J. Lipid Res.  Neurobiology of Lipids (ISSN1683-5506)
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Journal of Lipid Research, Vol. 42, 639-648, April 2001
Copyright © 2001 by Lipid Research, Inc.


Original Article

Composition and ultrastructure of size subclasses of normal human peripheral lymph lipoproteins: quantification of cholesterol uptake by HDL in tissue fluids

M. N. Nanjeea, C. J. Cookea, J. S. Wongb, R. L. Hamiltonb, W. L. Olszewskia,c, and N. E. Millera
a Department of Cardiovascular Biochemistry, St. Bartholomew's and the Royal London School of Medicine and Dentistry, Charterhouse Square, London EC1M 6BQ, UK
b Cardiovascular Research Institute and Department of Anatomy, University of California, San Francisco, CA 94143
c Department of Surgical Research and Transplantology, Medical Research Center, Polish Academy of Sciences 02-106, Warsaw, Pawinskiego, Poland

Correspondence to: N. E. Miller, To whom correspondence should be addressed., n.e.miller{at}mds.qmw.ac.uk (E-mail)

Peripheral lymph lipoproteins have been characterized in animals, but there is little information about their composition, and none about their ultrastructure, in normal humans. Therefore, we collected afferent leg lymph from 16 healthy males and quantified lipids and apolipoproteins in fractions separated by high performance-size exclusion chromatography. Apolipoprotein B (apoB) was found almost exclusively in low density lipoproteins. The distribution of apoA-I, particularly in lipoprotein A-I (LpA-I) without A-II particles, was shifted toward larger particles relative to plasma. The fractions containing these particles were also enriched in apoA-II, apoE, total cholesterol, and phospholipids and had greater unesterified cholesterol-to-cholesteryl ester ratios than their counterparts in plasma. Fractions containing smaller apoA-I particles were enriched in phospholipid. Most apoA-IV was lipid poor or lipid free. Most apoC-III coeluted with large apoA-I-containing particles. Electron microscopy showed that lymph contained discoidal particles not seen in plasma.

These findings support other evidence that high density lipoproteins (HDL) undergo extensive remodeling in human tissue fluid. Total cholesterol concentration in lymph HDL was 30% greater (P < 0.05) than could be explained by the transendothelial transfer of HDL from plasma, providing direct confirmation that HDL acquire cholesterol in the extravascular compartment. Net transport rates of new HDL cholesterol in the cannulated vessels corresponded to a mean whole body reverse cholesterol transport rate via lymph of 0.89 mmol (344 mg)/day. Nanjee, M. N., C. J. Cooke, J. S. Wong, R. L. Hamilton, W. L. Olszewski, and N. E. Miller. Composition and ultrastructure of size subclasses of normal human peripheral lymph lipoproteins: quantification of cholesterol uptake by HDL in tissue fluids. J. Lipid Res. 2001. 42: 639;–648.

Supplementary key words: tissue fluid, apolipoproteins, phospholipids, electron microscopy


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