Abstract
Monoclonal antibody P1H12 recognizes circulating endothelial cells and endothelia
of all sizes of blood vessels. To identify the protein recognized by P1H12, we expressed
a cDNA library in CHO cells and sequenced the cDNA from positive cells. The P1H12
sequence was identical, except at several bases, to that reported for melanoma cell
surface antigen MUC18/CD146. Aggregation assays demonstrated that CD146 mediates Ca++-independent homotypic endothelial cell adhesion. P1H12 mAb abrogated interactions
between human microvascular endothelial cells (HMVECs) but not between human umbilical
vein endothelial cells (HUVECs). P1H12 mAb abrogated P1H12-positive (CHOP1H12)-association with HMVECs or HUVECs. CD146 distribution is sparser on HUVECs than
on HMVECs. These data imply that HMVECs and HUVECs express the CD146 binding partner
but that CD146 is functional (or at sufficient density) only on HMVECs. HMVEC monolayers
treated with soluble P1H12 mAb showed increased permeability to albumin, with accompanying
changes in actin, paxillin, FAK, and caveolin distribution and changes in tyrosine
phosphorylation of FAK. Stimulation with P1H12 mAb led to redistribution of NF-κB
to the nucleus. P1H12 mAb bound to beads inhibited closure of wounded endothelial
monolayers. CD146 thus joins VE-cadherin and PECAM-1 as a molecule that mediates homotypic
endothelial cell adhesion. CD146 has both structural functions and signaling functions
important for endothelial monolayer integrity. (J Lab Clin Med 2001;138:322-31)
Abbreviations:
BSA (bovine serum albumin), CHOP1H12 (CHO, P1H12 positive), CHOwt (CHO, wild type), HUVEC (human umbilical vein endothelial cell), HMVEC (human microvascular endothelial cell), IgG (immunoglobulin G), mAb (monoclonal antibody), RT-PCR (reverse transcriptase–polymerase chain reaction)To read this article in full you will need to make a payment
Purchase one-time access:
Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online accessOne-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:
Subscribe to Translational ResearchAlready a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
References
- Vascular endothelial-cadherin is an important determinant of microvascular integrity in vivo.Proc Natl Acad Sci U S A. 1999; 96: 9815-9820
- Vascular-endothelial cadherin (CD144)—but not PECAM-1 (CD31)-based cell-to-cell contacts convey the maintenance of a quiescent endothelial monolayer.Int Arch Allerg Immunol. 1999; 120: 237-244
- Leukocyte-endothelial adhesion molecules.Blood. 1994; 84: 2068-2101
- Circulating activated endothelial cells in sickle cell anemia.N Engl J Med. 1997; 337: 1584-1590
- S-Endo-1, a pan-endothelial monoclonal antibody recognizing a novel human endothelial antigen.Tissue Antigens. 1996; 48: 531-539
- Tissue factor expression by endothelial cells in sickle cell anemia.J Clin Invest. 1998; 101: 1899-1904
- MUC18, a marker of tumor progression in human melanoma, shows sequence similarity to the neural cell adhesion molecules of the immunoglobin superfamily.Proc Natl Acad Sci USA. 1989; 86: 9891-9895
- MUC18: a cell adhesion molecule with a potential role in tumor growth and tumor cell dissemination.Curr Top Microbiol Immunol. 1996; 213: 95-105
- Direct correlation between MUC18 expression and metastatic potential of human melanoma cells.Melanoma Res. 1993; 3: 35-41
- MUC18, a melanoma-progression associated molecule, and its potential role in tumor vascularization and hematogenous spread.Cancer Res. 1994; 54: 5689-5694
- Functional aspects of three molecules associated with metastasis development in human malignant melanoma.Invasion Metastasis. 1989; 9: 338-350
- Melanoma cell-cell interactions are mediated through heterophilic Mel-CAM/ligand adhesion.Cancer Res. 1997; 57: 3835-3840
- Expression of MCAM/MUC18 by human melanoma cells leads to increased tumor x growth and metastasis.Cancer Res. 1997; 57: 2295-2303
- The role of CD146 (Mel-CAM) in biology and pathology.J Pathol. 1999; 189: 4
- Isolation and functional characterization of the A32 melanoma-associated antigen.Cancer Res. 1994; 54: 2514-2520
- Soluble P1H12/ CD146 is present in the plasma of sickle cell anemia patients.Blood. 1998; 92: 525a
- Synovial fluid CD146 (MUC18), a marker for sy-novial membrane angiogenesis in rheumatoid arthritis.Arthritis Rheum. 1999; 42: 622-630
- A novel technique for culture of human dermal microvascular endothelial cells under either serum-free or serum-supplemented conditions: isolation by panning and stimulation with vascular endothelial growth factor.Exp Cell Res. 1997; 230: 244-251
- Endothelial adherens junctions: implications in the control of vascular permeability and angiogenesis.J Clin Invest. 1997; 100: S7-S10
- Evans blue dye as a marker of albumin clearance in cultured endothelial monolayer and isolated lung.J Appl Physiol. 1992; 72: 865-873
- Genomic organization of the melanoma-associated glycoprotein MUC18: implications for the evolution of the immunoglobulin domains.Proc Natl Acad Sci U S A. 1993; 90: 8514-8518
- Vascular endothelial cell activation and permeability responses to thrombin.Blood Coagul Fibrinol. 1995; 6: 609-626
- Activation of human endothelial cells via S-endo-1 antigen (CD146) stimulates the tyrosine phosphorylation of focal adhesion kinase p125(FAK).J Biol Chem. 1998; 273: 26852-26856
- Expression of adhesion molecules on the endothelium of normal tissue vessels and vascular tumors.Lab Invest. 1993; 69: 322-328
- The biology of PECAM-1.J Clin Invest. 1997; 99: 3-8
- A new Mel-CAM (CD146)-specific monoclonal antibody, MN-4, on paraffin-embedded tissue.Mod Pathol. 1998; 11: 1098-1106
- Melanoma progression-associated glycoprotein MUC18/MCAM mediates homotypic cell adhesion through interaction with a heterophilic ligand.Int J Cancer. 1997; 73: 769-774
- Platelet endothelial cell adhesion molecule-1 (PECAM-1) homophilic adhesion is mediated by immuno-globulin-like domains 1 and 2 and depends on the cytoplasmic domain and the level of surface expression.J Biol Chem. 1996; 271: 18561-18570
- Cytoskeleton cross-talk during cell motility.FEBS Lett. 1999; 452: 96-99
- The mechanics of vascular cell motility.Microcirculation. 1998; 5: 239-257
- Vascular endothelial growth factor.Int J Biochem Cell Biol. 1998; 30: 1169-1174
- Integrin-mediated signal transduction pathways.Histol Histopathol. 1999; 14: 1001-1009
- Transcriptional regulation of endothelial cell adhesion molecules: NF-kappa B and cytokine-inducible enhancers.FASEB J. 1995; 9: 899-909
- P1H12 as an emryonic endothelial-endothelial adhesion molecule and possible hemangioblast marker.Blood. 1998; 92: 173a
- A common precursor for primitive erythropoiesis and definitive haematopoiesis.Nature. 1997; 386: 488-492
- A common precursor for hematopoietic and endothelial cells.Development. 1998; 125: 725-732
- Targeted null-mutation in the vascular endothelial-cadherin gene impairs the organization of vascular-like structures in embryoid bodies.Proc Natl Acad Sci USA. 1997; 94: 6273-6278
Article info
Publication history
Accepted:
July 11,
2001
Received in revised form:
June 22,
2001
Received:
January 4,
2001
Footnotes
☆Supported by the National Institutes of Health (HL30160 and DK56326).
☆☆Reprint requests: Robert P. Hebbel, MD, Mayo Mail Code 480, 420 Delaware Street SE, Minneapolis, MN 55455.
Identification
Copyright
© 2001 Mosby, Inc. Published by Elsevier Inc. All rights reserved.
