Cloning of rat HIV-1-chemokine coreceptor CKR5 from microglia and upregulation of its mRNA in ischemic and endotoxinemic rat brain.
Chemokine receptors play a crucial role in the recruitment of immune cells to sites of inflammation. Although chronic diseases of the brain are often accompanied by inflammatory events, there is presently no information about the occurrence and regulation of these receptors in the central nervous system (CNS). Moreover, one CC-chemokine receptor, CKR5, has recently been identified as coreceptor for HIV-1 entry into macrophages. HIV-1 target cells in brain are macrophage-related microglia, which suggests that they are infected by the same mechanism (He et al.,: Nature 385:645-649, 1997). Although rats are not susceptible to HIV-1 infection, they can be used to study chemokine receptor regulation in a variety of brain pathologies. After cloning CC-CKR5 and establishing reverse transcriptase polymerase chain reaction (RT-PCR) for its ligands macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and regulated on activation, normal T cell-expressed and secreted (RANTES), we studied expression of these four mRNAs in purified microglia and compared it with their expression in rat brain. Lipopolysaccharide (LPS)-treated microglia showed transiently increased mRNA levels of both CKR5 and its ligands. Similar data were obtained from brains of LPS-injected rats. In middle cerebral artery occluded (MCAO)-animals, RANTES mRNA was unaffected, whereas CKR5 mRNA showed a sustained rise until 96 hr after surgery. MIPs exogenously added to microglial cultures markedly reduced CKR5 mRNA expression, whereas RANTES did not. MIP mRNAs, in contrast to RANTES and CKR5 mRNAs, were undetectable in normal brain. RANTES appears to play a role distinct from MIPs in brain. In summary, upregulation of CC-chemokines and CKR5 in the CNS upon bacterial infection or in ischemia may impact on microglial activation stage and result in increased risk of HIV-1 infection.
Department of Psychiatry, University of Freiburg, Germany.
O
O
Spleiss
N
N
Gourmala
H W
HW
Boddeke
A
A
Sauter
B L
BL
Fiebich
M
M
Berger
P J
PJ
Gebicke-Haerter
D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000502:physiology; D001483:Base Sequence; D015152:Blotting, Northern; D002545:Brain Ischemia / Q000378:metabolism*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D019446:Endotoxemia / Q000378:metabolism*; D008297:Male; D017628:Microglia / Q000378:metabolism*; D008969:Molecular Sequence Data; D009474:Neurons / Q000502:physiology; D009841:Oligonucleotides / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis*; D051381:Rats; D019713:Receptors, CCR5 / Q000096:biosynthesis / Q000378:metabolism*; D015854:Up-Regulation / Q000502:physiology*
D009841:Oligonucleotides; D012333:RNA, Messenger; D019713:Receptors, CCR5