Lipopolysaccharide and splenic tumoricidal macrophage activation

Splenic macrophage tumoricidal activity was examined and a splenic macrophage tumoricidal assay was established. Initially, mixtures of lipopolysaccharide (LPS) and spleen single cell suspensions (SSCS) were cultured for 1–4 days. Adherent macrophages, washed free of nonadherent cells and LPS, were...

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Bibliographic Details
Published in:Journal of leukocyte biology 1994-12, Vol.56 (6), p.714-722
Main Authors: Verstovsek, Srdan, Zaleskis, Gintaras, Maccubbin, Darbie L., Mihich, Enrico, Ehrke, M. Jane
Format: Article
Language:English
Subjects:
Animals
autofluorescence
Cell Adhesion
Cell Death
Female
Flow Cytometry
Fluorescence
Humans
Immunotherapy, Adoptive
interferon‐γ
kinetics
Lipopolysaccharides - pharmacology
Macrophage Activation - drug effects
Macrophages - cytology
Macrophages - drug effects
Macrophages - immunology
Mast-Cell Sarcoma - therapy
Mice
Mice, Inbred C57BL
Mice, Inbred DBA
Phenotype
Spleen - cytology
Spleen - drug effects
Spleen - immunology
Stimulation, Chemical
Time Factors
Tumor Cells, Cultured
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Summary:Splenic macrophage tumoricidal activity was examined and a splenic macrophage tumoricidal assay was established. Initially, mixtures of lipopolysaccharide (LPS) and spleen single cell suspensions (SSCS) were cultured for 1–4 days. Adherent macrophages, washed free of nonadherent cells and LPS, were then examined and were found to lack tumoricidal activity in a standard 18‐h 51Cr release assay. However, tumoricidal activity was generated if LPS was added to the SSCS cultures at later time points during the 4‐day incubation period; maximal activity was seen when LPS was added on day 3. In parallel, significant changes in macrophage autofluorescence and morphology, but not phenotype, were observed. Next, SSCS were cultured for 1–4 days without stimulating agents. Adherent macrophages were then washed free of nonadherent cells and LPS was added. Significant tumoricidal activity developed in time‐ and LPS concentration‐dependent fashions. The presence of nonadherent spleen cells in physical contact with the macrophages during the SSCS culture was essential for the macrophages in the resultant monolayer to be responsive to LPS. Activated splenic macrophage‐mediated lysis of tumor cells was shown to depend on the contact between the two cells. J. Leukoc. Biol. 56: 714–722; 1994.
ISSN:0741-5400
1938-3673
DOI:10.1002/jlb.56.6.714