Measurment of rhesus monkey (Macaca mulatta) apolipoprotein B in serum by radioimmunoassay: comparison of immunoreactivities of rhesus and human low density lipoproteins

Abstract

A sensitive and specific double antibody radio-immunoassay for the major apolipoprotein (apoB) of rhesus (Macaca mulatta) serum very low density lipoprotein (VLDL) and low density lipoprotein (LDL) is described. The anti-serum was raised to LDL (d 1.030–1.040 g/ml) and the LDL2 (d 1.020–1.050 g/ml) was labeled with 125I by the chloramine-T or iodine monochloride method. The assay, which was sensitive to 0.02–0.5 µg of LDL2, had an inter-assay coefficient of variation of 4.5%. This assay was successfully used to measure apoB in the whole serum and low density lipoproteins of control monkeys maintained on a standard Purina monkey chow (PMC) diet and of three groups of monkeys fed atherogenic diets: an “average American diet,” a 25% peanut oil and 2% cholesterol-supplemented PMC diet, and a 25% coconut oil and 2% cholesterol-supplemented PMC diet. The control monkeys (n = 13) had a serum cholesterol of 146 ± 28 mg/dl and an apoB of 50 ± 18 mg/dl. In the monkeys maintained on the atherogenic diets the serum apoB was elevated: 103 ± 28 mg/dl (American), 102 ± 35 mg/dl (peanut oil), and 312 ± 88 mg/dl (coconut oil). The values for serum total cholesterol were 333 ± 65 mg/dl (American), 606 ± 212 mg/dl (peanut oil), and 864 ± 233 mg/dl (coconut oil) and were elevated relative to controls (P < 0.001). For each of the diets, total serum cholesterol correlated with serum apoB (P < 0.001). The slopes of the regression lines of serum apoB vs. cholesterol for the monkeys on the PMC, American, and coconut oil diets were similar (m = 0.531, 0.401, and 0.359, respectively), but differed from that of monkeys on the peanut oil diet (m = 0.121). The immunoreactivities of rhesus and human LDL were compared using specific antisera raised against these antigens. In homologous assay systems, monkey and human LDL exhibited unique immunological determinants. The same results were obtained with the delipidated preparations of the two LDLs using antisera raised against either monkey or human apoB. Crossover studies using a heterologous tracer with each anti-serum resulted in the selection of a specific population of antibodies directed against antigenic sites shared by these two LDL species.

Keywords

• low density lipoprotein radioimmunoassay
• atherosclerosis
• hypercholesterolemia