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LPS and Taxol Activate Lyn Kinase Autophosphorylation in Lpsn, but Not in Lpsd, Macrophages

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Abstract

Background

The anti-tumor agent, Taxol, has been shown in murine macrophages to stimulate tumor necrosis factor (TNF), modulate TNF receptors, induce a large panel of immediate-early genes, and induce protein tyrosine phosphorylation indistinguishably from LPS. These data, coupled with the finding that lipid A antagonists block Taxol-induced stimulation, support the hypothesis that these two structurally unrelated compounds activate a common, receptor-associated signaling apparatus. A very early event in LPS signaling of human monocytes is activation of lyn kinase activity. We therefore sought to evaluate the activation of lyn kinase by LPS and Taxol in LPS-responsive (Lpsn) and LPS-hyporesponsive (Lpsd) macrophages.

Materials and Methods

C3H/OuJ (Lpsn) and C3H/HeJ (Lpsd) macrophages were stimulated by LPS or Taxol. Cell lysates were subjected to immunoprecipitation with anti-lyn antibody, gel electrophoresis, and in vitro kinase assays. Autoradiography and Phosphor-Imager analysis were carried out to detect incorporation of 32P into lyn protein.

Results

Within seconds of stimulation, LPS and Taxol induce in Lpsn macrophages a depression of autophosphorylation, followed within minutes by autophosphorylation of both p53 and p56 lyn species. Lpsd macrophages respond to LPS and Taxol with the initial decrease in activity, but fail to respond to LPS with autophosphorylation, and respond only to a limited extent upon Taxol stimulation. Tyrosine phosphatase inhibitors exerted inhibitory effects on LPS stimulation of lyn autophosphorylation.

Conclusions

Decreased lyn kinase activity within seconds and autophosphorylation within minutes of LPS or Taxol stimulation in Lpsn macrophages strongly supports the hypothesis that LPS and Taxol share a common signaling pathway. The finding that C3H/HeJ macrophages respond to LPS and Taxol with a normal depression of lyn activity, but fail to autophosphorylate lyn normally in response to LPS or Taxol, suggests that the Lpsd defect is distal to LPS-receptor interaction. Finally, the inhibitory effect of tyrosine phosphatase inhibitors on LPS-induced lyn autophosphorylation suggests that tyrosine phosphatase(s) may participate in the regulation of lyn kinase activity.

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References

  1. (1991) Physician’s Desk Reference. Medical Economics Data, Montvale, NJ.

  2. Nightingale S. (1994) Paclitaxel approved for treatment of certain breast cancer patients. J.A.M.A. 271: 1818.

    Article  CAS  PubMed  Google Scholar 

  3. Rowinsky EK, Donehower RC, Jones RJ, Tucker RW. (1988) Microtubule changes and cytotoxicity in leukemic cell lines treated with taxol. Cancer Res. 48: 4093–4099.

    PubMed  CAS  Google Scholar 

  4. Ding AH, Porteu F, Sanchez E, Nathan CF. (1990) Shared actions of endotoxin and taxol on TNF receptors and TNF release. Science 248: 370–372.

    Article  CAS  PubMed  Google Scholar 

  5. Carboni JM, Singh C, Tepper MA. (1993) Taxol and lipopolysaccharide activation of a murine macrophage cell line and induction of similar tyrosine phosphoproteins. Monogr. Natl. Cancer Inst. 15: 95–101.

    Google Scholar 

  6. Manthey CL, Brandes ME, Perera P-Y, Vogel SN. (1992) Taxol increases steady-state levels of lipopolysaccharide-inducible genes and protein-tyrosine phosphorylation in murine macrophages. J. Immunol. 149: 2459–2465.

    PubMed  CAS  Google Scholar 

  7. Manthey CL, Qureshi N, Stütz PL, Vogel SN. (1993) Lipopolysaccharide antagonists block taxol-induced signaling in murine macrophages. J. Exp. Med. 178: 695–701.

    Article  CAS  PubMed  Google Scholar 

  8. Manthey CL, Perera P-Y, Salkowski CA, Vogel SN. (1994) Taxol provides a second signal for murine macrophage tumoricidal activity. J. Immunol 152: 825–831.

    PubMed  CAS  Google Scholar 

  9. Vogel SN. (1992) The Lps gene: Insights into the genetic and molecular basis of LPS responsiveness and macrophage differentiation. In: Beutler B (ed). Tumor Necrosis Factors: The Molecules and Their Emerging Role in Medicine. Raven Press, New York, pp. 485–513.

    Google Scholar 

  10. Stefanova I, Corcoran ML, Horak EM, Wahl LM, Bolen JB, Horak ID. (1993) Lipopolysaccharide induces activation of CD14-associates protein tyrosine kinase p53/56lyn. J. Biol. Chem. 268: 20725–20728.

    PubMed  CAS  Google Scholar 

  11. Henricson BE, Manthey CL, Perera P-Y, Hamilton TA, Vogel SN. (1993) Dissociation of lipopolysaccharide (LPS)-inducible gene expression in murine macrophages pretreated with smooth LPS versus monophosphoryl lipid A. Infect. Immun. 61: 2325–2333.

    PubMed  PubMed Central  CAS  Google Scholar 

  12. Mclntire FC, Sievert HW, Barlow GH, Finley RA, Lee WY. (1967) Chemical physical and biological properties of a lipopolysaccharide from Escherichia coli K235. Biochemistry 6: 2363–2372.

    Article  Google Scholar 

  13. Burkhardt AL, Bolen JB. (1993) Immune-complex assays for tyrosine protein kinases. In: Coligan JE, Kruisbeek AM, Margulies DH, Shevach EM, Strober W. eds. Current Protocols in Immunology. Greene Publishing Associates and Wiley-Interscience, New York, pp. 11.4.1–11.4.8.

    Google Scholar 

  14. Resh M. (1994) Myristylation and palmitylation of src family members: The fats of the matter. Cell 76: 411–413.

    Article  CAS  PubMed  Google Scholar 

  15. Cooper JA, Howell B. (1993) The when and how of src regulation. Cell 73: 1051–1054.

    Article  CAS  PubMed  Google Scholar 

  16. Hailman E, Lichenstein HS, Wurfel MM, et al. (1994) Lipopolysaccharide (LPS)-binding protein accelerates the binding of LPS to CD 14. J. Exp. Med. 179: 269–277.

    Article  CAS  PubMed  Google Scholar 

  17. Weinstein SL, June CH, DeFranco AL. (1993) Lipopolysaccharide-induced protein tyrosine phosphorylation in human macrophages is mediated by CD 14. J. Immunol. 151: 3829–3838.

    PubMed  CAS  Google Scholar 

  18. Jacobavits A, Sharon N, Zan-Bar I. (1982) Acquisition of mitogenic responsiveness by non-responding lymphocytes upon insertion of appropriate membrane components. J. Exp. Med. 156: 1274–1279.

    Article  Google Scholar 

  19. Geppert TD, Whitehurst CE, Thompson P, Beutler B. (1994) Lipopolysaccharide signals activation of tumor necrosis factor biosynthesis through the Ras/Raf-1/MEK/MAPK pathway. Mol. Med. 1: 93–103.

    PubMed  PubMed Central  CAS  Google Scholar 

  20. Girault J-A. (1994) Protein kinases and phosphatases. Neurotransmissions 10: 1–6.

    Google Scholar 

Download references

Acknowledgements

This work was supported by National Institutes of Health Grant AI-18797 (SNV). The opinions or assertions contained within are the private views of the authors and should not be construed as official or necessarily reflecting the views of USUHS or the Department of Defense. The experiments reported herein were conducted according to the principles set forth in the Guide for the Care and Use of Laboratory Animals, Institute of Laboratory Animal Resources, National Research Council DHEW Publication No. (NIH) 85-23. The authors are extremely thankful to Dr. Joseph Bolen for his very thoughtful discussions and critical review of this manuscript and the Drug Synthesis and Chemistry Branch, Developmental Therapeutics Program, Division of Cancer Treatment, National Cancer Institute, Bethesda, MD, U.S.A., for Taxol.

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Authors and Affiliations

  1. Department of Microbiology and Immunology, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, Maryland, 20814, USA

    Beth E. Henricson & Stefanie N. Vogel

  2. Department of Molecular Drug Mechanisms, Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, New Jersey, USA

    Joan M. Carboni

  3. Department of Molecular Biology, Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, New Jersey, USA

    Anne L. Burkhardt

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  1. Beth E. Henricson
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  2. Joan M. Carboni
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  3. Anne L. Burkhardt
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  4. Stefanie N. Vogel
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Henricson, B.E., Carboni, J.M., Burkhardt, A.L. et al. LPS and Taxol Activate Lyn Kinase Autophosphorylation in Lpsn, but Not in Lpsd, Macrophages. Mol Med 1, 428–435 (1995). https://doi.org/10.1007/BF03401580

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