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
    • Subscriptions
    • Permissions and Reprints
    • Reviewing
  • Articles
    • OnlineFirst
    • Current Issue
    • Past Issues
    • Meeting Abstracts
    • Collections
      • COVID-19 & Cancer Resource Center
      • Focus on Computer Resources
      • Highly Cited Collection
      • Editors' Picks
      • "Best of" Collection
  • For Authors
    • Information for Authors
    • Author Services
    • Early Career Award
    • Best of: Author Profiles
    • Submit
  • Alerts
    • Table of Contents
    • Editors' Picks
    • OnlineFirst
    • Citations
    • Author/Keyword
    • RSS Feeds
    • My Alert Summary & Preferences
  • News
    • Cancer Discovery News
  • COVID-19
  • Webinars
  • 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 Research
Cancer Research
  • Home
  • About
    • The Journal
    • AACR Journals
    • Subscriptions
    • Permissions and Reprints
    • Reviewing
  • Articles
    • OnlineFirst
    • Current Issue
    • Past Issues
    • Meeting Abstracts
    • Collections
      • COVID-19 & Cancer Resource Center
      • Focus on Computer Resources
      • Highly Cited Collection
      • Editors' Picks
      • "Best of" Collection
  • For Authors
    • Information for Authors
    • Author Services
    • Early Career Award
    • Best of: Author Profiles
    • Submit
  • Alerts
    • Table of Contents
    • Editors' Picks
    • OnlineFirst
    • Citations
    • Author/Keyword
    • RSS Feeds
    • My Alert Summary & Preferences
  • News
    • Cancer Discovery News
  • COVID-19
  • Webinars
  • Search More

    Advanced Search

Microenvironment and Immunology

Amplifying TLR-MyD88 Signals within Tumor-Specific T Cells Enhances Antitumor Activity to Suboptimal Levels of Weakly Immunogenic Tumor Antigens

Degui Geng, Liqin Zheng, Ratika Srivastava, Cruz Velasco-Gonzalez, Adam Riker, Svetomir N. Markovic and Eduardo Davila
Degui Geng
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Liqin Zheng
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Ratika Srivastava
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Cruz Velasco-Gonzalez
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Adam Riker
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Svetomir N. Markovic
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Eduardo Davila
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 10.1158/0008-5472.CAN-10-0247 Published October 2010
  • 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.

    Activating TLR2-MyD88 signals in tumor-reactive CD8 T cells lowers the activation threshold to a weakly immunogenic tumor antigen. Purified pmel, TLR2−/−pmel, and MyD88−/− pmel CD8 T cells were activated with MyD88−/− splenocytes pulsed with varying concentrations of the mgp100 peptide or plate-bound anti-CD3 antibody with or without TLR2 agonist. Four days later, cytokine levels determined were by ELISA, whereas proliferation was determined by [3H]thymidine uptake. D, the intracellular level of IFN-γ and granzyme B were determined by flow cytometry 4 d following activation of mgp100-pulsed APCs. Top right, the percentage of cytokine-positive cells. All data are representative of three or more independent experiments each yielding identical trends.

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

    TLR2 engagement on pmel CD8 T cells enhances cytolytic function against a weakly immunogenic melanoma tumor. A and B, purified pmel CD8 T cells were activated with mgp100-pulsed MyD88−/− splenocytes with or without TLR2 agonist and 5 d later, cytolytic activity against B16 melanoma cells was determined at different effector-to-target ratios (B) or against EL4 cells pulsed with varying concentrations of mgp100 peptide at a 3:1 effector-to-target ratio. C, in vivo cytotoxicity assays were conducted as described in Materials and Methods. Briefly, resting pmel, TLR2−/−pmel, or MyD88−/−pmel T cells were injected (i.v.) into MyD88−/− mice. One day later, mice received CFSEhigh mgp100 peptide–pulsed MyD88−/− target cells and CFSElow cells pulsed with an irrelevant H-2Db–restricted peptide with or without the TLR1/2 agonist. D, the percentage of targets recovered from spleens 24 h after adoptive transfer was evaluated by FACS analysis. Data are representative of two experiments (five mice per group). *, P ≤ 0.02, ANOVA; n.s. not significant.

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

    TLR2-MyD88 signals in pmel CD8 T cells enhance antitumor activity against an established melanoma tumor. A, WT BL6 or MyD88−/− mice (D) were injected (s.c.) with B16 melanoma cells and sublethally irradiated (400 cGy) when tumors reached a size of ∼50 mm2. One day later, mice were i.v. injected (1 × 106) with previously activated but resting pmel, TLR2−/−pmel, or MyD88−/−pmel CD8 T cells and i.p. injected with mgp10025-33 antigen and anti-CD40 antibody in each the absence or presence of TLR2 ligand. Mice received weekly peritumioral s.c. injections of TLR2 ligand or control saline. Tumor sizes were calculated by measuring perpendicular by longitudinal diameter. Data are compiled from four (B and C) or three independent (D) experiments, each yielding identical trends. *, P < 0.001.

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

    TLR2-MyD88 signals within tumor-specific T cells enhance antitumor activity by promoting T-cell longevity. A, mice were challenged with tumor and treated with ACT as described in Fig. 3. Twelve days after ACT, CD90.1+CD8+ T cells in the tumor and lymph nodes was determined by FACS. B, purified pmel, TLR2−/−pmel, and MyD88−/−pmel T cells were labeled with CFSE and injected (i.v.) into WT mice followed by injection with mgp100 peptide and TLR2 ligand (i.v.). Twelve days later, spleens were collected, and the number of transferred cells was determined by setting the instrument gates to count a set number of the CD45.1 cells. Representative plot for two experiments, five mice per group. C, purified T cells were labeled with CFSE and activated with mgp100-pulsed MyD88−/− splenocytes in the absence and presence of TLR2 ligand. Four days later, cells were stained with Annexin V. D, purified CD90.1+CD45.2+ pmel and CD90.2+CD45.2+ MyD88−/−pmel T cells were activated in vitro with mgp100 peptide–pulsed WT splenocytes. One day later, pmel and MyD88−/−pmel T cells were purified, mixed at a 1:1 ratio, and i.v. injected into CD45.1+ mice. The ratio of pmel to MyD88−/−pmel T cells was determined by staining cell suspensions with anti-CD90.1 and anti-CD45.2 antibodies. Data from one of two to three independent experiments, with three to five mice per group. *, P < 0.05, ANOVA.

  • Figure 5.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 5.

    Overexpressing TLR2 on tumor-reactive T cells enhances the therapeutic efficacy of adoptively T-cell transfer. A, pmel CD8 T cells transduced with RV-expressing TLR2 and GFP (pmel-RV-TLR2) or GFP only (pmel-RV-con) were sorted by FACS. Cytokine production was determined by ELISA. T cells were activated with MyD88−/− APCs pulsed with varying concentrations of antigen in the absence or presence of TLR2 ligand. B and C, BL6 mice bearing an established B16 tumor were treated with varying numbers of pmel-RV-TLR2 or pmel-RV-con with (B) or without (C) the TLR2 ligand. All mice were vaccinated with mgp100, anti-CD40 antibody, and TLR2 ligand, as described in Fig. 3. Data are compiled from three independent experiments, with each experiment yielding similar results. *, P < 0.001, MIXED procedure of SAS.

  • Figure 6.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 6.

    TLR2 stimulation on melanoma-specific patient T cells lowers the activation threshold to subdominant tumor antigens. Tumor antigen-specific CD8 T cells from melanoma patients were expanded and sorted as described in Materials and Methods. Cytokine production, proliferation, and cytolytic activity of Mart1-, gp100-, and tyrosinase-specific CD8 T cells were tested against peptide-pulsed HLA-A2+ A-375 melanoma target cells in the presence or absence of TLR2 ligand. The A375 melanoma cell line was pulsed with varying concentrations of peptides starting at 1 μg/mL and diluted 10-fold or pulsed with 0.001 μg/mL peptide for in vitro cytotoxicity assays. B, cytokine production was evaluated using a Milliplex cytokine array 48 h after stimulation, whereas T-cell proliferation was measured at the end of 3 days via [3H]thymidine incorporation. C, cytotoxicity was determined in a 6-h 51Cr release assay at an effector-to-target ratio of 1:1. D, alternatively, gp100-specific CD8 T cells were mixed with melanoma cells (SK-MEL-23) expressing endogenous mgp100 at varying effector-to-target ratios. Blood samples were collected from 10 melanoma patients at the time of diagnosis. The data presented are from three (B and C) and two (D) different donors from which the highest number of tumor-specific T cells could be generated. All experimental determinations were performed in triplicate; averages ± SDs were consistently within 15% of the mean. Error bars represent mean ± SD of triplicate samples. *, P < 0.01, ANOVA.

Additional Files

  • Figures
  • Supplementary Data, Geng, et al.

    Files in this Data Supplement:

    • Supplementary Figures 1-3
PreviousNext
Back to top
Cancer Research: 70 (19)
October 2010
Volume 70, Issue 19
  • 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 Research 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.
Amplifying TLR-MyD88 Signals within Tumor-Specific T Cells Enhances Antitumor Activity to Suboptimal Levels of Weakly Immunogenic Tumor Antigens
(Your Name) has forwarded a page to you from Cancer Research
(Your Name) thought you would be interested in this article in Cancer Research.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
Amplifying TLR-MyD88 Signals within Tumor-Specific T Cells Enhances Antitumor Activity to Suboptimal Levels of Weakly Immunogenic Tumor Antigens
Degui Geng, Liqin Zheng, Ratika Srivastava, Cruz Velasco-Gonzalez, Adam Riker, Svetomir N. Markovic and Eduardo Davila
Cancer Res October 1 2010 (70) (19) 7442-7454; DOI: 10.1158/0008-5472.CAN-10-0247

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Share
Amplifying TLR-MyD88 Signals within Tumor-Specific T Cells Enhances Antitumor Activity to Suboptimal Levels of Weakly Immunogenic Tumor Antigens
Degui Geng, Liqin Zheng, Ratika Srivastava, Cruz Velasco-Gonzalez, Adam Riker, Svetomir N. Markovic and Eduardo Davila
Cancer Res October 1 2010 (70) (19) 7442-7454; DOI: 10.1158/0008-5472.CAN-10-0247
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
    • Materials and Methods
    • Results
    • Discussion
    • Disclosure of Potential Conflicts of Interest
    • Acknowledgments
    • Footnotes
    • References
  • Figures & Data
  • Info & Metrics
  • PDF
Advertisement

Related Articles

Cited By...

More in this TOC Section

  • TLR4-Mediated Inflammation Promotes Cellular Transformation
  • CD103 Signaling in Human TRM Cells
  • Expansion of Neoclonotypes and Anti–PD-1 Clinical Efficiency
Show more Microenvironment and Immunology
  • Home
  • Alerts
  • Feedback
  • Privacy Policy
Facebook  Twitter  LinkedIn  YouTube  RSS

Articles

  • Online First
  • Current Issue
  • Past Issues
  • Meeting Abstracts

Info for

  • Authors
  • Subscribers
  • Advertisers
  • Librarians

About Cancer Research

  • About the Journal
  • Editorial Board
  • Permissions
  • Submit a Manuscript
AACR logo

Copyright © 2021 by the American Association for Cancer Research.

Cancer Research Online ISSN: 1538-7445
Cancer Research Print ISSN: 0008-5472
Journal of Cancer Research ISSN: 0099-7013
American Journal of Cancer ISSN: 0099-7374

Advertisement