From this cohort, we have previously determined that high levels of anti-MDA-LDL, anti-OxLDL and anti-PC IgM (but not IgG) are SCR7 price protection factors for atherosclerosis development [21]. In the present study we explore the roles of anti-PC IgG1, IgG2 and IgA, different antibody idiotypes as well as the long term stability of anti-PC IgM levels. We have also studied combinations of anti-PC with anti-MDA-LDL or anti-oxLDL. Serum samples were obtained prior to enrollment and again at follow-up from 226 individuals with established hypertension (diastolic pressure >95 mm Hg) who participated in

the Swedish component of the European Lacidipine Study on Atherosclerosis (ELSA). During the admission process, information on age, gender, blood pressure, weight, height, smoking habits and previous medical history were recorded along with laboratory values of different parameters including creatinine, fasting glucose, total

cholesterol, high-density lipoprotein (HDL) cholesterol and triglycerides. For detailed information on ELSA, please refer to the original article [25]. The study was approved by the Ethics Committee of the Karolinska Hospital and was conducted in accordance with the Helsinki Declaration. All subjects gave informed consent. The mean of the maximum Intima-Media Thicknesses (IMT) in the far walls of common carotids and bifurcations (CBMmax) was determined by B-mode ultrasonography at the time of inclusion, and 4 years afterwards. All scans were performed with the Biosound 200 II device (All Imaging Systems inc., Dolutegravir in vivo Irvine CA, USA) and read at the Ultrasound Coordinating Center with quality assurance accomplished as reported. The levels of the different anti-PC antibody classes/subclasses at enrollment were evaluated with respect to increase or decrease

in IMT at the 4-year follow up. Pooled C-X-C chemokine receptor type 7 (CXCR-7) human IgG (Baxter, Deerfield IL, USA) and IgM (Sigma Aldrich, St. Louis MO, USA) was passed over PC-Sepharose columns (Biosearch technologies, Novato CA, USA). The columns were then washed with Phosphate Buffered Saline (PBS) pH 7.4 with Tween20 to remove non-bound immunoglobulins. These non-anti-PC antibodies were collected to be used as control antibodies (flowthrough immunoglobulins) in later experiments. The bound PC-specific antibodies were then eluted with 0.01 M acetic acid and concentrated/buffer exchanged to PBS pH 7.4 using Centricon Plus-70 centrifugation filter units (Millipore, Billerica MA, USA). Detection of IgM anti-PC antibodies was performed with an enzyme linked immunoassay (ELISA) using Athera CVDefineTM kit (Athera Biotechnologies AB, Stockholm, Sweden). The kit is based on PC covalently linked to bovine serum albumin (PC-BSA) coated onto 96-well Nunc Maxisorp micro-titer plates. The assay was carried out in accordance with the manufacturer’s recommendations.