Perforin mRNA in primary peritoneal exudate cytotoxic T lymphocytes.
Considerable evidence indicates that cloned CTL cell lines kill target cells by releasing toxic granules that contain a cytolytic protein, called perforin, and several serine esterases (granzymes A to F). However, primary CTL, such as the highly cytolytic peritoneal exudate lymphocyte (PEL) cell population, have been found by a hemolytic assay to have no perforin, or perhaps only borderline levels of that protein, suggesting that these cells use a different lytic mechanism. To determine whether or not primary CTL express the perforin gene, we have here compared mRNA from PEL CTL and from a cloned CTL cell line, 2C, by Northern blot analysis using a perforin cDNA probe. CD8+ PEL CTL contain approximately 30% of the amount of perforin message present in 2C. Moreover, depletion of CD8+ T cells from the total peritoneal exudate cell population removes both cytolytic activity and perforin message. We have previously shown that PEL CTL elicit the same changes in target cells as cloned CTL cell lines and are resistant to lysis by the toxic granules purified from these cells lines. Taken together these results are consistent with the view that primary CTL, as well as long term cloned CTL cell lines, exercise their cytolytic activity by means of perforin.
Nagler-Anderson
C
C
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Lichtenheld
M
M
Eisen
H N
HN
Podack
E R
ER
D000818:Animals; D015152:Blotting, Northern; D002478:Cells, Cultured; D003594:Cytoplasmic Granules / Q000032:analysis / Q000276:immunology; D003602:Cytotoxicity, Immunologic; D006461:Hemolysis; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000032:analysis* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D054353:Perforin; D010529:Peritoneal Cavity; D052899:Pore Forming Cytotoxic Proteins; D012333:RNA, Messenger / Q000302:isolation & purification*; D013602:T-Lymphocytes, Cytotoxic / Q000032:analysis* / Q000276:immunology
D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D052899:Pore Forming Cytotoxic Proteins; D012333:RNA, Messenger; D054353:Perforin