PMID- 8218939
OWN - NLM
STAT- MEDLINE
DCOM- 19931215
LR  - 20191101
IS  - 1043-4666 (Print)
IS  - 1043-4666 (Linking)
VI  - 5
IP  - 3
DP  - 1993 May
TI  - Studies of binding and internalization of human recombinant monocyte chemotactic 
      and activating factor (MCAF) by monocytic cells.
PG  - 264-75
AB  - Recombinant human monocyte chemotactic and activating factor (MCAF) was iodinated 
      and specific binding sites for this cytokine were detected on human peripheral 
      blood monocytes, the monocytic leukemia cell line THP-1, and on 
      PMA-differentiated HL60 and U937 cell lines. The binding sites were specific for 
      MCAF since other polypeptide cytokines and the chemotactic peptide 
      formyl-methionyl-leucyl-phenylalanine (fMLP) failed to compete for 125I-rhMCAF 
      binding. Steady-state binding experiments at 4 degrees C revealed the presence of 
      13,000 and 18,000 receptor sites/cell on monocytes and THP-1 cells with Kd values 
      of 22.5 nM and 25.7 nM, respectively. Compared to a human natural MCAF, rhMCAF 
      was less potent in inducing maximal monocyte migration. Human natural MCAF 
      similarly competed more efficiently for 125I-rhMCAF binding than unlabelled 
      rhMCAF. The ligand-receptor association was highly temperature-dependent, with 
      maximal ligand uptake at 37 degrees C accompanied by internalization of the 
      ligand-receptor complexes. The internalized 125I-MCAF was progressively degraded 
      and released into the culture medium starting at 30 min. Lysosomotropic ammonium 
      chloride could inhibit the degradation of this ligand suggesting the involvement 
      of lysosomal enzymes in the proteolytic digestion. Incubation with cycloheximide 
      did not block the rapid reappearance of MCAF receptors within 20 min on the cell 
      surface indicative of receptor recycling rather than new protein synthesis. These 
      data indicate that monocytic cells express specific receptors for rhMCAF which 
      can be dynamically regulated by MCAF.
FAU - Wang, J M
AU  - Wang JM
AD  - Laboratory of Molecular Immunoregulation, National Cancer Institute, Frederick 
      Cancer Research and Development Center, Maryland 21702.
FAU - Hishinuma, A
AU  - Hishinuma A
FAU - Oppenheim, J J
AU  - Oppenheim JJ
FAU - Matsushima, K
AU  - Matsushima K
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Cytokine
JT  - Cytokine
JID - 9005353
RN  - 0 (Chemokine CCL2)
RN  - 0 (Chemotactic Factors)
RN  - 0 (Cytokines)
RN  - 0 (Receptors, CCR2)
RN  - 0 (Receptors, Chemokine)
RN  - 0 (Receptors, Cytokine)
RN  - 0 (Recombinant Proteins)
RN  - 98600C0908 (Cycloheximide)
RN  - NI40JAQ945 (Tetradecanoylphorbol Acetate)
SB  - IM
MH  - Biological Transport
MH  - Cell Line
MH  - Chemokine CCL2
MH  - Chemotactic Factors/*metabolism
MH  - Cycloheximide/pharmacology
MH  - Cytokines/*metabolism
MH  - Humans
MH  - In Vitro Techniques
MH  - Kinetics
MH  - Leukemia, Monocytic, Acute
MH  - Monocytes/*metabolism
MH  - Neutrophils/*metabolism
MH  - Receptors, CCR2
MH  - *Receptors, Chemokine
MH  - Receptors, Cytokine/biosynthesis/*metabolism
MH  - Recombinant Proteins/metabolism
MH  - Tetradecanoylphorbol Acetate/pharmacology
MH  - Tumor Cells, Cultured
EDAT- 1993/05/01 00:00
MHDA- 1993/05/01 00:01
CRDT- 1993/05/01 00:00
PHST- 1993/05/01 00:00 [pubmed]
PHST- 1993/05/01 00:01 [medline]
PHST- 1993/05/01 00:00 [entrez]
AID - 1043-4666(93)90014-V [pii]
AID - 10.1016/1043-4666(93)90014-v [doi]
PST - ppublish
SO  - Cytokine. 1993 May;5(3):264-75. doi: 10.1016/1043-4666(93)90014-v.