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J Am Coll Cardiol, 2007; 49:1684-1692, doi:10.1016/j.jacc.2006.11.051 (Published online 5 April 2007).
© 2007 by the American College of Cardiology Foundation
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Fc{gamma} Receptors IIa on Cardiomyocytes and Their Potential Functional Relevance in Dilated Cardiomyopathy

Alexander Staudt, MD*, Petra Eichler, PhD{dagger}, Christiane Trimpert, Msc*, Stephan B. Felix, MD*,* and Andreas Greinacher, MD{dagger}

* Klinik für Innere Medizin B, Ernst-Moritz-Arndt-Universität, Greifswald, Germany
{dagger} Institut für Immunologie und Transfusionsmedizin, Ernst-Moritz-Arndt-Universität, Greifswald, Germany


Figure 1
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  		Figure 1 Effects of Intact IgG/Antibody Fragments on Isolated, Field-Stimulated Rat Cardiomyocytes

Changes of calcium transients and systolic cell shortening during superfusion (% changes from baseline; mean ± SEM; n = 6 experiments for each patient's/control sample) with: intact immunoglobulin (Ig)G from plasma of healthy blood donors (control IgG) and intact IgG from dilated cardiomyopathy (DCM) patients (DCM-IgG); F(ab')2 fragments of control IgG and of respective DCM-IgG; intact DCM-IgG after pre-incubation with the F(ab')2 fragments of control subjects and of respective DCM patients; intact DCM-IgG after pre-incubation of cardiomyocytes with human Fc fragments; and goat-anti-human-F(ab')-IgG after pre-incubation of cardiomyocytes with F(ab')2 fragments of control IgG and of respective DCM patients. +++p < 0.001 significantly different from control IgG; ***p < 0.001 significantly different from DCM-IgG.

 

Figure 2
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  		Figure 2 Summary of Functional Findings

Intact immunoglobulin (Ig)G of dilated cardiomyopathy (DCM) patients induced negative inotropic effects by binding via their Fab part to the antigenic epitope on cardiomyocytes and then via their Fc part to the Fc{gamma} receptor IIa (Fc{gamma}R) (A). The F(ab')2 fragments of these antibodies inhibited the effect of intact DCM IgG (B), as did Fc fragments of normal IgG (C). Reconstitution of Fc parts by sequential incubation of cardiomyocytes with DCM F(ab')2 fragments and goat-anti-human F(ab') IgG induced a negative inotropic effect (D) comparable to findings for intact DCM IgG. A detailed description of Fc receptor cross linking is in the Discussion section. AG = antigen.

 

Figure 3
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  		Figure 3 Fc{gamma} Receptor on Rat Cardiomyocytes

Immunofluorescent staining of isolated rat cardiomyocytes with a rhodamine-conjugated bovine anti-goat secondary antibody after pre-incubation with polyclonal goat primary antibodies against Fc{gamma} receptor I (CD64), Fc{gamma} receptor II (CD32), or Fc{gamma} receptor III (CD16); goat immunoglobulin (Ig)G and antibodies against troponin I, serving as negative and positive control subjects, respectively (counterstaining with 4',6-Diamidino-2-phenyindole, dilactate [DAPI]; representative figures of n = 6 experiments for each antibody).

 

Figure 4
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  		Figure 4 Fc{gamma} Receptor Type IIa on Rat Cardiomyocytes

Immunofluorescent staining of isolated rat cardiomyocytes with a rhodamine-conjugated bovine anti-goat secondary antibody after preincubation with polyclonal goat primary antibodies against Fc{gamma} receptor IIa, Fc{gamma} receptor IIb, or Fc{gamma} receptor IIb/c. Immunofluorescence of fibroblasts and rat leukocytes with a rhodamine-conjugated bovine anti-goat secondary antibody after preincubation with polyclonal goat primary antibodies against Fc{gamma} receptor IIa served as negative and positive control subjects, respectively (counterstaining with 4',6-Diamidino-2-phenyindole, dilactate [DAPI]; representative figures of n = 6 experiments for each antibody).

 

Figure 5
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  		Figure 5 Fc{gamma} Receptor Type IIa on Human Cardiomyocytes

Immunofluorescent staining of human cardiomyocytes isolated from endomyocardial biopsies with a rhodamine-conjugated bovine anti-goat secondary antibody after preincubation with polyclonal goat primary antibodies against Fc{gamma} receptor IIa, Fc{gamma} receptor IIb, or Fc{gamma} receptor IIc; goat immunoglobulin G and antibodies against troponin I, serving as negative and positive control subjects, respectively (representative figures of n = 6 experiments for each antibody).

 




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Copyright © 2007 by the American College of Cardiology Foundation.