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4 results on '"Itzia S, Gómez-Alonso"'

1. Low Concentration of the Neutrophil Proteases Cathepsin G, Cathepsin B, Proteinase-3 and Metalloproteinase-9 Induce Biofilm Formation in Non-Biofilm-Forming Staphylococcus epidermidis Isolates

Author

Itzia S. Gómez-Alonso, Sergio Martínez-García, Gabriel Betanzos-Cabrera, Esmeralda Juárez, María C. Sarabia-León, María Teresa Herrera, Fernando Gómez-Chávez, Luvia Sanchez-Torres, Sandra Rodríguez-Martínez, Mario E. Cancino-Diaz, Jorge Cancino, and Juan C. Cancino-Diaz

Subjects
Staphylococcus epidermidis, biofilm, proteinase-3, neutrophil, commensal, cathepsin G, Biology (General), QH301-705.5, Chemistry, QD1-999
Abstract

Neutrophils play a crucial role in eliminating bacteria that invade the human body; however, cathepsin G can induce biofilm formation in a non-biofilm-forming Staphylococcus epidermidis 1457 strain, suggesting that neutrophil proteases may be involved in biofilm formation. Cathepsin G, cathepsin B, proteinase-3, and metalloproteinase-9 (MMP-9) from neutrophils were tested on the biofilm induction in commensal (skin isolated) and clinical non-biofilm-forming S. epidermidis isolates. From 81 isolates, 53 (74%) were aap+, icaA−, icaD− genotype, and without the capacity of biofilm formation under conditions of 1% glucose, 4% ethanol or 4% NaCl, but these 53 non-biofilm-forming isolates induced biofilm by the use of different neutrophil proteases. Of these, 62.3% induced biofilm with proteinase-3, 15% with cathepsin G, 10% with cathepsin B and 5% with MMP -9, where most of the protease-induced biofilm isolates were commensal strains (skin). In the biofilm formation kinetics analysis, the addition of phenylmethylsulfonyl fluoride (PMSF; a proteinase-3 inhibitor) showed that proteinase-3 participates in the cell aggregation stage of biofilm formation. A biofilm induced with proteinase-3 and DNAse-treated significantly reduced biofilm formation at an early time (initial adhesion stage of biofilm formation) compared to untreated proteinase-3-induced biofilm (p < 0.05). A catheter inoculated with a commensal (skin) non-biofilm-forming S. epidermidis isolate treated with proteinase-3 and another one without the enzyme were inserted into the back of a mouse. After 7 days of incubation period, the catheters were recovered and the number of grown bacteria was quantified, finding a higher amount of adhered proteinase-3-treated bacteria in the catheter than non-proteinase-3-treated bacteria (p < 0.05). Commensal non-biofilm-forming S. epidermidis in the presence of neutrophil cells significantly induced the biofilm formation when multiplicity of infection (MOI) 1:0.01 (neutrophil:bacteria) was used, but the addition of a cocktail of protease inhibitors impeded biofilm formation. A neutrophil:bacteria assay did not induce neutrophil extracellular traps (NETs). Our results suggest that neutrophils, in the presence of commensal non-biofilm-forming S. epidermidis, do not generate NETs formation. The effect of neutrophils is the production of proteases, and proteinase-3 releases bacterial DNA at the initial adhesion, favoring cell aggregation and subsequently leading to biofilm formation.

Published
2022
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3. The Glycosyltransferases sdgA and sdgB Expression in Staphylococcus Epidermidis Depends On The Conditions of Biofilm Formation

Author

Itzia S. Gómez-Alonso, Ilse D. Estrada-Alemán, Sergio Martínez-García, Humberto Peralta, Erika T. Quintana, Claudia Guerrero-Barajas, Cipriano Chávez-Cabrera, Sandra Rodríguez-Martínez, Mario E. Cancino-Diaz, and Juan C. Cancino-Diaz

Abstract

The Staphylococcus aureus’ SdrG protein is glycosylated by SdgA and SdgB for their protection against its degradation by the neutrophil’s cathepsin G. So far, there is not information about the role of Staphylococcus epidermidis’ SdgA nor SdgB in the production of biofilm, therefore the main of this work was to determine the distribution and expression of sdrG, sdgA and sdgB genes in S. epidermidis in conditions of biofilm. The frequency of the genes sdrG, sdgA and sdgB were evaluated by PCR in a collection of 75 isolates. The isolates were grown in dynamic conditions (in agitation) or static conditions (biofilm productor: planktonic or sessile cells). The expression of sdrG, sdgA and sdgB were determined by RT-qPCR in cells grown under dynamic conditions (CGDC), as well as planktonic and sessile cells, and in cells adhered to a catheter (in vivo). The genes sdrG and sdgB were detected in 100% of isolates, meanwhile the gene sdgA was detected in 71% of the samples (psdrG, sdgA and sdgB mRNAs. The planktonic and sessile cells expressed sdrG and sdgB, and the same was seen in cells adhered to the catheter. In particular, one isolate, able to induce biofilm under cathepsin G treatment, expressed sdrG and sdgB in planktonic, sessile and in cells adhered to the catheter. This suggests that the state of cells adherence is an important factor for the transcription of sdgA, sdgB and sdrG.

Published
2021

4. The expression of glycosyltransferases sdgA and sdgB in Staphylococcus epidermidis depends on the conditions of biofilm formation

Author

Itzia S, Gómez-Alonso, Ilse D, Estrada-Alemán, Sergio, Martínez-García, Humberto, Peralta, Erika T, Quintana, Claudia, Guerrero-Barajas, Cipriano, Chávez-Cabrera, Sandra, Rodríguez-Martínez, Mario E, Cancino-Diaz, and Juan C, Cancino-Diaz

Subjects
Mice, Cathepsin G, Bacterial Proteins, Biofilms, Staphylococcus epidermidis, Animals, Glycosyltransferases, Staphylococcal Infections
Abstract

The Staphylococcus aureus SdrG protein is glycosylated by SdgA and SdgB for protection against its degradation by the neutrophil cathepsin G. So far, there is no information about the role of Staphylococcus epidermidis SdgA or SdgB in biofilm-forming; therefore, the focus of this work was to determine the distribution and expression of the sdrG, sdgA and sdgB genes in S. epidermidis under in vitro and in vivo biofilm conditions. The frequencies of the sdrG, sdgA and sdgB genes were evaluated by PCR in a collection of 75 isolates. Isolates were grown in dynamic (non-biofilm-forming) or static (biofilm-forming) conditions. The expression of sdrG, sdgA and sdgB was determined by RT-qPCR in cells grown under dynamic conditions (CGDC), as well as in planktonic and sessile cells from a biofilm and cells adhered to a catheter implanted in Balb/c mice. The sdrG and sdgB genes were detected in 100% of isolates, while the sdgA gene was detected in 71% of the sample (p 0.001). CGDC did not express sdrG, sdgA and sdgB mRNAs. Planktonic and sessile cells expressed sdrG and sdgB, and the same was observed in cells adhered to the catheter. In particular, one isolate, capable of inducing a biofilm under treatment with cathepsin G, expressed sdrG and sdgB in planktonic and sessile cells and cells adhering to the catheter. This suggests that bacteria require biofilm conditions as an important factor for the transcription of the sdgA, sdgB and sdrG genes.

Published
2021

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