Skip to main content
  • AACR Publications
    • Blood Cancer Discovery
    • Cancer Discovery
    • Cancer Epidemiology, Biomarkers & Prevention
    • Cancer Immunology Research
    • Cancer Prevention Research
    • Cancer Research
    • Clinical Cancer Research
    • Molecular Cancer Research
    • Molecular Cancer Therapeutics

AACR logo

  • Register
  • Log in
  • My Cart
Advertisement

Main menu

  • Home
  • About
    • The Journal
    • AACR Journals
    • Journal Sections
    • Subscriptions
    • Permissions and Reprints
  • Articles
    • OnlineFirst
    • Current Issue
    • Past Issues
    • Collections
      • COVID-19 & Cancer Resource Center
      • Precision Medicine and Therapeutic Resistance
      • Clinical Trials
      • Immuno-oncology
      • Editors' Picks
      • "Best of" Collection
  • For Authors
    • Information for Authors
    • Author Services
    • Best of: Author Profiles
    • Submit
  • Alerts
    • Table of Contents
    • Editors' Picks
    • OnlineFirst
    • Citation
    • Author/Keyword
    • RSS Feeds
    • My Alert Summary & Preferences
  • News
    • Cancer Discovery News
    • Journal Press Releases
  • COVID-19
  • Webinars
  • 10th Anniversary
  • Search More

    Advanced Search

  • AACR Publications
    • Blood Cancer Discovery
    • Cancer Discovery
    • Cancer Epidemiology, Biomarkers & Prevention
    • Cancer Immunology Research
    • Cancer Prevention Research
    • Cancer Research
    • Clinical Cancer Research
    • Molecular Cancer Research
    • Molecular Cancer Therapeutics

User menu

  • Register
  • Log in
  • My Cart

Search

  • Advanced search
Cancer Discovery
Cancer Discovery
  • Home
  • About
    • The Journal
    • AACR Journals
    • Journal Sections
    • Subscriptions
    • Permissions and Reprints
  • Articles
    • OnlineFirst
    • Current Issue
    • Past Issues
    • Collections
      • COVID-19 & Cancer Resource Center
      • Precision Medicine and Therapeutic Resistance
      • Clinical Trials
      • Immuno-oncology
      • Editors' Picks
      • "Best of" Collection
  • For Authors
    • Information for Authors
    • Author Services
    • Best of: Author Profiles
    • Submit
  • Alerts
    • Table of Contents
    • Editors' Picks
    • OnlineFirst
    • Citation
    • Author/Keyword
    • RSS Feeds
    • My Alert Summary & Preferences
  • News
    • Cancer Discovery News
    • Journal Press Releases
  • COVID-19
  • Webinars
  • 10th Anniversary
  • Search More

    Advanced Search

Research Briefs

The Vigorous Immune Microenvironment of Microsatellite Instable Colon Cancer Is Balanced by Multiple Counter-Inhibitory Checkpoints

Nicolas J. Llosa, Michael Cruise, Ada Tam, Elizabeth C. Wicks, Elizabeth M. Hechenbleikner, Janis M. Taube, Richard L. Blosser, Hongni Fan, Hao Wang, Brandon S. Luber, Ming Zhang, Nickolas Papadopoulos, Kenneth W. Kinzler, Bert Vogelstein, Cynthia L. Sears, Robert A. Anders, Drew M. Pardoll and Franck Housseau
Nicolas J. Llosa
1Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Michael Cruise
2Department of Pathology, Johns Hopkins University, Baltimore, Maryland.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Ada Tam
3Flow Cytometry Core, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Elizabeth C. Wicks
4Department of Surgery, Johns Hopkins University, Baltimore, Maryland.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Elizabeth M. Hechenbleikner
4Department of Surgery, Johns Hopkins University, Baltimore, Maryland.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Janis M. Taube
2Department of Pathology, Johns Hopkins University, Baltimore, Maryland.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Richard L. Blosser
3Flow Cytometry Core, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Hongni Fan
1Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Hao Wang
5Department of Oncology Biostatistics and Bioinformatics, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Brandon S. Luber
5Department of Oncology Biostatistics and Bioinformatics, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Ming Zhang
6Ludwig Center for Cancer Genetics and Therapeutics, Howard Hughes Medical Institute, and the Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Nickolas Papadopoulos
6Ludwig Center for Cancer Genetics and Therapeutics, Howard Hughes Medical Institute, and the Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Kenneth W. Kinzler
6Ludwig Center for Cancer Genetics and Therapeutics, Howard Hughes Medical Institute, and the Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Bert Vogelstein
6Ludwig Center for Cancer Genetics and Therapeutics, Howard Hughes Medical Institute, and the Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Cynthia L. Sears
1Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland.
7Department of Medicine, Johns Hopkins University, Baltimore, Maryland.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Robert A. Anders
2Department of Pathology, Johns Hopkins University, Baltimore, Maryland.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Drew M. Pardoll
1Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland.
2Department of Pathology, Johns Hopkins University, Baltimore, Maryland.
7Department of Medicine, Johns Hopkins University, Baltimore, Maryland.
8Department of Molecular Biology and Genetics, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: fhousse1@jhmi.edu Dpardol1@jhmi.edu
Franck Housseau
1Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: fhousse1@jhmi.edu Dpardol1@jhmi.edu
DOI: 10.1158/2159-8290.CD-14-0863 Published January 2015
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • Additional Files
  • Figure 1.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 1.

    Geographic distribution in situ of MSI and MSS colorectal cancer–infiltrating lymphocytes. Formalin-fixed, paraffin-embedded tissue sections were characterized by IHC for CD4+, CD8+, and FOXP3+ cell infiltration. Three distinct histologic areas designated as TIL, tumor stroma, and invasive front (where tumor invaded normal lamina propria) were histologically identified and separately analyzed. Invasive front (A) and TIL/stroma (B) areas of representative MSS (bottom) and MSI (top) specimens are shown (×20 magnification). Dashed lines in A delineate the invasive front with the tumor tissue on the top side and the normal tissue on the bottom side. Red stars and blue arrows in B indicate the tumor stroma and tumor epithelium-infiltrating immune cells, respectively. Scale bars, 100 μm. C, cell density was scored in 14 MSS (blue) and 9 MSI (red) specimens by determining the average number of stained cells in 5 distinct hpf (0.0028 mm2/hpf). The graphs display the mean for each group; *,statistically significant differences between MSS and MSI (P < 0.05, using Mann–Whitney U test).

  • Figure 2.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 2.

    Th1 and CTL-based immune signature and elevated checkpoint expression in MSI colorectal cancer. RNA was extracted from tissue samples laser-microdissected representing TIL in tumor nests (A), stroma surrounding tumor (B), and invasive front (C) areas of MSS (blue squares) and MSI (red circles) colorectal cancer specimens. Immune-related gene expression profiles were assessed using TaqMan-based qRT-PCR for selected genes. Sets of genes were defined by functional relevance (Th1/Tc1, CTL, Th17, Treg, proinflammation, immune checkpoints, and metabolism). The Y-axis represents an arbitrary unit of expression 2−ΔCt, ΔCt representing cycle threshold (Ct) of the gene of interest normalized by Ct of ubiquitous genes (GUSB and GAPDH). The graphs display the geometric means. Their differential representation between MSS and MSI specimens was analyzed using adjusted Wilcoxon–Mann–Whitney test as described in Methods. *, Wilcoxon P < 0.05. D, gene group comparison in TIL, tumor stroma, and invasive front areas between MSS and MSI specimens. Permutation test results based on the maximum Wilcoxon–Mann–Whitney test statistic within the gene groups Th1/Tc1, CTL, Th17, and immune checkpoints. *, statistically significant differences between MSS and MSI (P < 0.05).

  • Figure 3.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 3.

    PD-1 and LAG-3 expression in MSI and MSS colorectal cancer specimens. IHC analysis of PD-1 and LAG-3 expression in invasive front (A) and TIL/stroma (B) areas was performed on formalin-fixed, paraffin-embedded tissue sections of a representative set of MSI (top) and MSS (bottom) colorectal cancer specimens. Magnification, ×20; scale bars, 100 μm; red stars and blue arrows in B indicate the tumor stroma and tumor epithelium-infiltrating immune cells, respectively.

  • Figure 4.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 4.

    MSI colorectal cancers are characterized by IFNγ-producing PD1hi TIL and PDL-1+ tumor-infiltrating myeloid cells. A, freshly dissociated MSS and MSI colon tumors (T) as well as patient-matched normal tissue (N) were assessed by MFC for the expression of PD-1 on infiltrating CD4+ and CD8+ T cells. PD-1 expression in tumor was normalized to the normal tissue run simultaneously and both histograms were aligned to delineate in tumor samples the PD1hi cells when compared with normal tissue. B, proportion of PD-1hi CD4+ and CD8+ cells among CD3+ lymphocytes infiltrating MSS (blue squares) and MSI (red circles) specimens. In each group the mean is indicated; *, statistically significant differences between MSS and MSI (*, P < 0.05; ****, P < 0.0001; nonparametric Mann–Whitney U test). C, representative ICS for IFNγ production by in vitro phorbol-12-myristate-13-acetate/ionomycin–activated T cells (3 hours). The dot plots show the coexpression of PD-1 and IFNγ in CD4+ T cells and CD8+ T cells in a representative set of MSS (left) and MSI (right) colorectal cancer (top) and patient-matched normal (bottom) specimens. The gates delineate PD1hi and PD1lo cells. D, colocalization of CD163 and PD-L1 expression in invasive front (left) and TIL/stroma (right) areas of a representative set of MSS (bottom) and MSI (top) colorectal cancer specimens were assessed by IHC; ×20 magnification. Scale bars, 100 μm. Red stars indicate the tumor stroma. E, PD-L1 expression scores in 7 MSS (blue) and 7 MSI (red) colorectal cancer specimens (average of 5 hpf/sample). F, MFC analysis of PD-L1 expression on MSI colorectal cancer–infiltrating myeloid cells. Dot plots represent the expression of myeloid-associated markers on CD11b+HLA-DR−/low cells. Infiltrating myeloid cells were characterized as CD15−CD14+CD33+CD11c+ cells. PD-L1 expression (dark gray) is overlaid with corresponding isotype control (light gray).

Additional Files

  • Figures
  • Supplementary Data

    Files in this Data Supplement:

    • Supplementary Figure S1 - Supplementary Figure S1. Laser capture microdissection of FFPE tissue sections.
    • Supplementary Figure S2 - Supplementary Figure S2. Absolute numbers of CD3 cells per gram of tumor and normal colon tissue collected from MSS and MSI human CRC patients.
    • Supplementary Figure S3 - Supplementary Figure S3. Characterization of immune infiltration in MSS CRC outlier.
    • Supplementary Figure S4 - Supplementary Figure S4. PD-L1 induction on human CRC cell lines.
    • Supplementary Table S1 - Supplementary Table S1. Patient Demographics.
    • Supplementary Table S2 - Supplementary Table S2. Exome sequencing of MSS tumor outlier.
    • Supplementary Table S3 - Supplementary Table S3. Primers/probes used for Taqman PCR array.
PreviousNext
Back to top
Cancer Discovery: 5 (1)
January 2015
Volume 5, Issue 1
  • Table of Contents
  • Table of Contents (PDF)
  • About the Cover

Sign up for alerts

View this article with LENS

Open full page PDF
Article Alerts
Sign In to Email Alerts with your Email Address
Email Article

Thank you for sharing this Cancer Discovery article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
The Vigorous Immune Microenvironment of Microsatellite Instable Colon Cancer Is Balanced by Multiple Counter-Inhibitory Checkpoints
(Your Name) has forwarded a page to you from Cancer Discovery
(Your Name) thought you would be interested in this article in Cancer Discovery.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
The Vigorous Immune Microenvironment of Microsatellite Instable Colon Cancer Is Balanced by Multiple Counter-Inhibitory Checkpoints
Nicolas J. Llosa, Michael Cruise, Ada Tam, Elizabeth C. Wicks, Elizabeth M. Hechenbleikner, Janis M. Taube, Richard L. Blosser, Hongni Fan, Hao Wang, Brandon S. Luber, Ming Zhang, Nickolas Papadopoulos, Kenneth W. Kinzler, Bert Vogelstein, Cynthia L. Sears, Robert A. Anders, Drew M. Pardoll and Franck Housseau
Cancer Discov January 1 2015 (5) (1) 43-51; DOI: 10.1158/2159-8290.CD-14-0863

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Share
The Vigorous Immune Microenvironment of Microsatellite Instable Colon Cancer Is Balanced by Multiple Counter-Inhibitory Checkpoints
Nicolas J. Llosa, Michael Cruise, Ada Tam, Elizabeth C. Wicks, Elizabeth M. Hechenbleikner, Janis M. Taube, Richard L. Blosser, Hongni Fan, Hao Wang, Brandon S. Luber, Ming Zhang, Nickolas Papadopoulos, Kenneth W. Kinzler, Bert Vogelstein, Cynthia L. Sears, Robert A. Anders, Drew M. Pardoll and Franck Housseau
Cancer Discov January 1 2015 (5) (1) 43-51; DOI: 10.1158/2159-8290.CD-14-0863
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
    • Abstract
    • Introduction
    • Results
    • Discussion
    • Methods
    • Disclosure of Potential Conflicts of Interest
    • Authors' Contributions
    • Grant Support
    • Acknowledgments
    • Footnotes
    • References
  • Figures & Data
  • Info & Metrics
  • PDF
Advertisement

Related Articles

Cited By...

More in this TOC Section

  • Fas Mediates Bystander Tumor Killing by T Cells
  • Immune Checkpoint Inhibitor Myocarditis in Mice
  • Genetic Ancestry Impacts Somatic Alterations in Lung Cancers
Show more Research Briefs
  • Home
  • Alerts
  • Feedback
  • Privacy Policy
Facebook   Twitter   LinkedIn   YouTube   RSS

Articles

  • OnlineFirst
  • Current Issue
  • Past Issues

Info For

  • Authors
  • Subscribers
  • Advertisers
  • Librarians

About Cancer Discovery

  • About the Journal
  • Editors
  • Journal Sections
  • Permissions
  • Submit a Manuscript
AACR logo

Copyright © 2021 by the American Association for Cancer Research.

Cancer Discovery
eISSN: 2159-8290
ISSN: 2159-8274

Advertisement