Macrophage precursor cells produce perforin and perform Yac-1 lytic activity in response to stimulation with interleukin-2.
Macrophage precursor cells, derived from mouse bone marrow culture with granulocyte-macrophage colony-stimulating factor or colony-stimulating factor 1 (CSF-1) as growth factor and interleukin-2 (IL-2) as stimulating factor, were activated by IL-2 to exert strong cytolytic activity against Yac-1 cells. In response to IL-2 stimulation these bone marrow macrophage precursor cells produced perforin as lytic molecules. The purity of the precursor cells for the study was proved as homogeneous positivity for Mac-1, NK-1.1 and negativity for Lyt 1 and 2. The cells express CSF-1 receptors on their surface, are able to proliferate and differentiate into typical macrophages when stimulated with CSF-1, and are therefore members of the macrophage lineage. Perforin transcripts were identified by Northern blot analysis of IL-2-treated macrophage precursor cells, and the presence of perforin protein in the cytoplasmic granules was demonstrated by immunohistochemical staining using a monoclonal antiperforin antibody. In addition, the biological activity of the perforin contained in the macrophage precursor's granules could be documented as calcium-dependent lytic activity using Yac-1 and sheep red blood cells as targets. The results presented in this paper imply the existence of a bipotent precursor cell, which can mature into a typical macrophage if CSF-1 or phorbol 12-myristate 13-acetate is supplied as differentiation stimulating factor but develops into an NK/LAK cell when early activation with IL-2 is provided.
Li
H
H
Department of Immunobiology, Fraunhofer Institute of Toxicology, Hannover, Germany.
Pohler
U
U
Strehlow
I
I
Hertig
S
S
Baccarini
M
M
Emmendörffer
A
A
Tschopp
J
J
Lohmann-Matthes
M L
ML
D000818:Animals; D000941:Antigens / Q000032:analysis; D015152:Blotting, Northern; D003602:Cytotoxicity, Immunologic; D018076:DNA, Complementary / Q000235:genetics; D006412:Hematopoietic Stem Cells / Q000187:drug effects* / Q000502:physiology*; D007376:Interleukin-2 / Q000494:pharmacology*; D007694:Killer Cells, Natural / Q000276:immunology / Q000502:physiology; D008223:Lymphoma / Q000473:pathology; D008264:Macrophages / Q000187:drug effects* / Q000502:physiology*; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D016184:Receptors, Colony-Stimulating Factor / Q000235:genetics; D013268:Stimulation, Chemical; D014407:Tumor Cells, Cultured
D000941:Antigens; D018076:DNA, Complementary; D007376:Interleukin-2; D008562:Membrane Glycoproteins; D052899:Pore Forming Cytotoxic Proteins; D012333:RNA, Messenger; D016184:Receptors, Colony-Stimulating Factor; D054353:Perforin