Fas-Mediated Cytotoxicity Remains Intact in Perforin and Granzyme B Antisense Transfectants of a Human NK-like Cell Line

Angela Horn Montel, Markian R. Bochan, W. Goebel, Zacharie Brahmi

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Abstract

Cell-mediated cytotoxicity (CMC) has traditionally been thought to involve the release of granule components, including perforin and granzymes, from the effector cell (EC) onto the target cell (TC) membrane. Recently, a granule-independent cytolytic mechanism involving the interaction of Fas antigen (CD95) with Fas ligand has been described. We have generated antisense perforin (YT-xP1) and granzyme B (YT-xGrB) transfectants of the human NK-like cell line YT-INDY. These transfectants have greatly reduced cytolytic ability when compared to the vector-transfected control cell line (YT-neo). In this study, however, we demonstrate that the antisense transfectants retain the ability to lyse Fas+ TC. Fas-mediated lysis is Ca2+- independent and is inhibited by a monoclonal anti-Fas blocking Ab, M3. By RT-PCR, we detect message for FasL in unstimulated YT-xP1 and YT-xGrB transfectants, as well as in unstimulated YT-neo. By flow cytometry, we show that YT-neo, YT-xGrB, and YT-xP1 constitutively express surface FasL. These data indicate that in a human NK-like cell line, similar to the murine system, the granule and Fas pathways of cytotoxicity function independently of one another. At least with the TC tested, our data also indicate that the granule and Fas pathways together account for nearly 100% of the cytolytic ability of YT-INDY.

Original languageEnglish
Pages (from-to)312-317
Number of pages6
JournalCellular Immunology
Volume165
Issue number2
DOIs
StatePublished - Oct 15 1995

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Granzymes
Natural Killer Cells
Cell Line
Perforin
CD95 Antigens
Fas Ligand Protein
Flow Cytometry
Cell Membrane
Polymerase Chain Reaction
perforin-granzyme B

ASJC Scopus subject areas

  • Immunology
  • Cell Biology

Cite this

Fas-Mediated Cytotoxicity Remains Intact in Perforin and Granzyme B Antisense Transfectants of a Human NK-like Cell Line. / Montel, Angela Horn; Bochan, Markian R.; Goebel, W.; Brahmi, Zacharie.

In: Cellular Immunology, Vol. 165, No. 2, 15.10.1995, p. 312-317.

Research output: Contribution to journalArticle

Montel, Angela Horn ; Bochan, Markian R. ; Goebel, W. ; Brahmi, Zacharie. / Fas-Mediated Cytotoxicity Remains Intact in Perforin and Granzyme B Antisense Transfectants of a Human NK-like Cell Line. In: Cellular Immunology. 1995 ; Vol. 165, No. 2. pp. 312-317.
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