PMID- 11278864
OWN - NLM
STAT- MEDLINE
DCOM- 20010823
LR  - 20220330
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 276
IP  - 29
DP  - 2001 Jul 20
TI  - Role of phosphoinositide 3-kinase in monocyte recruitment under flow conditions.
PG  - 26846-51
AB  - Chemokines such as the monocyte chemol attractant protein-1 (MCP-1) convert 
      monocyte rolling to firm arrest under physiological flow conditions via integrin 
      activation and simultaneously activate phosphoinositide 3-kinase (PI3K). Here we 
      used adenoviral gene transfer and biochemical inhibitors to manipulate 
      PI3K-dependent pathways in human monocytes. In in vitro lipid kinase assays from 
      purified human monocytes, we showed that MCP-1 activates the "classical" 
      PI3Kalpha pathway and not PI3Kgamma, a PI3K isoform thought to be activated only 
      by the betagamma complex of heterotrimeric G proteins. The activity of PI3Kalpha 
      in purified human monocytes was evident within 30 s. MCP-1-induced monocyte 
      arrest was significantly inhibited both by wortmannin (n = 4; p < 0.01) and 
      LY294002 (n = 4; p < 0.01) with restoration of the rolling phenotype (p < 0.05 
      for both inhibitors, compared with rolling of control monocytes after MCP-1 
      treatment). To test the hypothesis that activation of PI3K is sufficient to 
      induce monocyte adhesion, we transduced the monocytic THP-1 cell line with a 
      recombinant adenovirus (Ad) carrying a constitutively active mutant of PI3K 
      (Ad.BD110). We examined the ability of these cells to adhere to human vascular 
      endothelium (HUVEC) transduced with adenoviruses carrying E-selectin, 
      intercellular adhesion molecule-1 (ICAM-1), and VCAM-1. Under flow conditions, 
      ICAM-1- and VCAM-1-dependent firm adhesion of Ad.BD110-transduced THP-1 cells was 
      enhanced compared with THP-1 cells infected with control Ad (n = 4; p < 0.01 for 
      both). Adhesion augmented by constitutive PI3K activation was entirely abrogated 
      by pretreatment with wortmannin (n = 3; p < 0.01). In contrast, a constitutively 
      active Akt construct had no effect on THP-1 adhesion (n = 3; p = NS). We conclude 
      that PI3K activation is necessary and sufficient to enhance monocytic adhesion 
      under physiological flow conditions. BD110-expressing THP-1 cells should provide 
      a useful tool for identifying the signaling pathways downstream of PI3K that are 
      necessary for monocyte recruitment relevant to a variety of human vascular 
      pathologies.
FAU - Gerszten, R E
AU  - Gerszten RE
AD  - Program in Cardiovascular Gene Therapy, Cardiovascular Research Center and the 
      Center for Immunology and Inflammatory Diseases, Massachusetts General Hospital, 
      Charlestown 02129, USA. gerszten@cvrc.mgh.harvard.edu
FAU - Friedrich, E B
AU  - Friedrich EB
FAU - Matsui, T
AU  - Matsui T
FAU - Hung, R R
AU  - Hung RR
FAU - Li, L
AU  - Li L
FAU - Force, T
AU  - Force T
FAU - Rosenzweig, A
AU  - Rosenzweig A
LA  - eng
GR  - AI40970/AI/NIAID NIH HHS/United States
GR  - DK50282/DK/NIDDK NIH HHS/United States
GR  - HL03348/HL/NHLBI NIH HHS/United States
GR  - HL54202/HL/NHLBI NIH HHS/United States
GR  - HL59521/HL/NHLBI NIH HHS/United States
GR  - HL61688/HL/NHLBI NIH HHS/United States
GR  - HL65584/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
DEP - 20010301
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Chemokine CCL2)
RN  - EC 2.7.1.- (Phosphatidylinositol 3-Kinases)
SB  - IM
MH  - Cell Line
MH  - Chemokine CCL2/pharmacology
MH  - Humans
MH  - Monocytes/*cytology/drug effects/enzymology
MH  - Phosphatidylinositol 3-Kinases/*metabolism
EDAT- 2001/03/30 10:00
MHDA- 2001/08/24 10:01
CRDT- 2001/03/30 10:00
PHST- 2001/03/30 10:00 [pubmed]
PHST- 2001/08/24 10:01 [medline]
PHST- 2001/03/30 10:00 [entrez]
AID - S0021-9258(20)89796-8 [pii]
AID - 10.1074/jbc.M011235200 [doi]
PST - ppublish
SO  - J Biol Chem. 2001 Jul 20;276(29):26846-51. doi: 10.1074/jbc.M011235200. Epub 2001 
      Mar 1.