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FMLP-, thapsigargin-, and H2O2-evoked changes in intracellular free calcium concentration in lymphocytes and neutrophils of type 2 diabetic patients

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Abstract

Type 2 diabetic (T2DM) patients are immune-compromised having a higher susceptibility to infections and long-term complications in different parts of the body contributing to increased morbidity and mortality. A derangement in the homeostasis of intracellular free calcium concentration [Ca2+]i is known to be associated with several diseases in the body including T2DM. Both neutrophils and lymphocytes play active protective roles in host immune response to infection showing impairment in microbicidal functions including phagocytosis and chemotaxis which are calcium-dependent processes. This study evaluated the process of [Ca2+]i mobilization from both neutrophils and lymphocytes taken from blood of both T2DM patients and healthy age-matched control subjects investigating the effect of N-formyl-methionyl-leucyl-phenylalanine (fMLP), thapsigargin (TG), and hydrogen peroxide (H2O2) on [Ca2+]i homeostasis. This study employed isolated peripheral blood neutrophils and lymphocytes from 24 T2DM patients and 24 healthy volunteers. Either neutrophils or lymphocytes were stimulated separately with fMLP, TG, or H2O2. Induced changes in [Ca2+] in both neutrophils and lymphocytes were evaluated using spectrofluorometric methods. Stimulation of human neutrophils and lymphocytes with fMLP, TG, or H2O2 in the presence of [Ca2+]o resulted in significant decreases in [Ca2+]i mobilization from T2DM patients compared with healthy controls. These data indicate that neutrophils and lymphocytes from T2DM patients are less responsive to calcium mobilizing agents compared with granulocytes from healthy controls and this is possibly due to the hyperglycemia. The results suggest that agonist-evoked decrease in [Ca2+]i in immune cells might be one of the possible mechanisms of impaired immunity in diabetic patients.

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Acknowledgments

The authors are indebted to all the research staff members of Department of Physiology, University of Extremadura, Badajoz, Spain and University of Central Lancashire, Lancashire Teaching Hospitals NHS Trust, UK for their support. All the Type 2 Diabetic patients and controls who have given their blood samples for this work are greatly acknowledged. Dr J. Espino is a recipient of a research grant from the Ministerio de Educación, Cultura y Deporte (AP2009-0753).

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Authors and Affiliations

  1. School of Forensic and Investigative Sciences, University of Central Lancashire Preston, Lancashire, PR1 2HE, UK

    Shanti S. Kappala, Arati Iyengar & Jaipaul Singh

  2. Department of Physiology, Faculty of Science, University of Extremadura, Badajoz, Spain

    Javier Espino, Jose A. Pariente & Ana B. Rodriguez

  3. Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Centre, Omaha, NE, USA

    Keshore R. Bidasee

  4. Department of Diabetology and Endocrinology, Chorley General Hospital, LTHTR, Chorley, Lancashire, UK

    Satyan Rajbhandari

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  1. Shanti S. Kappala
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  2. Javier Espino
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Correspondence to Jaipaul Singh.

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Shanti S. Kappala and Javier Espino have contributed equally to this manuscript.

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Kappala, S.S., Espino, J., Pariente, J.A. et al. FMLP-, thapsigargin-, and H2O2-evoked changes in intracellular free calcium concentration in lymphocytes and neutrophils of type 2 diabetic patients. Mol Cell Biochem 387, 251–260 (2014). https://doi.org/10.1007/s11010-013-1890-5

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