Polymorphism of class A scavenger receptors in C57BL/6 mice.
Scavenger receptors class A (SR-A) have been hypothesized to regulate the development of atherosclerotic lesions through recognition of modified low density lipoprotein (LDL) and macrophage adhesion to substrata. Supporting data have been collected from studies using the monoclonal antibody 2F8, an antibody developed from the BALB/c strain-derived macrophage cell line, RAW.264. Although 2F8 immunostained both cultured peritoneal macrophages (MPM) and thymic macrophages from Swiss, BALB/c, and DBA/2 mice, no immunostaining was detected in cells and tissues from C57BL/6 mice, one of the most commonly used atherosclerosis-susceptible mouse strains. Similarly, 2F8 detected SR-A protein in MPM by Western blotting in all strains except C57BL/6. However, a guinea pig antiserum developed to a fusion protein of the extracellular SR-A domain detected appropriately sized bands in all strains. Incubation with 2F8 antagonized acetylated low-density lipoprotein (AcLDL)-induced cholesterol esterification in MPM from BALB/c, Swiss, and DBA/2 strains but had no effect on MPM from C57BL/6 mice. Sequencing of SR-A cDNA from C57BL/6 mice demonstrated complete identity with published sequence in the collagen-like domain. However, four single-residue substitutions were noted in the alpha-helical coiled-coil domain. Site-directed mutagenesis demonstrated that a single substitution (L168S) in this domain accounted for the loss of 2F8 immunoreactivity. Differing reactivities toward a commonly used monoclonal antibody were used to identify polymorphism of SR-A in C57BL/6 mice.
Daugherty
A
A
Gill Heart Institute Atherosclerosis Research Group, Division of Cardiovascular Medicine, University of Kentucky, Lexington, KY 40536, USA. adaugh@pop.uky.edu
Whitman
S C
SC
Block
A E
AE
Rateri
D L
DL
D000818:Animals; D000911:Antibodies, Monoclonal; D001057:Apolipoproteins E / Q000235:genetics; D001161:Arteriosclerosis / Q000209:etiology; D001483:Base Sequence; D004195:Disease Models, Animal; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D005188:False Negative Reactions; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL / Q000235:genetics*; D008811:Mice, Inbred DBA; D018345:Mice, Knockout / Q000235:genetics; D008969:Molecular Sequence Data; D011110:Polymorphism, Genetic; D011971:Receptors, Immunologic / Q000235:genetics* / Q000276:immunology / Q000502:physiology; D051116:Receptors, Scavenger; D015203:Reproducibility of Results; D051117:Scavenger Receptors, Class A; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D000911:Antibodies, Monoclonal; D001057:Apolipoproteins E; D000939:Epitopes; C495140:Msr1 protein, mouse; D011971:Receptors, Immunologic; D051116:Receptors, Scavenger; D051117:Scavenger Receptors, Class A