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Interferon production in murine macrophage-like cell lines
Abstract:
Five murine macrophage (M phi)-like cell lines were examined to determine their suitability for the characterization of M phi interferons (IFNs). The J774A.1, RAW 309 Cr.1, and RAW 264.7 cell lines produced 30-800 international units (IU)/10(6) cells when treated with 5-200 micrograms of bacterial lipopolysaccharide (LPS). No IFN was detected in LPS-treated P388D1 or PU5-1.8 cell cultures. All cell lines produced IFN when inoculated with Newcastle disease virus (NDV); however, only 15 IU/10(6) cells of acid stable IFN were produced in PU5-1.8 cell cultures in comparison to 4.2 X 10(3)-1.7 X 10(4) IU/10(6) cells in the other cell lines. Most of the IFN was produced within 4 h in LPS-treated cell cultures and within 12 h in NDV-infected cell cultures. All IFNs were stable at pH 2.0 and were neutralized with antiserum against mouse L cell IFN. These cell lines appear competent for use in studying the synthesis, molecular weights, and regulatory functions of M phi IFNs.
Insights
Researchers evaluated macrophage cell lines for interferon (IFN) characterization. J774A.1, RAW 309 Cr.1, and RAW 264.7 cells effectively produced macrophage interferons upon stimulation.
Area of Science:
- Immunology
- Cell Biology
- Virology
Background:
- Macrophage-derived interferons (IFNs) play a crucial role in innate immunity.
- Characterizing macrophage (M phi) IFNs requires suitable cell models.
- Previous studies have not comprehensively evaluated M phi cell lines for IFN production.
Purpose of the Study:
- To assess the suitability of five murine M phi-like cell lines for characterizing M phi IFNs.
- To compare IFN production levels and kinetics in response to different stimuli.
Main Methods:
- Five murine M phi-like cell lines (J774A.1, RAW 309 Cr.1, RAW 264.7, P388D1, PU5-1.8) were treated with bacterial lipopolysaccharide (LPS) or Newcastle disease virus (NDV).
- Interferon production was quantified using international units (IU)/10(6) cells.
- IFN stability (pH 2.0) and neutralization with anti-mouse L cell IFN antiserum were assessed.
Main Results:
- J774A.1, RAW 309 Cr.1, and RAW 264.7 cells produced significant levels of IFN (30-800 IU/10(6) cells) upon LPS treatment.
- P388D1 and PU5-1.8 cells showed no detectable IFN production with LPS.
- All cell lines produced IFN upon NDV inoculation, with PU5-1.8 producing significantly lower levels (15 IU/10(6) cells) of acid-stable IFN compared to others (4.2 x 10(3)-1.7 x 10(4) IU/10(6) cells).
Conclusions:
- J774A.1, RAW 309 Cr.1, and RAW 264.7 cell lines are suitable for studying M phi IFN synthesis, molecular weights, and regulatory functions.
- PU5-1.8 cells are less suitable for general M phi IFN studies due to low production, but may be useful for specific investigations of acid-stable IFN.
- IFN production kinetics vary, with peak production occurring within 4 hours for LPS and 12 hours for NDV stimulation.