Volume 156, Issue 1, Pages 185-192 (May 2001) Anti-cholesterol antibodies (ACHA) in patients with different atherosclerotic vascular diseases and healthy individuals. Characterization of human ACHA  Anna Horváth, George Füst, István Horváth, Gábor Vallus, Jenö Duba, Péter Harcos, Zoltán Prohászka, Éva Rajnavölgyi, Lı́via Jánoskuti, Margit Kovács, Albert Császár, László Romics, István Karádi  Atherosclerosis  Volume 156, Issue 1, Pages 185-192 (May 2001) DOI: 10.1016/S0021-9150(00)00630-4

Fig. 1 Binding of cholesterol-specific antibodies to solid phase cholesteol. Cholesterol-coated plates were incubated with different dilutions of IVIG (Sandoglobulin) and with NHS (Immunoglobulin concentration) in triplicates. Optical density measured at 492 nm. Background binding of IgG to uncoated plates was subtracted. Mean±S.D. of OD values are shown. Representative of five experiments with identical results. Atherosclerosis 2001 156, 185-192DOI: (10.1016/S0021-9150(00)00630-4)

Fig. 2 Inhibition of ACHA with lipoprotein preparations. Inhibition of the ACHA binding to the solid phase cholesterol by different concentrations of a human LDL preparation (A) and VLDL/LDL preparation (B). IVIG at concentrations of 1 mg/ml was mixed with different concentrations of a purified human LDL preparation (A), or a preparation containing VLDL and LDL (B) and with 0.1% casein as control and incubated at 37°C for 60 min. Different dilutions were made and the IgG binding to solid phase cholesterol was measured by HRPO-conjugated anti-human IgG and OPD substrate. Background binding of IgG to uncoated plates was subtracted. Mean±S.D. of OD values are shown. Atherosclerosis 2001 156, 185-192DOI: (10.1016/S0021-9150(00)00630-4)

Fig. 3 Study of the binding of ACHA to different sterols. ELISA plates were coated with 10 μg/ml cholesterol, coprostane and ergosterol (provitamine D2). The plates were incubated at 37°C for 60 min with different dilutions of IVIG and IgG binding was measured by HRPO-conjugated anti-human IgG and OPD substrate. Background binding of IgG to uncoated plates was subtracted. Mean±S.D. of OD values are shown. Atherosclerosis 2001 156, 185-192DOI: (10.1016/S0021-9150(00)00630-4)

Fig. 4 Study of IgG subclass distribution of ACHA in individual human sera. ELISA plates were coated with cholesterol as described in Section 2. The plates were incubated with serum samples from ten healthy individuals and ten CHD patients at a dilution of 1 to 100. After incubation and washing the plates were treated with antibodies to γ1, γ2, γ3, and γ4 heavy chain. Background binding to uncoated plates was subtracted. Data obtained as optical density values were expressed in arbitrary units per milliliter (AU/ml) related to the standard curve. Atherosclerosis 2001 156, 185-192DOI: (10.1016/S0021-9150(00)00630-4)

Fig. 5 Binding of IgG to solid phase cholesterol from different dilutions of pooled sera of healthy control and CHD patients. ELISA plates were coated with cholesterol were incubated with the pooled sera of 10 healthy individuals sera (□) and 10 CHD patients’ pooled sera (■), and the binding of IgG ACHAs were measured by specific antibodies. ACHA levels are presented as mean±S.D. Experiment was repeated three times. Atherosclerosis 2001 156, 185-192DOI: (10.1016/S0021-9150(00)00630-4)