Exosomes Present in the Human Ovarian Tumor Microenvironments Rapidly Arrest T Cells
Paul K. Wallace Ph.D. - Roswell Park Cancer Institute
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Introduction
Why do Tumors Progress?
Ovarian Tissue – Plenty of T cells
Ascites – More T cells Than Tumor
Monitor and Quantify T Cell Function
TCR Signaling in T Cells from Healthy Donor & Cancer Patient’s PBL
Confocal
Normal Human PBL T Cells - Not Activated
Normal Human PBL T Cells - Activated
T Cells from Primary and Metastatic Ovarian Tumors
Nuclear Translocation on the ImageStream
T Cells from Primary and Metastatic Ovarian Tumors
Similar results obtained on the ImageStream
Why Are the Tumor-Associated T Cells Non-Functional?
Ovarian Ascites Decreases the Translocation of NFκB in a Dose Dependent Manner
Tumor Ascites Inhibits CD4 & CD8 T Cell Activation
What Is the Immunosuppressive Factor(s) Present in Ovarian Tumor Ascites Fluids?
Characteristics of Immunosuppressive Factor(s) Present in Ovarian Ascites
Ether Extraction of Ascites Fluid Removes Inhibitory Activity
Hexane Isopropanol Extraction of Multiple Ascites Samples
Liposomes Containing PS/PC But Not PC Only
In What Form Is PS Expressed in Tumor Ascites Fluids
Ultracentrifugation of Tumor Ascites Fluids Removes Inhibitory Activity
Electron Microscopy of Unfixed Ovarian Ascites Fluid Pellet Reveal Exosomes
Exosomes From Ultracentrifuged Tumor Ascites Pellet Inhibit T Cell Activation and Inhibition is Reversed by Anti-PS Antibody
Release of Microvegetables (aka Exosomes) from Normal and Neoplastic Cells
Exosome Background
Phenotypic Analysis of Exosomes by Flow Cytometry
NovoCyte Microvesicle Analysis
Exosomes Bind PE Labeled Annexin V and Binding Is Inhibited by Unlabeled Annexin V
Exosome Phenotype by Western Array
Exosomes Inhibit CD69 Upregulation and CD107a Mobilization by T cells
Exosomes Inhibit IL-2 and IFNy Production by T cells
Antigen-Specific NFAT Signaling Inhibition by Exosomes
(DGK) Phosphorylates (DAG) to An Inactive Phosphatidic Acid
Inhibition of Diacylglycerol Kinase
Conclusions: What We Know
Conclusions: What We Want to Know
Acknowledgements
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