Transcriptome analysis of Streptococcus pneumoniae treated with the designed antimicrobial peptides, DM3

The antimicrobial peptide human Beta defensin 3 (hBD3) is an essential part of the innate immune system and is involved in protection against respiratory pathogens by specifically permeabilizing bacterial membranes. The Gram-positive bacterium Streptococcus pneumoniae causes serious diseases including pneumonia, meningitis, and septicemia, despite being frequently exposed to human defense molecules, including hBD3 during colonization and infection. Thus, the question arises how pneumococci adapt to stress caused by antimicrobial peptides. We addressed this subject by analyzing the proteome of S. pneumoniae after treatment with hBD3 and compared our data with the proteomic changes induced by LL-37, another crucial antimicrobial peptide present in the human respiratory tract. As antimicrobial peptides usually cause membrane perturbations, the response to the membrane active cationic detergent cetyltrimethylammonium bromide (CTAB) was examined to assess the specificity of the pneumococcal response to antimicrobial peptides. In brief, hBD3 and LL-37 induce a similar response in pneumococci and especially, changes in proteins with annotated transporter and virulence function have been identified. However, LL-37 causes changes in the abundance of cell surface modification proteins that cannot be observed after treatment with hBD3. Interestingly, CTAB induces unique proteomic changes in S. pneumoniae. Though, the detergent seems to activate a two-component system that is also activated in response to antimicrobial peptide stress (TCS 05). Overall, our data represent a novel resource on pneumococcal adaptation to specific cell surface stresses on a functional level. This knowledge can potentially be used to develop strategies to circumvent pneumococcal resistance to antimicrobial peptides.

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The place of endogenous antimicrobial peptides in the pathogenetic mechanisms of the development of community-acquired pneumonia caused by Streptococcus pneumoniae among infants

CHILD`S HEALTH ◽

10.22141/2224-0551.12.4.2017.107626 ◽

2017 ◽

Vol 12 (4) ◽

pp. 459-464

Author(s):

G.O. Lezhenko ◽

O.E. Pashkova ◽

H.V. Kraynya

Keyword(s):

Streptococcus Pneumoniae ◽

Antimicrobial Peptides ◽

Community Acquired Pneumonia ◽

Pathogenetic Mechanisms

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De Novo Transcriptome Analysis and Detection of Antimicrobial Peptides of the American Cockroach Periplaneta americana (Linnaeus)

PLoS ONE ◽

10.1371/journal.pone.0155304 ◽

2016 ◽

Vol 11 (5) ◽

pp. e0155304 ◽

Cited By ~ 17

Author(s):

In-Woo Kim ◽

Joon Ha Lee ◽

Sathiyamoorthy Subramaniyam ◽

Eun-Young Yun ◽

Iksoo Kim ◽

...

Keyword(s):

Antimicrobial Peptides ◽

Transcriptome Analysis ◽

Periplaneta Americana ◽

De Novo ◽

American Cockroach ◽

De Novo Transcriptome

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Generic and Specific Adaptive Responses of Streptococcus pneumoniae to Challenge with Three Distinct Antimicrobial Peptides, Bacitracin, LL-37, and Nisin

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00769-09 ◽

2009 ◽

Vol 54 (1) ◽

pp. 440-451 ◽

Cited By ~ 50

Author(s):

Joanna A. Majchrzykiewicz ◽

Oscar P. Kuipers ◽

Jetta J. E. Bijlsma

Keyword(s):

Streptococcus Pneumoniae ◽

Antimicrobial Peptides ◽

Abc Transporters ◽

Defense Mechanisms ◽

Negative Regulator ◽

Adaptive Responses ◽

Hoechst 33342 ◽

Novel Proteins ◽

Immunity Genes ◽

Increased Sensitivity

ABSTRACT To investigate the response of Streptococcus pneumoniae to three distinct antimicrobial peptides (AMPs), bacitracin, nisin, and LL-37, transcriptome analysis of challenged bacteria was performed. Only a limited number of genes were found to be up- or downregulated in all cases. Several of these common highly induced genes were chosen for further analysis, i.e., SP0385-SP0387 (SP0385-0387 herein), SP0912-0913, SP0785-0787, SP1714-1715, and the blp gene cluster. Deletion of these genes in combination with MIC determinations showed that several putative transporters, i.e., SP0785-0787 and SP0912-0913, were indeed involved in resistance to lincomycin and LL-37 and to bacitracin, nisin, and lincomycin, respectively. Mutation of the blp bacteriocin immunity genes resulted in an increased sensitivity to LL-37. Interestingly, a putative ABC transporter (SP1715) protected against bacitracin and Hoechst 33342 but conferred sensitivity to LL-37. A GntR-like regulator, SP1714, was identified as a negative regulator of itself and two of the putative transporters. In conclusion, we show that resistance to three different AMPs in S. pneumoniae is mediated by several putative ABC transporters, some of which have not been associated with antimicrobial resistance in this organism before. In addition, a GntR-like regulator that regulates two of these transporters was identified. Our findings extend the understanding of defense mechanisms of this important human pathogen against antimicrobial compounds and point toward novel proteins, i.e., putative ABC transporters, which can be used as targets for the development of new antimicrobials.

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Differential Expression of Antimicrobial Peptides in Streptococcus pneumoniae Keratitis and STAT3-Dependent Expression of LL-37 by Streptococcus pneumoniae in Human Corneal Epithelial Cells

Pathogens ◽

10.3390/pathogens8010031 ◽

2019 ◽

Vol 8 (1) ◽

pp. 31 ◽

Cited By ~ 2

Author(s):

Prerana Sharma ◽

Natalia Sharma ◽

Priyasha Mishra ◽

Joveeta Joseph ◽

Dilip K. Mishra ◽

...

Keyword(s):

Streptococcus Pneumoniae ◽

Epithelial Cells ◽

Antimicrobial Peptides ◽

Differential Expression ◽

Human Monocyte ◽

Mitogen Activated Protein Kinase ◽

Human Cornea ◽

Corneal Epithelial Cells ◽

Corneal Epithelial ◽

Human Corneal Epithelial Cells

Streptococcus pneumoniae is the leading cause of bacterial keratitis in the developing world with a growing trend of acquiring resistance against various antibiotics. In the current study, we determined the expression of different antimicrobial peptides (AMPs) in response to S. pneumoniae in patients, as well as in primary and immortalized human corneal epithelial cells. We further focused on LL-37 and determined its expression in human cornea infected with S. pneumoniae and studied the killing ability of LL-37 against S. pneumoniae. The expression of AMPs was determined by quantitative PCR and the phosphorylation of signaling proteins was evaluated by immunoblot analysis. LL-37 expression was also determined by immunofluorescence and Western blot method and the killing ability of LL-37 against S. pneumoniae was determined by colony-forming units. Differential expression of antimicrobial peptides was observed in patients with S. pneumoniae keratitis. Although S. pneumoniae induced expression of the AMPs in human corneal epithelial cells (HCEC), it did not induce AMP expression in U937, a human monocyte cell line. S. pneumoniae also caused activation of nuclear factor kappa-light-chain enhancer of activated B cells (NF-κB)and mitogen activated protein kinase (MAPK) pathways in corneal epithelial cells. LL-37 was found to be effective against both laboratory and clinical strains of S. pneumoniae. LL-37 induction by S. pneumoniae in human corneal epithelial cells was mediated by signal transducer and activator of transcription 3 (STAT3) activation, and inhibition of STAT3 activation significantly reduced LL-37 expression. Our study determines an extensive profile of AMPs expressed in the human cornea during S. pneumoniae infection, and suggests the potential of LL-37 to be developed as an alternative therapeutic intervention to fight increasing antibiotic resistance among bacteria.

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