C Protein-Induced Myocarditis and Subsequent Dilated Cardiomyopathy: Rescue from Death and Prevention of Dilated Cardiomyopathy by Chemokine Receptor DNA Therapy

Abstract

Severe experimental autoimmune myocarditis and subsequent dilated cardiomyopathy (DCM) were successfully produced in Lewis rats by immunization with recombinant cardiac C protein. Seventy-five percent of immunized rats died between days 15 and 49 postimmunization, and all of the survived rats showed typical DCM characterized by the presence of ventricular dilatation and extensive fibrosis. Immunopathological and chemokine analysis during the acute phase revealed that there were marked macrophage infiltration with myocyte necrosis and up-regulation of MCP-1 and IFN-γ-inducible protein-10 (IP-10). Based on these findings, we prepared plasmid DNAs encoding the binding site of CCR2 and CXCR3, which are receptors for MCP-1 and IP-10, respectively. The culture supernatant of cells transfected with these DNAs inhibited the migration of T cells and macrophages induced by MCP-1 and IP-10. Remarkably, administration of the DNAs to C protein-immunized rats prevented the disease progression and rescued animals from death. The present study has demonstrated for the first time that gene therapy targeting the chemokine receptor could be a powerful tool for the control of experimental autoimmune myocarditis and DCM.

Chagas Heart Disease Pathogenesis: One Mechanism or Many?

“…myosin binding protein C (MyBPc), Cha antigen, desmin, actin, myoglobin, tubulin, and the human β1 adrenergic receptor) as a result of T. cruzi infection ( [61][62][63][64] and K. M. Bonney, unpublished results). Observations that autoimmune responses are also initiated against these antigens in susceptible mouse strains immunized with purified cardiac myosin or myosin peptides in various models of experimental autoimmune myocarditis (EAM) support this hypothesis [65,66]. Serological analysis of infected mice also indicates responsiveness to a multitude of other yet unidentified cardiac antigens [51].…”

T-bet Controls Pathogenicity of CTLs in the Heart by Separable Effects on Migration and Effector Activity

“…Studies of Th cell subset differentiation indicate that conditions that favor Th1 differentiation, such as the presence of IL-12 during initial activation of naive CD4 ϩ T cells, also favor the expression of functional selectin ligands (8 -12) and certain chemokine receptors, especially CXCR3 and CCR5 (13). Blockage of CXCR3 prevents disease progression in a model of experimental autoimmune myocarditis (14). It was recently demonstrated that T-bet is required for the expression of CXCR3 and functional P-selectin ligands on CD4 ϩ Th cells in vitro (15).…”

Matrix Metalloproteinase (MMP)-9, but Not MMP-2, Is Involved in the Development and Progression of C Protein-Induced Myocarditis and Subsequent Dilated Cardiomyopathy

“…Experimental autoimmune carditis (EAC) that is induced in Lewis rats by immunization with C protein is a T cell-mediated disease, but various types of autoantibodies are involved for the shift from EAC to DCM (10). In addition, it was shown that several chemokines play an important role in disease progression (9).…”

“…In previous studies, we have demonstrated that cardiac C protein localizes in thick filaments of cardiac muscles (8) and has a strong carditis-inducing ability (9,10). Experimental autoimmune carditis (EAC) that is induced in Lewis rats by immunization with C protein is a T cell-mediated disease, but various types of autoantibodies are involved for the shift from EAC to DCM (10).…”

“…The preparation of recombinant C protein was precisely described previously (9). PCR products corresponding to cardiac C protein fragment 2 (CC2) were inserted into a cloning vector, pCR4 Blunt-TOPO, in the Zero Blunt TOPO Kit (Invitrogen), and clones with correct sequences were subcloned into the pQE30 expression vector (Qiagen).…”