Atherosclerosis Atherosclerosis, arteriosclerotic vascular disease (ASVD) Atherosclerosis is an inflammatory disease characterized by endothelial activation and dysfunction, lipid accumulation, monocyte infiltration and differentiation, T-cell infiltration and activation, foam cell formation, and fibrosis in the lesion area. Two of its common final results, coronary and cerebrovascular disease, are the major causes of morbidity worldwide. Nowadays, atherosclerosis is increasingly considered an immune-mediated inflammatory process of the vasculature in which intense immunological activity is occurring. Hypertension, hyperlipidemia, hyperglycemia, diabetes, obesity, cigarette smoking, age, and sex are well-established risk factors for atherosclerosis (Jan et al., 2010). Baboon Papio cynocephalus 9556 Bank vole Clethrionomys glareolus 447135 Bear Ursus americanus 9643 Birds Passeroidea 175121 Brown Trout Salmo trutta 8032 Buffalo Bison bison 9901 Carnivores Vulpes 9625 Cat Felis catus 9685 Catfishes Siluriformes 7995 Cattle Bos taurus 9913 Chicken Gallus gallus 9031 Chimpanzee Pan troglodytes 9598 chinchillas Chinchillidae 10150 Copper Pheasant Syrmaticus soemmerringii 9067 Deer Cervus elaphus 9860 Deer mouse Peromyscus maniculatus 10042 Dog Canis familiaris 9615 Ducks Anas 8835 Ferret Mustela putorius furo 9669 Fish Hyperotreti 117565 Gerbil Gerbillina 10045 Goat Capra hircus 9925 Gray wolf Canis lupus 9612 Guinea pig Cavia porcellus 10141 Hamster Mesocricetus auratus 10036 Horse Equus caballus 9796 Human Homo sapiens 9606 Macaque Macaca fascicularis 9541 Mongolian Gerbil Meriones unguiculatus 10047 Monkey Platyrrhini 9479 Mouse Mus musculus 10090 None None Parrot Psittacidae 9224 Pig Sus scrofa 9823 Rabbit Oryctolagus cuniculus 9986 Rainbow trout Oncorhynchus mykiss 8022 Rat Rattus 10114 Raven Corvus corax 56781 sei whale Balaenoptera borealis 9768 Sheep Ovis aries 9940 Squirrel Spermophilus richardsonii 37591 Tree shrew Tupaiidae 9393 Trouts, salmons & chars Salmoninae 504568 Turkey Meleagris gallopavo 9103 Vole Microtus ochrogaster 79684 Water buffalo Bubalus bubalis 391902 human apo B-100 peptide-2 protein vaccine VO_0004275 Subunit vaccine Research subcutaneous injection Peptide antigen was prepared using Imject SuperCarrier EDC kit (Pierce, Rockford, IL) according to manufacturer’s instruction with minor modification. One milligram peptide in 500 μl conjugation buffer was mixed with 2 mg carrier in 200 μl deionized water. This mixture was then incubated with 1 mg conjugation reagent (EDC, 1-ethyl-3-[3-dimethylaminopropyl]-carbodiimide HCl) at room temperature for 2 h followed by dialysis against 0.083 M sodium phosphate, 0.9 M sodium chloride, pH 7.2, solution overnight at 4 °C. The dialyzed conjugate was diluted with Imject dry blend purification buffer to a final volume of 1.5 ml. Alum was used as immunoadjuvant and mixed with peptide conjugate with 1:1 dilution in volume. The amount of peptide in each immunization was 33 μg/100 μl per injection (Chyu et al., 2005). subcutaneous injection Peptide-2, a peptide taken from the sequence of human apo B-100, with an amino acid sequence of ATRFKHLRKYTYNYEAESSS (Chyu et al., 2005). male apo E (−/−) mice Peptide immunization For early immunization study, male apo E (−/−) mice, maintained on normal chow diet, received subcutaneous primary immunization with peptide-1 (n = 10), peptide-2 (n = 10) or alum alone (n = 9) at 6–7 weeks of age, followed by an intra-peritoneal booster 3 weeks later. Two weeks after booster, diet was switched to high cholesterol chow (0.15% cholesterol, 21% fat, TD 88137, Harlan-Teklad) and continued until sacrifice at the age of 25 weeks. Blood samples were collected by retro-orbital bleeding 2 weeks after booster and at the time of sacrifice. For late immunization study, male apo E (−/−) mice were kept on high cholesterol diet since 6–7 weeks of age. Since peptide-1 immunization did not reduce atherosclerosis (see Result below), peptide-1 was omitted in this part of the study. At 16 weeks of age, mice received subcutaneous primary immunization with peptide-2 or alum alone, followed by an intra-peritoneal booster 3 weeks later. Mice were sacrificed at age of 30 weeks. A separate group of mice were kept on high cholesterol diet and sacrificed at 16 weeks of age to serve as baseline control (Chyu et al., 2005). Peptide-2 immunization reduced aortic atherosclerosis by 40% and plaque inflammation by 80% compared to controls without a reduction in circulating cholesterol levels whereas Peptide-1 immunization had no effect. Peptide-2 immunization also reduced the progression of aortic lesions when mice were immunized at 16 weeks of age, suggesting the possibility of immuno-modulation in treating established atherosclerosis (Chyu et al., 2005). MDA-modified human apo B-100 peptide Vaccine VO_0004276 Subunit vaccine Research Intramuscular injection (i.m.) Intramuscular injection (i.m.) MDA-modified apo B-100 (Fredrikson et al., 2005). Male apo E0 mice on C57BL/6 background Mice (n = 8–10 per group) were given a first injection with a MDA modified peptide conjugated to the carrier or the carrier alone at 6 weeks of age and booster injections 3 and 5 weeks later. Each injection contained 50 mg of the MDA-modified peptide conjugated to 50mg of the carrier cBSA (cationized bovine serum albumin) dissolved in 0.083M sodium phosphate 0.9M NaCl pH 7.2, according to the manufacture’s protocol (No. 77652, Pierce, Rockford, IL, USA), and with Alum (aluminum hydroxide, Pierce) included in all injections as adjuvant. The mice were fed a cholesterol diet (0.15% cholesterol, 21% fat; Lactamin AB, Kimstad, Sweden) from the age of 10 weeks (Fredrikson et al., 2005). Immunization with fragment P45 reduced atherosclerosis with 48%; fragment P74with 31; whereas immunization with P240 had no effect (Fredrikson et al., 2005). Chyu et al., 2005 journal Chyu KY, Zhao X, Reyes OS, Babbidge SM, Dimayuga PC, Yano J, Cercek B, Fredrikson GN, Nilsson J, Shah PK 2005 338 4 1982-1989 Biochemical and biophysical research communications Fredrikson et al., 2005 journal Fredrikson GN, Andersson L, Söderberg I, Dimayuga P, Chyu KY, Shah PK, Nilsson J 2005 38 2 171-179 Autoimmunity Jan et al., 2010 journal Jan M, Meng S, Chen NC, Mai J, Wang H, Yang XF 2010 2010 459798 Journal of biomedicine & biotechnology