dudley primer revised slides...microsoft powerpoint - dudley primer revised slides.pptm author...
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Adoptive Immunotherapy
Mark E. Dudley, PhDCell Production FacilitySurgery Branch, NCI
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Cell Therapy•Red cells•Stem cells•Platelets
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Adoptive T Cell Transfer• Potent therapy • Unique research tool
– Host lymphodepletion– Lymphocyte attributes– Tumor antigen targets
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Examples of Successful Adoptive Immunotherapy
Target T cells Reference
PTLD Viral antigen specific Heslop, 2009
Viral infection Viral specific (uncultured) Feuchtinger et al, 2010
B cell tumor Genetically retargeted Kalos et al 2011
Melanoma Tumor infiltrating lymphocytes Rosenberg et al 2011
Sarcoma Genetically retargeted Robbins et al 2011
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Adoptive Cell Therapy with TIL for Melanoma
Dudley et al, Nat Rev Ca, 2003
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ACT with Lymphodepletion• Expand T cells to large numbers in the absence
of potentially suppressive tumor environment• Selection of optimal T cell attributes • Manipulation of host immune system without
damaging the effector T cells
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All cells undergo extensive safety and quality testing to produce a certificate of analysis (COA) for each
final product prior to infusion
COA Criteria:
- Viable cell count- Tumor recognition- Gram stain- Fungal testing- Mycoplasma testing- Bacterial testing- Endotoxin testing- Cytological analysis- TCR expression- RCR
Ensuring a Safe Product for ACT
www.piercenet.com/media/cytokine_kit.gifwww.daigger.com http://isips.org/products/graphics/12a.gif
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C.K. (200cGy) Pre 12 days
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L.R.
1200
cGy
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• ACT with lymphodepletion– Responses can be rapid– No relation between bulk of disease and
response– Impact metastatic disease at any site
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Adoptive T Cell Transfer• Potent therapy • Unique research tool
– Host lymphodepletion– Lymphocyte attributes– Tumor antigen target
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Adoptive Cell TherapySelected T cells
Lymphodepletion
Transient Lymphopenia
0
300
600
900
1200
1500
1800
-9 -6 -3 0 3 6 9 12
AL
C (l
ymph
ocyt
es /
u
Bulk TIL
-14 -7
-5
Cy Fludarabine IL-2
Rapid Expansionstart harvest
TIL infusion
13
Days
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Chemotherapy orRadiation therapy
Mechanisms underlying the impact of lymphodepletion on adoptively transferred T cells
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Lymphodepletion•Elimination of TREG
•Increased homeostatic cytokines•APC activation through TLR•Stem cell “facilitation”
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pre
pt 7TIL
Spec
ific
lysi
s
Effector: target ratio
0.1 1 10 100
T cells are active in vivo
Patient 10938 mel (HLA-A2-
)
526 mel (HLA-A2+)
Specifically lytic Phenotypically active
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Persistence of Transferred TIL
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Transferred T cells up-regulate expression of HLA by tumor cells in vivo
Pre
trea
tmen
t
CD8 MHC Class IIMHC Class I
56 d
ays
post
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Impact of Lymphodepletion
• Functional and phenotypic activation of transferred T cells• Proliferation and persistence• Traffic to tumor and up-regulation of HLA on tumor cells• Regression of bulky tumor masses
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Adoptive immunotherapy –a unique research tool
• Host Conditioning • T cell attributes• Target selection
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Attributes of T cells Associated with Response
• Minimum time in culture• Persistence in vivo after transfer• Replicative potential
– Long telomeres• Less differentiated phenotype
– High expression of CD27, CD28
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Potent immune responses require CD8+ and CD4+ cells
APCAPCTh
CTL
Th
ILIL--22CTL
CTLCTL
ILIL--22
ILIL--22• Antigen Presenting Cells (APC) activate both
CD8+ CTL and CD4+ Thelper cells
• At a site of inflammation, Thelper cells secrete IL-2 to further activate and cause proliferation of CTL
• TREG dampen immune responses and inactivate CTL
• In ACT, chemotherapy eliminates endogenous T cells. Tumor reactive CTL are administered with exogenous IL-2
TREG
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What is the role of CD4 cells
• CD4+ T-regs limit CD8+ ACT effectiveness in mouse models • Anecdotal examples of CD4+ T cells correlated potent in vivo
tumor rejection• CD8+ TIL number is associated with response
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Young TIL
CD8 enrichment of TIL for Therapy
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Optimized Clinical scale CD8+ enrichment of TIL using CliniMACS
Bulk TIL Enriched Depleted
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E.H.
YoungTIL;
CD8
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Patient 6
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CD8+ Cells can Mediate Tumor Regression• 56 patients treated
– 33 with chemotherapy lymphodepletion– 23 with TBI and chemo lymhodepletion
• 29 (52%) objective responders
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Adoptive immunotherapy –a unique research tool
• Host Conditioning • T cell attributes• Target selection
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TCR-Transduced Peripheral Blood Lymphocytes (PBL)
• HLA-A2 patients only• Uses PBMC pheresis prior to start• Defective virus “infects” patient lymphocytes• Engineered genes insert into lymphocyte genome• New protein receptors are expressed (TCR) • T cells start hunting tumor cells
Transduce cells Expand and testPBMC Pheresis
http://flickr.com/photos/jepoirrier/385575933/
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Development of TCR gene therapy
From, Rosenberg et al, Nat. Rev. Cancer April 2008
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Anti-tumor antigen receptor containing retroviral vectorsTarget Tumor Antigen
2A TCR betaTCR alphaLTR LTRsd sa
MART-1 (DMF5)
IRES TCR betaTCR alphaLTR LTRsd sa
gp100:154
TCR betaTCR alphaLTR LTRsd sa
NY-ESO-12A
Johnson LA, et al 2009, Blood 114:535-46
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MART-1 and GP100• Melanocyte differentiation antigens• Expressed by tumors and normal melanocytes
– Skin, eyes, ears• TIL – low/no toxicity• Moderate affinity “F4” TCR – no toxicity
• “F5” and g154 TCRs selected for high affinity antigen recognition
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Day 8: CD8 positive cells
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Melan-A positive control
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Day 8: Melan-A (rare specific staining)
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Day +12 Day +26
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SB Clinical Trials with MDA TCR
20-30% objective Response~70% Grade 2-3 Toxicity
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Anti-tumor antigen receptor containing retroviral vectorsTarget Tumor Antigen
2A TCR betaTCR alphaLTR LTRsd sa
MART-1 (DMF5)
IRES TCR betaTCR alphaLTR LTRsd sa
gp100:154
TCR betaTCR alphaLTR LTRsd sa
NY-ESO-12A
Robbins PF et al, 2011, J Clin. Oncol. 29:917-24
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NY-ESO-1 TCR• NY-ESO-1 – Cancer/Testes antigen• Not expressed in normal adult tissues except
testes (no class I expression)• Expressed by 10-50% of tumors of multiple
histologies including melanoma, breast, prostate, thyroid and ovarian.
• Expressed by ~90% of synovial cell sarcomas
• Eso TCR has an alpha chain CDR3 modification
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Patients and outcomes for Eso TCR therapy
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ESO TCR• 5/11 responses in melanoma• 4/6 responses in synovial sarcoma• No toxicity related to cells
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Adoptive T Cell Transfer
• Potent therapy – Clinical responses in refractory disease– Break tolerance to self antigens
• Unique research tool– Patient, T cells, tumor