Background
Methods
Cell Lines and Cell Culture
Tumor-Associated MDSC Generation Protocol
i. Induction
ii. MDSC Isolation
iii. Suppression Assay
Characterization of myeloid suppressor cells
i. Morphology of MDSC
ii. Flow cytometry analyses of cell phenotypes
iii. Real-time RT-PCR for gene expression of myeloid suppressor cells and tumor cell lines
iv. Measurement of tumor-derived factors by ELISA
v. Functional studies
vi. Immunohistochemistry
Statistical analysis
Results
Induction of tumor-associated human myeloid suppressor cells
Inducing Tumor Cell Line | Mean Percent Suppression | SEM | Inducing Tumor Cell Line | Mean Percent Suppression | SEM |
---|---|---|---|---|---|
Controls | Cervical/Endometrial (4/5) | ||||
T cells alone | 0.00 |
** HeLa
|
68.35
|
5.36
| |
Medium only | -2.35 | 0.86 |
** ME-180
|
75.24
|
3.83
|
Lung Fibroblasts | -1.03 | 0.96 |
** SIHA
|
54.49
|
8.66
|
Ditt Fibroblasts | -0.13 | 2.91 |
** RL95-2
|
52.11
|
3.84
|
** GM-CSF + IL-6
|
56.30
|
5.01
| SW 756 | -83.60 | 2.18 |
HNSCC (6/8)
|
Ovarian (6/9)
| ||||
** SCCL-MT1
|
91.83
|
0.82
|
** A2780
|
64.46
|
5.33
|
Irradiated
|
89.18
|
0.20
|
** ES-2
|
63.62
|
5.17
|
T cell Depl. |
81.49
|
4.98
|
** TOV-21G
|
52.86
|
11.37
|
** USC-HN2
1
|
87.97
|
ND
|
** SK-OV-3
|
51.44
|
9.81
|
** SCC-4
|
65.72
|
2.08
|
* NIHOVCAR-3
|
47.89
|
1.08
|
** CAL-27
|
66.26
|
6.21
|
* SW 626
|
46.54
|
4.07
|
** SW 451
|
59.49
|
9.59
| HOC-7 | 41.77 | 19.15 |
* FaDu
|
30.98
|
4.45
| HEY | 22.20 | 3.87 |
RPMI 2650 | 17.46 | 5.01 | Caov-3 | -146.53 | 2.69 |
SW 2224 | -13.48 | 11.21 |
Breast (0/9)
| ||
Thyroid (1/2)
| MCF-7 | 16.95 | 0.39 | ||
** SW 579
|
68.97
|
3.41
| 734B | 16.72 | 2.32 |
SW 1949 | 43.90 | 13.68 | T47D | 8.47 | 1.23 |
Brain (2/9)
| BT-474 | 0.83 | 11.53 | ||
** NU-04
|
69.41
|
4.02
| SKBR3 | -0.09 | 13.53 |
** U118MG
|
51.96
|
1.48
| MDA-MB-468 | -3.46 | 0.25 |
SW 598 | 14.29 | 4.14 | GI-101 | -6.41 | 0.92 |
A172 | 2.26 | 4.97 | SV-BR-1 | -8.00 | 1.75 |
IMR-5 | -1.23 | 3.09 | MDA231 | -16.21 | 2.60 |
IMR-32 | -3.16 | 7.48 |
Bladder (1/3)
| ||
TE 671 | -12.23 | 4.29 |
** T24
|
53.89
|
3.97
|
Y79 | -72.63 | 5.58 | SW 780 | 8.10 | 10.01 |
BM-166 | -83.22 | 0.05 | SW 733 | -54.63 | 0.45 |
Melanoma (1/3)
|
Prostate (2/3)
| ||||
** A375
|
56.16
|
0.64
|
** DU 145
|
54.73
|
2.07
|
CaCl74-36 | 17.26 | 6.83 |
* LNCaP
|
29.09
|
2.78
|
Colo 38 | 15.83 | 1.49 | PC3 | 15.12 | 9.09 |
Sarcomas (4/9)
|
Renal (3/6)
| ||||
** 4-998
|
58.31
|
0.82
|
** 786-O
|
75.91
|
6.06
|
Irradiated
|
52.10
|
0.44
|
** CAKI-1
|
64.94
|
3.70
|
T cell Depl. |
65.23
|
8.17
|
** CAKI-2
|
63.62
|
5.17
|
* Rh30
|
44.63
|
2.51
| SW 156 | 36.51 | 10.69 |
* HOS
|
42.58
|
4.86
| ACHN | 9.85 | 0.20 |
* SW 1353
|
42.22
|
4.42
| SK-NEP-1 | 0.00 | 1.82 |
HT 1080 | 19.37 | 5.92 |
Gastric (1/2)
| ||
SA-4 | 12.53 | 1.05 |
** SW 1961
|
64.55
|
3.04
|
HS 919 | 3.01 | 5.31 | KATO-III | 7.65 | 2.16 |
SW 80 | -5.00 | 3.93 |
Colorectal (5/6)
| ||
HS 913T | -56.35 | 1.45 |
** SW 732
|
69.19
|
1.29
|
Lung (4/11)
|
** DLD-1
|
65.59
|
3.19
| ||
Non-small cell (2/7)
|
** SW 608
|
53.11
|
5.15
| ||
* A427
|
27.71
|
6.87
|
** SW 707
|
52.38
|
0.64
|
* SW 1573
|
21.47
|
1.64
|
* HT-29
|
38.37
|
4.91
|
NCI-H292 | 8.23 | 2.89 | LS147T | 13.62 | 3.87 |
NCI-H1650 | 6.67 | 4.18 |
Pancreatic (3/10)
| ||
SK-MES-1 | 4.31 | 6.03 |
** SW 1990
|
78.15
|
1.21
|
NCI-H125 | 1.54 | 3.69 |
* Panc 2.03
|
22.28
|
4.37
|
NCI-H1975 | 0.28 | 2.89 |
* Panc 4.14
|
21.82
|
2.60
|
Small Cell 2/3)
| Panc 9.6.94 | 27.28 | 8.47 | ||
** NCI-H464
|
63.96
|
6.00
| Panc-1 | 7.82 | 3.69 |
* NCI-H60
|
47.79
|
7.71
| Panc 3.27 | 6.98 | 5.44 |
NCI-HUT 69C | -24.28 | 16.75 | ASPC-1 | 3.09 | 2.36 |
Mesothelioma (0/1)
| CAPAN-1 | -1.34 | 2.27 | ||
SW 1503 | 1.93 | 2.02 | Panc 2.5 | -1.79 | 6.08 |
Liver (2/5)
| MIA PaCa-2 | -4.38 | 1.94 | ||
* HA 22T
|
44.01
|
4.22
|
Epidermoid (0/1)
| ||
* HEP 3B
|
23.52
|
7.44
| A431 | -31.12 | 8.55 |
PLC | 22.05 | 5.22 | |||
MAH | 11.87 | 4.72 | |||
HEP-G2 | -2.10 | 9.40 |