Dendritic cell subsets and CD4 T cell immunity in Melanoma. Ben Wylie 1 st year PhD Candidate
|
|
- Nora Barrett
- 6 years ago
- Views:
Transcription
1 Dendritic cell subsets and CD4 T cell immunity in Melanoma Ben Wylie 1 st year PhD Candidate
2 Melanoma Melanoma is the 4 th most common cancer in Australia. Current treatment options are ineffective resulting in a poor prognosis. Novel T cell based therapies have been shown to significantly prolong survival in some patients.
3 T cell Immunity Dendritic cells (DCs) are the master regulators of T cell immunity. Adapted from Lake and Robinson Nat Revs. Cancer. 5(5):
4 T cell Immunity Dendritic cells (DCs) are the master regulators of T cell immunity. Adapted from Lake and Robinson Nat Revs. Cancer. 5(5):
5 T cell Immunity Dendritic cells (DCs) are the master regulators of T cell immunity. Adapted from Lake and Robinson Nat Revs. Cancer. 5(5):
6 T cell Immunity Dendritic cells (DCs) are the master regulators of T cell immunity. Adapted from Lake and Robinson Nat Revs. Cancer. 5(5):
7 T cell Immunity Dendritic cells (DCs) are the master regulators of T cell immunity. Adapted from Lake and Robinson Nat Revs. Cancer. 5(5):
8 Tumour Immunity DCs acquire melanoma antigens and traffic to skin draining lymph nodes to present antigen. Adapted from Munz et al J Exp. Med. 202:
9 CD4 + T cell Response Presentation of cognate antigen primes a T cell response leading to differentiation and proliferation of CD4 + T cells. Th0
10 CD4 + T cell Response Presentation of cognate antigen primes a T cell response leading to differentiation and proliferation of CD4 + T cells. Th0 Th1 Th2 Treg
11 CD4 + T cell Response Presentation of cognate antigen primes a T cell response leading to differentiation and proliferation of CD4 + T cells. Th0 Th1 Th2 Treg
12 CD4 + T cell Response Presentation of cognate antigen primes a T cell response leading to differentiation and proliferation of CD4 + T cells. Th0? Th1? Th2? Treg
13 CD4 + T cell Subsets Different subsets of CD4 + T cells have distinct cytokine profiles and function. Th Subtype Cytokines/Markers Th1 Th2 Th9 Th17 Th22 Cytotoxic CD4 + Treg IFNy IL-4, IL-5, IL-13 IL-9 IL-17A, IL-17F, IL-21 IL-22 CD107a, Granzyme B Foxp3, CD25, IL-10
14 CD4 + T cell Response in Melanoma Multiple groups using the OVA model system have reported that the CD4 T cell compartment remains ignorant to melanoma-derived antigen.* OT-II (CD4) T cells reported to be 100X less senitive to OVA antigen than OT-I (CD8) T cells.** *Preynat-Seauve O, et al Cancer Res. 67: *Stoitzner P, et al Cancer Immunol Immunother. 57: *Gerner MY, et al J Immunol. 181: *DiLillo DJ, et al J Immunol. 184: **Li et al J Immunol. 166:
15 B16.gD Melanoma Model gd-specific TCR transgenic mouse MHC Class II restricted for the glycoprotein D (gd) epitope of HSV CD4 + T cells recognise and proliferate in response to gd B16 melanoma cell line transduced to express the HSV-derived gd epitope and a GFP reporter gene.
16 B16.gD Melanoma Model gd-specific TCR transgenic mouse MHC Class II restricted for the glycoprotein D (gd) epitope of HSV CD4 + T cells recognise and proliferate in response to gd B16 melanoma cell line transduced to express the HSV-derived gd epitope and a GFP reporter gene. Parental Transduced GFP
17 B16.gD Melanoma Model Purified CD4 + T cells CFSE Label gdt-ii Mouse (CD45.1) B16.gD Tumour Bearing Mouse (CD45.2) gd Harvest tdln & Spleen
18 B16.gD Melanoma Model Purified CD4 + T cells CFSE Label gdt-ii Mouse (CD45.1) B16.gD Tumour Bearing Mouse (CD45.2) gd Harvest tdln & Spleen
19 B16.gD Melanoma Model Purified CD4 + T cells CFSE Label gdt-ii Mouse (CD45.1) B16.gD Tumour Bearing Mouse (CD45.2) gd Harvest tdln & Spleen
20 B16.gD Melanoma Model Purified CD4 + T cells CFSE Label gdt-ii Mouse (CD45.1) B16.gD Tumour Bearing Mouse (CD45.2) gd CD45-1 CD4 Harvest tdln & Spleen
21 Antigen Specific CD4 + T cell Proliferation Tumour Draining Node Spleen CFSE Dilution CFSE Dilution Inoculate B16-gD cutaneous I.V. 5 x 10 5 CFSE gdt-ii Harvest/Analysis Day -x Day 0 Day 2.5
22 Antigen Specific CD4 + T cell Proliferation Tumour Draining Node Spleen CFSE Dilution CFSE Dilution Inoculate B16-gD cutaneous I.V. 5 x 10 5 CFSE gdt-ii Harvest/Analysis Day -x Day 0 Day 2.5
23 CD4 + T cell Phenotyping Aim 1: Phenotyping the CD4 + T cell response generated to melanoma derived antigens. Th1 Th2 Th9 Th17 Th22 Cytotoxic Treg IFN-γ TNFα IL-4 IL-9 IL-17 IL-22 Granzyme B IL-10 FOXP3
24 Experimental Plan Harvest tumour draining lymph node cells and restimulate for intracellular cytokine staining. Adoptive transfer of antigen specific CD4 + T cells. PMA/Ionomycin Restim + BFA gd Harvest tdln 60 hours later 5 hours Stain for subset specific intracellular cytokines
25 T cell Staining Panels Core Stain CD4 CD45-1
26 T cell Staining Panels Th1/Th17/Th22 IFNy IL-17A IL-22 Core Stain CD4 CD45-1
27 T cell Staining Panels Th1/Th17/Th22 IFNy IL-17A IL-22 Core Stain CD4 CD45-1 Th2/Th9 IL-4 IL-13 IL-9
28 T cell Staining Panels Th1/Th17/Th22 IFNy IL-17A IL-22 Treg Foxp3 IL-10 CD25 Core Stain CD4 CD45-1 Th2/Th9 IL-4 IL-13 IL-9
29 T cell Staining Panels Th1/Th17/Th22 IFNy IL-17A IL-22 Treg Foxp3 IL-10 CD25 Core Stain CD4 CD45-1 Th2/Th9 IL-4 IL-13 IL-9 Cytotoxic Granzyme B CD107a CD25
30 RESULTS HSV Control Th1 Panel: gdt-ii response (white open) vs. endogenous (grey shaded) IFNγ IL-17α IL-22
31 RESULTS HSV Control Th2 Panel: gdt-ii response (white open) vs. endogenous (grey shaded) IL-4 IL-9 CD44
32 Cytotoxic Panel: RESULTS HSV Control gdt-ii response (white open) vs. endogenous (grey shaded) CD107a Granzyme B CD25
33 RESULTS HSV Control Treg Panel: gdt-ii response (white open) vs. endogenous (grey shaded) Endogenous Transferred CD25 Foxp3 IL-10
34 Optimisation Panel Design Markers and colours Positive Controls In vitro T cell skewing Gating FMOs, isotypes or single stains
35 Aim 2: Dendritic Cell Subsets Determine the ability of dendritic cell subsets to drive unique and more effective T cell responses to melanoma. It has been shown recently that different DC subsets are able to promote unique T cell responses. Langerhans cells required for development of Th17 cells during Candida albicans infection.* *(Igyarto B. Z Immunity. 35: )
36 Aim 2: Dendritic Cell Subsets Determine the ability of dendritic cell subsets to drive unique and more effective T cell responses to melanoma. Hypothesis: Different subsets of DCs may be able to drive more productive CD4 + T cell responses against melanoma.
37 Ex vivo Proliferation Assay Sort DC subsets from tumour bearing mice and coculture with specific CD4 + T cells ex vivo. B16.gD Tumour C T cell Transgenic Mouse gd Sorted DCs from tdlns CFSE labelled CD4 T cells Analyse CFSE Dilution
38 Mouse Dendritic Cell Subsets Mouse DC Phenotyping: Subtype Markers Location Plasmacytoid B220 + PDCA1 + Circulating Resident CD8 + CD4 - Lymph/Spleen Resident CD4 + CD8 - Lymph/Spleen Resident CD8 - CD4 - Lymph/Spleen Migratory CD11b + Dermis Migratory CD103 + Dermis Migratory CD326 + Epidermis
39 Mouse Dendritic Cell Subsets Mouse DC Phenotyping: Subtype Markers Location Plasmacytoid B220 + PDCA1 + Circulating Resident CD8 + CD4 - Lymph/Spleen Resident CD4 + CD8 - Lymph/Spleen Resident CD8 - CD4 - Lymph/Spleen Migratory CD11b + Dermis Migratory CD103 + Dermis Migratory CD326 + Epidermis
40 Mouse Dendritic Cell Subsets Mouse DC Phenotyping: Subtype Markers Location Plasmacytoid B220 + PDCA1 + Circulating Resident CD8 + CD4 - Lymph/Spleen Resident CD4 + CD8 - Lymph/Spleen Resident CD8 - CD4 - Lymph/Spleen Migratory CD11b + Dermis Migratory CD103 + Dermis Migratory CD326 + Epidermis
41 Mouse Dendritic Cell Subsets To further segregate DCs: XCR1 (cross-presenting DCs) Clec9A (cross-presenting DCs and pdcs) SIRPα (CD8 - resident and migratory DCs) DEC205 (CD8 + resident and migratory DCs) CD86 (co-stimulation marker)
42 Mouse Dendritic Cell Subsets To further segregate DCs: XCR1 (cross-presenting DCs) Clec9A (cross-presenting DCs and pdcs) SIRPα (CD8 - resident and migratory DCs) DEC205 (CD8 + resident and migratory DCs) CD86 (co-stimulation marker) CD86 SIRPα XCR1
43 Sorting DC Subsets Sort DC subsets from tumour bearing mice and coculture with specific CD4 + T cells ex vivo. Live Cells
44 Sorting DC Subsets Sort DC subsets from tumour bearing mice and coculture with specific CD4 + T cells ex vivo. Live Cells DCs
45 Sorting DC Subsets Sort DC subsets from tumour bearing mice and coculture with specific CD4 + T cells ex vivo. Live Cells DCs CD8a DCs
46 Sorting DC Subsets Sort DC subsets from tumour bearing mice and coculture with specific CD4 + T cells ex vivo. Live Cells DCs CD8a DCs CD103 DCs Langerhans Cells
47 Sorting DC Subsets Sort DC subsets from tumour bearing mice and coculture with specific CD4 + T cells ex vivo. Live Cells DCs CD8a DCs CD103 DCs CD11b DCs* Langerhans Cells CD11c DCs*
48 Mouse Dendritic Cell Subsets Mouse DC Phenotyping: Subtype Markers Location Plasmacytoid B220 + PDCA1 + Circulating Resident CD8 + CD4 - Lymph/Spleen Resident CD4 + CD8 - Lymph/Spleen Resident CD8 - CD4 - Lymph/Spleen Migratory CD11b + Dermis Migratory CD103 + Dermis Migratory CD326 + Epidermis
49 Ex vivo Proliferation DC subsets from tumour bearing mice co-cultured with antigen specific CD4 + T cells ex vivo.
50 Ex vivo Proliferation DC subsets from tumour bearing mice co-cultured with antigen specific CD4 + T cells ex vivo.
51 Recombinant Mouse Models Interrogate antigen presentation by DC subsets in vivo by using recombinant mice. Mouse/DC LCs CD11b + DC CD103 + DC CD8 + DC Migratory Migratory Migratory LNresident LNresident CD4 + DC CD11c.DTR CCR7 o/o Clec9A.dTR hulangerin.dta Batf3 o/o CD11c.DTR B6 or IAE -/- Chimaeras +/
52 Recombinant Mouse Models Interrogate antigen presentation by DC subsets in vivo by using recombinant mice. Mouse/DC LCs CD11b + DC CD103 + DC CD8 + DC Migratory Migratory Migratory LNresident LNresident CD4 + DC CD11c.DTR CCR7 o/o Clec9A.dTR hulangerin.dta Batf3 o/o CD11c.DTR B6 or IAE -/- Chimaeras +/
53 Recombinant Mouse Models Interrogate antigen presentation by DC subsets in vivo by using recombinant mice. Mouse/DC LCs CD11b + DC CD103 + DC CD8 + DC Migratory Migratory Migratory LNresident LNresident CD4 + DC CD11c.DTR CCR7 o/o Clec9A.dTR hulangerin.dta Batf3 o/o CD11c.DTR B6 or IAE -/- Chimaeras +/
54 Recombinant Mouse Models Interrogate antigen presentation by DC subsets in vivo by using recombinant mice. Mouse/DC LCs CD11b + DC CD103 + DC CD8 + DC Migratory Migratory Migratory LNresident LNresident CD4 + DC CD11c.DTR CCR7 o/o Clec9A.dTR hulangerin.dta Batf3 o/o CD11c.DTR B6 or IAE -/- Chimaeras +/
55 Recombinant Mouse Models Interrogate antigen presentation by DC subsets in vivo by using recombinant mice. Mouse/DC LCs CD11b + DC CD103 + DC CD8 + DC Migratory Migratory Migratory LNresident LNresident CD4 + DC CD11c.DTR CCR7 o/o Clec9A.dTR hulangerin.dta Batf3 o/o CD11c.DTR B6 or IAE -/- Chimaeras +/
56 Flow Cytometry Intracellular Cytokine Staining: T cell phenotyping Sorting: DC subsets Proliferation Assays: CFSE dilution
57 Acknowledgements Supervisors: Dr. Jason Waithman Prof. Prue Hart Group Members: Dr. Vanessa Fear Dr. Elke Seppanen Sarah Lacey Rachael Zemek This work is supported by grants from Cancer Australia and Cure Cancer Australia.
SUPPLEMENTARY INFORMATION
doi:1.138/nature1554 a TNF-α + in CD4 + cells [%] 1 GF SPF 6 b IL-1 + in CD4 + cells [%] 5 4 3 2 1 Supplementary Figure 1. Effect of microbiota on cytokine profiles of T cells in GALT. Frequencies of TNF-α
More informationSupplemental Figure 1
Supplemental Figure 1 1a 1c PD-1 MFI fold change 6 5 4 3 2 1 IL-1α IL-2 IL-4 IL-6 IL-1 IL-12 IL-13 IL-15 IL-17 IL-18 IL-21 IL-23 IFN-α Mut Human PD-1 promoter SBE-D 5 -GTCTG- -1.2kb SBE-P -CAGAC- -1.kb
More informationSupplemental Figure 1. Signature gene expression in in vitro differentiated Th0, Th1, Th2, Th17 and Treg cells. (A) Naïve CD4 + T cells were cultured
Supplemental Figure 1. Signature gene expression in in vitro differentiated Th0, Th1, Th2, Th17 and Treg cells. (A) Naïve CD4 + T cells were cultured under Th0, Th1, Th2, Th17, and Treg conditions. mrna
More informationDendritic cells in cancer immunotherapy Aimin Jiang
Dendritic cells in cancer immunotherapy Aimin Jiang Feb. 11, 2014 Dendritic cells at the interface of innate and adaptive immune responses Dendritic cells: initiators of adaptive immune responses Dendritic
More informationSupplementary Information. Tissue-wide immunity against Leishmania. through collective production of nitric oxide
Supplementary Information Tissue-wide immunity against Leishmania through collective production of nitric oxide Romain Olekhnovitch, Bernhard Ryffel, Andreas J. Müller and Philippe Bousso Supplementary
More informationNature Medicine: doi: /nm.3922
Title: Glucocorticoid-induced tumor necrosis factor receptor-related protein co-stimulation facilitates tumor regression by inducing IL-9-producing helper T cells Authors: Il-Kyu Kim, Byung-Seok Kim, Choong-Hyun
More informationHua Tang, Weiping Cao, Sudhir Pai Kasturi, Rajesh Ravindran, Helder I Nakaya, Kousik
SUPPLEMENTARY FIGURES 1-19 T H 2 response to cysteine-proteases requires dendritic cell-basophil cooperation via ROS mediated signaling Hua Tang, Weiping Cao, Sudhir Pai Kasturi, Rajesh Ravindran, Helder
More informationRegulation of anti-tumor immunity through migration of immune cell subsets within the tumor microenvironment Thomas F. Gajewski, M.D., Ph.D.
Regulation of anti-tumor immunity through migration of immune cell subsets within the tumor microenvironment Thomas F. Gajewski, M.D., Ph.D. Professor, Departments of Pathology and Medicine Program Leader,
More informationSupplementary Figure 1. mrna expression of chitinase and chitinase-like protein in splenic immune cells. Each splenic immune cell population was
Supplementary Figure 1. mrna expression of chitinase and chitinase-like protein in splenic immune cells. Each splenic immune cell population was sorted by FACS. Surface markers for sorting were CD11c +
More informationBlocking antibodies and peptides. Rat anti-mouse PD-1 (29F.1A12, rat IgG2a, k), PD-
Supplementary Methods Blocking antibodies and peptides. Rat anti-mouse PD-1 (29F.1A12, rat IgG2a, k), PD- L1 (10F.9G2, rat IgG2b, k), and PD-L2 (3.2, mouse IgG1) have been described (24). Anti-CTLA-4 (clone
More informationSupplementary Figure 1. Characterization of basophils after reconstitution of SCID mice
Supplementary figure legends Supplementary Figure 1. Characterization of after reconstitution of SCID mice with CD4 + CD62L + T cells. (A-C) SCID mice (n = 6 / group) were reconstituted with 2 x 1 6 CD4
More information% of live splenocytes. STAT5 deletion. (open shapes) % ROSA + % floxed
Supp. Figure 1. a 14 1 1 8 6 spleen cells (x1 6 ) 16 % of live splenocytes 5 4 3 1 % of live splenocytes 8 6 4 b 1 1 c % of CD11c + splenocytes (closed shapes) 8 6 4 8 6 4 % ROSA + (open shapes) % floxed
More informationCanberra, Australia). CD11c-DTR-OVA-GFP (B6.CD11c-OVA), B6.luc + and. Cancer Research Center, Germany). B6 or BALB/c.FoxP3-DTR-GFP mice were
Supplemental Materials and Methods Mice Female C57BL/6 (B6, I-E null, H-2 b ), BALB/c (H-2 d ) + ), FVB/N (H-2 q, I-E null, CD45.1 + ), and B6D2F1 (H-2 b/d ) mice were purchased from the Animal Resources
More informationECM1 controls T H 2 cell egress from lymph nodes through re-expression of S1P 1
ZH, Li et al, page 1 ECM1 controls T H 2 cell egress from lymph nodes through re-expression of S1P 1 Zhenhu Li 1,4,Yuan Zhang 1,4, Zhiduo Liu 1, Xiaodong Wu 1, Yuhan Zheng 1, Zhiyun Tao 1, Kairui Mao 1,
More informationSupplementary Figure 1. IL-12 serum levels and frequency of subsets in FL patients. (A) IL-12
1 Supplementary Data Figure legends Supplementary Figure 1. IL-12 serum levels and frequency of subsets in FL patients. (A) IL-12 serum levels measured by multiplex ELISA (Luminex) in FL patients before
More informationSUPPLEMENTARY INFORMATION
Complete but curtailed T-cell response to very-low-affinity antigen Dietmar Zehn, Sarah Y. Lee & Michael J. Bevan Supp. Fig. 1: TCR chain usage among endogenous K b /Ova reactive T cells. C57BL/6 mice
More informationCD4 + T cells recovered in Rag2 / recipient ( 10 5 ) Heart Lung Pancreas
a CD4 + T cells recovered in Rag2 / recipient ( 1 5 ) Heart Lung Pancreas.5 1 2 4 6 2 4 6 Ctla4 +/+ Ctla4 / Ctla4 / Lung Ctla4 / Pancreas b Heart Lung Pancreas Ctla4 +/+ Ctla4 / Ctla4 / Lung Ctla4 / Pancreas
More informationIL-34 is a tissue-restricted ligand of CSF1R required for the development of Langerhans cells and microglia
Supplementary Figures IL-34 is a tissue-restricted ligand of CSF1R required for the development of Langerhans cells and microglia Yaming Wang, Kristy J. Szretter, William Vermi, Susan Gilfillan, Cristina
More informationSupplementary Fig. 1 p38 MAPK negatively regulates DC differentiation. (a) Western blot analysis of p38 isoform expression in BM cells, immature DCs
Supplementary Fig. 1 p38 MAPK negatively regulates DC differentiation. (a) Western blot analysis of p38 isoform expression in BM cells, immature DCs (idcs) and mature DCs (mdcs). A myeloma cell line expressing
More informationSupplemental Table I.
Supplemental Table I Male / Mean ± SEM n Mean ± SEM n Body weight, g 29.2±0.4 17 29.7±0.5 17 Total cholesterol, mg/dl 534.0±30.8 17 561.6±26.1 17 HDL-cholesterol, mg/dl 9.6±0.8 17 10.1±0.7 17 Triglycerides,
More informationDynamic Interplay among Monocyte-Derived, Dermal, and Resident Lymph Node Dendritic Cells during the Generation of Vaccine Immunity to Fungi
Article Dynamic Interplay among Monocyte-Derived, Dermal, and Resident Lymph Node Dendritic Cells during the Generation of Vaccine Immunity to Fungi Karen Ersland, 1 Marcel Wüthrich, 2 and Bruce S. Klein
More informationSupplementary Figure 1. Example of gating strategy
Supplementary Figure 1. Example of gating strategy Legend Supplementary Figure 1: First, gating is performed to include only single cells (singlets) (A) and CD3+ cells (B). After gating on the lymphocyte
More informationNK cell flow cytometric assay In vivo DC viability and migration assay
NK cell flow cytometric assay 6 NK cells were purified, by negative selection with the NK Cell Isolation Kit (Miltenyi iotec), from spleen and lymph nodes of 6 RAG1KO mice, injected the day before with
More informationNature Immunology: doi: /ni Supplementary Figure 1. Examples of staining for each antibody used for the mass cytometry analysis.
Supplementary Figure 1 Examples of staining for each antibody used for the mass cytometry analysis. To illustrate the functionality of each antibody probe, representative plots illustrating the expected
More informationSupplemental Materials
Supplemental Materials Programmed death one homolog maintains the pool size of regulatory T cells by promoting their differentiation and stability Qi Wang 1, Jianwei He 1, Dallas B. Flies 2, Liqun Luo
More informationSupplementary Figures
Supplementary Figures Supplementary Figure 1. NKT ligand-loaded tumour antigen-presenting B cell- and monocyte-based vaccine induces NKT, NK and CD8 T cell responses. (A) The cytokine profiles of liver
More informationSUPPLEMENTARY INFORMATION
doi: 1.138/nature775 4 O.D. (595-655) 3 1 -ζ no antibody isotype ctrl Plated Soluble 1F6 397 7H11 Supplementary Figure 1 Soluble and plated anti- Abs induce -! signalling. B3Z cells stably expressing!
More informationSupplemental Figure Legends
Supplemental Figure Legends Supplemental Figure 1. SemaB / mice have normal immune cell populations. Cells were prepared from the spleens of WT and SemaB / mice, stained with various antibodies and then
More informationL-selectin Is Essential for Delivery of Activated CD8 + T Cells to Virus-Infected Organs for Protective Immunity
Cell Reports Supplemental Information L-selectin Is Essential for Delivery of Activated CD8 + T Cells to Virus-Infected Organs for Protective Immunity Rebar N. Mohammed, H. Angharad Watson, Miriam Vigar,
More informationFluorochrome Panel 1 Panel 2 Panel 3 Panel 4 Panel 5 CTLA-4 CTLA-4 CD15 CD3 FITC. Bio) PD-1 (MIH4, BD) ICOS (C398.4A, Biolegend) PD-L1 (MIH1, BD)
Additional file : Table S. Antibodies used for panel stain to identify peripheral immune cell subsets. Panel : PD- signaling; Panel : CD + T cells, CD + T cells, B cells; Panel : Tregs; Panel :, -T, cdc,
More informationILC1 and ILC3 isolation and culture Following cell sorting, we confirmed that the recovered cells belonged to the ILC1, ILC2 and
Supplementary Methods and isolation and culture Following cell sorting, we confirmed that the recovered cells belonged to the, ILC2 and subsets. For this purpose we performed intracellular flow cytometry
More informationCytotoxicity assays. Rory D. de Vries, PhD 1. Viroscience lab, Erasmus MC, Rotterdam, the Netherlands
Cytotoxicity assays Rory D. de Vries, PhD 1 1 Viroscience lab, Erasmus MC, Rotterdam, the Netherlands Anti-influenza immunity Humoral / CD4+ / CD8+ / NK? Function of CTL Elimination of virus-infected cells?
More informationMATERIALS AND METHODS. Neutralizing antibodies specific to mouse Dll1, Dll4, J1 and J2 were prepared as described. 1,2 All
MATERIALS AND METHODS Antibodies (Abs), flow cytometry analysis and cell lines Neutralizing antibodies specific to mouse Dll1, Dll4, J1 and J2 were prepared as described. 1,2 All other antibodies used
More informationSynergistic effects of indoleamine 2,3-dioxygenase (IDO) inhibitor drugs and chemotherapy in melanoma.
Synergistic effects of indoleamine 2,3-dioxygenase (IDO) inhibitor drugs and chemotherapy in melanoma. Madhav D. Sharma Cancer Immunology, Inflammation, and Tolerance Program, Department of Pediatrics,
More informationSupplemental Information. Aryl Hydrocarbon Receptor Controls. Monocyte Differentiation. into Dendritic Cells versus Macrophages
Immunity, Volume 47 Supplemental Information Aryl Hydrocarbon Receptor Controls Monocyte Differentiation into Dendritic Cells versus Macrophages Christel Goudot, Alice Coillard, Alexandra-Chloé Villani,
More informationMicro 204. Cytotoxic T Lymphocytes (CTL) Lewis Lanier
Micro 204 Cytotoxic T Lymphocytes (CTL) Lewis Lanier Lewis.Lanier@ucsf.edu Lymphocyte-mediated Cytotoxicity CD8 + αβ-tcr + T cells CD4 + αβ-tcr + T cells γδ-tcr + T cells Natural Killer cells CD8 + αβ-tcr
More informationSupplemental materials
Supplemental materials 1 Supplemental Fig. 1 Immunogram This immunogram summarizes patient clinical data and immune parameters at corresponding time points for Patient IMF-32. The top panel illustrates
More informationSupplemental Information. Human CD1c + Dendritic Cells Drive. the Differentiation of CD103 + CD8 + Mucosal Effector T Cells via the Cytokine TGF-
Immunity, Volume 38 Supplemental Information Human CD1c + Dendritic Cells Drive the Differentiation of CD103 + CD8 + Mucosal Effector T Cells via the Cytokine TGF- Chun I. Yu Christian Becker Yuanyuan
More informationCELLULAR AND MOLECULAR REQUIREMENTS FOR REJECTION OF B16 MELANOMA IN THE SETTING OF REGULATORY T CELL DEPLETION AND HOMEOSTATIC PROLIFERATION
CELLULAR AND MOLECULAR REQUIREMENTS FOR REJECTION OF B16 MELANOMA IN THE SETTING OF REGULATORY T CELL DEPLETION AND HOMEOSTATIC PROLIFERATION Justin Kline 1, Long Zhang 1, and Thomas F. Gajewski 1,2 Departments
More informationAkt and mtor pathways differentially regulate the development of natural and inducible. T H 17 cells
Akt and mtor pathways differentially regulate the development of natural and inducible T H 17 cells Jiyeon S Kim, Tammarah Sklarz, Lauren Banks, Mercy Gohil, Adam T Waickman, Nicolas Skuli, Bryan L Krock,
More informationSupplementary Figure 1: TSLP receptor skin expression in dcssc. A: Healthy control (HC) skin with TSLP receptor expression in brown (10x
Supplementary Figure 1: TSLP receptor skin expression in dcssc. A: Healthy control (HC) skin with TSLP receptor expression in brown (10x magnification). B: Second HC skin stained for TSLP receptor in brown
More informationSupplementary Figure 1. Efficient DC depletion in CD11c.DOG transgenic mice
Supplementary Figure 1. Efficient DC depletion in CD11c.DOG transgenic mice (a) CD11c.DOG transgenic mice (tg) were treated with 8 ng/g body weight (b.w.) diphtheria toxin (DT) i.p. on day -1 and every
More informationB220 CD4 CD8. Figure 1. Confocal Image of Sensitized HLN. Representative image of a sensitized HLN
B220 CD4 CD8 Natarajan et al., unpublished data Figure 1. Confocal Image of Sensitized HLN. Representative image of a sensitized HLN showing B cell follicles and T cell areas. 20 µm thick. Image of magnification
More informationInstitute t of Experimental Immunology University of Bonn, Germany
How T cells recognize antigen Christian Kurts Institute t of Experimental Immunology University of Bonn, Germany When do T cells recognize antigen? Antigen uptake Activation phase 1st recognition Effector
More informationTITLE: MODULATION OF T CELL TOLERANCE IN A MURINE MODEL FOR IMMUNOTHERAPY OF PROSTATIC ADENOCARCINOMA
AD Award Number: DAMD17-01-1-0085 TITLE: MODULATION OF T CELL TOLERANCE IN A MURINE MODEL FOR IMMUNOTHERAPY OF PROSTATIC ADENOCARCINOMA PRINCIPAL INVESTIGATOR: ARTHUR A HURWITZ, Ph.d. CONTRACTING ORGANIZATION:
More informationAlessandra Franco, M.D., Ph.D. UCSD School of Medicine Department of Pediatrics Division of Allergy, Immunology and Rheumatology
Natural regulatory T cells recognize the heavy constant region (Fc) of immunoglobulins: a novel mechanism for IVIG immunotherapy in Pediatric Immune-mediated diseases Alessandra Franco, M.D., Ph.D. UCSD
More informationSUPPLEMENT Supplementary Figure 1: (A) (B)
SUPPLEMENT Supplementary Figure 1: CD4 + naïve effector T cells (CD4 effector) were labeled with CFSE, stimulated with α-cd2/cd3/cd28 coated beads (at 2 beads/cell) and cultured alone or cocultured with
More informationSupplementary Figure 1. Deletion of Smad3 prevents B16F10 melanoma invasion and metastasis in a mouse s.c. tumor model.
A B16F1 s.c. Lung LN Distant lymph nodes Colon B B16F1 s.c. Supplementary Figure 1. Deletion of Smad3 prevents B16F1 melanoma invasion and metastasis in a mouse s.c. tumor model. Highly invasive growth
More informationCombined Rho-kinase inhibition and immunogenic cell death triggers and propagates immunity against cancer
Supplementary Information Combined Rho-kinase inhibition and immunogenic cell death triggers and propagates immunity against cancer Gi-Hoon Nam, Eun-Jung Lee, Yoon Kyoung Kim, Yeonsun Hong, Yoonjeong Choi,
More informationSUPPLEMENTARY FIGURES
SUPPLEMENTARY FIGURES Supplementary Figure 1: Chemokine receptor expression profiles of CCR6 + and CCR6 - CD4 + IL-17A +/ex and Treg cells. Quantitative PCR analysis of chemokine receptor transcript abundance
More informationKrishnamoorthy et al.,
Krishnamoorthy et al., c d e ND ND Supplementary Figure 1 RSV-induces inflammation even in the asence of allergen. Tolerized pups were either infected with RSV or not. The mice were sacrificed a week following
More informationW/T Itgam -/- F4/80 CD115. F4/80 hi CD115 + F4/80 + CD115 +
F4/8 % in the peritoneal lavage 6 4 2 p=.15 n.s p=.76 CD115 F4/8 hi CD115 + F4/8 + CD115 + F4/8 hi CD115 + F4/8 + CD115 + MHCII MHCII Supplementary Figure S1. CD11b deficiency affects the cellular responses
More informationNature Immunology: doi: /ni Supplementary Figure 1. Id2 and Id3 define polyclonal T H 1 and T FH cell subsets.
Supplementary Figure 1 Id2 and Id3 define polyclonal T H 1 and T FH cell subsets. Id2 YFP/+ (a) or Id3 GFP/+ (b) mice were analyzed 7 days after LCMV infection. T H 1 (SLAM + CXCR5 or CXCR5 PD-1 ), T FH
More informationAntibodies for human plasmacytoïd dendritic cells studies Dendritics SAS, 60 avenue Rockefeller, F Lyon
Antibodies for human plasmacytoïd dendritic cells studies Dendritics SAS, 60 avenue Rockefeller, F-69008 Lyon www.dendritics.net Human plasmacytoïd dendritic cells (PDCs) are considered the main sentinels
More informationIRF5 distinguishes severe asthma in humans and drives Th1 phenotype and airway hyperreactivity in mice
IRF5 distinguishes severe asthma in humans and drives Th1 phenotype and airway hyperreactivity in mice Timothy B. Oriss,, Prabir Ray, Anuradha Ray JCI Insight. 2017;2(10):e91019.. Research Article Pulmonology
More informationControl of intestinal inflammation by regulatory T cells
Control of intestinal inflammation by regulatory T cells Fiona Powrie Sir William Dunn School of Pathology University of Oxford fiona.powrie@path.ox.ac.uk Regulatory T cells prevent immune pathology in
More informationSupplemental Materials for. Effects of sphingosine-1-phosphate receptor 1 phosphorylation in response to. FTY720 during neuroinflammation
Supplemental Materials for Effects of sphingosine-1-phosphate receptor 1 phosphorylation in response to FTY7 during neuroinflammation This file includes: Supplemental Table 1. EAE clinical parameters of
More informationSupplementary Figure 1 Protease allergens induce IgE and IgG1 production. (a-c)
1 Supplementary Figure 1 Protease allergens induce IgE and IgG1 production. (a-c) Serum IgG1 (a), IgM (b) and IgG2 (c) concentrations in response to papain immediately before primary immunization (day
More informationAdaptive (acquired) immunity. Professor Peter Delves University College London
Adaptive (acquired) immunity Professor Peter Delves University College London p.delves@ucl.ac.uk Haematopoiesis Haematopoiesis Lymphocytes = adaptive response Recognition of pathogens by adaptive cells,
More informationVISTA, a novel immune checkpoint protein ligand that suppresses anti-tumor tumor T cell responses. Li Wang. Dartmouth Medical School
VISTA, a novel immune checkpoint protein ligand that suppresses anti-tumor tumor T cell responses Li Wang Dartmouth Medical School The B7 Immunoglobulin Super-Family immune regulators APC T cell Co-stimulatory:
More informationExpression of indoleamine 2,3-dioxygenase by plasmacytoid dendritic cells in tumor-draining lymph nodes
Research article Expression of indoleamine 2,3-dioxygenase by plasmacytoid dendritic cells in tumor-draining lymph nodes David H. Munn, 1,2,3 Madhav D. Sharma, 1,2 Deyan Hou, 1,2 Babak Baban, 2 Jeffrey
More informationD CD8 T cell number (x10 6 )
IFNγ Supplemental Figure 1. CD T cell number (x1 6 ) 18 15 1 9 6 3 CD CD T cells CD6L C CD5 CD T cells CD6L D CD8 T cell number (x1 6 ) 1 8 6 E CD CD8 T cells CD6L F Log(1)CFU/g Feces 1 8 6 p
More informationSUPPORTING INFORMATIONS
SUPPORTING INFORMATIONS Mice MT/ret RetCD3ε KO α-cd25 treated MT/ret Age 1 month 3 mnths 6 months 1 month 3 months 6 months 1 month 3 months 6 months 2/87 Survival 87/87 incidence of 17/87 1 ary tumor
More informationNature Medicine doi: /nm.3957
Supplementary Fig. 1. p38 alternative activation, IL-21 expression, and T helper cell transcription factors in PDAC tissue. (a) Tissue microarrays of pancreatic tissue from 192 patients with pancreatic
More informationUnderstanding the T cell response to tumors using transnuclear mouse models
Understanding the T cell response to tumors using transnuclear mouse models Stephanie Dougan Dana-Farber Cancer Institute Boston, MA Presenter Disclosure Information Stephanie Dougan The following relationships
More informationTCR, MHC and coreceptors
Cooperation In Immune Responses Antigen processing how peptides get into MHC Antigen processing involves the intracellular proteolytic generation of MHC binding proteins Protein antigens may be processed
More informationExamples of questions for Cellular Immunology/Cellular Biology and Immunology
Examples of questions for Cellular Immunology/Cellular Biology and Immunology Each student gets a set of 6 questions, so that each set contains different types of questions and that the set of questions
More informationMURDOCH RESEARCH REPOSITORY
MURDOCH RESEARCH REPOSITORY This is the author s final version of the work, as accepted for publication following peer review but without the publisher s layout or pagination. The definitive version is
More informationExcerpt from MACS&more Vol 17 1/2016. Kenji Miki¹, Koji Nagaoka¹, Hermann Bohnenkamp², Takayuki Yoshimoto³, Ryuji Maekawa¹*, and Takashi Kamigaki⁴
Excerpt from MCS&more Vol 17 1/2016 Dendritic cells pulsed with induce both OV-specific CD4 + and CD8 + T cells and cause antitumor effects in a mouse model of lymphoma Kenji Miki¹, Koji Nagaoka¹, Hermann
More informationScott Abrams, Ph.D. Professor of Oncology, x4375 Kuby Immunology SEVENTH EDITION
Scott Abrams, Ph.D. Professor of Oncology, x4375 scott.abrams@roswellpark.org Kuby Immunology SEVENTH EDITION CHAPTER 11 T-Cell Activation, Differentiation, and Memory Copyright 2013 by W. H. Freeman and
More informationSupplementary Figure S1: Alignment of CD28H. (a) Alignment of human CD28H with other known B7 receptors. (b) Alignment of CD28H orthologs.
Supplementary Figure S1: Alignment of CD28H. (a) Alignment of human CD28H with other known B7 receptors. (b) Alignment of CD28H orthologs. Predicted CD28H protein sequences from human, chimpanzee (pan
More informationMCSF drives regulatory DC development in stromal co-cultures supporting hematopoiesis
Petvises et al. BMC Immunology (2018) 19:21 https://doi.org/10.1186/s12865-018-0255-y RESEARCH ARTICLE Open Access MCSF drives regulatory DC development in stromal co-cultures supporting hematopoiesis
More informationSupplementary Figure 1. Efficiency of Mll4 deletion and its effect on T cell populations in the periphery. Nature Immunology: doi: /ni.
Supplementary Figure 1 Efficiency of Mll4 deletion and its effect on T cell populations in the periphery. Expression of Mll4 floxed alleles (16-19) in naive CD4 + T cells isolated from lymph nodes and
More informationOptimizing Intracellular Flow Cytometry:
Optimizing Intracellular Flow Cytometry: Simultaneous Detection of Cytokines and Transcription Factors An encore presentation by Jurg Rohrer, PhD, BD Biosciences 10.26.10 Outline Introduction Cytokines
More informationSupplementary Materials for
www.sciencetranslationalmedicine.org/cgi/content/full/8/352/352ra110/dc1 Supplementary Materials for Spatially selective depletion of tumor-associated regulatory T cells with near-infrared photoimmunotherapy
More informationSupplementary Figure 1. Double-staining immunofluorescence analysis of invasive colon and breast cancers. Specimens from invasive ductal breast
Supplementary Figure 1. Double-staining immunofluorescence analysis of invasive colon and breast cancers. Specimens from invasive ductal breast carcinoma (a) and colon adenocarcinoma (b) were staining
More informationwell for 2 h at rt. Each dot represents an individual mouse and bar is the mean ±
Supplementary data: Control DC Blimp-1 ko DC 8 6 4 2-2 IL-1β p=.5 medium 8 6 4 2 IL-2 Medium p=.16 8 6 4 2 IL-6 medium p=.3 5 4 3 2 1-1 medium IL-1 n.s. 25 2 15 1 5 IL-12(p7) p=.15 5 IFNγ p=.65 4 3 2 1
More informationLineage relationship and protective immunity of memory CD8 T cell subsets
23 Nature Publishing Group http://www.nature.com/natureimmunology Lineage relationship and protective immunity of memory CD8 T cell subsets E. John Wherry 1 *,Volker Teichgräber 1 *,Todd C. Becker 1,David
More informationDendritic cells and their role in immunity
PART I Dendritic cells and their role in immunity CHAPTER 1 Subpopulations and differentiation of mouse dendritic cells Carlos Ardavín Universidad Autónoma 1.1 DENDRITIC CELL SUBPOPULATIONS Dendritic cells
More informationDURACLONE IF BE CERTAIN ABOUT THE RESPONSE. l res. a il n c n. For Research Use Only - Not for use in Diagnostic procedures
DURACLONE IF earch tria l res lc a om ic il n c n nio pa Yo ur BE CERTAIN ABOUT THE RESPONSE For Research Use Only - Not for use in Diagnostic procedures BE CERTAIN ABOUT THE RESPONSE The sensitive and
More informationB6/COLODR/SPL/11C/83/LAP/#2.006 B6/COLODR/SPL/11C/86/LAP/#2.016 CD11C B6/COLODR/SPL/11C/80/LAP/#2.011 CD11C
CD3-specific antibody-induced immune tolerance and suppression of autoimmune encephalomyelitis involves TGF-β production through phagocytes digesting apoptotic T cells Sylvain Perruche 1,3, Pin Zhang 1,
More informationThe Use of Human Dendritic Cell
The Use of Human Dendritic Cell Subsets in Cancer Vaccines Jolien Sweere Student #: 3145298 Master Thesis Infection and Immunity Supervisor: Dr. Jeanette Leusen Immunotherapy University Medical Centre
More informationImmune Cells in Atherosclerosis Regulatory vs Inflammatory T cells Göran K Hansson
Immune Cells in Atherosclerosis Regulatory vs Inflammatory T cells Göran K Hansson Center for Molecular Medicine Karolinska Institute Stockholm, Sweden DECLARATION OF CONFLICT OF INTEREST Goran Hansson
More informationSupplemental Figure 1. Cell-bound Cetuximab reduces EGFR staining intensity. Blood
Antibody-mediated depletion of CD19-CAR T cells Supplemental 1 Supplemental Materials Supplemental Figure 1. Supplemental Figure 1. Cell-bound Cetuximab reduces EGFR staining intensity. Blood cells were
More informationSupplementary Figure 1
Supplementary Figure 1 Identification of IFN-γ-producing CD8 + and CD4 + T cells with naive phenotype by alternative gating and sample-processing strategies. a. Contour 5% probability plots show definition
More informationndln NK Cells (x10 3 ) Days post-infection (A/PR/8) *** *** *** Liver NK Cells (x10 4 ) Days post-infection (MCMV)
A mln NK Cells(x ) 6 1 * ndln NK Cells (x ) ns C Lung NK Cells(x ) 1 1 7 * D LN NK Cells (x ) 1 7 1 7 Days post-infection (A/PR/8) * * E Liver NK Cells (x ) 1 7 Days post-infection (A/PR/8) * * * 1 7 Days
More informationNaive and memory CD8 T cell responses after antigen stimulation in vivo
University of Iowa Iowa Research Online Theses and Dissertations Summer 2011 Naive and memory CD8 T cell responses after antigen stimulation in vivo Matthew David Martin University of Iowa Copyright 2011
More informationSignal 3 requirement for memory CD8 1 T-cell activation is determined by the infectious pathogen
3176 DOI 1.12/eji.21141537 Eur. J. Immunol. 211. 41: 3176 3186 Signal 3 requirement for memory CD8 1 T-cell activation is determined by the infectious pathogen Selina J. Keppler 1,2 and Peter Aichele 1
More informationTable S1. Viral load and CD4 count of HIV-infected patient population
Table S1. Viral load and CD4 count of HIV-infected patient population Subject ID Viral load (No. of copies per ml of plasma) CD4 count (No. of cells/µl of blood) 28 7, 14 29 7, 23 21 361,99 94 217 7, 11
More informationpplementary Figur Supplementary Figure 1. a.
pplementary Figur Supplementary Figure 1. a. Quantification by RT-qPCR of YFV-17D and YFV-17D pol- (+) RNA in the supernatant of cultured Huh7.5 cells following viral RNA electroporation of respective
More informationSupplemental Information. CD4 + CD25 + Foxp3 + Regulatory T Cells Promote. Th17 Cells In Vitro and Enhance Host Resistance
Immunity, Volume 34 Supplemental Information D4 + D25 + + Regulatory T ells Promote Th17 ells In Vitro and Enhance Host Resistance in Mouse andida albicans Th17 ell Infection Model Pushpa Pandiyan, Heather
More informationInterferon-dependent IL-10 production by Tregs limits tumor Th17 inflammation
Interferon-dependent IL-10 production by Tregs limits tumor Th17 inflammation C. Andrew Stewart,, Werner Müller, Giorgio Trinchieri J Clin Invest. 2013;123(11):4859-4874. https://doi.org/10.1172/jci65180.
More informationGeneration of ST2-GFP reporter mice and characterization of ILC1 cells following infection
Supplementary Figure 1 Generation of ST2-GFP reporter mice and characterization of ILC1 cells following infection with influenza virus. (a) ST2-GFP reporter mice were generated as described in Methods.
More informationSupplemental Information. T Cells Enhance Autoimmunity by Restraining Regulatory T Cell Responses via an Interleukin-23-Dependent Mechanism
Immunity, Volume 33 Supplemental Information T Cells Enhance Autoimmunity by Restraining Regulatory T Cell Responses via an Interleukin-23-Dependent Mechanism Franziska Petermann, Veit Rothhammer, Malte
More informationPeli1 negatively regulates T-cell activation and prevents autoimmunity
Peli1 negatively regulates T-cell activation and prevents autoimmunity Mikyoung Chang 1,*, Wei Jin 1,5,*, Jae-Hoon Chang 1, Yi-chuan Xiao 1, George Brittain 1, Jiayi Yu 1, Xiaofei Zhou 1, Yi-Hong Wang
More informationSupplementary Figure 1. Normal T lymphocyte populations in Dapk -/- mice. (a) Normal thymic development in Dapk -/- mice. Thymocytes from WT and Dapk
Supplementary Figure 1. Normal T lymphocyte populations in Dapk -/- mice. (a) Normal thymic development in Dapk -/- mice. Thymocytes from WT and Dapk -/- mice were stained for expression of CD4 and CD8.
More informationSupplementary Fig. 1 No relative growth advantage of Foxp3 negative cells.
Supplementary Fig. 1 Supplementary Figure S1: No relative growth advantage of Foxp3 negative cells. itreg were induced from WT (A) or FIR (B) CD4 + T cells. FIR itregs were then removed from the TCR signal
More informationNaive, memory and regulatory T lymphocytes populations analysis
Naive, memory and regulatory T lymphocytes populations analysis Jaen Olivier, PhD ojaen@beckmancoulter.com Cellular Analysis application specialist Beckman Coulter France Introduction Flow cytometric analysis
More informationand follicular helper T cells is Egr2-dependent. (a) Diagrammatic representation of the
Supplementary Figure 1. LAG3 + Treg-mediated regulation of germinal center B cells and follicular helper T cells is Egr2-dependent. (a) Diagrammatic representation of the experimental protocol for the
More informationThe Good, the Bad and the Ugly: Clinical trials which assess vaccine characteristics. ISBT Meeting, San Francisco, CA November 4-8, 2004
The Good, the Bad and the Ugly: Clinical trials which assess vaccine characteristics ISBT Meeting, San Francisco, CA November 4-8, 2004 Ideal cancer vaccine trial 1. An informative immune assay 2. Ability
More information