Session Type: ePosters
Session Title: ePosters
Authors(s): D. Bongiorno (1), N. Musso (2), C. Giuseppe (3), L. Lorenzo Mattia (1), B. Dalida (1), G. Margherita (3), C. Filippo (3), C. Floriana (1), S. Stefani (1)
Authors Affiliations(s): (1) Department Of Biomedical And Biotechnological Sciences, Section Of Microbiology, University Of Catania, Italy, (2) Department Of Biomedical And Biotechnological Sciences, Section Of Microbiology, University Of Catania - Catania (Italy), Italy, (3) Department of Drug and Healt Sciences, University of Catania, 95125 Catania, Italy., Italy
Background:
The ability of S. aureus to infect bone and osteoblasts is correlated to its incredible virulence armamentarium that can mediate the invasion/internalization process, cytotoxicity, membrane damage and intracellular persistence. Using an ex-vivo model of human MG-63 osteoblasts infected by two S. aureus clinical strains belonging to ST239-SCCmecIII-t037 and ST228-SCCmecI-t041 clones, and ATCC12598 ST30-t076 as control strain, we analyzed the interaction, persistence and modulation of expression of selected bacterial genes, cell viability and their influence on human gene expression of pro-inflammatory cytokines and oxidative stress phenomena.
Methods:Intracellular frequency of the bacterial strains was evaluated with Immaging Flow Cytometry (IFC) at a multiplicity of infection (MOI) of 100:1. The evaluation of internalization and persistence on the viability of MG-63 cells was performed by MTT assay. Gene expression analysis was performed by quantitative Real-Time PCR. All the experiments were performed at 3 and 24 hours p.i. with bacterial strains.
Results:Using IFC analysis, we found that strains differently invaded osteoblasts after 3h and 24h: ATCC12598 internalized in 70% and 50% of cells, ST239-SCCmecIII in 50% and 45% and ST228-SCCmecI in 30% and 20%, respectively. ST239-III exerted a significative cytotoxic activity due to the over-expression of virulence and adhesion genes. ST239-III was able to significantly increase the gene expression of pro-inflammatory cytokines in MG-63 osteoblast cells at both time points; and determine a strong up-regulation of cytokines mRNAs 24 hours p.i. The lower invasiveness of ST228-I was associated with non-cytotoxic activity inside osteoblasts.
Conclusions:Our results, can open a new way of considering therapies, going in the direction of an individualized therapeutic strategies that should take into account the difference existing between MSSA and MRSA as well as the distinctive features of the different clones. Such strategies rely on the study of the genetic and biochemical basis of both pathogen and host.
Keyword(s): MRSA, Internalization, Host-Phatogene interactionCOI Institutional Grants: Yes
Session Type: ePosters
Session Title: ePosters
Authors(s): D. Bongiorno (1), N. Musso (2), C. Giuseppe (3), L. Lorenzo Mattia (1), B. Dalida (1), G. Margherita (3), C. Filippo (3), C. Floriana (1), S. Stefani (1)
Authors Affiliations(s): (1) Department Of Biomedical And Biotechnological Sciences, Section Of Microbiology, University Of Catania, Italy, (2) Department Of Biomedical And Biotechnological Sciences, Section Of Microbiology, University Of Catania - Catania (Italy), Italy, (3) Department of Drug and Healt Sciences, University of Catania, 95125 Catania, Italy., Italy
Background:
The ability of S. aureus to infect bone and osteoblasts is correlated to its incredible virulence armamentarium that can mediate the invasion/internalization process, cytotoxicity, membrane damage and intracellular persistence. Using an ex-vivo model of human MG-63 osteoblasts infected by two S. aureus clinical strains belonging to ST239-SCCmecIII-t037 and ST228-SCCmecI-t041 clones, and ATCC12598 ST30-t076 as control strain, we analyzed the interaction, persistence and modulation of expression of selected bacterial genes, cell viability and their influence on human gene expression of pro-inflammatory cytokines and oxidative stress phenomena.
Methods:Intracellular frequency of the bacterial strains was evaluated with Immaging Flow Cytometry (IFC) at a multiplicity of infection (MOI) of 100:1. The evaluation of internalization and persistence on the viability of MG-63 cells was performed by MTT assay. Gene expression analysis was performed by quantitative Real-Time PCR. All the experiments were performed at 3 and 24 hours p.i. with bacterial strains.
Results:Using IFC analysis, we found that strains differently invaded osteoblasts after 3h and 24h: ATCC12598 internalized in 70% and 50% of cells, ST239-SCCmecIII in 50% and 45% and ST228-SCCmecI in 30% and 20%, respectively. ST239-III exerted a significative cytotoxic activity due to the over-expression of virulence and adhesion genes. ST239-III was able to significantly increase the gene expression of pro-inflammatory cytokines in MG-63 osteoblast cells at both time points; and determine a strong up-regulation of cytokines mRNAs 24 hours p.i. The lower invasiveness of ST228-I was associated with non-cytotoxic activity inside osteoblasts.
Conclusions:Our results, can open a new way of considering therapies, going in the direction of an individualized therapeutic strategies that should take into account the difference existing between MSSA and MRSA as well as the distinctive features of the different clones. Such strategies rely on the study of the genetic and biochemical basis of both pathogen and host.
Keyword(s): MRSA, Internalization, Host-Phatogene interactionCOI Institutional Grants: Yes
Session Type: ePosters
Session Title: ePosters
Authors(s): D. Bongiorno (1), N. Musso (2), C. Giuseppe (3), L. Lorenzo Mattia (1), B. Dalida (1), G. Margherita (3), C. Filippo (3), C. Floriana (1), S. Stefani (1)
Authors Affiliations(s): (1) Department Of Biomedical And Biotechnological Sciences, Section Of Microbiology, University Of Catania, Italy, (2) Department Of Biomedical And Biotechnological Sciences, Section Of Microbiology, University Of Catania - Catania (Italy), Italy, (3) Department of Drug and Healt Sciences, University of Catania, 95125 Catania, Italy., Italy
Background:
The ability of S. aureus to infect bone and osteoblasts is correlated to its incredible virulence armamentarium that can mediate the invasion/internalization process, cytotoxicity, membrane damage and intracellular persistence. Using an ex-vivo model of human MG-63 osteoblasts infected by two S. aureus clinical strains belonging to ST239-SCCmecIII-t037 and ST228-SCCmecI-t041 clones, and ATCC12598 ST30-t076 as control strain, we analyzed the interaction, persistence and modulation of expression of selected bacterial genes, cell viability and their influence on human gene expression of pro-inflammatory cytokines and oxidative stress phenomena.
Methods:Intracellular frequency of the bacterial strains was evaluated with Immaging Flow Cytometry (IFC) at a multiplicity of infection (MOI) of 100:1. The evaluation of internalization and persistence on the viability of MG-63 cells was performed by MTT assay. Gene expression analysis was performed by quantitative Real-Time PCR. All the experiments were performed at 3 and 24 hours p.i. with bacterial strains.
Results:Using IFC analysis, we found that strains differently invaded osteoblasts after 3h and 24h: ATCC12598 internalized in 70% and 50% of cells, ST239-SCCmecIII in 50% and 45% and ST228-SCCmecI in 30% and 20%, respectively. ST239-III exerted a significative cytotoxic activity due to the over-expression of virulence and adhesion genes. ST239-III was able to significantly increase the gene expression of pro-inflammatory cytokines in MG-63 osteoblast cells at both time points; and determine a strong up-regulation of cytokines mRNAs 24 hours p.i. The lower invasiveness of ST228-I was associated with non-cytotoxic activity inside osteoblasts.
Conclusions:Our results, can open a new way of considering therapies, going in the direction of an individualized therapeutic strategies that should take into account the difference existing between MSSA and MRSA as well as the distinctive features of the different clones. Such strategies rely on the study of the genetic and biochemical basis of both pathogen and host.
Keyword(s): MRSA, Internalization, Host-Phatogene interactionCOI Institutional Grants: Yes
Session Type: ePosters
Session Title: ePosters
Authors(s): D. Bongiorno (1), N. Musso (2), C. Giuseppe (3), L. Lorenzo Mattia (1), B. Dalida (1), G. Margherita (3), C. Filippo (3), C. Floriana (1), S. Stefani (1)
Authors Affiliations(s): (1) Department Of Biomedical And Biotechnological Sciences, Section Of Microbiology, University Of Catania, Italy, (2) Department Of Biomedical And Biotechnological Sciences, Section Of Microbiology, University Of Catania - Catania (Italy), Italy, (3) Department of Drug and Healt Sciences, University of Catania, 95125 Catania, Italy., Italy
Background:
The ability of S. aureus to infect bone and osteoblasts is correlated to its incredible virulence armamentarium that can mediate the invasion/internalization process, cytotoxicity, membrane damage and intracellular persistence. Using an ex-vivo model of human MG-63 osteoblasts infected by two S. aureus clinical strains belonging to ST239-SCCmecIII-t037 and ST228-SCCmecI-t041 clones, and ATCC12598 ST30-t076 as control strain, we analyzed the interaction, persistence and modulation of expression of selected bacterial genes, cell viability and their influence on human gene expression of pro-inflammatory cytokines and oxidative stress phenomena.
Methods:Intracellular frequency of the bacterial strains was evaluated with Immaging Flow Cytometry (IFC) at a multiplicity of infection (MOI) of 100:1. The evaluation of internalization and persistence on the viability of MG-63 cells was performed by MTT assay. Gene expression analysis was performed by quantitative Real-Time PCR. All the experiments were performed at 3 and 24 hours p.i. with bacterial strains.
Results:Using IFC analysis, we found that strains differently invaded osteoblasts after 3h and 24h: ATCC12598 internalized in 70% and 50% of cells, ST239-SCCmecIII in 50% and 45% and ST228-SCCmecI in 30% and 20%, respectively. ST239-III exerted a significative cytotoxic activity due to the over-expression of virulence and adhesion genes. ST239-III was able to significantly increase the gene expression of pro-inflammatory cytokines in MG-63 osteoblast cells at both time points; and determine a strong up-regulation of cytokines mRNAs 24 hours p.i. The lower invasiveness of ST228-I was associated with non-cytotoxic activity inside osteoblasts.
Conclusions:Our results, can open a new way of considering therapies, going in the direction of an individualized therapeutic strategies that should take into account the difference existing between MSSA and MRSA as well as the distinctive features of the different clones. Such strategies rely on the study of the genetic and biochemical basis of both pathogen and host.
Keyword(s): MRSA, Internalization, Host-Phatogene interactionCOI Institutional Grants: Yes