100 60 10 0 year cancer/ neuronal disorder infection/development 1 tumor immunology bc yang expected...
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100 60 10 0 year
Cancer/ Neuronal disorder Infection/Development
1
Tumor immunologyBC Yang
Expected life-span
2010
衛生署資料,台灣人十大死因2
排名 年度 88 89 90 91 92 93 94 95 96
1 惡性腫瘤 135.32 142.23 147.68 152.88 156.01 160.54 163.8 166.5 175.9
2 腦血管疾病 57.39 60.1 58.82 53.46 54.98 54.48 57.8 55.2 56.2
3 心臟疾病 51.33 47.56 49.25 50.93 52.23 56.79 57.1 53.8 56.7
4 糖尿病 40.99 42.6 40.79 39.26 44.38 40.58 46.2 42.5 44.6
5 事故傷害 58.88 47.4 42.58 37.79 36.3 37.33 36.8 35.1 31.1
6 慢性病跟肝硬化 23.53 23.32 23.45 21.35 22.98 23.63 24.7 22.1 22.5
7 肺炎 18.2 14.88 16.77 20.17 22.6 24.44 25.0 23.6 25.7
8 腎病徵候群及腎病性病 15.78 17.45 18.15 18.55 19.08 20.67 21.2 20.6 22.2
9 自殺 10.36 11.14 12.45 13.59 14.16 15.31 18.8 19.3 17.2
10 高血壓性疾病 8.43 -- 7.9 8.67 8.17 7.97 8.3 8.0 8.6
98 年 癌症、心臟疾病(不含高血壓)、腦血管疾病、肺炎、糖尿病、事故傷害、慢性下呼吸道疾病、慢性肝病及肝硬化,自殺、腎臟疾病。
THE JOURNAL OF EXPERIMENTAL BIOLOGY 207 (23): 4013-4014
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There are very few scientific papers that continue to have an impact on research more than 50 years after their initial publication. A paper published in J. Exp. Biol. in 1951 by Rupert Billingham and Peter Medawar is one such paper.
THE BIRTH OF TRANSPLANTATION IMMUNOLOGY: THEBILLINGHAM–MEDAWAR EXPERIMENTS AT BIRMINGHAM UNIVERSITY AND UNIVERSITY COLLEGE LONDON
(The technique of free skin grafting in mammals)
Evidence for active host defense against cancer
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80 years of immunotherapy. (Currie GA, 1972, Br. J. Cancer 26: 141)
A critique of the evidence for active host defense against cancer, based on personal studies of 27 murine tumors of spontaneous origin. (Hewitt HB, et al. 1976, Br. J. Cancer 33:241)
Circumvent Evidence
The high frequency of cancers in immunosuppressed patients Extremes of age Immunosuppressive drugs
Tumors that are infiltrated by T cells have an improved prognosis
Spontaneous regression occurs Melanoma, breast, lung cancers, etc
Circulating tumor antibodiesMHC-I down-regulation
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Full immunity needed to fight cancer9
CD4+ T cells contribute to the remodeling of the microenvironment required for sustained tumor regression upon oncogene inactivation (Cancer Cell, 2010, 18:485-498)
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Immune selection in the development of cancer: no two tumors are alike
Initiation, proliferation diversification
Microevolution, selection of immune resistance
Fey MF & Tobler A 1996
Immune escape and unchecked proliferation
Use of tumor cell lines17
Commonly derived from advanced tumors
Retain the genetic instability and lose the ability of adaptations
introduced in large numbers (more than 106 cells)
Immune therapy (T cell-based)19
Recombinant and synthetic vaccinationCytokine treatments (IL-2; GM-CSF; IFN)Cellular therapy with tumor-specific CTLEngineered macrophages (Th1…?)Antigen-pulsed macrophages or dendritic
cells
Tumor associated antigens20
Tumor-specific shared antigens: restricted in expression to tumors and immune privilege sites.
Tissue-specific differentiation antigens: tyrosinase. (melanoma)
Tumor-specific antigens: mutated, tranlocated genes.
Ubiquitous antigens with over-expression in tumors.
Rosenberg SA. 1999, Immunity 10:281
Examples
Down-regulation of the class I presentation pathway
Resistance to killingAntigen-specific
mechanisms (Treg?)Global mechanisms
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Antigen-specific mechanisms27
Tumor antigen-loss variants
Loss of the melanoma tumor-associated antigen in patients with recurrent metastatic melanoma (J Clin Invest 1996, 98:1633)
B cell tumors expressing class II induced a rapid tolerance of cognate CD4 T cell carrying a transgenic TCR. (PNAS, 1998, 95:1178)
Tolerance
Down regulation of the MHC class presentation pathway
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Downregulation of MHC class I expression is frequently seen in human tumors.
Loss of MHC-I as a mechanism for tumor escape from CTL-mediated elimination (longitudinal study of melanoma patients)
Five major HLA altered phenotypes found in tumor tissues (Human Immunol. 2000, 61:65)
The five altered phenotypes 29
Normal A1A2B8B35Cw7Cw4
1. Total loss -2. Haplotype loss A1B8Cw7
3. Locus loss A1A2B8B35
4. Allelic loss A2B8B35Cw7Cw4
5. Compound phenotype A1
(Human Immunol. 2000, 61:65)
Resistance to killing30
Defective Fas pathwayResistance to Granzyme B
Fas-L and Neutrophil
Cytotoxic T cells
Innate immunity
Key person in Fas signal study32
Shigekazu Nagata
For a man who spends most of his time thinking about death, Shigekazu Nagata is remarkably upbeat. For over a decade, he has been making a name for himself with research on apoptosis, the mechanism of programmed cell death, and during that time he has watched the field come alive.Nature Medicine 7, 759 (2001)
Peter H. Krammer
BC Yang 2008/2/20
Loss sensitivity to Fas-mediated apoptosis
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FLICE-like inhibitory proteinsBcl-2, Bcl-XLdefected sphingomyelinase
activation (ceramide)decoy receptor 3 (DcR3), soluble
Fas
Fas-L+-melanoma cells are relatively resistant to killing of neutrophils
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B16F10
Chen YL, et al J. Immunol. 171:1183-1192.
Global Mechanisms35
TGF-betaIL-10Growth in immune privilege
sitesMucin production: interfering
intercellular adhesionFas-L? (Fas counterattack)Tumor extracellular matrix
Shaping T cell functions by extracellular matrix.
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NPC Done by 張逸如 (1998)
Hematoxylin/eosin staining CD3+ cells
Immune cells are there! But, in peripheral stroma area (cell cuffing)
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How can tumor cells highjack immune cells?
Extracellular matrix shapes Fas-mediated apoptosis.
Protective: U-118, Huh-7, HeLa, A549, NIH3T3
Non-protective: MCF-7, HepG2
Su CC, et al. 2007 Phosphatidylinositol 3-Kinase/Akt activation by integrin-tumor matrix interaction suppresses Fas-mediated apoptosis in T cells. J Immunol 179:4589-4597.
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Coculture with tumor cells inhibits the Fas-mediated apoptosis.
0.0
10.0
20.0
30.0
40.0
50.0
60.0
J CH-11 U118V U118V+CH-11
U118R U118R+CH-11
HeLa HeLa+CH-11
Huh-7 Huh-7 +CH-11
A549 A549 +CH-11
Apo
ptos
is (%
)
24 h 48 h
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0
10
20
30
40
50
60
None Z-vade (10μm)
SP600125 (10μm)
PD98059 (10μm)
SB202190 (5μm)
SN-50 (10μm)
KT5720 (2.5μm)
HSP70 (10μm)
Ap
opto
sis
(%)
Jurkat Jurkat + CH-11 Jurkat + U-118MG Jurkat + U-118MG + CH-11
Fas-mediated death is not suppressed by components of MAPK, NFkB, PKA, HSP70.
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Fas
Fas-LImmune cells
Tumor
Tissue environment ?
Cytokines ?
What will happen when Fas-stimulated immune cells resist to die?
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J Immunol, 2003, 171: 3947–3954.
Jurkat and Molt-4 cells were cultured alone (lane 1) or in the presence of U118(V), U118(R), U373(V), or U373(R) (lanes 2, 3, 4, and 5, respectively) for 24 h.
T cells in tumor microenvironment
林亨楷
Regulatory T cells: the third man46
Shimon Sakaguchi (2000) Regulatory T Cells Key Controllers of Immunologic Self-Tolerance Cell 101: 455-458
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Joe O’connell, 2002, Fas ligand and the fate of antitumorur cytotoxic T lymphocytes. Immunology 105:263-266.
In vivo consequences of Fas-L expression by tumors(using over-expression system)
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Renca, MH134, L5178Y, B16-BL6, CT26*
Note: *:syngenic, nude, SCID #: allogenic, lpr
Ref: Lejeune FJ et al., 1998, Curr. Op. Immunol.
Enhanced rejection
Delayed rejection
CT26#B16-F10
Distinct structure of tumor mass
Glioma in Nude mice
May be Fas-L associated?
Why TIL in particular sites?
Penetration of tumor by immune cells
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Fas-LR
Control
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Granulocytes mediates the Fas-L-associated apoptosis during lung metastasis of melanoma that determines the metastatic behavior (B16F10 in C57BL/6) Br J Cancer, 87: 359 (2002)
i.v. 5 x 105 cells /mouse; Observed at day 14; (bar = 1 cm)
Kang, S. M. et al. 1997. Fas ligand expression in islets of Langerhans does not confer immune privilege and instead targets them for rapid destruction. Nature Med. 3:738.
FasL reverse signaling55
Suzuki I. and Fink PJ.. Maximal Proliferation of Cytotoxic T lymphocytes Requires Reverse Signaling Through Fas Ligand. J. Exp. Med., 187:123, 1998.
Boursalian TE, Fink PJ, Mutation in Fas Ligand Impairs Maturation of Thymocytes Bearing Moderate Affinity T Cell Receptors, J. Exp. Med., 198, 349, 2003.
Sun M, Ames KT, Suzuki I,. Fink PJ, The Cytoplasmic Domain of Fas Ligand Costimulates TCR Signals, J. Immunol., 177, 1481, 2006.
Pamela J. Fink, h.D.
Department of Immunology
University of Washington
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Fas ligand (FasL/CD95L) is best known for its role in delivering apoptotic signals through its receptor, Fas (APO-1/CD95). In this study, we present evidence that FasL has a second role as a signaling receptor. …….(abstract)
Suzuki I. and Fink PJ. (1998) J. Exp. Med., 187:123.
Depressed proliferation and cell recovery of FasL-deficient CTLs relative to FasL+ CTLs over the course of an MLC. (a and b) [3H]TdR uptake and viable cell counts over days 1-5 from long-term CTL lines from B6, B6.lpr, and B6.gld mice cultured with allogeneic H-2k splenocytes. (c) CTL responders were purified CD8+ T cells derived from B6 and B6.gld mice cultured with H-2bm1 splenocytes. (d) [3H]TdR uptake by CTL responders used in a, cultured with C3H.gld splenocytes.
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Fas ligand (FasL/CD95L) is best known for its role in delivering apoptotic signals through its receptor, Fas (APO-1/CD95). In this study, we present evidence that FasL has a second role as a signaling receptor. …….(abstract)
Suzuki I. and Fink PJ. (1998) J. Exp. Med., 187:123.
Ectopic expression of FasL in NIH3T3
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(A). Full-length and truncated FasL. Adapted from Orlinick JR, et al. J Biol Chem 1997, 272:32221-29. Jodo S, et al. J Immunol 2005, 174:4470-4474. FasL-
70: deleting the N-terminal 2-70 aa; FasL- 33: deleting the N-terminal 2-33 aa. △ △
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(A). Nude mice were injected with 5x105 cells of NIH3T3 cells carrying FasL as indicated via tail vein. After 4 weeks, spontaneous transformed NIH3T3 tumor nodules formed in the lung. Ling sections were stained with H/E. Tumor nodules show dark purple stain. (B). FasL constructs were tranfected into B16F10 cells and injected into B6 mice. After 2 weeks, tumor nodules on the lung surface were counted. N1: B16F10-transfected with plasmid vector, Robozyme: B16F10-transfected with FasL-ribozyme plasmid.
Full-length FasL suppresses lung metastasis, while truncated FasL promotes tumor formation.
A B
Alternative paradigm62
A tumor is a local growth of abnormal tissue consisting of genetic-altered transformed cells and a number of other cell types and connective tissue components characteristic of each tumor type.
No host, as a tissue, no fighting.
Seljelid R et al 1999, Anticancer Res 19:4809
Tumor Stroma63
Fibroblasts
Macrophages
mast cells
Endothelium
lymphoid cells (T, B, granulocytes, NK cells)
intercellular substance; extracellular matrix
Ctrl.
shCol
Fibroblast attraction
林若曦
Effects of ECM on the tumor microenviroment:Collagen and tumor fibroblast
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