Blockade of T cell Costimulatory Signals using Adenovirus Vectors Prevents both the Induction and the Progression of Experimental Autoimmune Myocarditis

doi:10.1006/jmcc.2001.1511

Journal of Molecular and Cellular Cardiology

Volume 34, Issue 3, March 2002, Pages 279-295

https://doi.org/10.1006/jmcc.2001.1511 Get rights and content

Abstract

Experimental autoimmune myocarditis (EAM) has been used as a model for human myocarditis in relation to the autoimmune mechanism and proved to be a T cell-mediated autoimmune disease. Interactions of T cell surface receptors CD28 and CD40L with their ligands B7 and CD40, respectively, on APCs are critical for antigen-specific T cell activation under physiological and pathological conditions. To achieve effective inhibition of these interactions, we have constructed adenovirus vectors containing CTLA4Ig (AdexCTLA4Ig) and CD40Ig (AdexCD40Ig) and examined the effects of these adenovirus vectors in preventing EAM. AdexLacZ as a control, or AdexCTLA4Ig and/or AdexCD40Ig were injected intravenously into rats on day 0 or 14 after immunization to study the preventive effects on EAM in the T cell activation phase or inflammatory phase. Disease severity was estimated by the macroscopic and microscopic findings of the heart, heart weight to body weight ratios, and cellular and humoral immune responses on day 21. The onset of EAM after AdexCTLA4Ig or AdexCD40Ig treatment on day 0 was completely inhibited and antigen-specific lymphocyte proliferation was significantly reduced in those adenovirus-treatment groups, suggesting that those therapies induce antigen-specific T cell anergy. Moreover, significant reduction in disease severity was achieved after the adenovirus vector treatment even on day 14 compared with EAM rats. This study indicates the therapeutic potential of costimulatory pathway blockade by gene-transfer in myocarditis.

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Citation Excerpt :
Circulating IL-1β and TNF-α, which activate cytotoxic reactions, are markedly increased in patients with myocarditis [151,154,155], and the levels of IL-1β and TNF-α are correlated with the severity of the disease [156]. Inhibition of these cytokines can prevent the development of EAM [155–157]. IL-1β has multiple effects on cardiac function.
Interleukin-1β (IL-1β) belongs to IL-1 family and is a potent pro-inflammatory cytokine. It is known to be also involved in a variety of cellular activities, including cell proliferation, differentiation and apoptosis. In addition to its pathophysiologic role in host protection, IL-1β promotes the progression of a number of autoimmune diseases. Most of such diseases can be controlled by anti-IL-1β treatment. This review discusses the contribution of IL-1β to the course of autoimmune diseases, such as rheumatic diseases, uveitis, autoimmune thyroid diseases (AITD), insulin-dependent diabetes mellitus (IDDM), autoimmune inner ear disease (AIED), multiple sclerosis (MS), myocarditis, hepatitis and kidney diseases. The critical involvement of IL-1β in the pathogenesis of autoimmune diseases provides targets for developing therapeutic treatment.
CTLA4-Ig Relieves Inflammation in Murine Models of Coxsackievirus B3-Induced Myocarditis
2012, Canadian Journal of Cardiology
Citation Excerpt :
CTLA-4, a structural homolog for CD28, provides a competing inhibitory signal. Furthermore, CTLA4-Ig was applied in autoimmunity myocarditis and both improved cardiac function and alleviated autoimmunity myocardial damage.14,15 The recovery effects have been verified in our study.
T-cell–mediated cellular immunity is one of the most important factors in viral myocarditis. As an important costimulatory molecule, cytotoxic T-lymphocyte–associated antigen 4 (CTLA4) alleviates autoimmunity by influencing the balance of helper T cell (T_H) subtype 1 (T_H1) to T_H2 in autoimmune diseases. The effects and mechanisms of CTLA4 fusion protein (CTLA4-Ig) in mice with coxsackievirus B3 (CVB₃)–induced myocarditis were investigated.
BALB/c mice were randomly divided into a CVB₃ group, an IgG group, a CTLA4-Ig group, and a group of healthy control mice. Mice were humanely killed on day 7 post CVB₃ inoculation, then CVB₃, IFN-γ, mouse IL-4 (mIL-4), and mouse IL-2 (mIL-2) expression in myocardium were examined by real-time quantitative polymerase chain reaction, and the serum concentrations of IFN-γ, mIL-4, and mIL-2 were measured by enzyme-linked immunosorbent assay.
IFN-γ expression was significantly higher and mIL-4 levels in serum were lower in the CVB₃ group when compared with those in the healthy control group (P < 0.01). In the CTLA4-Ig group, the mouse mortality and CVB₃ mRNA in myocardium were reduced compared with those in the CVB₃ group. Furthermore, IFN-γ expression was lower, and mIL-4 was significantly higher compared with those values in the CVB₃ and the IgG groups. The levels of mIL-2 in all groups showed no statistical difference (P > 0.05).
T_H1 cytokines were predominant in the acute phase of viral myocarditis. CTLA4-Ig relieves myocardial inflammation, virus replication, and mouse mortality, probably by influencing the balance of T_H1 to T_H2.
L'immunité à médiation cellulaire est l'un des facteurs les plus importants de la myocardite virale. En tant qu'importante molécule costimulatoire, l'antigène 4 des lymphocytes T cytotoxiques (CTLA4) apaise l'auto-immunité en influençant l'équilibre des sous-types 1 et 2 des lymphocytes T auxiliaire (T_h) dans les maladies auto-immunes. Les effets et les mécanismes de la protéine de fusion CTLA4 (CTLA4-Ig) chez la souris ayant une myocardite induite par le virus Coxsackie B3 (CVB₃) ont été examinés.
Les souris BALB/c ont été réparties au hasard dans un groupe CVB₃, un groupe IgG, un groupe CTLA4-Ig et un groupe de souris témoins en santé. Les souris ont été tuées euthanasiées au 7^e jour après l'inoculation du CVB₃, puis l'expression du CVB₃, de l'INF- γ, de l'IL-4 chez la souris (mIL-4) et de l'IL-2 (mIL-2) chez la souris dans le myocarde a été étudiée par la réaction en chaîne par polymérase quantitative en temps réel, et les concentrations sériques de l'INF-γ, la mIL-4, et la mIL-2 ont été mesurées par titrage avec immunoabsorbant lié à une enzyme.
L'expression de l'INF-γ a été significativement plus élevée, et les niveaux sériques de la mIL-4 ont été plus bas dans le groupe CVB₃ comparativement à ceux du groupe témoin en santé (P < 0,01). Dans le groupe CTLA4-Ig, la mortalité chez la souris et l'ARN messager CVB₃ dans le myocarde ont été réduits comparativement au groupe CVB₃. Par ailleurs, l'expression de l'INF-γ a été plus faible, et la sIL-4 a été significativement plus élevée comparativement aux valeurs des groupes CVB₃ et IgG. Les niveaux de la mIL-2 de tous les groupes n'ont montré aucune différence statistique (P > 0,05).
Les cytokines T_h1 étaient prédominantes à la phase aiguë de la myocardite virale. Le CTLA4-Ig soulage l'inflammation du myocarde, la réplication virale et la mortalité chez la souris, probablement en influençant l'équilibre de T_h1 à T_h2.
Blocking the CD40-CD40L interaction by CD40-Ig reduces disease progress in murine myocarditis induced by CVB3
2010, Cardiovascular Pathology
CD40 and CD40L, as costimulatory molecules, are reported to play a critical role for cytokine production and antigen-specific T-cell activation in acute viral myocarditis (VMC). The purpose of this study was to probe the therapeutic effect of CD40-Ig in coxsackievirus B3 (CVB3)-induced myocarditis.
Adolescent male Balb/c mice were infected with cardiovirulent CVB3 and then injected with CD40-Ig (0.1 mg/kg) at 6 h and 3 days postinfection (pi), with IgG (0.1 mg/kg) as control. The surviving mice were sacrificed on the seventh day. The copies of CVB3 mRNA in the myocardium were detected by real-time PCR and the expression of CD40 protein and CD4⁺ T cell was assessed by immunohistochemistry method. The serum level of IL-4 and IFN-γ and their transcriptions in the myocardium were determined by ELISA and real-time PCR.
The mortality was low in VMC model mice receiving CD40-Ig treatment with relived inflammation and reduced transcription of CVB3 in the myocardium. The expression pattern of IFN-γ (Th1-related cytokine) and IL-4 (Th2-related cytokine) was altered to Th1 prevalence in the acute phase of VMC. Intervention of CD40-Ig switched the balance of Th1/Th2.
Our findings suggest that intervention of CD40-Ig can relieve the myocardial injury and inhibit viral replication of CVB3 in VMC. Its mechanism may involve blocking the interaction between CD40 and CD40L and regulating the Th1/Th2 balance.
Attenuation of experimental autoimmune myocarditis by blocking T cell activation through 4-1BB pathway
2009, Journal of Molecular and Cellular Cardiology
4-1BB, a member of the tumor necrosis factor receptor (TNFR) family, binds the 4-1BB ligand (4-1BBL), works as a costimulatory molecule, and regulates T cell-mediated immune responses. Although inflammation is an essential pathological feature of myocarditis, the role of 4-1BB in experimental autoimmune myocarditis (EAM) remains unclear. Lewis rats were immunized on day 0 with purified porcine cardiac myosin to establish EAM. 4-1BB-immunoglobulin (4-1BBIg) was administered intraperitoneally (n = 6) a total of 9 times (3 times per week). Rats were killed on day 21 to study effects of 4-1BB/4-1BBL pathway blockade. For controls, isotype-matched human IgG was administered in other EAM rats (n = 6). Histologic and echocardiographic examination showed development of EAM attenuated by 4-1BBIg. Suppression of mRNA expression for IL-1α, IL-1β, IL-4, IL-6, and TNF-α was noted in the heart tissue treated with 4-1BBIg. Treatment with 4-1BBIg reduced production of Th1-type cytokines, and inhibited T cell proliferation in vitro. In the 4-1BB signaling pathway in splenocytes, 4-1BBIg suppressed JNK, p38, and IκB activity but not that of ERK1/2. Blockade of T cell activation through the 4-1BB/4-1BBL pathway regulates development of EAM; therefore, 4-1BB may be an effective target for treating myocarditis.
Role of CD28 in fatal autoimmune disorder in scurfy mice
2007, Blood
Scurfy mice develop CD4 T-cell–mediated lymphoproliferative disease leading to death within 4 weeks of age. The scurfy mutation causes loss of function of the foxp3 gene (foxp3^sf), which is essential for development and maintenance of naturally occurring regulatory CD4 T cells (nTregs). In humans, mutations of the foxp3 gene cause immune dysregulation, polyendocrinopathy, enteropathy, and X-linked syndrome (IPEX). In most patients with IPEX and also in scurfy mice, T cells show hyperreactivity and levels of Th1- and Th2-associated cytokines are substantially elevated. We report that removal of CD28 expression rescued scurfy mice from early death. Longer-term surviving CD28-deficient scurfy mice still had lymphoproliferative disorder, but their CD4 T cells showed decreased interferon-γ and no sign of interleukin-4 or interleukin-10 hyperproduction. Furthermore, injection of CTLA4-Ig to block CD28-B7 interactions substantially improved the survival of scurfy mice by blocking effector T-cell differentiation. These data support the hypothesis that CD28-B7 interactions play a critical role in the etiology of lethal autoimmune disease in scurfy mice by stimulating the differentiation of antigen-activated naive T cells into effector T cells.
Prevention of experimental autoimmune myocarditis by hydrodynamics-based naked plasmid DNA encoding CTLA4-Ig gene delivery
2005, Journal of Cardiac Failure
Citation Excerpt :
This technology remedies the shortcoming of gene transfection using adenovirus, as outlined previously. Matsui et al demonstrated therapy with adenovirus vectors containing CTLA4-Ig to EAM.10 In their study, blockade of T-cell costimulation by CTLA4-Ig prevented the induction and progression of EAM, as shown in their histologic findings, HW/BW ratio, and cellular and hormonal immune response.
Rat experimental autoimmune myocarditis (EAM) is a T cell–mediated disease that resembled the giant cell myocarditis seen in humans. Soluble CTLA4 improves some autoimmune diseases by blocking costimulatory signals on T cell. We investigated the effect of hydrodynamics-based naked plasmid DNA encoding CTLA4-immunoglobulin (Ig) gene delivery.
Lewis rats were immunized with cardiac myosin and treated with hydrodynamic-based transfection, namely a rapid tail vein injection of a large volume of pCAGGS encoding CTLA4-Ig chimera solution on Day 0. The vector-derived CTLA4-Ig mRNA expressions were mainly detected in the liver and plasma CTLA4-Ig protein levels were maintained at about 2 μg/mL during the experiment period. On Day 17, the ratio of heart to body weight, the amount of mRNA of atrial natriuretic peptide, and the inflammatory areas in CTLA4 group were significantly lower than in the control group treated with empty plasmid. Maximum rate of intraventricular pressure rise and decline (dP/dT), minimum dP/dT, left ventricular end-diastolic pressure, and central venous pressure improved significantly after treatment with CTLA4-Ig. On Day 14, expressions of IL-2 in popliteal lymph nodes in the CTLA4-Ig group were significantly lower than in the control group.
Hydrodynamics-based transfection of plasmid encoding CTLA4-Ig chimera dramatically prevented EAM.

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^f1: Please address all correspondence to: Hiroshi Okamoto, M.D. Department of Cardiovascular Medicine, Hokkaido University Graduate School of Medicine, Kita-15, Nishi-7, Kita-ku, Sapporo 060-8638, Japan. Tel: 81-11-716-1161; Fax: 81-11-706-7874.E-mail: [email protected]

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Regular ArticlesBlockade of T cell Costimulatory Signals using Adenovirus Vectors Prevents both the Induction and the Progression of Experimental Autoimmune Myocarditis

Abstract

Am J Cardiol

J Am Coll Cardiol

Immunity

J Mol Cell Cardiol

J Neuroimmunol

J Mol Cell Cardiol

J Mol Cell Cardiol

Immunol Today

Am J Pathol

Am Heart J

Immunity

J Am Coll Cardiol

Viral myocarditis. A review

Am J Pathol

Nitric oxide contributes to the progression of myocardial damage in experimental autoimmune myocarditis in rats

Circulation

Rat dilated cardiomyopathy after autoimmune giant cell myocarditis

Circ Res

In vivo lymphocyte-mediated myocardial injuries demonstrated by adoptive transfer of experimental auto- immune myocarditis

Circulation

The role of T-cell costimulatory activation pathways in transplant rejection

N Engl J Med

CD40 and CD154 in cell-mediated immunity

Annu Rev Immunol

Immunosuppression in vivo by a soluble form of the CTLA4 T cell activation molecule

Science

Persistent and secondary adenovirus-mediated hepatic gene expression using adenovirus vector containing CTLA4IgG

Hum Gene Ther

Cardiac myosin from pig heart ventricle. Purification and enzymatic properties

J Biochem (Tokyo)

Regular Articles
Blockade of T cell Costimulatory Signals using Adenovirus Vectors Prevents both the Induction and the Progression of Experimental Autoimmune Myocarditis