Logo of jexpmedHomeThe Rockefeller University PressEditorsContactInstructions for AuthorsThis issue
J Exp Med. Mar 1, 1988; 167(3): 1003–1015.
PMCID: PMC2188880

A monoclonal antibody (3G5)-defined ganglioside antigen is expressed on the cell surface of microvascular pericytes

Abstract

The identification of microvascular pericytes in vitro relies principally on morphological characteristics and growth dynamics, as there is a paucity of immunochemical markers for these cells. Consequently, an attempt was made to identify mAb reagents that would aid in both the rapid identification and enrichment of retinal capillary pericytes in vascular cell cultures. A panel of mAbs raised by xenogeneic immunization of mice with various tissues was screened for immunoreactivity with dissociated cultures of bovine retinal capillary pericytes. Two antibodies from the panel (3G5 and HISL-8) were seen to react with pericytes by indirect immunofluorescence. The mAb 3G5 was selected for further study. mAb 3G5 did not react with dissociated cultures of smooth muscle cells, endothelial cells, or retinal pigmented endothelial cells. The pericyte 3G5 antigen was insensitive to the action of trypsin; therefore, mAb 3G5 was used to selectively purify pericytes from trypsinized mixed retinal cell cultures by flow cytometry. 3G5+ pericytes (representing 8% of cells in a mixed retinal cell culture) were enriched at least nine-fold to represent greater than 70% of cells. The mAb 3G5 stained retinal capillaries in vivo with a fluorescence distribution consistent with pericyte staining. The 3G5 antigen of cultured pericytes was found to be a glycolipid of mobility intermediate between ganglioside markers GM1 and GM2.

Full Text

The Full Text of this article is available as a PDF (872K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Rhodin JA. Ultrastructure of mammalian venous capillaries, venules, and small collecting veins. J Ultrastruct Res. 1968 Dec;25(5):452–500. [PubMed]
  • Buzney SM, Massicotte SJ, Hetu N, Zetter BR. Retinal vascular endothelial cells and pericytes. Differential growth characteristics in vitro. Invest Ophthalmol Vis Sci. 1983 Apr;24(4):470–480. [PubMed]
  • Cohen MP, Frank RN, Khalifa AA. Collagen production by cultured retinal capillary pericytes. Invest Ophthalmol Vis Sci. 1980 Jan;19(1):90–94. [PubMed]
  • Frank RN, Kinsey VE, Frank KW, Mikus KP, Randolph A. In vitro proliferation of endothelial cells from kitten retinal capillaries. Invest Ophthalmol Vis Sci. 1979 Nov;18(11):1195–1200. [PubMed]
  • Gitlin JD, D'Amore PA. Culture of retinal capillary cells using selective growth media. Microvasc Res. 1983 Jul;26(1):74–80. [PubMed]
  • Herman IM, D'Amore PA. Microvascular pericytes contain muscle and nonmuscle actins. J Cell Biol. 1985 Jul;101(1):43–52. [PMC free article] [PubMed]
  • Joyce NC, DeCamilli P, Boyles J. Pericytes, like vascular smooth muscle cells, are immunocytochemically positive for cyclic GMP-dependent protein kinase. Microvasc Res. 1984 Sep;28(2):206–219. [PubMed]
  • Joyce NC, Haire MF, Palade GE. Contractile proteins in pericytes. I. Immunoperoxidase localization of tropomyosin. J Cell Biol. 1985 May;100(5):1379–1386. [PMC free article] [PubMed]
  • Joyce NC, Haire MF, Palade GE. Contractile proteins in pericytes. II. Immunocytochemical evidence for the presence of two isomyosins in graded concentrations. J Cell Biol. 1985 May;100(5):1387–1395. [PMC free article] [PubMed]
  • Powers AC, Rabizadeh A, Akeson R, Eisenbarth GS. Characterization of monoclonal antibody 3G5 and utilization of this antibody to immobilize pancreatic islet cell gangliosides in a solid phase radioassay. Endocrinology. 1984 Apr;114(4):1338–1343. [PubMed]
  • Rabinowe SL, Nayak RC, Krisch K, George KL, Eisenbarth GS. Aging in man. Linear increase of a novel T cell subset defined by antiganglioside monoclonal antibody 3G5. J Exp Med. 1987 May 1;165(5):1436–1441. [PMC free article] [PubMed]
  • Maciag T, Cerundolo J, Ilsley S, Kelley PR, Forand R. An endothelial cell growth factor from bovine hypothalamus: identification and partial characterization. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5674–5678. [PMC free article] [PubMed]
  • Carson MP, Haudenschild CC. Microvascular endothelium and pericytes: high yield, low passage cultures. In Vitro Cell Dev Biol. 1986 Jun;22(6):344–354. [PubMed]
  • Eisenbarth GS, Oie H, Gazdar A, Chick W, Schultz JA, Scearce RM. Production of monoclonal antibodies reacting with rat islet cell membrane antigens. Diabetes. 1981 Mar;30(3):226–230. [PubMed]
  • Srikanta S, Eisenbarth GS. Islet cell antigens. Initial studies of their biology and function. Mol Biol Med. 1986 Apr;3(2):113–127. [PubMed]
  • Srikanta S, Telen M, Posillico JT, Dolinar R, Krisch K, Haynes BF, Eisenbarth GS. Monoclonal antibodies to a human islet cell surface glycoprotein: 4F2 and LC7-2. Endocrinology. 1987 Jun;120(6):2240–2244. [PubMed]
  • KUWABARA T, COGAN DG. Studies of retinal vascular patterns. I. Normal architecture. Arch Ophthalmol. 1960 Dec;64:904–911. [PubMed]
  • Svennerholm L, Fredman P. A procedure for the quantitative isolation of brain gangliosides. Biochim Biophys Acta. 1980 Jan 18;617(1):97–109. [PubMed]
  • Magnani JL, Brockhaus M, Smith DF, Ginsburg V. Detection of glycolipid ligands by direct binding of carbohydrate-binding proteins to thin-layer chromatograms. Methods Enzymol. 1982;83:235–241. [PubMed]
  • Cogan DG, Kuwabara T. The mural cell in perspective. Arch Ophthalmol. 1967 Aug;78(2):133–139. [PubMed]
  • KUWABARA T, COGAN DG. Retinal vascular patterns. VI. Mural cells of the retinal capillaries. Arch Ophthalmol. 1963 Apr;69:492–502. [PubMed]
  • Frank RN, Dutta S, Mancini MA. Pericyte coverage is greater in the retinal than in the cerebral capillaries of the rat. Invest Ophthalmol Vis Sci. 1987 Jul;28(7):1086–1091. [PubMed]
  • Eimoto T. Ultrastructure of an infantile hemangiopericytoma. Cancer. 1977 Nov;40(5):2161–2170. [PubMed]
  • Page IH, Green JG, Robertson AL. The physician's incompleat guide to atherosclerosis. Ann Intern Med. 1966 Jan;64(1):189–203. [PubMed]
  • Tilton RG, Kilo C, Williamson JR, Murch DW. Differences in pericyte contractile function in rat cardiac and skeletal muscle microvasculatures. Microvasc Res. 1979 Nov;18(3):336–352. [PubMed]
  • Hakomori S. Glycosphingolipids in cellular interaction, differentiation, and oncogenesis. Annu Rev Biochem. 1981;50:733–764. [PubMed]
  • Nayak RC, Omar MA, Rabizadeh A, Srikanta S, Eisenbarth GS. "Cytoplasmic" islet cell antibodies. Evidence that the target antigen is a sialoglycoconjugate. Diabetes. 1985 Jun;34(6):617–619. [PubMed]
  • COGAN DG, TOUSSAINT D, KUWABARA T. Retinal vascular patterns. IV. Diabetic retinopathy. Arch Ophthalmol. 1961 Sep;66:366–378. [PubMed]
  • Engerman R, Bloodworth JM, Jr, Nelson S. Relationship of microvascular disease in diabetes to metabolic control. Diabetes. 1977 Aug;26(8):760–769. [PubMed]
  • Speiser P, Gittelsohn AM, Patz A. Studies on diabetic retinopathy. 3. Influence of diabetes on intramural pericytes. Arch Ophthalmol. 1968 Sep;80(3):332–337. [PubMed]
  • SVENNERHOLM L. CHROMATOGRAPHIC SEPARATION OF HUMAN BRAIN GANGLIOSIDES. J Neurochem. 1963 Sep;10:613–623. [PubMed]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...