CD8+CD28− Regulatory T Lymphocytes Prevent Experimental Inflammatory Bowel Disease in Mice

doi:10.1053/j.gastro.2006.09.008

Gastroenterology

Volume 131, Issue 6, December 2006, Pages 1775-1785

https://doi.org/10.1053/j.gastro.2006.09.008 Get rights and content

Background & Aims: Immune responses to innocuous intestinal antigens appear tightly controlled by regulatory T lymphocytes. While CD4⁺ T lymphocytes have recently attracted the most attention, CD8⁺ regulatory T-cell populations are also believed to play an important role in control of mucosal immunity. However, CD8⁺ regulatory T-cell function has mainly been studied in vitro and no direct in vivo evidence exists that they can control mucosal immune responses. We investigated the capacity of CD8⁺CD28⁻ T cells to prevent experimental inflammatory bowel disease (IBD) in mice. Methods: CD8⁺CD28⁻ regulatory T cells were isolated from unmanipulated mice and tested for their capacity to inhibit T-cell activation in allogeneic mixed lymphocyte cultures in vitro and to prevent IBD induced by injection of CD4⁺CD45RB^high cells into syngeneic immunodeficient RAG-2 mutant mice. Results: CD8⁺CD28⁻ T lymphocytes inhibited proliferation and interferon gamma production by CD4⁺ responder T cells in vitro. CD8⁺CD28⁻ regulatory T cells freshly isolated from spleen or gut efficiently prevented IBD induced by transfer of colitogenic T cells into immunodeficient hosts. Regulatory CD8⁺CD28⁻ T cells incapable of producing interleukin-10 did not prevent colitis. Moreover, IBD induced with colitogenic T cells incapable of responding to transforming growth factor β could not be prevented with CD8⁺CD28⁻ regulatory T cells. CD8⁺CD28⁺ T cells did not inhibit in vitro or in vivo immune responses. Conclusions: Our findings show that naturally occurring CD8⁺CD28⁻ regulatory T lymphocytes can prevent experimental IBD in mice and suggest that these cells may play an important role in control of mucosal immunity.

Section snippets

Mice

All mice (females) were used at 6–10 weeks of age except where indicated differently. C57BL/6 and DBA/2 mice were purchased from Janvier (Le Genest St Isle, France). RAG-2–deficient and major histocompatibility complex (MHC)-deficient (IAβ°β2m°) C57BL/6 mice were bred in our specific pathogen-free animal facility and were originally obtained from the Centre de Développement des Technologies Advancées–Centre National de la Recherche Scientifique (Orléans, France). IL-10–deficient C57BL/6 mice

Phenotypic Analysis of CD8⁺CD28⁻ T Lymphocytes

To assess the relation of CD8⁺CD28⁻ T cells to other previously reported CD8⁺ regulatory T lymphocytes, we analyzed the phenotype of these cells by flow cytometry (Figure 1). C57BL/6 splenocytes were stained with antibodies specific for CD4, CD8, and CD28 or an isotype-matched control antibody (Figure 1A). CD8⁺ T cells generally expressed slightly lower levels of CD28 than CD4⁺ cells. However, no clear CD8⁺CD28⁻ population could be distinguished. CD8⁺CD28⁻ cells were therefore defined as those

Discussion

Here we have shown that CD8⁺CD28⁻ T lymphocytes from unmanipulated wild-type mice efficiently inhibited proliferation and IFN-γ production by CD4⁺ responder T cells in allogeneic mixed lymphocyte cultures. Naive CD8⁺CD28⁻ regulatory T cells, isolated from spleen or intestines, efficiently inhibited IBD induced by transfer of CD4⁺CD45RB^high cells into immunodeficient mice. This in vivo immunosuppression required IL-10 production by regulatory T cells and responsiveness to TGF-β of colitogenic

References (52)

C.A. Piccirillo et al.
Naturally-occurring CD4+CD25+ immunoregulatory T cells: central players in the arena of peripheral tolerance
Semin Immunol
(2004)
M. Terabe et al.
Immunoregulatory T cells in tumor immunity
Curr Opin Immunol
(2004)
J. Maul et al.
Peripheral and intestinal regulatory CD4+ CD25(high) T cells in inflammatory bowel disease
Gastroenterology
(2005)
M.V. Dhodapkar et al.
Antigen-bearing immature dendritic cells induce peptide-specific CD8+ regulatory T cells in vivo in humans
Blood
(2002)
E. Xystrakis et al.
Identification of a novel natural regulatory CD8 T-cell subset and analysis of its mechanism of regulation
Blood
(2004)
M. Allez et al.
Expansion of CD8+ T cells with regulatory function after interaction with intestinal epithelial cells
Gastroenterology
(2002)
L. Gorelik et al.
Abrogation of TGFbeta signaling in T cells leads to spontaneous T cell differentiation and autoimmune disease
Immunity
(2000)
K.A. Hogquist et al.
Central tolerance: learning self-control in the thymus
Nat Rev Immunol
(2005)
R.R. Hardy et al.
B cell development pathways
Annu Rev Immunol
(2001)
B. Stockinger
T lymphocyte tolerance: from thymic deletion to peripheral control mechanisms
Adv Immunol
(1999)

D. Hu et al.

Analysis of regulatory CD8 T cells in Qa-1-deficient mice

Nat Immunol

(2004)

Cited by (96)

CD8+ T cells: The past and future of immune regulation
2020, Cellular Immunology
Citation Excerpt :
Loss of the prototypical co-stimulatory molecule CD28 has been seen to occur in multiple species and is associated with an anergic or senescent phenotype [49,50]. CD28- CD8reg have been implicated as regulatory agents in infection [51], transplantation [52], tumor settings, and autoimmunity [21,53,54]. In human autoimmune disease, it was reported that individuals with T1D or MS had decreased levels of these CD8reg in peripheral blood when compared to healthy controls, suggesting that deficiency of these cells may contribute to disease [55].
Regulation of the adaptive immune response is critical for health. Regulatory activity can be found in multiple components of the immune system, however, the focus on particular components of the immune regulatory network has left many aspects of this critical immune component understudied. Here we review the evidence for activities of CD8+ T cells in immune homeostasis and regulation of autoimmune reactivity. The heterogeneous nature of identified CD8+ cell types are examined, and common phenotypes associated with functional activities are defined. The varying types of antigen signal crucial for CD8+ T cell regulatory activity are identified and the implications of these activation pathways for control of adaptive responses is considered. Finally, the promising capacity for transgenic antigen receptor directed cytotoxicity as a mechanism for modulation of autoimmunity is detailed.
CTLA-4: From mechanism to autoimmune therapy
2020, International Immunopharmacology
CD28 and CTLA-4 are both important stimulatory receptors for the regulation of T cell activation. Because receptors share common ligands, B7.1 and B7.2, the expression and biological function of CTLA-4 is important for the negative regulation of T cell responses. Therefore, elimination of CTLA-4 can result in the breakdown of immune tolerance and the development of several diseases such as autoimmunity. Inhibitory signals of CTLA-4 suppress T cell responses and protect against autoimmune diseases in many ways. In this review, we summarize the structure, expression and signaling pathway of CTLA-4. We also highlight how CTLA-4 defends against potentially self-reactive T cells. Finally, we discuss how the CTLA-4 regulates a number of autoimmune diseases that indicate manipulation of this inhibitory molecule is a promise as a strategy for the immunotherapy of autoimmune diseases.
Anti-LFA-1 induces CD8 T-cell dependent allograft tolerance and augments suppressor phenotype CD8 cells
2018, Cellular Immunology
Citation Excerpt :
While others have found an increase in CD4+CD25+FoxP3+ T-regulatory cells when using LFA-1 blockade in a skin graft model [2], we were unable to find similar increases in the cardiac allograft model. In fact, depleting CD25+ cells had no effect, which is consistent with a role for CD8+CD28− suppressor cells as they are CD25− [45–48]. The increase in CD8+CD28− cells in association with αLFA-1 may help explain why αLFA-1 and CTLA4-Ig have been reported to be synergistic in transplant models [2,49,50].
The induction of tolerance to transplanted organs is a major objective in transplantation immunology research. Lymphocyte function-associated antigen-1 (LFA-1) interactions have been identified as a key component of the T-cell activation process that may be interrupted to lead to allograft tolerance. In mice, αLFA-1 mAb is a potent monotherapy that leads to the induction of donor-specific transferable tolerance. By interrogating important adaptive and innate immunity pathways, we demonstrate that the induction of tolerance relies on CD8⁺ T-cells. We further demonstrate that αLFA-1 induced tolerance is associated with CD8⁺CD28⁻ T-cells with a suppressor phenotype, and that while CD8 cells are present, the effector T-cell response is abrogated. A recent publication has shown that CD8⁺CD28⁻ cells are not diminished by cyclosporine or rapamycin, therefore CD8⁺CD28⁻ cells represent a clinically relevant population. To our knowledge, this is the first time that a mechanism for αLFA-1 induced tolerance has been described.
Adaptive and innate immunoregulatory cells
2018, Dubois' Lupus Erythematosus and Related Syndromes
In systemic lupus erythematosus (SLE), the impaired immune homeostasis is associated with clinical manifestations that generally affect multiple tissues and organs. The inability to properly suppress proinflammatory events that lead to tissue damage and subsequent loss of organ function is significantly influenced by an ongoing dysfunction of immunoregulatory cell activities (Table 11.1). This is because different types of immune cells display functional abnormalities in SLE, including those endowed with immunoregulatory functions. Indeed, the immune dysfunction in SLE affects checkpoints that are involved in both the effector and regulatory immune responses because of an impairment of the mechanisms of maintenance and/or establishment of immune tolerance. In normal conditions, immunoregulatory cells suppress proinflammatory and effector responses to maintain immune homeostasis (Fig. 11.1). However, in SLE this important mechanism of peripheral immune tolerance to self becomes insufficient in adequately controlling autoimmune reactivity either because of numeric and/or functional deficits of the immunoregulatory cells or because of an acquired inadequacy of these cells to inhibit local mediators and immune cells that sustain and/or amplify inflammation. Different studies have implicated dysfunctional immune regulation at the level of the adaptive and innate immune systems in SLE, and defined phenotypic and functional abnormalities which will be the focus of this chapter. We will also include considerations that while the suppressive activity of immunoregulatory cells is crucial for the inhibition of autoimmune responses and inflammation, additional factors (such as stage of disease) need to be taken into account in the facilitation or reduction of the activity of these cells in SLE.
CD8+CD28+/CD8+CD28− T cell equilibrium can predict the active stage for patients with inflammatory bowel disease
2017, Clinics and Research in Hepatology and Gastroenterology
The balance of blood CD8⁺CD28⁺/CD8⁺CD28⁻ T cells has been verified to be vital for patients with ulcerative colitis (UC), but their role in inflammatory bowel disease (IBD) remains unknown. This investigation aimed to evaluate the efficiency of the balance in predicting the active stage in IBD patients.
Fifty-three IBD subjects, including 31 UC and 22 Crohn's disease (CD) patients, were enrolled, and their peripheral blood CD8⁺CD28⁺ and CD8⁺CD28⁻ T cell levels were tested using flow cytometry. The risk factors related to prognosis were compared between UC and CD patients. A 1-year follow-up was performed for all the IBD patients, and the CD8⁺ T cells and their ratio were compared at the 3rd, 6th, 9th, and 12th months during follow-up. The sensitivity and specificity of the CD8⁺ T cell level and balance were analyzed through receiver operator characteristic (ROC) curves. The cumulative remission lasting rates (CRLRs) under the different factors were analyzed using the Kaplan–Meier method.
Higher prescription rates of immunosuppressants, steroids, probiotics, and biological agents (BAs) were found in CD subjects in comparison to UC subjects (P = 0.005, 0.024, 0.034, and 0.001), as was a higher active rate during follow-up (95.5% of CD patients vs 67.7% of UC patients, P = 0.035). The CD8⁺CD28⁺ T cell level and the CD8⁺CD28⁺/CD8⁺CD28⁻ T cell ratio were significantly higher in UC patients than in CD patients, but the reverse was true for CD8⁺CD28⁻ T cells during follow-up at the 9th and 12th month (all P < 0.05). The diagnostic models of the initial CD8⁺CD28⁺ and CD8⁺CD28⁻ T cell numbers and the CD8⁺CD28⁺/CD8⁺CD28⁻ T cell ratio in predicting the active stage were found to be significant, with areas under the curves (AUCs) of 0.883, 0.098, and 0.913 for UC subjects (with 95% CI: 0.709–0.940, 0.009–0.188, and 0.842–1.003; P = 0.001, 0.00, and 0.000) and 0.812, 0.078, and 0.898 for CD subjects (with 95% CI: 0.683–0.957, 0.003–0.158, and 0.837–0.998; P = 0.003, 0.00, and 0.000). The cut-off values showed that when the ratios were 1.30 for UC and 1.22 for CD patients, the best sensitivity and specificity were observed, with 91.6% and 89.0% for UC and 88.5% and 85.1% for CD, respectively. The CRLRs were significantly higher in female, non-BA-treated, non-surgical IBD subjects when compared to male, BA-treated, surgical subjects (P = 0.031, 0.000, and 0.000). The number of CD8⁺CD28⁺ and CD8⁺CD28⁻ T cells and the CD8⁺CD28⁺/CD8⁺CD28⁻ T cell ratio were correlated with BA treatment and surgery (all P < 0.05).
The CD8⁺CD28⁺/CD8⁺CD28⁻ T cell balance, expected to be a novel immunologic marker, presented a satisfactory efficiency with high sensitivity and specificity in predicting the active stage in UC and CD patients, and the balance was closely related to the use of BAs and surgery.
CD8+ T activation attenuates CD4+ T proliferation through dendritic cells modification
2015, Cellular Immunology
Citation Excerpt :
CD8+ T cells are generally recognized as a cytotoxic T-cell population, which plays a critical role in anti-viral infection, tumor cell elimination, and allograft rejection. In contrast, accumulating CD8+ T subsets with immunoregulatory/immunosuppressive capacities have been reported in CD4+ T-associated autoimmune and inflammatory diseases [1], including asthma [2], systemic lupus erythematosus (SLE) [3], inflammatory bowel disease (IBD) [4,5], and experimental autoimmune encephalomyelitis (EAE) [6,7]. Several potential mechanisms for the immunoregulation of these CD8+ T on CD4+ T-mediated immunity have been explored, such as immune regulatory cytokines secretion (e.g., IL-10, TGF-β, IFN-γ) [8], cell–cell direct contact [9,10], and cytotoxic lysis of APCs in a perforin-dependent or Fas-dependent manner [11,12].
Emerging evidence has suggested that CD8⁺ T had modulatory function on CD4⁺ T mediated autoimmune and inflammatory diseases. However, the underlying mechanisms remain unclear. In this study, we found that CD8⁺ T activation inhibited OVA_323–339 antigen specific CD4⁺ T cells proliferation in vitro and in vivo. Further investigation demonstrated that this immunosuppression largely depended on the soluble factor from activated CD8⁺ T to modify the phenotype and functions of DCs. Moreover, not only the inhibitors for IDO or iNOS, but also IFN-γ neutralization markedly reversed this immunosuppression on OVA_323–339 antigen specific CD4⁺ T cells proliferation. Interestingly, CD8⁺ T cells absence aggravated the pathological damage in lung in OVA-induced asthma model, but alleviated by CD8⁺ T transfer and activation. Thus, these findings suggested that activated CD8⁺ T population exerted feedback regulation in DCs modification, and then attenuated CD4⁺ T mediated immune response.

View all citing articles on Scopus

: Supported by grants from the Association François Aupetit (2001, 2002) and from the Association pour la Recherche sur le Cancer (to I.M.-M.).

View full text

Basic–alimentary tractCD8+CD28− Regulatory T Lymphocytes Prevent Experimental Inflammatory Bowel Disease in Mice

Section snippets

Mice

Phenotypic Analysis of CD8+CD28− T Lymphocytes

Discussion

Semin Immunol

Curr Opin Immunol

Gastroenterology

Blood

Blood

Gastroenterology

Immunity

Central tolerance: learning self-control in the thymus

Nat Rev Immunol

B cell development pathways

Annu Rev Immunol

T lymphocyte tolerance: from thymic deletion to peripheral control mechanisms

Adv Immunol

Naturally arising CD4+ regulatory T cells for immunologic self-tolerance and negative control of immune responses

Annu Rev Immunol

Regulatory T cells: friend or foe in immunity to infection?

Nat Rev Immunol

Regulatory T cells mediate maternal tolerance to the fetus

Nat Immunol

Regulatory T cells and intestinal homeostasis

Immunol Rev

Defects in CD8+ regulatory T cells in the lamina propria of patients with inflammatory bowel disease

J Immunol

Experimental models of inflammatory bowel disease reveal innate, adaptive, and regulatory mechanisms of host dialogue with the microbiota

Immunol Rev

Regulatory interactions between CD45RBhigh and CD45RBlow CD4+ T cells are important for the balance between protective and pathogenic cell- mediated immunity

J Exp Med

An essential role for interleukin 10 in the function of regulatory T cells that inhibit intestinal inflammation

J Exp Med

T cells that cannot respond to TGF-β escape control by CD4(+)CD25(+) regulatory T cells

J Exp Med

A CD4+ T-cell subset inhibits antigen-specific T-cell responses and prevents colitis

Nature

License to heal: bidirectional interaction of antigen-specific regulatory T cells and tolerogenic APC

J Immunol

Regulatory functions of CD8+CD28– T cells in an autoimmune disease model

J Clin Invest

Oral tolerance

Immunol Rev

Essential roles of CD8+CD122+ regulatory T cells in the maintenance of T cell homeostasis

J Exp Med

Cutting edge: CD8+CD122+ regulatory T cells produce IL-10 to suppress IFN-gamma production and proliferation of CD8+ T cells

J Immunol

Analysis of regulatory CD8 T cells in Qa-1-deficient mice

Nat Immunol

Basic–alimentary tract
CD8⁺CD28⁻ Regulatory T Lymphocytes Prevent Experimental Inflammatory Bowel Disease in Mice

Phenotypic Analysis of CD8⁺CD28⁻ T Lymphocytes

Regulatory interactions between CD45RB^high and CD45RB^low CD4+ T cells are important for the balance between protective and pathogenic cell- mediated immunity