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Role of intracellular calcium in priming of human peripheral blood monocytes by bacterial lipopolysaccharide

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Abstract

To determine the role of intracellular calcium ([Ca2+]i) in the priming of monocytes (MΦ) by bacterial lipopolysaccharide (LPS), the membrane expression of two functional proteins and phagocytosis and respiratory burst were examined by microfluorimetry. LPS induced a significant increase in HLA-DR and C3bi receptor (CR3) expression within 2 h of its addition to whole blood. The enhanced expression of both antigens by LPS was dose-dependent, with concentrations as low as 0.1 ng/ ml producing a response. The involvement of [Ca2+]i was demonstrated by loading isolated MΦ with the intracellular calcium chelator quin-2 or the inhibitor of intracellular calcium redistribution TMB-8 prior to addition of LPS. Both compounds inhibited the LPS-induced increase in HLA-DR and CR3 expression. No role for extracellular calcium, for calcium slow channel flux, or for the calcium-calmodulin complex in LPS priming was demonstrated when LPS was added in the presence of EGTA, trifluperazine (TFP), or verapamil. The addition of the calcium ionophores A23187 or ionomycin failed to increase expression of either antigen. Prior exposure to LPS primed MΦ for enhanced phagocytosis and respiratory burst activity. These functions were inhibited by TMB-8, but not by TFP or verapamil. Addition of LPS to isolated MΦ increased [Ca2+]i by 23% at 30 sec and 42% at 5 min, as measured by the calcium-sensitive, intracellular probe indo-1. These results suggest that intracellular Ca2+ mobilization is necessary, but not sufficient, for LPS-induced priming of human peripheral blood monocytes.

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Authors and Affiliations

  1. Departments of Medicine, University of Louisville School of Medicine, 40292, Louisville, Kentucky

    Kenneth R. McLeish, Samuel R. Wellhausen & Gregory T. Stelzer

  2. Departments of Biochemistry, University of Louisville School of Medicine, 40292, Louisville, Kentucky

    William L. Dean

Authors
  1. Kenneth R. McLeish
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  2. William L. Dean
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  3. Samuel R. Wellhausen
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  4. Gregory T. Stelzer
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Additional information

This work was supported in part by grant HL36303 (Dr. Dean) from the National Institutes of Health, and from the Cecil E. and Mamie M. Bales Medical Research Fund, University of Louisville (Dr. McLeish).

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McLeish, K.R., Dean, W.L., Wellhausen, S.R. et al. Role of intracellular calcium in priming of human peripheral blood monocytes by bacterial lipopolysaccharide. Inflammation 13, 681–692 (1989). https://doi.org/10.1007/BF00914312

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