Cancer Immunology, Immunotherapy

, Volume 49, Issue 2, pp 85–93 | Cite as

Antigenicity of human melanoma cells transfected to express the B7-1 co-stimulatory molecule (CD80) varies with the level of B7-1 expression

  • Donna O. McCarthy
  • Nicole Glowacki
  • Kathleen Schell
  • Carol A. Emler
  • Mark R. Albertini
ORIGINAL ARTICLE

Abstract

 The aim of this study was to compare the antigenicity of human melanoma cells molecularly modified by particle-mediated gene transfer to have transient or stable expression of the B7-1 co-stimulatory molecule (CD80). The unmodified melanoma cells (mel5, m21) had no constitutive expression of B7-1, but 22%–28% of cells had transient B7-1 expression 24 h following transfection with cDNA for B7-1 (mel5-B7, m21-B7). In addition, 85%–90% of cells had stable B7-1 expression following transfection with cDNA for B7-1 and in vitro culture under selection conditions (mel5-B7neo, m21-B7neo). Allogeneic HLA-unmatched normal donor peripheral blood mononuclear cells (PBMC) secreted greater amounts of granulocyte/macrophage-colony-stimulating factor (GM-CSF) when incubated for 3 days with m21-B7neo than did PBMC incubated with m21-B7, which, in turn, secreted greater amount of GM-CSF than PBMC incubated with m21. Similarly, cell-mediated cytotoxicity against unmodified melanoma cells by PBMC co-cultured for 5 days with the modified or unmodified melanoma cells was proportional to the level of B7-1 expression on the stimulating cells. This cytolytic activity had both an HLA-class-I-restricted and an HLA-class-I-unrestricted component. Following 5 days of co-culture, PBMC expression of CD28, the ligand for B7-1, was down-regulated in proportion to the level of B7-1 expression on the stimulating melanoma cells. Thus, particle-mediated gene delivery of cDNA for B7-1 into human melanoma cells increased expression of functional B7-1 and enhanced the antigenicity of the gene-modified cells in proportion to their level of B7-1 expression.

Key words Human melanoma B7-1 co-stimulatory molecule Particle-mediated gene transfer Cytotoxicity Flow cytometry 

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Copyright information

© Springer-Verlag Berlin Heidelberg 2000

Authors and Affiliations

  • Donna O. McCarthy
    • 1
  • Nicole Glowacki
    • 1
  • Kathleen Schell
    • 1
  • Carol A. Emler
    • 2
  • Mark R. Albertini
    • 1
  1. 1.K4/414 Clinical Science Center, University of Wisconsin-Madison, 600 Highland Avenue, Madison, WI 53792, USA Tel.: +1608-263-0117US
  2. 2.Agracetus Inc., Middleton, Wisconsin 53562, USAUS

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