Lipoproteins in Primary Biliary Cirrhosis 1

Abstract

Evidence gradually accumulated by moving boundary electrophoresis (1-4), chemical fractionation, (5-7) using method of Cohn and his associates (8) biliary cirrhotic ACD 2 plasma and 9 ml. of normal ACD2 plasma were dialyzed against several changes of citrate-sodium chloride reagent8 for 48 hours at 0°C. Densities of the dialyzed solutions were adjusted to approximately 1.065 with 4.0 M NaCl and distilled water until a final volume of 11.0 ml. was obtained for the biliary cirrhotic and 13.0 ml. for the normal plasma. Nineml. aliquots of each were placed in 13 X 85 mm. plastic centrifuge tubes and ultracentrifuged at 30,000 rpm (80,000 G) for 18 to 20 hours at 50 C. The top 3.0 cm. of solution were separated with a tube-slicing device. From the upper and lower cuts containing low and high density lipoproteins, respectively, 100 X samples were removed for determination of solution density. The upper cut of the biliary cirrhotic plasma was diluted to 6.0 ml. with 2.3 M NaCl and from the normal plasma with 2.0 M NaCl. An aliquot of 4.5 ml. was removed from each into 13 X 85 mm. plastic tubes for density gradient studies (described presently); the remainder was dialyzed together with both lower cuts (high density lipoproteins) against 0.15 M NaCl for 24 hours at 0°C. The dialyzed solutions were then analyzed for lipids, nitrogen, and dry weight.A linear density gradient as described by LinderstromLang and Lanz (13,14) and Anfinsen (15) was made of the diluted upper cuts as follows: 4.5 ml. 0.1 M NaCl were layered over the 4.5-ml. aliquot of low density lipoproteins that had been previously placed in a 13 X 85 mm. plastic tube. A simply-constructed wire stirrer with a spiral button tip was gently inserted to the interface of the two solutions, from which point gentle vertical strokes of increasing 0.75 cm. distances were made, keeping the plane of the spiral tip as level as possible. The manipulation usually required four strokes each down and up from the interface. The tubes were then ultracentrifuged at 30,000 rpm for 18 to 20 hours at 5°C. Lipoproteins concentrate in such gradients at layers where the lipoprotein density is equal to the density of the sodium chloride solution. The resulting gradients were partitioned into 8 or 9 cuts, 0.75 cm. each, either by slicing with a tube-slicing device or by aspirating with a finetipped pipette. From the volume of each cut (approximately 1.0 ml.) a 100 X sample was removed for determination of solution density. To the remainder of each cut was added an equal volume of 0.15 M NaCl; each 2ACD (acid citrate-dextrose) is the special anticoagulant solution used when fractionating plasma proteins by the Cohn method (8).8 Citrate-sodium chloride reagent consisted of 0.02 M trisodium citrate and 0.07 M sodium chloride.133

Abnormalities of High Density Lipoproteins in Abetalipoproteinemia*

“…Total HDL is absent in Tangier disease (29), absent or reduced in chronic biliary obstruction (30,31), and decreased in liver disease (32), in some patients with the nephrotic syndrome (30,33,34), and in certain hyperlipemias (30,35). In most of these conditions, however, there are accompanying changes in the very low density (D < 1.019) and low density (D 1.019 to 1.063) lipoproteins, with an apparent inverse relationship be-tween very low density and high density lipoprotein concentrations (36,37).…”

Formation of lipoprotein-X. Its relationship to bile compounds.

“…Received for publication 19 May 1975 and in revised form 29 December 1975. designated "lipoprotein X" (LP-X)1 (1)(2)(3)(4)(5)(6)(7)(8)(9). One important and characteristic feature of LP-X is its mobility towards the cathode on agar gel electrophoresis.…”

The Lipid Composition of Serum in Patients With Liver Disease*

“…filter paper electrophoresis ( 16,17) and ultracentrifugation (17)(18)(19). Abnormalities in these lipoprotein fractions, moreover, have been demonstrated by both chemical ( 15,17,19) and immunological means (20). By ultracentrifugal analysis, Havel, Eder and Bragdon (19) In the present investigation, a chromatographic technique for the estimation of the main individual phospholipids of serum (21,22) was employed in order to characterize further the serum lipid abnormalities of patients with liver disease.…”

“…Marked increases in the Sf6 and Sf8 with varying increases in the Sf10-17 ultracentrifugal classes of lipoproteins have been reported in patients with biliary cirrhosis (13). In patients with hepatitis, increases in the SA12, [12][13][14][15][16][17][18][19][20], and 20-100 classes, which correlated positively with the icteric index, and decreases in the Sf00-400 class, which correlated negatively with the icteric index, have been found (14). Increased concentrations of /3-lipoprotein and decreased-to-absent concentrations of a-lipoprotein have been observed with zone electrophoresis of serum from patients with jaundice (15).…”