Jenni Loyall

Jacobs (United States)

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Richard B. Bankert

University at Buffalo, State University of New York

Raymond J. Kelleher

Harvard University

Kunle Odunsi

University of Chicago Medicine Comprehensive Cancer Center

Sathy V. Balu‐Iyer

University at Buffalo, State University of New York

Gautam N. Shenoy

University at Buffalo, State University of New York

Vandana Iyer

University at Buffalo, State University of New York

Paul K. Wallace

Fugro (United States)

Charles S. Berenson

VA Western New York Healthcare System

Anas M. Fathallah

Halozyme Therapeutics (United States)

Joseph D. Tario

Roswell Park Comprehensive Cancer Center

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University at Buffalo, State University of New York

Roswell Park Comprehensive Cancer Center

University of Rochester Medical Center

New York University

University of Pennsylvania

University of Rochester

Jacobs (United States)

Weatherford College

The University of Texas Southwestern Medical Center

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Supplemental Figure S1 from Extracellular Vesicles Present in Human Ovarian Tumor Microenvironments Induce a Phosphatidylserine-Dependent Arrest in the T-cell Signaling Cascade

Raymond J. Kelleher Sathy V. Balu‐Iyer Jenni Loyall Anthony J. Sacca Gautam N. Shenoy

Fig S1. Delipidation of ascites fluid.

Extracellular Vesicles

10.1158/2326-6066.22537408

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Germ line tumor-associated immunoglobulin VH region peptides provoke a tumor-specific immune response without altering the response potential of normal B cells

Blood (2004)

Qiang Lou Raymond J. Kelleher Alessandro Sette Jenni Loyall Scott Southwood

10.1182/blood-2004-01-0105

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Supplemental Figure S2 from Extracellular Vesicles Present in Human Ovarian Tumor Microenvironments Induce a Phosphatidylserine-Dependent Arrest in the T-cell Signaling Cascade

Raymond J. Kelleher Sathy V. Balu‐Iyer Jenni Loyall Anthony J. Sacca Gautam N. Shenoy

Fig S2. PS/PC liposome titration.

Extracellular Vesicles

Extracellular vesicles

10.1158/2326-6066.22537405

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Supplementary Table 1 from Exosomes Associated with Human Ovarian Tumors Harbor a Reversible Checkpoint of T-cell Responses

Gautam N. Shenoy Jenni Loyall Orla Maguire Vandana Iyer Raymond J. Kelleher

List of reagents used in flow cytometry and fluorescence microscopy

Table (database)

10.1158/2326-6066.22538890.v1

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Supplementary Figures 1 - 4 from Exosomes Associated with Human Ovarian Tumors Harbor a Reversible Checkpoint of T-cell Responses

Gautam N. Shenoy Jenni Loyall Orla Maguire Vandana Iyer Raymond J. Kelleher

S1: Exosome derived from different ascites fluids show varying degrees of inhibition. S2: Exosomes do not affect T cell death following activation. S3: Exosomes inhibit IL-2 production by T cells. S4: Exosomes inhibit IFN-gamma production by T cells.

Exosome

10.1158/2326-6066.22538893

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Exosomes Represent an Immune Suppressive T Cell Checkpoint in Human Chronic Inflammatory Microenvironments

Immunological Investigations (2020)

Gautam N. Shenoy Maulasri Bhatta Jenni Loyall Raymond J. Kelleher Joel M. Bernstein

Background: T cells present in chronic inflammatory tissues such as nasal polyps (from chronic rhinosinusitis patients) have been demonstrated to be hypo-responsive to activation via the TCR, similar to tumor-specific T cells in multiple different human tumor microenvironments. While immunosuppressive exosomes have been known to contribute to the failure of the tumor-associated T cells to respond optimally to activation stimuli, it is not known whether they play a similar role in chronic inflammatory microenvironments. In the current study, we investigate whether exosomes derived from chronic inflammatory microenvironments contribute to the immune suppression of T cells. Methods: Exosomes were isolated by ultracentrifugation and characterized by size and composition using nanoparticle tracking analysis, scanning electron microscopy, antibody arrays and flow exometry. Immunosuppressive ability of the exosomes was measured by quantifying its effect on activation of T cells, using nuclear translocation of NFκB as an activation endpoint. Results: Exosomes were isolated and characterized from two different types of chronic inflammatory tissues - nasal polyps from chronic rhinosinusitis patients and synovial fluid from rheumatoid arthritis patients. These exosomes arrest the activation of T cells stimulated via the TCR. This immune suppression, like that which is seen in tumor microenvironments, is dependent in part upon a lipid, ganglioside GD3, which is expressed on the exosomal surface. Conclusion: Immunosuppressive exosomes present in non-malignant chronic inflammatory tissues represent a new T cell checkpoint, and potentially represent a novel therapeutic target to enhance the response to current therapies and prevent disease recurrences.

10.1080/08820139.2020.1748047

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Data from Exosomes Associated with Human Ovarian Tumors Harbor a Reversible Checkpoint of T-cell Responses

Gautam N. Shenoy Jenni Loyall Orla Maguire Vandana Iyer Raymond J. Kelleher

<div>AbstractNano-sized membrane-encapsulated extracellular vesicles isolated from the ascites fluids of ovarian cancer patients are identified as exosomes based on their biophysical and compositional characteristics. We report here that T cells pulsed with these tumor-associated exosomes during TCR-dependent activation inhibit various activation endpoints including translocation of NFκB and NFAT into the nucleus, upregulation of CD69 and CD107a, production of cytokines, and cell proliferation. In addition, the activation of virus-specific CD8+ T cells that are stimulated with the cognate viral peptides presented in the context of class I MHC is also suppressed by the exosomes. The inhibition occurs without loss of cell viability and coincidentally with the binding and internalization of these exosomes. This exosome-mediated inhibition of T cells was transient and reversible: T cells exposed to exosomes can be reactivated once exosomes are removed. We conclude that tumor-associated exosomes are immunosuppressive and represent a therapeutic target, blockade of which would enhance the antitumor response of quiescent tumor-associated T cells and prevent the functional arrest of adoptively transferred tumor-specific T cells or chimeric antigen receptor T cells. Cancer Immunol Res; 6(2); 236–47. ©2018 AACR.</div>

NFAT

Internalization

Exosome

10.1158/2326-6066.c.6548932.v1

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Supplemental Methods and Figure Legends from Extracellular Vesicles Present in Human Ovarian Tumor Microenvironments Induce a Phosphatidylserine-Dependent Arrest in the T-cell Signaling Cascade

Raymond J. Kelleher Sathy V. Balu‐Iyer Jenni Loyall Anthony J. Sacca Gautam N. Shenoy

Description of additional methods and figure legends for Supplemental Figures

Extracellular Vesicles

10.1158/2326-6066.22537387

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