Basic Science Reports

Induction of a Regulatory T Cell Type 1 Response Reduces the Development of Atherosclerosis in Apolipoprotein E–Knockout Mice

Ziad Mallat, Andrea Gojova, Valérie Brun, Bruno Esposito, Nathalie Fournier, Françoise Cottrez, Alain Tedgui, Hervé Groux
https://doi.org/10.1161/01.CIR.0000089083.61317.A1
Circulation. 2003;108:1232-1237
Originally published September 8, 2003

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Abstract

Background— T helper type 1 (Th1) response plays a permissive role in atherosclerosis. We hypothesized that adoptive transfer of a novel subtype of T lymphocytes called regulatory T cells type 1 (Tr1) would inhibit Th1 responses by inducing a bystander immune suppression and therefore limit the development of atherosclerosis.

Methods and Results— Clones of ovalbumin (OVA)–specific Tr1 cells expanded in vitro were administered intraperitoneally (106 cells per mouse) with their cognate antigen (50 μg of OVA subcutaneously in complete Freund’s adjuvant [CFA]) to female apolipoprotein E–knockout mice. A group of mice received only (OVA/CFA) immunization without Tr1 cells. Two other control groups received no immunization and were injected with either Tr1 cells or saline. After 9 weeks of treatment, mice injected with (OVA/CFA)+OVA-specific Tr1 cells showed a significant decrease in Th1 responses, as revealed by a decrease in OVA-specific IgG2a serum levels (P<0.0001), a decrease in the production of interferon-γ (P<0.001), and an increase in interleukin-10 production (P<0.001) by cultured spleen and lymph T cells compared with controls. In addition, cytokine production by concanavalin A–stimulated spleen cells showed a clear switch to a regulatory immune response in mice treated with (OVA/CFA)+Tr1. This was associated with a significant reduction in atherosclerotic lesion size in both the thoracic aorta and aortic sinus of mice treated with (OVA/CFA)+Tr1 compared with controls (P=0.002 to P<0.0001). Plaques of mice injected with (OVA/CFA)+Tr1 showed significantly lower accumulation of macrophages and T cells than plaques of control mice.

Conclusions— Tr1-type regulatory immune response reduces the development of experimental atherosclerosis.

Received June 3, 2003; de novo received July 9, 2003; accepted July 21, 2003.

Atherosclerosis is a chronic disease of the arterial wall in which both innate and adaptive immunoinflammatory mechanisms are involved.1–3 The profile of the T-cell response in atherosclerosis is of the T helper type 1 (Th1),3 whose proatherogenic potential has been demonstrated in experimental models of atherosclerosis.4–10 As in other chronic immunoinflammatory diseases, it has been suggested that the development and progression of atherosclerosis may result from an imbalance between Th1 and Th2 responses.3,11 However, evidence is now accumulating for a novel functionally distinct subpopulation of T cells, called regulatory T cells, that exert important regulatory functions in various immunoinflammatory diseases (see Groux et al,12 Roncarolo et al,13 and McGuirk et al14 for review). Several subsets of regulatory T (Tr) cells with distinct phenotypes and distinct mechanisms of action have now been identified. These include Tr1 cells,15–20 which secrete high levels of interleukin (IL)-10 and low to moderate levels of transforming growth factor (TGF)-β; Th3 cells,21,22 which secrete primarily TGF-β; and CD4+CD25+ T cells, which inhibit immune responses through cell-to-cell contact.23 We have shown that repeated stimulation of naive T cells from ovalbumin (OVA) T-cell receptor–transgenic mice with OVA and IL-10 results in the generation of T-cell clones with a unique cytokine profile distinct from that of Th0, Th1, or Th2 cells.15 These Tr1 cells produce IL-10 and some IL-5 and interferon-γ (IFN-γ) with or without TGF-β but with little or no IL-2 or IL-4 and proliferate poorly after polyclonal T-cell receptor–mediated activation. Functional studies on Tr1 cells have indicated that Tr1 cells have immunosuppressive properties and have been shown to prevent the development of Th1-mediated autoimmune diseases.15 Coculture of naive CD4+ T cells and human Tr1 clones in the presence of allogenic antigen-presenting cells results in the suppression of proliferative responses.15 Similarly, Tr1 clones specific for filamentous hemagglutinin from Bordetella pertussis suppress proliferation and cytokine production by a Th1 clone against an unrelated antigen, influenza virus hemagglutinin.20 In both cases, the suppressive effects of Tr1-cell clones are reversed by neutralizing IL-10, suggesting that regardless of their antigen specificities, Tr1 cell suppression is a bystander effect mediated through the production of IL-10. Here, we tested the hypothesis that transfer of such Tr1 cells to apolipoprotein (apo) E–knockout mice will inhibit pathogenic Th responses and reduce the development of atherosclerosis.

Methods

Generation, Expansion, and Transfer of OVA-Specific Tr1 Clones

The mouse T-cell clones were obtained from DO11-10 BALB/c mice after in vitro differentiation as described previously.15 Naive (MEL-14bright) CD4+, KJ-1.26+ cells were stimulated repeatedly with OVA peptide 323–339 every week for 3 weeks in the presence of IL-10. T-cell clones were generated as described previously.15 Clones were expanded and analyzed for cytokine secretion after activation with antigen-presenting cells and OVA peptide. Selected clones were then stimulated with irradiated splenocytes and OVA peptide every 2 weeks and further expanded with IL-2 (R&D Systems). T-cell clones were used at least 10 days after the last stimulation. Here, we used clone A-10-9, which has been described previously and shown to prevent Th1- and Th2-mediated immune responses.15,24

Fifteen-week-old female apoE-knockout C57Bl/6 mice (Charles River, Orleans, France) were immunized subcutaneously with either saline (n=11) or 50 μg of OVA in complete Freund’s adjuvant (CFA) (both from Sigma) (n=18) every 12 days for 9 weeks. The saline-treated mice were then divided into 2 groups that at day 0 received, in addition to subcutaneous saline, a unique intraperitoneal injection (200 μL) of either saline (n=5) or 106 OVA-specific Tr1 cells without OVA/CFA (n=6). The OVA/CFA-immunized group was divided into 2 other groups of mice that at day 0 received an intraperitoneal injection (200 μL) of either saline (OVA/CFA group, n=9) or 106 OVA-specific Tr1 cells (OVA/CFA+Tr1 group, n=9).

Cytokine Assays

In brief, ELISA plates (Polylabo) were coated with the appropriate coating anti-cytokine monoclonal antibodies (11B11, 2A5, TRFK4, and XGM-1 for IL-4, IL-10, IL-5, and IFN-γ, respectively) in carbonate buffer and incubated at 4°C overnight. Sandwich ELISAs were performed as described previously.24 Plates were read on an ELISA reader at a wavelength of 405 nm after color development (Labsystems iEMS reader).

Analysis of OVA-Specific Serum IgE, IgG1, and IgG2a

OVA-specific Ig levels were measured by use of a 2-step sandwich ELISA as described.24 To detect IgG1 and IgG2a, we used biotinylated rat monoclonal antibody (A85-1 and R19-15 for IgG1 and IgG2a, respectively; Becton Dickinson) at 2 μg/mL. Standards for OVA-specific IgG1 were pooled sera from hyperimmunized BALB/c mice.

Purification and Culture of Spleen and Lymph Node Cells

T cells were purified from spleen or draining lymph nodes by negative selection with anti-CD11b (M1/70), anti-B220, and anti-NK cells (DX5) followed by depletion with a mixture of magnetic beads coated with anti-rat Ig (Dynal).

For cytokine measurements, purified T cells (105) were mixed with irradiated splenic antigen-presenting cells (4×105) and OVA/CFA (0.25 mg/mL) concanavalin A (conA) (2 μg/mL) in 96-well plates. Supernatants were collected at 24 hours (for IL-4 measurements) and at 48 hours (for IL-5, IL-10, and IFN-γ measurements) and assayed for cytokine levels by ELISA as described previously.24

For the cell proliferation assay, purified T cells were mixed with irradiated splenic cells and conA or OVA. [3H]-Thymidine (1 μCi; Perkin Elmer) was added for the last 12 hours of cell culture.

Analysis of Atherosclerotic Plaque Size and Composition

Mice were killed at 24 weeks of age. Plasma total and HDL cholesterol were measured with a commercially available cholesterol kit (Sigma). Morphometric and immunohistochemical studies were performed in the aortic sinus and the thoracic aorta (spanning from the brachiocephalic artery to the renal arteries and including the first 3 mm2 of the brachiocephalic artery) as described previously.9

Collagen fibers were stained with Sirius red. Immunohistochemical analysis was performed as described previously.9 The following primary antibodies were used: MOMA-2 (BioSource International) as a specific marker for macrophages; anti-mouse CD3-θ (Santa Cruz), anti–α-smooth muscle actin, alkaline phosphatase conjugate, clone 1A4 (Sigma), and anti–IL-10 antibody (Santa Cruz). At least 4 sections per animal were analyzed for each immunostaining. Morphometric analysis was performed with an automated image processor (Histolab, Microvision) as described previously.9

Statistical Analysis

The effects of treatment on lesion area, plaque composition, serum Ig levels, and cytokine production by spleen and lymph T cells were evaluated by ANOVA and Bonferroni/Dunn’s test or Student’s t test. A value of P<0.05 was considered to be statistically significant.

Results

Transfer of Tr1 Cells With Their Cognate Antigen Induces Immune Suppression

As expected (Table 1), treatment with OVA in CFA resulted in elevated serum levels of OVA-specific antibodies, predominantly IgG2a, and to a much lesser extent IgE, in comparison with the saline-treated group, in which OVA-specific IgGs were undetectable. Treatment of mice with OVA/CFA induced a Th1-like response, as revealed by the production of high levels of IFN-γ (Th1 cytokine) but low levels of IL-10 and no IL-4 (Th2 cytokines) by OVA-stimulated lymph T cells (Table 1). Injection of regulatory Tr1 cells alone did not alter the immune response (Table 1). However, administration of Tr1 cells to OVA/CFA-immunized animals significantly inhibited the secretion of OVA-specific IgG2a antibodies, as reported previously in a model using alum as an adjuvant.24 Moreover, the suppressive effect of Tr1 cells was confirmed by the marked decrease in IFN-γ production and proliferative response observed in the (OVA/CFA)+Tr1 group compared with the OVA/CFA after in vitro recall response of T cells (P<0.001). This was associated with enhanced IL-10 production by T cells stimulated with OVA in vitro (P<0.001), suggesting that the transferred OVA-specific Tr1 cells were functionally active (Table 1). TGF-β in the supernatants was below detectable levels.

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TABLE 1. Proliferation and Cytokine Production by OVA-Stimulated Lymph Node T Cells and Serum Levels of OVA-Specific IgG2a and IgE

IL-10 is a cytokine with important immunosuppressive effect in vivo. Therefore, to analyze the consequence of high IL-10 secretion in mice transferred with Tr1 cells and treated with OVA/CFA, we analyzed the cytokine response of T cells after polyclonal activation with conA. As expected, conA stimulation induced a Th0 type response in T cells from saline- or Tr1-treated mice and a Th1-type response in mice treated with OVA/CFA (Table 2). In contrast, in mice treated with regulatory Tr1 cells and OVA/CFA, we observed a marked decrease in IFN-γ production and an increase in IL-10 production by conA-stimulated T cells (Table 2). Interestingly, the ratio of INF-γ to IL-10 was reduced by 3- to 4-fold in the (OVA/CFA)+Tr1 group compared with the other groups of mice, suggesting that the transfer of regulatory Tr1 cells followed by a systemic delivery of their specific antigen induced a bystander nonspecific regulatory immune response as a result of the chronic stimulation of Tr1 cells with OVA/CFA. However, as shown previously in other inflammatory models, the inhibitory function of Tr1 cells is local,15,25 and chronic stimulation of Tr1 cells did not result in systemic secretion of IL-10 in the serum, because <30 pg/mL of IL-10 was measured in the serum of the 4 different groups of mice.

View this table:

TABLE 2. Cytokine Production by ConA-Stimulated Splenic T Cells

Transfer of Tr1 Cells With Their Cognate Antigen Reduces Atherosclerotic Lesion Size in ApoE-Deficient Mice

Serum total cholesterol and HDL levels did not differ between groups (Table 3). Atherosclerotic lesion size in the thoracic aorta did not differ between the saline and Tr1 groups (Table 3). There was a modest but not statistically significant reduction in lesion size in the thoracic aorta of mice treated with OVA/CFA in comparison with the saline or Tr1 group (P=0.09 and P=0.07, respectively). In contrast, there was a marked and highly significant reduction in lesion size in the (OVA/CFA)+Tr1 group in comparison with either OVA/CFA (42% reduction, P=0.008), Tr1 alone, or saline (55% reduction, P≤0.001) (Table 3).

View this table:

TABLE 3. Serum Cholesterol Levels and Atherosclerotic Lesion Size and Composition in the Aortic Sinus of ApoE-Deficient Mice

Atherosclerotic lesion size in the aortic sinus did not differ between saline-, Tr1-, and OVA/CFA-treated animals. However, we found a highly significant decrease of plaque size in the (OVA/CFA)+Tr1 group compared with either the saline (P=0.002), Tr1 (P<0.0001), or OVA/CFA group (P=0.001) (Table 3 and Figure 1). Despite the significant decrease in lesion size, plaques of mice treated with (OVA/CFA)+Tr1 showed a marked decrease in relative macrophage infiltration (32% to 46% reduction, Table 3 and Figure 1) and T-cell accumulation (37% to 50% reduction, Table 3), with no change in smooth muscle cell or collagen content (Table 3). In addition, we found intense IL-10 staining in the plaques of mice treated with (OVA/CFA)+Tr1, whereas no or barely detectable levels of IL-10 were found in plaques from the other 3 groups (Figure 2). IL-10 staining was also detected in the adventitia of all study groups but was clearly enhanced in the (OVA/CFA)+Tr1 group (Figure 2). Overall, these results suggest that a regulatory T-cell response is associated with a less pronounced inflammatory plaque phenotype.

Figure1

Figure 1. Representative photomicrographs show sections of advanced atherosclerotic plaques (oil red O staining) from aortic sinus of mice treated with Tr1, OVA/CFA, or (OVA/CFA)+Tr1. There was a substantial reduction in lesion size in OVA/CFA+Tr1 group. There was also a reduction in macrophage infiltration (red MOMA-2 staining) within lesions of mice treated with OVA/CFA+Tr1 compared with either Tr1- or OVA/CFA-treated mice. Magnification ×40 for oil red O staining; ×400 for MOMA-2 staining.

Figure2

Figure 2. Representative photomicrographs show IL-10 red staining (arrows) in sections of advanced atherosclerotic plaques from aortic sinus of mice treated with OVA/CFA (a and c) or (OVA/CFA)+Tr1 (b and d). Intense IL-10 staining (arrows in b and d) was found in plaques of mice treated with OVA/CFA+Tr1. Magnification: a and b, ×200; c and d, ×400.

Discussion

A substantial body of evidence suggests an important role for Th1-mediated immunoinflammatory responses in the development and progression of atherosclerotic lesions, at least in experimental models of human-like atherosclerosis.3–10 There are also reports from human studies suggesting a strong association between Th1-related responses and plaque instability.9,26–29 Therefore, much of the recent research in the field of atherosclerosis, based on current dogma suggesting that the immunoinflammatory response in atherosclerosis is controlled by distinct Th1 and Th2 subpopulations of T cells, has focused on the potential role of Th2-mediated responses in the modulation of the atherosclerotic process. The finding by several independent groups that IL-10, a Th2-related cytokine, exerts major antiatherogenic effects has particularly reinforced the Th1/Th2 paradigm in immunity to atherosclerosis.11,30–33 However, IL-10 is not specific to Th2 cells. Interestingly, another subtype of T cells, Tr1, with cytokine profiles distinct from either Th1 or Th2 cells, has been identified and shown to play an important role in the regulation of and protection against various Th1- and Th2-mediated immunoinflammatory diseases (see References 12–14 for review). Therefore, we hypothesized that in atherosclerosis, there may be an imbalance between the effector and the regulatory (for instance, Tr1) arms of the immune response and suggested that supplementation with Tr1 cells may lead to the induction of a regulatory immune phenotype and a reduction in Th1- and Th2-mediated responses, ultimately altering plaque development and/or composition.

Here, we showed that these regulatory Tr1 cells, when transferred into mice with their cognate antigen, induced a significant suppression of Th1-mediated responses and led to an increase in IL-10 production by stimulated peripheral T cells. Tr1-mediated immune suppression was observed in response to stimulation with either OVA or conA, suggesting the induction of a systemic bystander immune suppression, which most likely resulted from repeated administration of OVA/CFA. Interestingly, the induction of Tr1 responses was associated with a significant reduction in atherosclerotic plaque size and a marked reduction in the relative accumulation of inflammatory macrophages and T lymphocytes with preservation of smooth muscle cell and collagen contents. It is noteworthy that the transfer and in vivo activation of Tr1 cells led to a profound reduction in the ratio of IFN-γ to IL-10 compared with the other groups (Table 2) and with the appearance of intense IL-10 staining within the plaques, suggesting that this change in the balance between proatherogenic and antiatherogenic mediators may have contributed to the alterations in lesion development and composition. These results show that modulation of the immune response is achievable by transfer and activation of Tr1 cells and leads to limitation of the development of atherosclerosis in apoE-deficient mice.

It could be argued that our data may simply be compatible with a systemic effect of circulating IL-10 and that such an effect may be independent of any regulatory or immunosuppressive function. Although it is very difficult to separate the regulatory functions of Tr1 clones from their capacity to produce IL-10 (several studies have established the critical contribution of the immunomodulatory cytokine IL-10 to the regulatory potential of Tr1; for review, see References 12 and 13), we believe that several data argue against such a hypothesis. First, we have been unable to detect any substantial production or difference in serum IL-10 levels between the various study groups, whereas IL-10 protein accumulation was increased locally within the plaques of the OVA/CFA+Tr1 group. Second, not all IL-10–producing T cells are regulatory T cells. Pinderski et al33 have recently shown that overexpression of IL-10 by T lymphocytes (IL-10 was placed under the control of the IL-2 promoter) led to a switch toward a Th2 phenotype that is phenotypically and functionally distinct from all Tr1 clones. Unlike the results obtained with the overexpression of IL-10 in the latter study,33 we showed in our work that appropriate activation of Tr1 led to significant reductions in both Th1- (IFN-γ) and Th2-related (IL-4 and IL-5) cytokines and immunoglobulins (IgG2a and IgE, respectively) (Tables 1 and 2⇑). This is consistent with previous studies showing that only regulatory T cells, like Tr1, are able to suppress both Th1- and Th2-related responses.12,13,20,24

Another potential limitation to our study is the confounding effect of CFA on atherosclerotic lesion development. Indeed, previously published studies have reported a reduction in atherosclerosis with CFA/IFA.34,35 These reductions in the aortic arch or the thoracic aorta were moderate (18% and 19%) and were accompanied by changes in total and/or HDL cholesterol levels that may have contributed to limitation of plaque progression. It is noteworthy that in our study, the OVA/CFA group showed a reduction in atherosclerotic lesion size in the thoracic aorta (23%) similar to that reported in studies using CFA/IFA, even in the absence of changes in cholesterol levels. However, in our study, the additional transfer of Tr1 cells resulted in a much more important and marked 55% reduction in atherosclerosis.

In conclusion, our results provide the first proof of the concept that cell transfer of a novel subpopulation of T cells with regulatory functions, here Tr1 cells, is able to modulate the in vivo immune response and leads to substantial limitation of plaque size while inducing changes in plaque composition indicative of a less inflammatory plaque phenotype. This work paves the way for the development and testing of novel therapeutic strategies based on the enhancement and/or transfer of specific regulatory immune cells (ie, Tr1 cells specific for a plaque-derived antigen) to limit the development and prevent the complications of atherosclerosis.

Acknowledgments

This work was supported by Action Concertée Incitative Jeunes Chercheurs, Ministère de la Recherche, France (to Dr Mallat), and Fondation de France.

Footnotes

  • This article originally appeared Online on August 11, 2003 (Circulation. 2003;108:r47–r52).

References

  1. ↵
    Lusis AJ. Atherosclerosis. Nature. 2000; 407: 233–241.
    OpenUrlCrossRefPubMed
  2. ↵
    Glass CK, Witztum JL. Atherosclerosis: the road ahead. Cell. 2001; 104: 503–516.
    OpenUrlCrossRefPubMed
  3. ↵
    Hansson GK. Immune mechanisms in atherosclerosis. Arterioscler Thromb Vasc Biol. 2001; 21: 1876–1890.
    OpenUrlAbstract/FREE Full Text
  4. ↵
    Gupta S, Pablo AM, Jiang XC, et al. IFN-gamma potentiates atherosclerosis in apoE knock-out mice. J Clin Invest. 1997; 99: 2752–2761.
    OpenUrlCrossRefPubMed
  5. ↵
    Whitman SC, Ravisankar P, Elam H, et al. Exogenous interferon-gamma enhances atherosclerosis in apolipoprotein E−/− mice. Am J Pathol. 2000; 157: 1819–1824.
    OpenUrlCrossRefPubMed
  6. ↵
    Lee TS, Yen HC, Pan CC, et al. The role of interleukin 12 in the development of atherosclerosis in apoE-deficient mice. Arterioscler Thromb Vasc Biol. 1999; 19: 734–742.
    OpenUrlAbstract/FREE Full Text
  7. ↵
    Zhou X, Nicoletti A, Elhage R, et al. Transfer of CD4(+) T cells aggravates atherosclerosis in immunodeficient apolipoprotein E knockout mice. Circulation. 2000; 102: 2919–2922.
    OpenUrlAbstract/FREE Full Text
  8. ↵
    Laurat E, Poirier B, Tupin E, et al. In vivo downregulation of T helper cell 1 immune responses reduces atherogenesis in apolipoprotein E-knockout mice. Circulation. 2001; 104: 197–202.
    OpenUrlAbstract/FREE Full Text
  9. ↵
    Mallat Z, Corbaz A, Scoazec A, et al. Interleukin-18/interleukin-18 binding protein signaling modulates atherosclerotic lesion development and stability. Circ Res. 2001; 89: E41–E45.
    OpenUrlCrossRefPubMed
  10. ↵
    Whitman SC, Ravisankar P, Daugherty A. Interleukin-18 enhances atherosclerosis in apolipoprotein E(−/−) mice through release of interferon-gamma. Circ Res. 2002; 90: E34–E38.
    OpenUrlCrossRefPubMed
  11. ↵
    Daugherty A, Rateri DL. T lymphocytes in atherosclerosis: the yin-yang of Th1 and Th2 influence on lesion formation. Circ Res. 2002; 90: 1039–1040.
    OpenUrlFREE Full Text
  12. ↵
    Groux H. An overview of regulatory T cells. Microbes Infect. 2001; 3: 883–889.
    OpenUrlCrossRefPubMed
  13. ↵
    Roncarolo MG, Bacchetta R, Bordignon C, et al. Type 1 T regulatory cells. Immunol Rev. 2001; 182: 68–79.
    OpenUrlCrossRefPubMed
  14. ↵
    McGuirk P, Mills KH. Pathogen-specific regulatory T cells provoke a shift in the Th1/Th2 paradigm in immunity to infectious diseases. Trends Immunol. 2002; 23: 450–455.
    OpenUrlCrossRefPubMed
  15. ↵
    Groux H, Bigler M, O’Garra A, et al. A CD4+ T-cell subset inhibits antigen-specific T cell responses and prevents colitis. Nature. 1997; 389: 737–742.
    OpenUrlCrossRefPubMed
  16. ↵
    Doetze A, Satoguina J, Burchard G, et al. Antigen-specific cellular hyporesponsiveness in a chronic human helminth infection is mediated by T(h)3/T(r)1-type cytokines IL-10 and transforming growth factor-beta but not by a T(h)1 to T(h)2 shift. Int Immunol. 2000; 12: 623–630.
    OpenUrlAbstract/FREE Full Text
  17. ↵
    Cavani A, Nasorri F, Prezzi C, et al. Human CD4+ T lymphocytes with remarkable regulatory functions on dendritic cells and nickel-specific Th1 immune responses. J Invest Dermatol. 2000; 114: 295–302.
    OpenUrlCrossRefPubMed
  18. ↵
    Levings MK, Sangregorio R, Galbiati F, et al. IFN-alpha and IL-10 induce the differentiation of human type 1 T regulatory cells. J Immunol. 2001; 166: 5530–5539.
    OpenUrlAbstract/FREE Full Text
  19. ↵
    MacDonald AJ, Duffy M, Brady MT, et al. CD4 T helper type 1 and regulatory T cells induced against the same epitopes on the core protein in hepatitis C virus-infected persons. J Infect Dis. 2002; 185: 720–727.
    OpenUrlAbstract/FREE Full Text
  20. ↵
    McGuirk P, McCann C, Mills KH. Pathogen-specific T regulatory 1 cells induced in the respiratory tract by a bacterial molecule that stimulates interleukin 10 production by dendritic cells: a novel strategy for evasion of protective T helper type 1 responses by Bordetella pertussis. J Exp Med. 2002; 195: 221–231.
    OpenUrlAbstract/FREE Full Text
  21. ↵
    Fukaura H, Kent SC, Pietrusewicz MJ, et al. Induction of circulating myelin basic protein and proteolipid protein-specific transforming growth factor-beta1-secreting Th3 T cells by oral administration of myelin in multiple sclerosis patients. J Clin Invest. 1996; 98: 70–77.
    OpenUrlCrossRefPubMed
  22. ↵
    Chen Y, Kuchroo VK, Inobe J, et al. Regulatory T cell clones induced by oral tolerance: suppression of autoimmune encephalomyelitis. Science. 1994; 265: 1237–1240.
    OpenUrlAbstract/FREE Full Text
  23. ↵
    Shevach EM. Regulatory T cells in autoimmunity. Annu Rev Immunol. 2000; 18: 423–449.
    OpenUrlCrossRefPubMed
  24. ↵
    Cottrez F, Hurst SD, Coffman RL, et al. T regulatory cells 1 inhibit a Th2-specific response in vivo. J Immunol. 2000; 165: 4848–4853.
    OpenUrlAbstract/FREE Full Text
  25. ↵
    Barrat FJ, Cua DJ, Boonstra A, et al. In vitro generation of interleukin 10-producing regulatory CD4(+) T cells is induced by immunosuppressive drugs and inhibited by T helper type 1 (Th1)- and Th2-inducing cytokines. J Exp Med. 2002; 195: 603–616.
    OpenUrlAbstract/FREE Full Text
  26. ↵
    Liuzzo G, Kopecky SL, Frye RL, et al. Perturbation of the T-cell repertoire in patients with unstable angina [see comments]. Circulation. 1999; 100: 2135–2139.
    OpenUrlAbstract/FREE Full Text
  27. ↵
    Liuzzo G, Goronzy JJ, Yang H, et al. Monoclonal T-cell proliferation and plaque instability in acute coronary syndromes. Circulation. 2000; 101: 2883–2888.
    OpenUrlAbstract/FREE Full Text
  28. ↵
    Liuzzo G, Vallejo AN, Kopecky SL, et al. Molecular fingerprint of interferon-gamma signaling in unstable angina. Circulation. 2001; 103: 1509–1514.
    OpenUrlAbstract/FREE Full Text
  29. ↵
    Mallat Z, Henry P, Fressonnet R, et al. Increased plasma concentrations of interleukin-18 in acute coronary syndromes. Heart. 2002; 88: 467–469.
    OpenUrlAbstract/FREE Full Text
  30. ↵
    Mallat Z, Besnard S, Duriez M, et al. Protective role of interleukin-10 in atherosclerosis. Circ Res. 1999; 85: e17–e24.
    OpenUrlAbstract/FREE Full Text
  31. ↵
    Pinderski Oslund LJ, Hedrick CC, Olvera T, et al. Interleukin-10 blocks atherosclerotic events in vitro and in vivo. Arterioscler Thromb Vasc Biol. 1999; 19: 2847–2853.
    OpenUrlAbstract/FREE Full Text
  32. ↵
    von der Thusen JH, Kuiper J, Fekkes ML, et al. Attenuation of atherogenesis by systemic and local adenovirus-mediated gene transfer of interleukin-10 in LDLr−/− mice. FASEB J. 2001; 15: 2730–2732.
    OpenUrlFREE Full Text
  33. ↵
    Pinderski LJ, Fischbein MP, Subbanagounder G, et al. Overexpression of interleukin-10 by activated T lymphocytes inhibits atherosclerosis in LDL receptor-deficient mice by altering lymphocyte and macrophage phenotypes. Circ Res. 2002; 90: 1064–1071.
    OpenUrlAbstract/FREE Full Text
  34. ↵
    Hansen PR, Chew M, Zhou J, et al. Freunds adjuvant alone is antiatherogenic in apoE-deficient mice and specific immunization against TNFalpha confers no additional benefit. Atherosclerosis. 2001; 158: 87–94.
    OpenUrlCrossRefPubMed
  35. ↵
    Binder CJ, Horkko S, Dewan A, et al. Pneumococcal vaccination decreases atherosclerotic lesion formation: molecular mimicry between Streptococcus pneumoniae and oxidized LDL. Nat Med. 2003; 9: 736–743.
    OpenUrlCrossRefPubMed
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Circulation
September 9, 2003, Volume 108, Issue 10

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  • Induction of a Regulatory T Cell Type 1 Response Reduces the Development of Atherosclerosis in Apolipoprotein E–Knockout Mice
    Ziad Mallat, Andrea Gojova, Valérie Brun, Bruno Esposito, Nathalie Fournier, Françoise Cottrez, Alain Tedgui and Hervé Groux
    Circulation. 2003;108:1232-1237, originally published September 8, 2003
    https://doi.org/10.1161/01.CIR.0000089083.61317.A1
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