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. 2007 Jun 1;109(11):4882-9.
doi: 10.1182/blood-2006-10-043463. Epub 2007 Jan 23.

CD137 stimulation delivers an antigen-independent growth signal for T lymphocytes with memory phenotype

Yuwen Zhu  1 , Gefeng Zhu, Liqun Luo, Andrew S Flies, Lieping Chen
Affiliations

CD137 stimulation delivers an antigen-independent growth signal for T lymphocytes with memory phenotype

Yuwen Zhu et al. Blood. .

Abstract

CD137 has long been recognized as a costimulatory receptor for growth and functional maturation of recently activated T cells in the presence of T-cell receptor signal. In this report, we present the fact that, in the absence of MHC and antigen, triggering of CD137 by an agonist monoclonal antibody induces vigorous growth of both CD8(+) and CD4(+) T cells with memory phenotype, whereas it does not affect naive T cells. Moreover, T cells with memory phenotype accumulate progressively in transgenic mice overexpressing CD137 ligand. CD137-mediated proliferation of memory T cells is directly through CD137 on T cells and does not require IL-15 and IFN-gamma. Our results define a new role of CD137 signal in the growth of memory T cells.

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Figures

Figure 1
Figure 1
CD137 stimulation selectively stimulates proliferation of memory T cells. B6 mice were injected with 100 μg control rat IgG or 2A on day 0 and 2 and fed with drinking water with 0.8 mg/mL BrdU from day 3 to 7. Splenocytes (A) or intrahepatic lymphocytes (B) were harvested on day 7 and stained for CD8, CD4, CD44, and BrdU by flow cytometry. Data were presented by gating on CD8 or CD4. Total numbers of CD44hi or CD122+ cells in CD8+ or CD4+ T-cell subsets in spleens (C) are shown on days 5 and 8, respectively, after antibody treatment. Error bars indicate SD. The number of total intrahepatic lymphocytes as well as CD4+ and CD8+ subsets on day 7 are also shown (D). The results are from one representative of 3 independent experiments with similar results. (E) Accumulation of memory phenotype T cells in CD137L transgenic mice. The phenotype of spleen or LN T cells from 1-month-old and 5-month-old CD137L transgenic together with age-matched littermate mice was determined by flow cytometry. Data shown are gated on CD3+ (1-month-old), CD8+, or CD4+ (5-month-old). The percentages of CD44hi and CD62Llo cells are indicated. Results are one representative of at least 3 independent experiments. *P < .05, **P < .001.
Figure 2
Figure 2
Generation and characterization of CD137 KO mice. (A) The targeting map of the CD137 genomic locus. The signal peptide with the ATG starting code and first 6 exons encoding extracellular and transmembrane regions of murine CD137 were replaced with the Neo cassette. A short open bar labeled “Probe” indicates the position of the 3′ end probe for screening of ES cells, and PCR indicates the position of PCR products in screening of CD137-deficient mice using primers. Shaded boxes represent exons within the murine CD137 open reading frame. (B) Southern blotting of heterozygous and homozygous CD137 mutants in the gDNA from targeted ES cells. The upper band (6691 bp) shows the targeted fragment, and the lower one (6147 bp) represents the one from the normal genome. (C) Splenocytes from WT B6 or CD137 KO mice were activated by Con A for 24 hours, and live cells were stained for CD137 or PD-1 gated on CD3+ cells by specific mAbs and subsequently analyzed by flow cytometry. Shaded histograms indicate isotype control; open histograms, anti-CD137 or anti-PD1. (D) Whole splenocytes or purified T cells from CD137KO or WT control mice and were activated by Con A or plate-bound CD3 mAb at the indicated concentrations. [3H]TdR was included in the cultures 16 hours before harvesting. The results are from one representative of 2 independent experiments with similar results. (E) Splenocytes from untreated WT or CD137 KO mice were stained for CD44 and CD62L, gated on CD8+ or CD4+ cells, respectively. The results are from one representative of 2 independent experiments with similar results.
Figure 3
Figure 3
CD137 on T cells is essential and sufficient for the anti-CD137 mAb effect. (A) Purified T cells from WT or CD137KO mice were adoptively transferred into WT B6/Thy1.1 mice. Mice treated with rat IgG or 2A (shown) were fed with BrdU as described. The data are gated on spleen CD8+ CD44hi cells and show the BrdU incorporation from host versus donor origin distinguished by Thy1.2 staining. The results are from one representative of 2 independent experiments with 3 mice each. (B) WT B6 or CD137KO mice injected with PBS, poly I:C or 2A on day 0 were fed with BrdU. Spleen cells were harvested and stained for CD8, CD44, and anti-BrdU. Data shown were gated on CD8+ cells. The results are from one representative of 2 independent experiments with 3 mice. (C) Purified T cells from WT B6/Thy1.1 mice were transferred into WT or CD137KO mice. Mice were treated as described in panel A. The data are gated on spleen CD8+ CD44hi cells and show the BrdU incorporation from host versus donor origin distinguished by Thy1.1 staining.
Figure 4
Figure 4
CD137 stimulation induces proliferation of memory but not naive T cells. In vitro activated OT-1 cells were adoptively transferred into naive B6 mice and rested for more than 40 days to generate memory OT-1 cells. B6 mice containing naive (upper panels) or memory OT-1 × RAG1KO TCR transgenic T cells (lower panels) were subsequently treated with rat IgG or 2A. The mice were fed with BrdU-containing drinking water for 5 days. Spleen cells were harvested and stained for CD8, OT-1 tetramer, and anti-BrdU. Data shown were gated on CD8+ cells. The results are from one representative of 2 independent experiments with similar results.
Figure 5
Figure 5
Effects of CD137 mAb on naive T-cell homeostasis in lymphopenic mice. CFSE-labeled naive OT-1 × RAG1 KO T cells (1 × 106) were adoptively transferred into sublethally irradiated B6 mice. Then, 100 μg rat IgG or 2A was injected intraperitoneally on day 0 (upper panels) or day 7 (lower panels) after cell transfer. Spleen cells were prepared at day 6 after antibody treatment and analyzed by flow cytometry. Histogram plots of CFSE intensity of transferred OT-1 cells (gated on CD8+ OT-1 tetramer-positive cells) in the spleen are shown. The results are from one representative of 3 independent experiments with similar results.
Figure 6
Figure 6
Memory T-cell proliferation induced by CD137 stimulation is antigen and MHC independent. (A) Memory OT-1 cells generated as described Figure 4 were labeled with CFSE and adoptively transferred into H-2Kb KO mice and subsequently treated with CD137 mAb or control antibody on day 1 and 3. On day 7, spleen cells were harvested and stained for CD8, OT-1 tetramer. CFSE intensity of transferred memory OT-1 cells was shown by flow cytometry gated on CD8 and OT-1 tetramer-positive cells. (B) B6 mice transferred with naive OT-1 T cells were preimmunized with OVA plus poly I:C to generate memory OT-1 cells. Fifty days later, mice were injected with anti–H-2Kb blocking mAb on day −1 and 2. On day 0 and day 2, mice were treated with CD137 mAb or control mAb, respectively, and fed with PBS containing BrdU as shown previously. Spleen cells were prepared on day 7 after treatment and analyzed by flow cytometry. Histogram plots of CFSE intensity of transferred OT-1 cells (gated on CD8+ OT-1 tetramer-positive cells) in the spleen are shown. The results are from one representative of 2 independent experiments with 3 mice in each group.
Figure 7
Figure 7
IL-15 and IFN-γ are not required for CD137-induced memory T-cell proliferation. IL-15 KO and IFN-γ KO mice were treated with the indicated mAb and fed with BrdU as shown in Figure 1. Data shown are percentages of BrdU+ cells gated on the CD8+CD44hi portion. The results are from one representative of 2 independent experiments with similar results. Error bars indicate SD. No significant difference was found among groups (P > .05).
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References

    1. Marrack P, Kappler J. Control of T cell viability. Annu Rev Immunol. 2004;22:765–787. - PubMed
    1. Fearon DT, Manders P, Wagner SD. Arrested differentiation, the self-renewing memory lymphocyte, and vaccination. Science. 2001;293:248–250. - PubMed
    1. Schluns KS, Lefrancois L. Cytokine control of memory T-cell development and survival. Nat Rev Immunol. 2003;3:269–279. - PubMed
    1. Goldrath AW, Sivakumar PV, Glaccum M, et al. Cytokine requirements for acute and basal homeostatic proliferation of naive and memory CD8+ T cells. J Exp Med. 2002;195:1515–1522. - PMC - PubMed
    1. Becker TC, Wherry EJ, Boone D, et al. Interleukin 15 is required for proliferative renewal of virus-specific memory CD8 T cells. J Exp Med. 2002;195:1541–1548. - PMC - PubMed

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