Analysis of specific Th1/Th2 helper cell responses and IgG subtype antibodies in anti-CD4 monoclonal antibody treated mice with autoimmune cardiomyopathy

Abstract

The cytokine repertoire of ADP/ATP carrier-specific humoral immune responses and the cytokine-dependent anti-ADP/ATP carrier antibody IgG subclasses were examined in a cohort of ADP/ATP carrier-immunized BALB/c mice treated with anti-CD4 monoclonal antibody. Eighteen male BALB/c mice (6–8 weeks old) were randomized into 3 groups: dilated cardiomyopathy (DCM) group, DCM-tolerance (Tol) group and control group. The mice in DCM group were immunized with the peptides derived from human ADP/ATP carrier protein for 6 months and mice in the control group were sham-immunized, while the mice in DCM-Tol group were immunized with ADP/ATP carrier protein and anti-CD4 McAb simultaneously. Serum autoantibody against ADP/ATP carrier and IgG subclasses were measured by ELISA, intracellular cytokines IFN-γ and IL-4 of Th cells were monitored with flow cytometry, and splenic T cell cytokines IFN-γ, IL-2, IL-4 and IL-6 were detected by using real-time fluorescent quantitative PCR. The results showed that the autoantibody against ADP/ATP carrier was found in all mice in DCM group, and the antibody level, serum IgG1 and IgG2a subclasses, cytokines in T cells and Th cells were all elevated in DCM group, as compared with those in control group (P<0.01). On the other hand, in DCM-Tol group, the autoantibody level and contents of all the cytokines were significantly different from those in DCM group (P<0.01), and were close to those in control group. And the levels of IgG1, IgG2a, IgG2b and IgG3 were influenced, to varying degrees, by anti-CD4 McAb as compared with those in DCM group. All these four types of IgG subclasses were substantially decreased in DCM-Tol group as compared with DCM group. It is concluded that the treatment with anti-CD4 McAb could prevent the activation of T cells, reverse the abnormal secretion of cytokines and the imbalance between Th1/Th2 cell subsets and abnormal production of autoantibody against ADP/ATP carrier, and eventually avoid myocardial injuries.

Keywords

CD4 / monoclonal antibody / autoimmunity / Th1/Th2 immune response / ADP/ATP carrier peptides

References

[1] KawaiC.. From myocarditis to cardiomyopathy: mechanisms of inflammation and cell death: learning from the past for the future. Circulation, 1999, 99(8): 1091-1100

[2] SiguschH. H., LehmannM. H., SchnittlerU., et al.. Tumour necrosis factor-alpha expression in idiopathic dilated cardiomyopathy: correlation to myocardial inflammatory activity. Cytokine, 2000, 12(8): 1261-1266

[3] IwataM., YoshikawaT., BabaA., et al.. Autoantibodies against the second extracellular loop of beta1-adrenerrgic receptors predict ventricular tachycardia and sudden death in patients with idiopathic dilated cardiomyopathy. J Am Coll Cardiol, 2001, 37: 418-424

[4] RatcliffeN. R., HutchinsJ., BarryB., et al.. Chronic myocar-ditis induced by T cells reactive to a single cardiac my-osin peptide: persistent inflammation, cardiac dilatation, myocardial scarring and continuous myocyte apoptosis. J Autoimmun, 2000, 15: 359-367

[5] BozkurtA., CanatarogluA., CetinerS., et al.. Lymphocyte subsets in patients with idiopathic dilated cardio-myopathy. Anadolu Kardiyol Derg, 2001, 1: 98-100

[6] MozaffariF., HanssonL., KiaiiS., et al.. Signaling molecules and cytokine production in T cells of multi-ple myeloma increased abnormalities with advancing stage. Br J Haematol, 2004, 124: 315-324

[7] FuM., MatsuiS.. Is cardiomyopathy an autoimmune disease?. Keio J Med, 2002, 51: 208-212

[8] LiaoY. H., FuM.. Autoimmunity in the pathogenesis of cardiomyopathy. J Autoimmun, 2000, 16: 1-2

[9] FuseK., KodamaM., ItoM., et al.. Polarity of helper T cell subsets represents disease nature and clinical course of experimental autoimmune myocarditis in rats. Clin Exp Immunol, 2003, 134(3): 403-408

[10] HimmelrichH., LaunoisR., MaillardI., et al.. In BALB/c mice, IL-4 production during the initial phase of infection with Leishmania major is necessary and sufficient to instruct Th2 cell development resulting in progressive disease. J Immuol, 2000, 164: 4819-4825

[11] AtanasyevaM., WangY., KayaZ., et al.. Experimental autoimmune myocarditis in A/J mice is an interleukin-4-dependent disease with a Th2 phenotype. Am J Pathol, 2001, 159: 193-203

[12] CunninghamM. W.. Cardiac myosin and the TH1/TH2 paradigm in autoimmune myocarditis. Am J Pathol, 2001, 159: 5-12

[13] WangQ. F., LiaoY. H., GongF. L.. A study of Th cell subsets in patients with dilated cardiomyopathy. J Clin Cardiol (Chinese), 1999, 15: 100-102

[14] StaudtA., BohmM., KnebelF., et al.. Potential role of autoantibodies belonging to the immunogolbulin G-3 subclass in cardiac dysfunction among patients with dilated cardiomyopathy. Circlulation, 2002, 106: 2448-2453

[15] LiaoY. H., YuanJ., WangZ. H., et al.. Infectious tolerance to ADP/ATP carrier peptides induced by anti-L3T4 monoclonal antibody in dilated cardiomyopathy. J Clin Immunol, 2005, 25: 376-384

[16] SchulzeK., BeckerB. F., SchauerR., et al.. Antibodies to ADP-ATP carrier—an autoantigen in myocarditis and dilated cardiomyopathy—impair cardiac function. Circulation, 1990, 81: 959-969

[17] SchwimmbeckP. L., BlandN. K., SchulthesisH. P., et al.. The possible value of synthetic peptides in the diagnosis and therapy of myocarditis and dilated cardiomyopathy. Eur Heart J, 1991, 12(SupplD): 76-80

[18] UlettG. C., KetheesanN., HirstR. G.. Cytokine gene expression in innately susceptible BALB/c mice and relatively resistant C57BL/6 mice during infection with virulent Burkholderia pseudomallei. Infect Immun, 2000, 68(4): 2034-2042

[19] LiaoY. H., ChengL. X., DaiS. P., et al.. Autoantibodies against ADP/ATP carrier from patients with dilated cardiomyopathy increase activity of voltage-dependent Ca channels in isolated cardiac myocytes. Blood Press, 1996, 3(Suppl): 41-44

[20] CaoW., ChenY., AlkanS., et al.. Human T helper (Th) cell lineage commitment is not directly linked to the secretion of IFN-gamma or IL-4: characterization of Th cells isolated by FACS based on IFN-gamma and IL-4 secretion. Eur J Immunol, 2005, 35(9): 2709-17

[21] CaforioA. L., GoldmanJ. H., BaigM. K., et al.. Cardiac autoantibodies in dilated cardiomyopathy become undetectable with disease progression. Heart, 1997, 77(1): 62-77

[22] WarraichR. S., DunnM. J., YacoubM. H.. Subclass specificity of autoantibodies against myosin in patients with idiopathic dilated cardiomyopathy: Pro-inflammatory antibodies in DCM patients. Biochem Biophy Res Commun, 1999, 259: 255-261

[23] WarraichR. S., NoutsiasM., KasacI. K., et al.. Immunoglobulin G3 cardiac myosin autoantibodies correlate with left ventricular dysfunction in patients with dilated cardiomyopathy: Immunoglobulin G3 and clinical correlates. Circulation, 2002, 143: 1076-1084

[24] DeKruyffR. H., JuS. T., MosmannT., et al.. Induction of antigen-specific antibody responses in primed and unprimed B cell: functional heterogeneity among Th1 and Th2 T cell clones. J Immunol, 1989, 142(8): 2575-2582

[25] MolnarI.. The balance shift in Th1/Th2 related IL-12/IL-5 cytokines in Graves’ disease during methimazole therapy. Autoimmunity, 2007, 40(1): 31-37