (23%) |
cT2 | 51 (59%) |
>cT2 | 14 (16%) |
Unknown | 2 (2%) |
T stadium post-NACT | |
ypT0, ypTis | 18 (21%) |
ypT1a | 11 (13%) |
ypT1b, ypT1c | 18 (21%) |
ypT2 | 26 (30%) |
>ypT2 | 10 (12%) |
Unknown | 4 (5%) |
N stadium pre-NACT | |
cN0 | 41 (47%) |
cN1 | 40 (46%) |
cN2, cN3 | 5 (6%) |
Unknown | 1 (1%) |
N stadium post-NACT | |
yN0 | 47 (54%) |
yN1 | 29 (33%) |
yN2/N3 | 8 (9%) |
Unknown | 3 (4%) |
Tumor type | |
Ductal invasive | 60 (69%) |
Lobular invasive | 11 (13%) |
Other | 14 (16%) |
Unknown | 2 (2%) |
Tumor grade | |
G1 | 4 (5%) |
G2 | 40 (46%) |
G3 | 42 (48%) |
Unknown | 1 (1%) |
Estrogen receptor (IHC)a | |
Positive | 68 (78%) |
Negative | 19 (22%) |
Progesterone receptor (IHC)a | |
Positive | 58 (67%) |
Negative | 29 (33%) |
HER2 a | |
Positive | 23 (26%) |
Negative | 64 (74%) |
Tumor subtype | |
ER-, PR-, HER2- | 14 (16%) |
ER-, PR-, HER2+ | 4 (5%) |
ER+/PR+, HER2- | 50 (58%) |
ER+, PR+, HER2+ | 19 (22%) |
NACT regimen | |
CTX | 53 (61%) |
CTX + Trastuzumab | 14 (16%) |
CTX + Avastin | 4 (5%) |
CTX + Lapatinib + Trastuzumab | 6 (7%) |
HTX | 9 (10%) |
Unknown | 1 (1%) |
Trastuzumab treatment | |
Yes | 20 (23%) |
No | 66 (76%) |
Unknown | 1 (1%) |
Clinical response | |
pCR | 15 (17%) |
pPR | 62 (71%) |
pNC | 6 (7%) |
Unknown | 4 (5%) |
Histological tumor regression gradeb | |
0 | 6 (7%) |
1 | 32 (37%) |
2 | 22 (25%) |
3 | 5 (6%) |
4 | 12 (14%) |
Unknown | 10 (12%) |
Local treatment | |
Mastectomy | 56 (64%) |
Breast conservation surgery | 26 (30%) |
Unknown | 5 (6%) |
DTC positive | |
pre-NACT | 16 (18%) |
post-NACT | 13 (15%) |
Collection and analysis of DTCs from the bone marrow
Pathology assessment and immunohistochemistry
Assessment of tumor-infiltrating immune cells
Statistical analysis
Results
Frequency distribution of total TILs and immune cell subtypes in pre-NACT core biopsies
Differential density distribution of total TILs and TIL subtypes at the ITF and in the TC
Interrelationship between TILs and TIL subtype infiltration at the ITF and in the TC
A: Correlation TILs ITF-ITF
| |||||||
ITF
| |||||||
ITF
|
TILs
|
CD3
|
CD20
|
CD68
|
CD4
|
CD8
| |
TILs
| 7.92 E-20 (0.83) | 3.50 E-7 (0.68) | 0.2508 (0.32) | 0.0224 (0.54) | 0.0002 (0.51) | ||
CD3
| 7.92 E-20 (0.83) | 1.39 E-5 (0.64) | 1 (0.23) | 5.35 E-5 (0.63) | 0.0054 (0.57) | ||
CD20
| 3.50 E-7 (0.68) | 1.39 E-5 (0.64) | 1 (−0.02) | 1 (0.26) | 0.0178 (0.40) | ||
CD68
| 0.2508 (0.32) | 1 (0.23) | 1 (−0.02) | 0.3366 (0.32) | 1 (0.01) | ||
CD4
| 0.0224 (0.54) | 5.35 E-5 (0.63) | 1 (0.26) | 0.3366 (0.32) | 0.0112 (0.42) | ||
CD8
| 0.0002 (0.51) | 0.0054 (0.57) | 0.0178 (0.40) | 1 (0.01) | 0.0112 (0.42) | ||
B: Correlation TILs TC-TC
| |||||||
TC
| |||||||
TC
|
TILs
|
CD3
|
CD20
|
CD68
|
CD4
|
CD8
| |
TILs
| 6.6 E-17 (0.78) | 0.0018 (0.44) | 0.0011 (0.45) | 0.0086 (0.4) | 5.08 E-5 (0.61) | ||
CD3
| 6.6 E-17 (0.78) | 7.26 E-5 (0.5) | 1 (0.1) | 0.0290 (0.37) | 2.05 E-7 (0.67) | ||
CD20
| 0.0018 (0.44) | 7.26 E-5 (0.5) | 1 (−0.02) | 1 (0.21) | 0.0051 (0.55) | ||
CD68
| 0.0011 (0.45) | 1 (0.1) | 1 (−0.02) | 0.099 (0.34) | 1 (0.1) | ||
CD4
| 0.0086 (0.4) | 0.0290 (0.37) | 1 (0.21) | 0.099 (0.34) | 0.0568 (0.36) | ||
CD8
| 5.08 E-5 (0.61) | 2.05 E-7 (0.67) | 0.0051 (0.55) | 1 (0.1) | 0.0568 (0.36) | ||
C: Correlation TILs ITF-TC
| |||||||
ITF
| |||||||
TC
|
TILs
|
CD3
|
CD20
|
CD68
|
CD4
|
CD8
| |
TILs
| 0.0005 (0.48) | 0.0003 (0.49) | 0.0251 (0.39) | 0.924 (0.27) | 0.0023 (0.45) | 0.1254 (0.35) | |
CD3
| 0.0013 (0.46) | 0.0005 (0.49) | 0.0013 (0.46) | 1 (0.08) | 0.0011 (0.48) | 0.0086 (0.42) | |
CD20
| 1 (0.12) | 1 (0.08) | 1 (0.23) | 1 (−0.19) | 1 (0.08) | 1 (0.08) | |
CD68
| 1 (0.22) | 1 (0.23) | 1 (0.03) | 0.0026 (0.45) | 0.0653 (0.37) | 1 (0.03) | |
CD4
| 1 (0.1) | 1 (0.16) | 1 (0.05) | 1 (0.25) | 0.0012 (0.47) | 1 (0.12) | |
CD8
| 1 (0.2) | 1 (0.24) | 0.495 (0.30) | 1 (−0.09) | 1 (0.21) | 1 (0.2) |
Association of TILs and TIL subtypes at the ITF and in the TC with clinicopathological parameters
p-value | |||
---|---|---|---|
Clinical parameter | TIL subtype | ITF | TC |
Tumor grade | TILs | 0.047a | 0.047a |
CD3 | 0.0407a | ||
cT stadium (pre-NACT) | TILs | 0.004d | |
CD3 | 0.004d | ||
CD4 | 0.0118b | ||
ypT stadium (post-NACT) | TILs | 0.0069c | 0.0014c |
CD3 | 0.0010c | 0.0069c | |
CD4 | 0.0014c | ||
CD8 | 0.0197c | ||
CD20 | 0.0151c | ||
IS CD3/CD8 | 0.0254c | ||
IS CD3/CD20 | 0.0144c | ||
clinical response | TILs | 0.0204b | |
CD3 | 0.0235b | ||
histological regression grade | TILs | 0.0007c | |
CD3 | 0.0308c | 0.0002c | |
CD4 | 0.037c | ||
CD8 | 0.0308c | 0.0008c | |
CD20 | 0.023c | 0.0196c | |
IS CD3/CD8 | 0.0093c | ||
IS CD3/CD20 | 0.0481c | ||
DTC presence post-NACT | CD4 | 0.0296c |