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Bactericidal effect of bovine lactoferrin and synthetic peptide lactoferrin chimera in Streptococcus pneumoniae and the decrease in luxS gene expression by lactoferrin

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Abstract

Streptococcus pneumoniae (pneumococcus) is responsible for nearly one million child deaths annually. Pneumococcus causes infections such as pneumonia, otitis media, meningitis, and sepsis. The human immune system includes antibacterial peptides and proteins such as lactoferrin (LF), but its activity against pneumococcus is not fully understood. The aim of this work was to evaluate the bactericidal effect of bovine lactoferrin (bLF) and the synthetic LF-peptides lactoferricin (LFcin17–30), lactoferrampin (LFampin265–284), and LFchimera against S. pneumoniae planktonic cells. The mechanism of damage was also investigated, as well as the impact of these peptides on the transcription levels of genes known to encode important virulence factors. S. pneumoniae planktonic cells were treated with bLF, LFcin17–30, LFampin265–284 and LFchimera at different time points. The viability of treated planktonic cells was assessed by dilution and plating (in CFU/ml). The interaction between LF and LF-peptides coupled to fluorescein was visualized using a confocal microscope and flow cytometry, whereas the damage at structural levels was observed by electron microscopy. Damage to bacterial membranes was further evaluated by membrane permeabilization by use of propidium iodide and flow cytometry, and finally, the expression of pneumococcal genes was evaluated by qRT-PCR. bLF and LFchimera were the best bactericidal agents. bLF and peptides interacted with bacteria causing changes in the shape and size of the cell and membrane permeabilization. Moreover, the luxS gene was down-regulated in bacteria treated with LF. In conclusion, LF and LFchimera have a bactericidal effect, and LF down-regulates genes involved in the pathogenicity of pneumococcus, thus demonstrating potential as new agents for the treatment of pneumococcal infections.

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Acknowledgments

We thank to BS Lourdes Rojas-Morales and BS Sirenia González-Pozos for their technical assistance in the Microscopy Unit of CINVESTAV-IPN, México. Authors also thank Gideon Matzkin, Emory University, for his valuable support in some laboratory procedures. This work was supported by grants from CONACYT (CB-2009-133677) and PROFAPI-UAS (2012/087; 2013/093).

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Authors and Affiliations

  1. Unidad de Investigación, Facultad de Medicina, Universidad Autónoma de Sinaloa, Cedros y Sauces, Fracc. Fresnos., C.P. 80246, Culiacán, Sinaloa, Mexico

    Nidia León-Sicairos, Uriel A. Angulo-Zamudio, Cynthia A. López-Torres, Ruth Reyes-Cortes & Adrian Canizalez-Román

  2. Hubert Department of Global Health, Rollins School of Public Health, Emory University, Atlanta, GA, USA

    Jorge E. Vidal

  3. Department of Oral Biochemistry ACTA, University of Amsterdam and VU University, Amsterdam, The Netherlands

    Jan G. M. Bolscher & Kamran Nazmi

  4. Departamento de Biología Celular, CINVESTAV-IPN, Mexico, D.F., Mexico

    Magda Reyes-López & Mireya de la Garza

  5. Departamento de Investigación, Hospital Pediátrico de Sinaloa, Culiacán, Sinaloa, Mexico

    Nidia León-Sicairos

Authors
  1. Nidia León-Sicairos
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  2. Uriel A. Angulo-Zamudio
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  3. Jorge E. Vidal
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  4. Cynthia A. López-Torres
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  5. Jan G. M. Bolscher
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  8. Magda Reyes-López
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  9. Mireya de la Garza
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Correspondence to Nidia León-Sicairos.

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León-Sicairos, N., Angulo-Zamudio, U.A., Vidal, J.E. et al. Bactericidal effect of bovine lactoferrin and synthetic peptide lactoferrin chimera in Streptococcus pneumoniae and the decrease in luxS gene expression by lactoferrin. Biometals 27, 969–980 (2014). https://doi.org/10.1007/s10534-014-9775-y

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