Antigenicity of human melanoma cells transfected to express the B7-1 co-stimulatory molecule (CD80) varies with the level of B7-1 expression |
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Authors: | Donna O McCarthy Nicole Glowacki Kathleen Schell Carol A Emler Mark R Albertini |
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Institution: | (1) K4/414 Clinical Science Center, University of Wisconsin-Madison, 600 Highland Avenue, Madison, WI 53792, USA Tel.: +1608-263-0117, US;(2) Agracetus Inc., Middleton, Wisconsin 53562, USA, US |
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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.
Received: 14 October 1999 / Accepted: 3 December 1999 |
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Keywords: | Human melanoma B7-1 co-stimulatory molecule Particle-mediated gene transfer Cytotoxicity Flow cytometry |
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