Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2007 Apr 17;104(16):6770-5.
doi: 10.1073/pnas.0604982104. Epub 2007 Apr 5.

Characterizing tumor-promoting T cells in chemically induced cutaneous carcinogenesis

Scott J Roberts  1 , Bernice Y Ng, Renata B Filler, Julia Lewis, Earl J Glusac, Adrian C Hayday, Robert E Tigelaar, Michael Girardi
Affiliations

Characterizing tumor-promoting T cells in chemically induced cutaneous carcinogenesis

Scott J Roberts et al. Proc Natl Acad Sci U S A. .

Abstract

There is a longstanding but poorly understood epidemiologic link between inflammation and cancer. Consistent with this, we previously showed that alphabeta T cell deficiency can increase resistance to chemical carcinogenesis initiated by 7,12-dimethylbenz[a]anthracene and promoted by phorbol 12-myristate 13-acetate. This provoked the hypothesis that alphabeta T cell deficiency removed T regulatory cells that limit the anti-tumor response or removed a specific tumor-promoting (T-pro) T cell population. Here we provide evidence for the latter, identifying a novel CD8(+) subset that is a candidate for T-pro cells. We demonstrate that CD8 cell-deficient mice show substantially less tumor incidence and progression to carcinoma, whereas susceptibility is restored by CD8(+) cell reconstitution. To characterize the putative T-pro cells, tumor-infiltrating lymphocytes were isolated from normal and CD4(-/-) mice, revealing an activated population of T cell receptor alphabeta(+)CD8(+)CD44(+)CD62L(-) cells expressing the inflammatory mediators IFNgamma, TNFalpha, and cyclooxygenase-2, but deficient in perforin, relative to recirculating cells of equivalent phenotype. This novel population of CD8(+) T cells has intriguing similarities with other lymphocytes that have been associated with tissue growth and invasiveness and has implications for inflammation-associated carcinogenesis, models of cancer immunosurveillance, and immunotherapeutic strategies.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Fig. 1.
Fig. 1.
Mice genetically deficient in CD8+ T cells demonstrate reduced tumor development and progression. Groups of mice (n = 10) subjected to DMBA/PMA were monitored weekly for tumors. (A) TCRβ−/− mice (deficient in both CD4+ and CD8+ T cells) and CD8−/− mice demonstrated significantly lower total tumor burden than strains in which the CD8 compartment was intact (FVB controls and CD4−/− mice). (B) Numbers of carcinomas were also substantially lessened in those mice (TCRβ−/− and CD8−/−) genetically lacking CD8+ T cells. ∗, P ≤ 0.05; ∗∗, P ≤ 0.01; ∗∗∗, P ≤ 0.005 (for CD4−/− or FVB vs. TCRβ−/− or CD8β−/−). (C) Progression to carcinoma, as exemplified by the carcinoma/papilloma (C/P) ratio at week 12, was significantly greater in the CD8-intact mice than in the CD8-deficient mice (P < 0.002 for CD4−/− or FVB vs. TCRβ−/− or CD8β−/−). Error bars represent standard error of the mean.
Fig. 2.
Fig. 2.
Reconstitution with CD8+ T cells restores the associated increased tumor susceptibility. Reconstitution of the CD8+ T cell compartment of TCRβ−/− mice (deficient in both CD4+ and CD8+ T cells) was performed in adult and neonatal recipients. (A and B) In adult TCRβ−/− mice reconstituted with CD8β+ SLN cells purified from normal FVB mice by using antibody-coated magnetic beads, tumors developed at a rate greater than that of unreconstituted TCRβ−/− mice and approximately equal to that of control CD4−/− mice. ∗, P ≤ 0.05; ∗∗, P ≤ 0.01; ∗∗∗, P ≤ 0.005 (for CD4−/− or TCRβ−/− plus CD8+ SLN vs. TCRβ−/−). (C and D) Similarly, TCRβ−/− mice reconstituted as neonates with FLHC progenitors from CD4−/− mice were more susceptible to carcinogenesis than unreconstituted TCRβ−/− mice. ∗, P ≤ 0.05; ∗∗, P ≤ 0.01. Error bars represent standard error of the mean.
Fig. 3.
Fig. 3.
CD8+ TIL (putative T-pro) are activated and produce inflammatory cytokines IFNγ and TNF��. (A) Flow cytometric analysis of lymph node (LN), PBL, and TIL from tumor-bearing FVB control mice (filled bars) and non-tumor-bearing naïve control mice (open bars) revealed the relative CD4/CD8 ratios in each population (n = 7). (B and C) Representative flow cytometric analysis of lymphocytes from FVB control mice demonstrates that the vast majority of CD8+ TIL are CD44+CD62L−, consistent with an activated effector/memory phenotype, in contrast to the peripheral blood, where only a small minority of CD8+ PBL are single-positive for CD44. (Numbers represent means of five cohorts of up to five mice.) (D and E) Cytometric bead array analysis of cytokine production by TIL from normal FVB mice demonstrated substantially higher secretion by CD8+, relative to equal numbers of CD4+, TIL of IFNγ and TNFα.
Fig. 4.
Fig. 4.
RT-PCR expression analysis comparison of TCRβ+CD8+CD44+CD62L− TIL vs. phenotypically equivalent PBL. Representative bands obtained from sqRT-PCR of mRNA isolated from FVB (A) and FVB.CD4−/− (B) mouse TIL and PBL (stained and sorted to >99.5% purity for TCRαβ+CD8+CD44+CD62L−) are shown for several inflammatory mediators (IFNγ, TNFα, and COX-2) and mediators involved in cellular killing (perforin, NKG2D, and FasL), along with control β-actin. (C) The mean relative expression reveals that the equivalent trends of expression (e.g., increased inflammatory mediators and decreased perforin) are consistent between PBL and TIL from FVB and FVB.CD4−/− mice (P ≤ 0.05 for all comparisons). (D) Real-time qRT-PCR fold expression differences are shown for IFNγ, TNFα, perforin, and COX-2 when TCRαβ+CD8+CD44+CD62L− TIL were compared with phenotypically equivalent PBL from tumor-bearing mice.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Mueller MM. Eur J Cancer. 2006;42:735–744. - PubMed
    1. Virchow R. Die Krankhaften Geschwülste. Berlin: von August Hirschwald; 1863. pp. 57–101.
    1. DePinho RA, Polyak K. Nat Genet. 2004;36:932–934. - PubMed
    1. Balkwill F, Mantovani A. Lancet. 2001;357:539–545. - PubMed
    1. Ikeda H, Old LJ, Schreiber RD. Cytokine Growth Factor Rev. 2002;13:95–109. - PubMed

Publication types

MeSH terms

Substances