Stanford researchers describe how their integration of CyTOF® mass cytometry analysis with other omic approaches revealed the existence of an immune clock during human pregnancy.

In this recorded webinar, Dr. Brice Gaudilliere, MD, PhD, and Nima Aghaeepour, PhD, both of Stanford University School of Medicine, describe how they characterized the molecular pacemakers of pregnancy by integrating CyTOF® mass cytometry analysis and other omic approaches on blood samples collected throughout pregnancy.

The maintenance of pregnancy relies on a finely tuned immune balance between tolerance to the fetal allograft and protective mechanisms against invading pathogens. Demonstrating the chronology of immune adaptations to a term pregnancy provides the framework for future studies examining deviations implicated in pregnancy-related pathologies including preterm birth and preeclampsia.

Gaudilliere presents data demonstrating that these adaptations are precisely timed, reflecting an immune clock of pregnancy in women delivering at term. An elastic net model with prior Bayesian distributions extracted from literature-based knowledge of the immune was used to develop a predictive model of interrelated immune events that accurately captured the chronology of pregnancy.

Aghaeepour describes how the immunological dataset derived from the mass cytometry analysis was integrated with other omic datasets collected simultaneously at each time point, including data from the transcriptome, microbiome, proteome and metabolome. He also presents a novel computational approach combining all available omic datasets into a predictive model that reveals unique interactions between the different pacemakers of pregnancy.

Presenters

	Dr. Brice Gaudilliere, MD, PhD Assistant Professor, Department of Anesthesiology Stanford University School of Medicine
	Nima Aghaeepour, PhD Assistant Professor Stanford University School of Medicine

For Research Use Only. Not for use in diagnostic procedures.

Watch Chris Linthwaite, President and CEO of Fluidigm, introduce the Hyperion™ Imaging System at the Future of Imaging announcement event held on October 24, 2017, in Toronto, Canada.

During this 90-minute program you will also hear three leading researchers share their insights and experiences with this revolutionary new approach to tissue imaging in the context of their own translational research programs.

Previously viewed live by researchers from around the world, this exciting program is now available to view on-demand.

The Hyperion Imaging System significantly expands the number of protein biomarkers that can be simultaneously detected from tissues and tumors as compared to results from standard fluorescent immunohistochemistry (IHC) methods. With the potential to revolutionize disease research and change the way diseases are treated and ultimately cured in the future, this new approach provides researchers with an unprecedented view of complex cellular phenotypes and their relationships in the context of the tissue microenvironment.

Presenters:

A New View:   
Empowering a Revolution in Tissue Imaging
Chris Linthwaite
President and CEO
Fluidigm

Uncovering Cellular Social Networks: 
Analysis of Tissue Ecosystems in Health 
and Disease by Imaging Mass Cytometry
Bernd Bodenmiller, PhD
Assistant Professor
Institute of Molecular Life Sciences
University of Zurich

Empowering Drug Discovery and Development:
Incorporating Imaging Mass Cytometry into a Multi-Platform Tissue Testing Workflow
Matt Silver, PhD
Principal Scientist
Global Clinical Biomarkers and Companion Diagnostics
EMD Serono

Advancing Translational and Clinical Research:
Imaging Mass Cytometry to Characterize the Tumor Immune Microenvironment   
Akil Merchant, MD
Assistant Professor of Medicine
Keck School of Medicine
University of Southern California

Learn more about how the Hyperion™ Imaging System significantly expands the number of protein markers that can be simultaneously detected in tumors and tissues by Imaging Mass Cytometry™.

More than 200 scientists from around the world gathered in Boston on June 9 for our 6th Annual Mass Cytometry Summit to share research findings, discuss advances and develop new collaborations. We are grateful to the researchers below who have graciously agreed to share their presentations with the mass cytometry community.

Register for the 7th Annual Mass Cytometry Summit.

New Applications

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A Mass Cytometry Surrogate for Light Scatter
Adeeb Rahman, PhD
Director of Technology Development Human Immune Monitoring Core
Icahn School of Medicine at Mount Sinai Hess Center for Science and Medicine

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Dynamics of T Lymphocyte Fate Specification Revealed by Tracing Cell Proliferative History
Zinaida Good, PhD
PhD Candidate, Computational and Systems Immunology
Garry P. Nolan and Sean C. Bendall labs Stanford University

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Extensive Characterization of Autoreactive CD8 T Cells in Type 1 Diabetes
Alice Wiedeman, PhD
Staff Scientist
Human Immunophenotyping Core
Benaroya Research Institute

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Best Practices and Data Analysis

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Panel Design Optimization through the Designed Distribution of Signal Interference
Bill O’Gorman, PhD
Genetech

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Computational Strategies for Mass Cytometry Data Analysis
Nikolay Samusik, PhD
Research Scientist Garry P. Nolan Lab
Stanford School of Medicine

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A New Analytic Approach for Live Singlet Identification
Bruce Bagwell, MD, PhD
President, Verity Software House

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Cytobank Solutions
Vinko Tosevki, PhD
Scientist, Head of Mass Cytometry Facility University of Zurich

Read more about the 6th Annual Mass Cytometry Summit

Want to learn more about mass cytometry?
Provide us with your contact information below.

For Research Use Only. Not for use in diagnostic procedures.

Enabling high-parameter studies, mass cytometry has quickly become a mainstay in basic and translational research laboratories around the world. In this video researchers share how mass cytometry is powering new insights in health and disease.

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Hear Researchers' Stories:

	Profiling Key Immune Cell Populations in Allergy, Autoimmunity and Infection Cheryl Kim  Director, Flow Cytometry Core Facility, La Jolla Institute for Allergy and Immunology
	Defining the Immune Microenvironment of AML Evan Lind Assistant Professor, Department of Molecular Microbiology and Immunolgy and Department of Cell, Development and Cancer Biology        Oregon Health & Science University
	Revealing the Immune Response to Tuberculosis Virginie Rozot Postdoctoral Fellow South African Tuberculosis Vaccine Initiative

Researchers identify a unique population of pathologic CD4+ T cells in both blood and synovial fluid using mass cytometry

In this video, Dr. Deepak Rao, MD, PhD, of Brigham and Women’s Hospital presents his recent research using mass cytometry and multidimensional flow cytometry to identify key pathologic T cell populations in the synovial fluid of subjects with rheumatoid arthritis.

Determining the pathologic functions of T cells that infiltrate target tissues remains a central challenge in autoimmune diseases. In this video, Dr. Deepak Rao, MD, PhD, of Brigham and Women’s Hospital describes recent work, published in Nature, identifying a unique population of pathologic CD4⁺ T cells, called T peripheral helper (T_PH) cells, that is markedly expanded in the joints of subjects with rheumatoid arthritis (RA). Rao and colleagues used mass cytometry and multidimensional flow cytometry to interrogate T cell populations in synovial tissue and blood from research subjects with RA, a chronic immune-mediated arthritis that affects up to 1% of the population.

Mass cytometric analysis of RA synovial tissue cells revealed a strikingly expanded population of PD-1^hiCXCR5^-CD4⁺ T cells, which constituted ~25% of synovial CD4⁺ T cells. Surprisingly, these cells are not exhausted, but instead highly express factors enabling B cell help, including IL-21, CXCL13, ICOS, SAP and MAF. Like T follicular helper (T_FH) cells, PD-1^hiCXCR5^- cells from synovium and blood induce plasma cell differentiation in vitro via IL 21. However, RNA-seq transcriptomics robustly separates PD-1^hiCXCR5^- cells from T_FH cells, with altered expression of Bcl6 and Blimp-1 and unique expression of chemokine receptors that direct migration to inflamed sites, such as CCR2, CX3CR1 and CCR5, in PD-1^hiCXCR5^- cells.

Rao and colleagues propose that PD-1^hiCXCR5^- T cells represent a T_PH cell population, analogous to T_FH cells, that supports B cell responses in pathologically inflamed nonlymphoid tissues. Given their marked expansion in RA joints, these cells may be important in driving pathologic B cell responses and autoantibody production within the inflamed target tissue.

Michelle Poulin, PhD, of Fluidigm provides a brief overview of mass cytometry, the high-parameter, single-cell analysis technology used by Rao in his research.

Speakers

Dr. Deepak Rao, MD, PhD   
Rheumatologist   
Co-Director, Human Immunology Center    
Brigham and Women's Hospital 
Boston, MA

Michelle Poulin, PhD
Applications Scientist
Fluidigm Canada Inc. 
Markham, ON, Canada

fluidigm

For Research Use Only. Not for use in diagnostic procedures.

Cheryl Kim of La Jolla Institute for Allergy and Immunology describes the value of mass cytometry for researchers at her institution.

Enabling multiparameter profiling of precious samples, mass cytometry has been instrumental in achieving research breakthroughs around the world. Hear Cheryl Kim, director of the Flow Cytometry Core Facility at La Jolla Institute for Allergy and Immunology, discuss the ways mass cytometry is helping researchers at her institution get the most information from limited samples.

Mass Cytometry in Focus

For Research Use Only. Not for use in diagnostic procedures.

Presented at the American Association for Cancer Research (AACR) 2017 meeting, this poster compares a targeted gene expression profiling approach using high-throughput qPCR readout and a nontargeted approach using mRNA sequencing.

 

Presented at the American Association for Cancer Research (AACR) 2017 meeting, this poster describes the use of the Clontech SMART-Seq v4 kit with the C1 system, and the resultant gene expression sensitivity and overall performance.

 

Evan Lind, PhD, of Oregon Health & Science University, explains how mass cytometry enables better understanding of immune systems

Enabling multiparameter profiling of precious samples, mass cytometry has been instrumental in achieving new research breakthroughs around the world. Hear how Evan Lind uses mass cytometry to research the leukemic microenvironment in acute myeloid leukemia (AML) patients in search of better targeted therapies.

Mass Cytometry in Focus

For Research Use Only. Not for use in diagnostic procedures.

Watch this video to learn about a novel approach to improving checkpoint blockade therapy in cancer under investigation in early-phase trials by Joshua Brody, MD, and colleagues at the Icahn School of Medicine at Mount Sinai.

Enhancing Checkpoint Blockade in Lymphoma with In Situ Vaccination

Checkpoint blockade therapy for cancer has had tremendous impact on clinical outcomes, yet only a subset of patients respond. Recent studies show that response to checkpoint blockade does not always correlate with tumor-associated antigen (TAA) load and so must be determined by factors beyond mutational burden. This suggests that checkpoint blockade is limited by suboptimal cross-presentation of TAA by activated dendritic cells (DC) and will be potentiated by recruitment, loading and activation of cross-presenting DC at the tumor site.

To test this hypothesis, an early-phase trial (NCT01976585) in low-grade lymphoma was carried out testing a unique in situ vaccine (ISV) combining: 1) fms-like tyrosine kinase 3 ligand (FLT3L) to recruit DC, 2) radiotherapy to load FLT3L-mobilized DC with TAA, and 3) toll-like receptor agonist (TLRa) to activate TAA-loaded DC for cross-presentation. Strikingly, Brody and colleagues found partial and complete systemic tumor regressions at distant, untreated tumors. They also found specific elimination of malignant B cells with sparing of healthy B cells, suggesting a systemic anti-tumor immune response.

This data prompted a new trial in which anti-PD1 monoclonal antibody (mAb) and ISV are combined with a novel immune-monitoring approach involving co-administration of "surrogate antigens (Ag)" at the ISV site. The approach is being applied to multiple cancer types, including breast, head and neck, melanoma and sarcoma.

In this webinar, Brody discusses these studies, including the key role mass cytometry played in defining the effects of treatment on the intratumoral and systemic immune repertoire with high resolution and in profiling the distinct checkpoint/co-activator molecules on TAA-specific, surrogate-Ag-specific and bulk CD8 T cells.

Michelle Poulin, PhD, Fluidigm Field Applications Scientist, gives a brief overview of mass cytometry, the high-parameter, single-cell technology used by Brody in his research.

Mass cytometry, or cytometry by time-of-flight, the basis of Fluidigm® CyTOF® technology, uniquely enables high-dimensional single-cell proteomic analysis for systems-level discovery and comprehensive functional profiling applications. This brief overview will describe the basic principles and workflow of the technology, including recent advancements.

Speakers

Joshua Brody, MD
Assistant Professor, Hematology and Medical Oncology
Director, Lymphoma Immunotherapy Program
Icahn School of Medicine at Mount Sinai
New York, NY

Michelle Poulin, PhD
Field Applications Scientist
Fluidigm Corporation
South San Francisco, CA

For Research Use Only. Not for use in diagnostic procedures.