Role of Dendritic Cells in a Mouse Model
of Lung Carcinoma
Rosalinda Sorrentino and Aldo Pinto
Department of Pharmacy (DIFARMA), University of
Salerno, Fisciano, 84084, Italy
Carcinogen, Genetic alterations, etc.
Lung cancer is one of the leading
causes of death worldwide.
Epithelial Cells
T cell
Macrophage
Dendritic Cell
Lung Injury/Chronic Inflammation
Chemotherapy:
Doxorubicin , Oxaliplatinum, Cisplatinum, Paclitaxel, Mitomycin C, etc.
Gefitinib, Crizotinib
…..however, low survival percentage is still counted…..
Aim: Can we pharmacologically manipulate DCs to counter Tumor Cell Growth???
Melief CJ. Nature 2005; 437, 41-42
IMMUNOEDITING STUDIES: TO EVALUATE THE ROLE OF DCS ON TUMOR OUTGROWTH
1. Tumor Cell Implantation
LLC1 (2x105 cells)
B16-F10 (1x105 cells)
Day 13
6-8 weeks old
Day 17
10 Days later
Sacrifice
i.p. Treatment
with TLR ligands
i.v. Injection
•H&E and immunohistochemical Analyses
•ELISAs on BAL and Lung Homogenates
2. Adoptive Transfer of Bone Marrow/Spleen/Lung-Derived Cells •Flow Cytometry Analyses
•Sorting Cells
7-Day Cell Culture
Naive Mice
i.v.
A)
BMDCs or
Spleen-derived
cells
B)
TLR ligand
Treatment O/N
Tumor-implanted Mice
Lung-derived DCs
Tumor-implanted Mice
Day 13
Day 10
Day 17
Sacrifice
Naive Mice
Sacrifice
24-72hours later
Day 10
i.v.
IMMUNOEDITING STUDIES: TO
EVALUATE THE ROLE OF
Week 1
i.t. NMU
50µg/mouse
6-12 weeks old
Week 2
i.t. NMU
10µg/mouse
DCS
ON
TUMOR ESTABLISHMENT/PROGRESSION
Sacrifice at:
3-7-30 days
Week 3
i.t. NMU
10µg/mouse
Chronic Inflammation/Lung Cancer establishment
Lung Cancer Progression
Healthy
***
0.20
0.15
**
0.10
0.05
0.00
3 Days
7 Days
30 Days
Tumor Mice
Naive
11
10
9
8
7
6
5
4
3
2
1
0
CD123-APC
**
***
0
3
6
9 12 15 18 21 24 27 30 33
Time (Days)
****
% B220+BDCA-2+CD123+
cells
% CD11c+B220+PDCA-1+ cells
Tumor lesions
(Ratio Tumor Area/Total Lung
Area)
B220-PE
LK
17.5
15.0
12.5
10.0
7.5
5.0
2.5
0.0
Healthy
Cancerous
Th1
polarization
A.
IFN I
IFNAR
mDCs
pDCs
NK cells
IL-12p70
CTL response
Stimulus
IFN I
B.
PLASMACELL
IFNAR
B cell
pDCs
IL-6R
IL-6
Stimulus
CD27
CD70
CD80 or
CD86
or PDL1
C.
pDCs
IDO
IMMUNOCOMPLEXES
T Reg-cell
Immune-suppression
CTLA-4
or PD-1
Stimulus
KYNURENINES
YY YYY
Y
T cell Death
pDCs
Th2 response
Tumor
Progression
Th1 response
Tc response
Tumor
Regression