Monoclonal anti-idiotypic antibodies regulate the expression of virus-induced murine myocarditis

Abstract

A monoclonal anti-idiotypic antibody (anti-Id), produced by electrofusion and designated anti-1d88, was able to modulate expression of murine autoimmune myocarditis mediated by coxsackievirus B3 (CVB3). The anti-Id was characterized as an immunoglobulin G2b species possessing K light chains and was able to reduce expression of inflammatory myocarditis in anti-Id-pretreated mice challenged with CVB3. Anti-Id88 was able to stimulate specific cell-mediated immunity against anti-ld88, as well as CVB3, and exerted a suppressive effect on the proliferation of mixed spleen cell populations from virus-exposed mice. Anti-Id stimulated an anti-anti-Id antibody 3 population able to bind antibody 2 F(ab')2 fragments or virus antigen in an indirect enzyme-linked immunosorbent assay. Western blot (immunoblot) analysis of anti-Id88 exhibited binding of syngeneic anti-Id antibody to idiotypes present on immunoglobulin G molecules from virus-immunized mice.

Adoptively transferred anti-idiotype pulsed B cells mediate autoimmune myocarditis

“…with 108 PFU of lethally UV-irradiated CVB3 virus in Freund's complete adjuvant, followed by weekly i.p. injections for at least 6 to 8 weeks of 107 PFU of virus (20)(21)(22)(23)(24). Syngeneic BALB/cJ donors of mouse anti-Ids were immunized subcutaneously with 50 ,ug of affinity column-purified, protein A-separated, anti-CVB3-purified IgG emulsified with Freund's complete adjuvant.…”

“…Serum was separated by utilizing Microtainer capillary blood separator tubes (Becton Dickinson, Rutherford, N.J.). ADOPTIVELY TRANSFERRED ANTI-ID PULSED B CELLS 3397 Monoclonal antibody used in these studies was prepared from a murine hybridoma developed and characterized as described in an earlier publication (22) and is designated as anti-Id88 (22). Anti-Id88, in terms of specificity, binds F(ab')2 of anti-CVB3 polyclonal antibodies, but not normal F(ab')2 fragments or F(ab')2 fragments from murine anti-DNP antibodies.…”

“…Previous studies from our laboratory have suggested the participation of anti-idiotypic antibodies (anti-Ids) in the induction and expression of autoimmune myocarditis induced by the enterovirus coxsackievirus B3 (CVB3) in a murine model (20). The array of immunobiological effects of polyclonal or monoclonal anti-Ids has included inhibition of myocarditis by pretreatment and/or immunization with anti-Ids in susceptible hosts (20), suppression of in vitro T-cell or splenocyte proliferative responses (22), specific modulation of macrophage chemotaxis in vitro (21), release of the lymphokine migration inhibition factor from sensitized T cells (19), and ability to bind selectively to myocyte target cell molecules and/or mimic solubilized heart-associated antigens (23). Anti-Ids were also demonstrated to induce specific, antiviral T-cell immunity (20,22,23) and stimulate secondary B-cell responses in viral or anti-Id-primed animals (20,22).…”

Polyclonal anti-idiotypic antibodies exhibit antigenic mimicry of limited type 1 fimbrial proteins of Escherichia coli

“…2,4-Dinitrophenol (DNP)ovalbumin was prepared by conjugation of 0.2 ml of dinitrofluorobenzene (DNFB) to 500 mg of ovalbumin (Sigma Chemical Co., St. Louis, Mo.) as described earlier (22,23). Briefly, ovalbumin was dissolved in 100 ml of 0.85 M saline with DNFB by stirring at 25°C for 30 min.…”

“…Fab preparation. F(ab')2 fragments of Ab-1 or anti-Ids (Ab-2) were prepared by methods described earlier (22,23). Briefly, serum IgG (3.0 mg) was dissolved in 0.14 M sodium acetate-sodium chloride buffer with 60 mg of pepsin at pH 4.0.…”