A monoclonal antibody, HCL-2, raised against human luteal cells reacts with apolipoprotein-B and detects the uptake of low density lipoprotein by luteinizing granulosa cells.
A monoclonal antibody, HCL-2, was raised by immunizing mice against human luteal cells. HCL-2 reacted with luteal cells and villous trophoblasts. The sodium dodecyl sulphate-polyacrylamide gel electrophoresis profile of immunopurified antigens from corpus luteum, chorionic villi, and placenta showed the same main protein band, the molecular mass of which is >200 kDa. The sequence of a portion of the N-terminal region of the antigenic protein purified from placenta was identical to that of apolipoprotein-B. The antigen purified from human serum and low density lipoprotein (LDL) using HCL-2 showed the same protein band as that from corpus luteum. Furthermore, the amino acid sequence (20 amino acids) of the protein purified from serum was also identical to that of apolipoprotein-B. Thus, we concluded that HCL-2 antigen is apolipoprotein-B. Human luteinizing granulosa cells isolated from the patients undergoing in-vitro fertilization treatment were cultured in the medium containing lipoprotein-deficient serum with or without supplementation of LDL. Using HCL-2, apolipoprotein-B was immunocytochemically detected on granulosa cells only in the presence of LDL. These findings showed that the uptake of LDL by granulosa cells was detected by immunocytochemical staining of apolipoprotein-B, indicating that HCL-2 is useful for analysing dynamic utilization of LDL by ovarian cells.
Yamada
S
S
Department of Gynecology and Obstetrics, Faculty of Medicine, Kyoto University, Japan.
Fujiwara
H
H
Ueda
M
M
Honda
T
T
Nakayama
T
T
Higuchi
T
T
Mori
T
T
Maeda
M
M
D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001055:Apolipoproteins B / Q000276:immunology* / Q000378:metabolism; D001769:Blood / Q000276:immunology; D002823:Chorion / Q000378:metabolism; D003338:Corpus Luteum / Q000166:cytology / Q000276:immunology* / Q000502:physiology*; D005260:Female; D006107:Granulosa Cells / Q000378:metabolism*; D006801:Humans; D007743:Labor, Obstetric / Q000378:metabolism; D008077:Lipoproteins, LDL / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D010053:Ovary / Q000378:metabolism; D011247:Pregnancy; D011261:Pregnancy Trimester, First / Q000378:metabolism; D011374:Progesterone / Q000378:metabolism
D000911:Antibodies, Monoclonal; D001055:Apolipoproteins B; D008077:Lipoproteins, LDL; D011374:Progesterone