1. Th2-biased anti-tumor immunity promotes glioma growth in a mouse model
Sami Sauma
April 12,2016

2. Background – Astrocytoma
Grade II
low grade astrocytoma
Grade III
anaplastic astrocytoma
Grade IV
glioblastoma multiforme (GBM)
Most common primary CNS tumor
Aggressive, short clinical history, less than 3 months
Aggressive; median survival 15 months

3. Background – Astrocytoma
AIII
CD163low CD163high
AII AIII GBM
Prosniak et al., 2013; Clin Cancer Res 19:

4. Background – The Immune System Helper T Cell
Zhu, 2008; Blood: 112 (5)

5. Background – The Immune System Alternatively Activated Monoctye
Zhu, 2008; Blood: 112 (5)

6. Background – The Immune System Tumor Associated Macrophages

7. Background – GL261 Glioma Cell Line
Commercialization
NCI Repository
Methylcholanthrene
Seligman and Shear, 1939
Ex vivo transplantation
Serial passaging
Orthotopic Model
Subcutaneous Model

8. Background – Antisense Oligodeoxynucleotides and the Tbet-/- Mouse
(AS ODN)
C57BL/6J
Widely used inbred strain of mice
Natural bias towards Th1 immunity
Refractory to tumor development
Antisense technology
Tbet-/-
C57BL/6 background
A targeted mutation of the T-box transcription factor Tbx21
CD4 T cells do not produce the Th1-type cytokine interferon-γ
Elevated levels of Th2-type cytokines

9. Background – Published Vaccine Data
GL261
GL261 + AS ODN
Gl261 (left flank) + AS ODN (right flank)
Harshyne et al., 2015; Cancer Immunol Immunother 64(3):299–309
Morin-Brureau et al., 2015; Cancer Immunol Immunother 64(4):447-57

10. Results – Subcutaneous Model (Vaccine)
GL261
GL261 + AS ODN
GL GL261 + AS ODN

11. Results – Intracranial Titration of GL261

12. Results – Orthotopic Model
Vaccine Intervention
PBS
GL261
+ AS ODN
PBS
GL261 +
AS ODN

13. Results – Orthotopic Model
Vaccine Intervention
PBS
GL261 chambered
+ AS ODN
PBS
GL261 ch.+
AS ODN

14. Results – Orthotopic Model
Vaccine Intervention
PBS
GL261 exosomes
+ AS ODN
PBS
GL261 exo +
AS ODN

15. Results – Orthotopic Model
PBS
GL261 exo + AS ODN
GL261 + AS ODN
qPCR analysis of left and right cortices, and tumor tissue reveals no significant difference between two vaccination groups and mock.
Elevated CD204 seen throughout.

16. Results – Subcutaneous Model
PBS
AS ODN
Results – Subcutaneous Model
AS ODN Intervention
Day 0 flank administration of untreated GL261 cells in C57 and Tbet-/- mice
Day 19/20 (dashed line) intraperitoneal injection of antisense or vehicle (PBS)

17. Results – Subcutaneous Model
AS ODN Intervention
PBS
AS ODN

18. Results – Orthotopic Model
AS ODN Intervention
PBS
AS ODN
PBS AS ODN

19. Results – Orthotopic Model
AS ODN Intervention
PBS
AS ODN
PBS
AS ODN

20. Results – Orthotopic Model
AS ODN Intervention
PBS
AS ODN
PBS
AS ODN
PBS
AS ODN
PBS
AS ODN
PBS
AS ODN
PBS
AS ODN
PBS
AS ODN