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The Effects of Bruton Tyrosine Kinase Inhibition on Chemotaxis and Superoxide Generation in Human Neutrophils

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Abstract

Purpose

The role of the Bruton tyrosine kinase (Btk) protein in neutrophil function has been evaluated using neutrophils from healthy volunteers after incubation with a Btk inhibitor, leflunomide metabolite analog (LFM-A13), suggesting an important role for Btk in neutrophil function. We sought to determine the role of Btk protein on neutrophil superoxide generation and chemotaxis stimulated by N-formyl-methionine-leucine-phenylalanine (fMLP).

Methods

Chemotaxis was assayed on agarose gel and superoxide generation by cytochrome C reduction. The affects of LFM-A13 on chemotaxis and superoxide generation in unstimulated and fMLP stimulated neutrophils were studied in Btk deficient neutrophils from XLA patients compared with matched controls analyzed simultaneously.

Results

Chemotaxis and stimulated superoxide production were similar in the normal and Btk deficient neutrophils and were similarly inhibited by LFM-A13. In one patient, LFMA13 had no effect on superoxide generation in Btk deficient neutrophils up to a concentration of 25 microM, while inhibited superoxide production by control neutrophils.

Conclusions

Our results suggest that Btk does not have a specific role in neutrophil fMLP-stimulated superoxide generation and chemotaxis since these activities were similarly inhibited by LFM-A13 in Btk deficient and normal neutrophils. The lack of superoxide generation following Btk inhibition by LFM-A13 in Btk deficient neutrophils from one patient may suggest some heterogeneity in the role of Btk in fMLP induced neutrophil superoxide generation.

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Acknowledgments

This study was supported by grants from Allergists for Israel (AFI), Jeffery Modell Foundation (JMF) and The Faculty of Health Sciences Ben-Gurion University of the Negev.

Conflict of Interest

The authors declare they have no conflict of interest.

Author information

Authors and Affiliations

  1. Pediatric Immunology Clinic, Soroka University Medical Center, POB 151, Beer-Sheva, Israel, 84101

    Arnon Broides & Jacov Levy

  2. Infectious Disease Laboratory, Department of Clinical Biochemistry, Soroka University Medical Center, Beer-Sheva, Israel

    Nurit Hadad & Rachel Levy

  3. Soroka University Medical Center, Beer-Sheva, Israel

    Arnon Broides, Nurit Hadad, Jacov Levy & Rachel Levy

  4. Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva, Israel

    Arnon Broides, Nurit Hadad, Jacov Levy & Rachel Levy

Authors
  1. Arnon Broides
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  2. Nurit Hadad
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  3. Jacov Levy
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  4. Rachel Levy
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Correspondence to Arnon Broides.

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Broides, A., Hadad, N., Levy, J. et al. The Effects of Bruton Tyrosine Kinase Inhibition on Chemotaxis and Superoxide Generation in Human Neutrophils. J Clin Immunol 34, 555–560 (2014). https://doi.org/10.1007/s10875-014-0046-z

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  • DOI: https://doi.org/10.1007/s10875-014-0046-z

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