multhoff [mode de compatibilité] · • release of interferones (i.e. ifn, tnf) • release of...
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Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Prof. Dr. Gabriele MulthoffKlinik für Strahlentherapie und Radiologische Onkologie Klinikum rechts der Isar, Technische Universität München
Immunomodulatory effects of irradiation
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Time scale of irradiation effects
Ionization, radicals, heat <sec
DNA damage, repair min - hours
Cell death hours - days
Immune effects hours - days
Mutations days - months
Cancer years - decades
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Major form of cell death after irradiation is apoptosis
2.5µm 2.5µm
Apoptosis Necrosis
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Apoptosis: Switch of phosphatidylserine (PS) from the inner to the outer membrane leaflet
0 Gy 2 Gy 10 Gy
outer leaflet
inner leaflet
NormalPS
PS
ApoptosisAnnexin V
green: Annexin V and PS antibodies detects PS in outer leafletblue: nucleus (DAPI)
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Characteristics of apoptosis and necrosis
Apoptosis NecrosisProgrammed cell death Accidental
Active Passive
DamageDNA
DamageMembrane
Cell shrinkage Cell swelling (osmosis)
Apoptotic bodies (Blebbing)Membrane intactPS in outer leaflet
Cell burstMembrane damaged PS in inner leaflet
No inflammation Inflammation
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
RadiationRadiationRadiationRadiation
DNADNADNADNA----DamageDamageDamageDamage
ProliferationProliferationProliferationProliferation
CellCellCellCell----CycleCycleCycleCycle
Apoptosis Apoptosis Apoptosis Apoptosis
Annexin VAnnexin VAnnexin VAnnexin VCaspases
RepairRepairRepairRepair
Hsp70
HMGB1
Danger signals of dying cells: HMGB1, survivin, Hsp70
Rödel, Frey, Multhoff, Gaipl 2013
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Hsp70 is also released by viable tumor cells in exosomes
Acetylcholinesterase (AChE) activity at characteristic density of 1.17g/ml
enzy
mat
ic a
ctiv
ity
0
100
200
AChE activity
[g/m
l]
1.08
1.11
1.14
1.15
1.17
1.19
1.21
1.22
1.24
1.08
1.11
1.14
1.15
1.17
1.19
1.21
1.22
1.24
CX+Colo+
silver stain
cyto
solTubulin
Hsp70
Hsc70
Bag-4
Colo - -+ +lys exos
CX - -+ +lys exos
Calnexin
Grp94 ER
Western Blot
Hsp70 in exosomes Gastpar Canc Res 2005
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Preparation of few ml of whole blood
Tumor patient for the
detection of
immune and
tumor markers
HSP70
HMGB1
Survivin
Monitoring of therapy response
Design of a kit
Clinical trial
Survivin
Hsp70
HMBG1
Danger signals serve as biomarkers in the serum of tumor patients to detect cancer and monitor outcome
• Gabriele Multhoff (TU München)
• Udo Gaipl (University Erlangen)• Franz Rödel (University Frankfurt)
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Non-targeted immunomodulatory effects induced by irradiation
• Bystander effects
• Abscopal effects Tumor
Irradiation
Abscopal
Effect
Bystander
Effect
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Principle of bystander effects
Bystander effects are defined as effects on tumor cells in close proximity to the irradiated tumor or to the tumor micromilieu that were not exposed to radiation
• Therapeutic intervention (irradiation) of tumors causes the release of “danger signals”, pro-inflammatory cytokines/chemokines, or reactive oxygen species in close proximity to the irradiated tumor
• These secreted factors induce locally a non-specific immune stimulation that can result in control of non-irradiated tumor or impact the tumor micromilieu
Tumor
Irradiation
Bystander
Effect
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Principle of abscopal effects
Abscopal effects are defined as systemic effects on tumors that are locally distinct from the irradiated tumor
• Therapeutic intervention (irradiation) of tumors causes the release of “danger signals” such as HMGB1, HSP, cell death signals, …,pro-inflammatory cytokines or reactive oxygen species
• These secreted signals derived from dying tumor cells can cause a stimulation of the immune system, predominately of the innate system
• Activated effector cells cause regression of distant non-irradiated tumors
Tumor
IrradiationAbscopal
Effect
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Summary: immune modulations caused by irradiationof the tumor
• DNA damage response• Induction of MHC and NKG2D ligands on tumor cells• Release of danger signals (i.e. HSP, HMGB1, survivin)• Release of interferones (i.e. IFN, TNF)• Release of inflammatory cytokines (i.e. IL1, IL6)• Release of T chemokines (i.e. CXCL9, 10, 11,16) ) that induce the expression of chemokine receptors (CXCR3) on T cells• Up-regulation of adhesion molecules on tumor ECs (i.e. ICAM1, VCAM, E-selectin)• Maturation of dendritic cells (DC)• Lymphocyte infiltration (T and NK cells)• Release of cytotoxic mediators (i.e. IFN, TNF, Granzyme, Perforin) by effector lymphocytes• Immune-mediated tumor cell kill
Burnette & Weichselbaumer Rad Oncol 2013
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
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• Monoclonal antibodies Gramatzki et Valerius1997 (i.e. anti-CD20 Maloney et al 1997, CEA Meyer et al 2009, Ras et al 1997, Kass et al 1999, EGFR Chung et al 2008, HER2 Goldenberg 1999,CO17-1A (EpCam) Riethmüller et al 1994, CTLA4 blockade Hodi et al 2003)
• Tumor vaccines Pardoll 1998 (i.e. irradiated tumor cells, tumor cell lysates Nestle
et al 1998, danger signals Matzinger 1994, Zitvogel et al 2008 i.e. HSP Multhoff et al 1995, HSP-peptide complexes Binder et Srivastava 2005, HMGB1 Gaipl et al 2011, Apetoh et al 2007, Annexin A5, survivin Rödel et al 2007, Kroemer et al 2005, mucin 1 Rammanthan et al 2005)
• Cell based therapies (i.e. T, NK cells Herberman et al 1989, Schaue et McBride 2010, Dendritic cells Schuler et al 1997, Banchereau et al 1998, Aicher et al 1997)
• Small molecule inhibitors (i.e. EGFR Krause et al 2005)
Immunological approaches that could be combined with irradiation (Oettgen et al 1991)
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Effects of Heat shock protein 90 (Hsp90) inhibition (Schilling et al 2008, Kabakov et al 2010, Camphausen et Tofilon 2007, Chiosi 2004, Neckers 1997)
• Degradation of oncogenic client proteins• EGFR endocytosis• Decrease in insulin-like growth factor 1(IGF-1R)• Cell cycle checkpoint activation (CDK4)• Tumor growth reduction• Blocking of DNA damage repair• Suppression of angiogenesis (HIF1, VEGF blocking)• Suppression of p-glycoprotein (MDR)• Decrease in S phaseBut• Induction of the expression of Hsp70 (inhibitor of apoptosis)
Clinical use of Hsp90 inhibitorsProstate, cervical, pancreatic, lung, ENT, breast, bladder, oesophagal carcinomas, glioblastoma , melanoma
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Multhoff JI 1997; Vega JI 2008; Gehrmann Plos One 2008; Falguieres MCB 2008, Staubacher Proteomics 2009
Normal
Tumor
Hsp70 Normal Tumor
Intracellular Low High
Membrane No Yes
Extracellular No Yes
Hsp70 localization in normal and tumor cells
Tumors differ from normal cells in their Hsp70 expression levels and localization
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
• AML, ALL, MDS (Yeh LR 2009, accepted)
• Squamous cell carcinoma lung (Pfister Cancer 2007)
• Lower rectal carcinoma (Pfister Cancer 2007)
• Colon carcinoma (Kocsis CSC 2009, accepted)
• Sarcoma (unpublished)
• Prostate carcinoma ?
Squamouscell carcinoma
Hsp70-FITC100 101 103102 104
Cou
nts
200
4060
Adenocarcinoma
Hsp70-FITC
100 101 103102 104
Cou
nts
20
104
68
Low rectal carcinoma
Hsp70-FITC
100 101 103102 104
Cou
nts
50
2510
1520
Coloncarcinoma
Cou
nts
100
2030
Hsp70-FITC
100 101 103102 104
Gastriccarcinoma
Hsp70-FITC
100 101 103102 104
50
1015
20
Cou
nts
Squamouscell carcinoma
Hsp70-FITC100 101 103102 104
Cou
nts
200
4060
Adenocarcinoma
Hsp70-FITC
100 101 103102 104
Cou
nts
20
104
68
Squamouscell carcinoma
Hsp70-FITC100 101 103102 104
Cou
nts
200
4060
Squamouscell carcinoma
Hsp70-FITC100 101 103102 104
Hsp70-FITC100 101 103102 104100 101 103102 104
Cou
nts
200
4060
Cou
nts
200
4060
200
4060
Adenocarcinoma
Hsp70-FITC
100 101 103102 104
Cou
nts
20
104
68
Adenocarcinoma
Hsp70-FITC
100 101 103102 104
Hsp70-FITC
100 101 103102 104100 101 103102 104
Cou
nts
20
104
68
20
104
68
Low rectal carcinoma
Hsp70-FITC
100 101 103102 104
Cou
nts
50
2510
1520
Coloncarcinoma
Cou
nts
100
2030
Hsp70-FITC
100 101 103102 104
Gastriccarcinoma
Hsp70-FITC
100 101 103102 104
50
1015
20
Cou
nts
Low rectal carcinoma
Hsp70-FITC
100 101 103102 104
Cou
nts
50
2510
1520
Coloncarcinoma
Cou
nts
100
2030
Hsp70-FITC
100 101 103102 104
Low rectalcarcinoma
Hsp70-FITC
100 101 103102 104
Cou
nts
50
2510
1520
Low rectalcarcinoma
Hsp70-FITC
100 101 103102 104
Hsp70-FITC
100 101 103102 104100 101 103102 104
Cou
nts
50
2510
1520
50
2510
1520
Coloncarcinoma
Cou
nts
100
2030
Hsp70-FITC
100 101 103102 104
Coloncarcinoma
Cou
nts
100
2030
100
2030
Hsp70-FITC
100 101 103102 104
Hsp70-FITC
100 101 103102 104100 101 103102 104
Gastriccarcinoma
Hsp70-FITC
100 101 103102 104
50
1015
20
Cou
nts
Gastriccarcinoma
Hsp70-FITC
100 101 103102 104
Hsp70-FITC
100 101 103102 104100 101 103102 104
50
1015
205
010
1520
Cou
nts
Multhoff Exp Hem 1999; Multhoff CSC 2001; Gastpar JI 2004; Staubacher Proteomics 2009
Membrane Hsp70 is a negative prognostic marker
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Hsp70-based therapeutic approaches
• Antibody (naked) therapy: Hsp70 antibody mediated ADCC
• Nanoparticle therapy: Doxorubicin-, anti-survivin loaded nanoparticles conjugated to Hsp70 antibody
• NK cell-based therapy: Hsp70 peptide activated NK cells
• Enzyme therapy: Granzyme B mediated apoptosis
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
1048
FACS ADCC assay
Antibody therapy: cmHsp70.1 mAb initiates ADCC in membrane Hsp70 positive CT26 tumor cells, in vitro
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Media 1xAb Ab+TKD
***
cmHsp70.1 mAb initiates ADCC in membrane Hsp70 positive CT26 tumor cells, in vivo
CT26 tumor reduction in BALB/c mice
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
p=0.310p<0.0001
days after Ab treatment days after Ab treatment
surv
ival
[%]
Hsp70 positive CT26 tumors
Hsp70 negative A20 lymphoma
Survival curves after 3 injections of cmHsp70.1 mAb
cmHsp70.1 mAb enhances survival of BALB/c mice bearing membrane Hsp70 positive CT26 tumors (i.p.)
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
4°C 37°C
cmHsp70.1-FITC cmHsp70.1-FITC
Membrane (4°C) Cytosol (37°C)
Proteomic and lipidomic profiling of Hsp70 associated compounds in lipid rafts
Subcellular distribution of Hsp70 in the endo-lysosomal compartment
Nanoparticle therapy: Elevated temperature results in uptake of Hsp70 from the membrane into the cytosol
Stangl et al JCMM 2011; Gaca et al JCR 2013
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
cmHsp70.1
PEG-linker
Nanoparticle unmodified(NP-PEG)
Adapted and modified from Spänkuch et. al., Neoplasia. 2008 Mar;10(3):223-34. ; Gaca et al JCR 2013
DoxorubicinAnti-survivin
Nanoparticle with Hsp70-mAb(NP-Hsp70)
Doxo-Nanoparticle unmodified(Doxo-NP-PEG)
Doxo-Nanoparticle with Hsp70-mAb
(Doxo-NP-Hsp70) cmHsp70.1
PEG-linker
PEG-linker
PEG-linker
Nanoparticle therapy: Hsp70 mAb-nanoparticle (NP)
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
NK cell therapy: TKD/IL-2 activates NK cells
Enhances the migratory capacity of NK cellsGastpar CR 2005
ctrl IL-2 IL-2/ TKD
CD94 177 198 707
NKG2D 72 86 208
NKp30 37 47 199
NKp44 1 132 399
NKp46 233 290 393
Up-regulates activating, NK receptorsGross CSC 2003, Gross Bio Chem 2003
Tumor
NK
NK
Cx+
20/110/
1 5/1 2/1
% s
peci
fic ly
sis
0
10
20
30
40
50
60
70
80
90controlHsp70 mAb
Cx-
20/110/
1 5/1 2/1
days after tumor injection
5 15 25 35 45 55 65 750 10 20 30 40 50 60 70su
rviv
al (
%)
0
20
40
60
80
100
control (n=21)T (IL-2/TKD) (n=14)NK (IL-2/TKD) (n=17)NK (IL-2) (n=5)
TKD/IL-2 activated NK cellsenhance survival of mice
Kill Hsp70 positive tumor cells, in vitroMulthoff Exp Hem 1999, Gastpar J Immunol 2004
Successful phase I clinical trialKrause Clin Can Res 2004
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Clinical phase II trial with IL-2/TKD activated NK cells after RCT
* Immunological assessment: 4 weeks after last treatment day of RCTTumor assessment: 4 weeks after last treatment day of RCT Eligible patients will be randomised to either study or control group after tumor assessment
Scr
eeni
ng fo
r H
sp70
RA
ND
OM
ISA
TIO
NRCT4 mo
4 wks*
CRSDPR
NK NK NK NK NK NK
0 1284 16 20 wks24
0 1284 16 20 wks24
NK Infusion of TKD/IL-2-activated NK cells
RCT4 mo
Radiotherapy: 60-66 Gy Chemotherapy:Cisplatin/ Vinorelbine
Study group: NK therapy Best Supportive Care
Control group: no NK therapy Best Supportive Care
Follow-up period(18 months after start of NK
treatment)
Hsp70 positive patients entering standard therapy
End of NK treatment
further cycles possible
Pre-study part Interventional study part
Start of NK treatment
Biological responses
Clinical responses (PFS, OS)
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Molecular therapy: granzyme B mediated tumor killing
Trapani Genome Biol 2(12):3014.1 (2001)
Granzyme B initiates perforin-independent apoptosis selectively in Hsp70 positive tumor cellsGross JBC 2003; Nylandsted J Exp Med 2004
Mammalian expression system for human granzyme B
Mechanism of killing by human granzyme B
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Induction of apoptosis in Hsp70+tumors by granzyme B
untre
ated
Granz
yme
BCam
ptot
hecin
Cas
pase
-3 p
ositi
ve c
ells
[%]
0
10
20
30
40
50
60
70
12h24h48h
Granzyme B
Camptothecin
Control
Granzyme B [µg]0.0 0.1 0.2 0.3 0.4 0.5 0.6
Abs
orpt
ion
[405
nm
]
0.0
0.1
0.2
0.3
0.4
0.5
# 2# 3# 4
Yield: 10-30 mg GrB per 2 l
supernatant with a purification yield
of 64%
Apoptosis induction in CT26 cells
Gehrmann JIM 2011
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
NK cell therapy
(IL-2/TKD)
Molecular therapy (Granzyme B)
NK
• DFG MU1238 7/2• Helmholtz Gemeinschaft “Präsidentenfonds“
• SFB-824• multimmune GmbH
• EU-CELLEUROPE• BMBF-Innovative Therapies
Antibody therapy (cmHsp70.1)
Radiation/ Hypoxia
Lipidomic and proteomic
profiling
Definition of tumor markers
in the serum
• BMBF Spitzencluster m4• TU-München
• Wilhelm Sander Stiftung• BMBF Kompetenzverbund Strahlenforschung
Funding
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Collaborations
Alexzander Asea (Scott & White Hospital Texas, USA)
Mike Atkinson, Michaela Nathrath (HMGU)
Günther Dollinger (UBW)
Anna Friedl , Kirsten Lauber (LMU)
Antonio DeMaio (UCSD, San Diego, USA)
Ralf Dressel, Frauke Alves, Christian Dullin (University Göttingen)
Franz Pfeifer (TUM)
Udo Gaipl, Benjamin Frey (University Erlangen)
Carmen Garrido (University Dijon), Marja Jäättelä (University Copenhagen)
Mike Horseman (University Aarhus)
Graham Pockley, Gemma Foulds (University Nottingham, UK)
Franz Rödel (University Frankfurt)
Gerd Schmitz (University Regensburg)
Arne Skerra, Lars Friedrich (WZW)
Vasilis Ntziachristos (HMGU)
Laszlo Vigh (University of Szeged)
Axel Walch, Isabel Winkelmann (HMGU)
Scientific team
Christine Bayer, PhD
Stephanie Ertl
Stephanie Erl
Mathias Gehrmann, PhD
Katharina Ilicic
Daniela Schilling, PhD
Thomas Schmid, PhD
Wolfgang Sievert
Stefan Stangl
Dieter Walsh
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Modified from Shresta et al. (1998), Curr. Opin. Immunol. 10: 581
NK cell therapy: NK cells kill tumor cells perforin-dependent / independent by granzyme B mediated apoptosis
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© MulthoffDaugaard et al., 2007; Fouchaque et al 1999
C-terminus (aa 540-646)
Substrate-bindingdomain (aa 385-540)
N-ATPasedomain (aa 1-385)
Multhoff Exp Hem 1999; Multhoff CSC 2001; Gastpar JI 2004; Staubacher Proteomics 2009
Heat shock protein 70 (Hsp70 / HSPA1A)
Hsp70
Major stress-inducible member of the HSP70 family
Stress factors: heat, ionizing irradiation, UV, chemotherapeutics, hypoxia, heavy metals, oxygen radicals, …
Localization: intracellular (cytosol, ER), membrane, extracellular
Gene: chromosome 6p (TNFα - Hsp70 - complement)
Protein: 641aa, 72 kDa, gobular, conserved, no transmembrane domain
Domains: ATPase (44kDa), substrate binding (18kDa), C-terminal helix (10kDa)
Moonlighting function: molecular chaperone (folding, unfolding, transport, anti-apoptotic, antigen processing)
3D structure
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
NK cells T cellsPeripheral blood 5-20% 60-80%
Origin Lymphoid progenitor in bone marrow
Lymphoid progenitor in bone marrow
Function First line of defenseViral/bacterial infectedcells, tumors
Adaptive immunityViral/bacterial infectedcells, tumors
Receptor Non-clonal, multiple Clonal
Phenotype TCR/CD3-, CD16/56+, KIR+, ILT+, Lectin-type+
TCR/CD3+, CD4/CD8
Killing ADCC, FAS/FASL, Granzymes, Perforin
FAS/FASL, Granzymes, Perforin
Cytokines IL-2, IL-15, IL-12, IL18 IL-2, IL-15, IL-12, IL18
Killer Immunoglobulin-like Receptors (KIRs)
Lectin-like receptors(CD94, NKG2A/C/D)
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Radiated
tumor
Gehrmann et al (2005) Cell Death Differentiation 12: 38-51; Gehrmann et al (2010) Int J CPT 48: 492
Irradiation increases the Hsp70 membrane expression
Therapies
•y-Irradiation
•Vincristin sulfate
•Taxoides
•Hsp90 inhibitors
•COX2 Inhibitors
Isotyp-FITC Hsp70-FITC
10 Gy
ControlFSC/SSC
Increase in membrane Hsp70 positive colon carcinoma cells following irradiation (10 Gy)
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
1863 Virchow „Tumors are infiltrated by lymphocytes“
1893 Coley „Severe bacterial infections result in tumor regression“
1909 Ehrlich „Virulent capacity of tumors“: Immune system restricts cancer growth
1958 Burnet „Hypothesis of immune surveillance of cancer“
1958 De Vries Adoptive transfer of allogeneic lymphocytes in tumor patients
1965 Mathe & Amiel Graft vs host disease (GvHD) and graft vs leukemia reactions (GvL)
20022007
Dunn, OchsenbeinSwann, Smyth
„Immune editing“: Elimination, Equilibration, Escape
2007 Stewart Innate immune system is the first line of defense: inflammation induced by NK cells, Mp, coordinated by DCs
2004 Dunn RAG-2 deficient mice (no T, B cells) develop epithelialtumors (50%)
2006 Malmberg, Ljunggren
IFNγ deficient mice develop tumors
Immune system and cancer
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
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2002 Diefenbach, Raulet NK cells, DCs, Makrophages (Mp), neutrophils secrete dangersignals to stimulate the innate immune system
2008 Lin Innate cytokines: IL-2, IL-12, IL-18, IL-23Innate danger signals: HSPs, HMGB1, Annexin A5
20032008
DiefenbachZitvogel
Innate ligands: NKG2D ligands (MICA/B, RAE-1, ULBP1,2,3), HSPs > activate NK cells
2008 Prestwich Secretion of pro-inflammatory cytokines by DCs
2007 Raman Innate chemokines: CXCL10, CXCL9, CXCL11
2004 Houghton Adaptive Immunity: T cells recognize TAA presented by DCs in context with co-stimulators: B7-1 (CD80), B7-2 (CD86), CD40/CD40L
19981999
LordNishimura
Th1: IL-2, IFN γ, TNFα, GM-CSF > Mp, T (CD8) cellsTh2: IL-4, IL-5, IL-10 > naive B cells
2004 Dranoff Kill mechanims of T cells: perforin, granzyme, Fas ligandT cell cytokines: IFNγ, TNFα, TNFβ activate Mp
Mechanisms of tumor elimination by the immune system
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Hypoxia
Resistance to therapy
Cell growth/differentiation
Genomicinstability
Programmed cell death/apoptosis
Tumor progression/ metastasis
Neoangiogenesis/vascularisation
Sutherland et al. 1998
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Arterial end
Höckel et al. 1999
Reoxygenation effects on overall survival of patients
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Indirect / direct effects of irradiation
Indirect: e- interacts with mediator (e.g. H2O)
H2O � H2O+ + e-
H2O+ + H2O�H3O+ + OH-
O2 fixation of DNA damage Production of RO2*
Direct: e- interacts directly with DNA
Abbreviations:e- : fast electron RO2* : Reactive Oxygen Species
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
Immune system
Granulocytespolymorphonuclearleukos
Agranulocytesmononuclearleukos
Neutrophils Lymphocytes(T, B, NK cells)
Basophils Monocytes
Eosinophils Macrophages / DCs
Multipotent Human Stem Cells (BM)
Myeloid Progenitors Lymphoid Progenitors
Mega- Erythrocyte Mastcell Myeloblast T cell B cell NK cellkaryocyte
Baso Neutro Eosino Monocyte Plasma cell
Klinik und Poliklinik für Strahlentherapie und Radiologische Onkologie
© Multhoff
• Interferones (IFN α, IFNγ, IFNβ, TNFα) Guilot et al 1997, Quesada et al 1984, Ludwig et al 1995
• Chemokines (CXCL8, CCl7, CCL8, CCl11, CCl13, CCL17, CCL19, CCL20)
• Cytokines (IL-2 Rosenberg et al 1998, Keilholz et al 1997, IL-15 Handgretinger)• Enzymes (granzyme, perforin)• Bacillus Calmette-Guerin (BCG) Herr et al 1992
Non-specific immunotherapeutic approaches (Oettgen et al 1991)