Expression and coreceptor function of APJ for primate immunodeficiency viruses.
APJ is a seven transmembrane domain G-protein-coupled receptor that functions as a coreceptor for some primate immunodeficiency virus strains. The in vivo significance of APJ coreceptor function remains to be elucidated, however, due to the lack of an antibody that can be used to assess APJ expression, and because of the absence of an antibody or ligand that can block APJ coreceptor activity. Therefore, we produced a specific monoclonal antibody (MAb 856) to APJ and found that it detected this receptor in FACS, immunofluorescence, and immunohistochemistry studies. MAb 856 also recognized APJ by Western blot, enabling us to determine that APJ is N-glycosylated. Using this antibody, we correlated APJ expression with coreceptor activity and found that APJ had coreceptor function even at low levels of expression. However, we found that APJ could not be detected by FACS analysis on cell lines commonly used to propagate primate lentiviruses, nor was it expressed on human PBMC cultured under a variety of conditions. We also found that some viral envelope proteins could mediate fusion with APJ-positive, CD4-negative cells, provided that CD4 was added in trans. These findings indicate that in some situations APJ use could render primary cell types susceptible to virus infection, although we have not found any evidence that this occurs. Finally, the peptide ligand for APJ, apelin-13, efficiently blocked APJ coreceptor activity.
Puffer
B A
BA
Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, 19104, USA.
Sharron
M
M
Coughlan
C M
CM
Baribaud
F
F
McManus
C M
CM
Lee
B
B
David
J
J
Price
K
K
Horuk
R
R
Tsang
M
M
Doms
R W
RW
D054392:Adipokines; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000073861:Apelin; D000075244:Apelin Receptors; D002352:Carrier Proteins / Q000378:metabolism; D002460:Cell Line; D065127:Dopamine D2 Receptor Antagonists; D015497:HIV-1 / Q000378:metabolism; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D007963:Leukocytes, Mononuclear; D051379:Mice; D008807:Mice, Inbred BALB C; D011323:Primates; D017448:Receptors, Dopamine D2 / Q000096:biosynthesis / Q000276:immunology / Q000502:physiology*; D043562:Receptors, G-Protein-Coupled; D015728:Receptors, HIV / Q000096:biosynthesis / Q000276:immunology / Q000502:physiology*; D011991:Receptors, Virus / Q000096:biosynthesis / Q000276:immunology / Q000502:physiology; D015302:Simian Immunodeficiency Virus / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology; D014162:Transfection
C115436:APLN protein, human; C104575:APLNR protein, human; D054392:Adipokines; D000911:Antibodies, Monoclonal; D000073861:Apelin; D000075244:Apelin Receptors; C402826:Apln protein, mouse; C491285:Aplnr protein, mouse; D002352:Carrier Proteins; D065127:Dopamine D2 Receptor Antagonists; D036341:Intercellular Signaling Peptides and Proteins; D017448:Receptors, Dopamine D2; D043562:Receptors, G-Protein-Coupled; D015728:Receptors, HIV; D011991:Receptors, Virus