Hypermucoviscous Carbapenem-Resistant Klebsiella pneumoniae ST25 Infect Human Intestinal Epithelial Cells and Induce Moderate Inflammation
Abstract
:1. Introduction
2. Results
2.1. K. pneumoniae ST25 Adhere and Infect Caco-2 Cells
2.2. K. pneumoniae ST25 Infection Affects Tight Junction Proteins Expression in Caco-2 Cells
2.3. K. pneumoniae ST25 Infection Affect COX-2 and iNOS Expression in Caco-2 Cells
2.4. K. pneumoniae ST25 Trigger Inflammatory Factors Production in Caco-2 Cells
2.5. K. pneumoniae ST25 Modulate TGF-β and TLL1 Expression in Caco-2 Cells
2.6. In Silico Analysis of Virulence Genes from K. pneumoniae ST25
3. Discussion
4. Materials and Methods
4.1. Strains
4.2. Cell Culture and Viability Assays
4.3. Lactate Dehydrogenase (LDH) Activity Assay
4.4. Adhesion, Invasion, and Permeability Experiments in Caco-2 Cells
4.5. RT-PCR
4.6. Cytokine Concentrations
4.7. Genomic Analysis
4.8. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Strain | Colbactin | Mucoviscosity Associated Protein | RmpA | TamA/TamB System | Type VI Secretion System Main Cluster (Genes ≥ 8) |
---|---|---|---|---|---|
KP28872 | - | - | - | + | tssA, tssE, tssF, tssG, tssI (vgrG), tssJ, tssk, tssL, tssM. |
LABACER01 | - | - | - | + | tssA, tssB (2 copies), tssC, tssD (hcp), tssF, tssG, tssH (clpv), tssI (vgrG), tssJ, tssk, tssL, tssM, evpJ (PAAR). |
LABACER27 | - | - | - | + | tssA, tssE, tssF, tssG, tssI (vgrG), tssJ, tssk, tssL, tssM. |
NTUH-K2044 | - | + | + | + | tssA, tssB (2 copies), tssC, tssD (hcp), tssF, tssG, tssH (clpv), tssI (vgrG), tssJ, tssk, tssL, tssM, evpJ (PAAR). |
SWU01 | - | - | - | + | tssA, tssB, tssC, tssD (hcp), tssF, tssG, tssH (clpv), tssI (vgrG), tssJ, tssk, tssL, tssM, evpJ (PAAR). |
Gene | Amplicon Size (pb) | Forward Primer (5′→3′) | Reverse Primer (5′→3′) |
---|---|---|---|
Occludin | 114 | GAGTTGTATCTGTTGTTGT | TTCGTGGTATAGCATTCT |
ZO-1 | 118 | GGTGAAGTGAAGACAATG | GGTAATATGGTGAAGTTAGAG |
Claudin-5 | 84 | TTAACAGACGGAATGAAGT | GAAGCGAAATCCTCAGTC |
COX-2 | 117 | GAGAGATGTATCCTCCCACAGTCA | GACCAGGCACCAGACCAAAG |
iNOS | 278 | CCTTACGAGGCGAAGAAGGACAG | CAGTTTGAGAGAGGAGGCTCCG |
TNF-α | 360 | GTCAGATCATCTTCTCGAACC | CAGATAGATGGGCTCATACC |
IL-6 | 211 | GACAGCCACTCACCTCTTCA | TTCACCAGGCAAGTCTCCTC |
MCP-1 | 93 | AGTCTCTGCCGCCCTTCT | GTGACTGGGGCATTGATTG |
IL-8 | 168 | ACTCCAAACCTTTCCACCC | CCCTCTTCAAAAACTTCTCCAC |
Strains | Assembly | Size | GC% | Scaffolds | CDS |
---|---|---|---|---|---|
KP28872 | GCA_013623415.1 | 5.9 | 56.9 | 364 | 5553 |
LABACER01 | GCA_013375175.1 | 5.6 | 57.1 | 61 | 5306 |
LABACER27 | GCA_013375185.1 | 5.6 | 57.1 | 51 | 5170 |
NTUH-K2044 | GCA_000009885.1 | 5.5 | 57.4 | 2 | 5028 |
SWU01 | GCA_001902475.1 | 5.7 | 57.4 | 2 | 5420 |
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Dentice Maidana, S.; Elean, M.; Fukuyama, K.; Imamura, Y.; Albarracín, L.; Saha, S.; Suda, Y.; Kurata, S.; Jure, M.Á.; Kitazawa, H.; et al. Hypermucoviscous Carbapenem-Resistant Klebsiella pneumoniae ST25 Infect Human Intestinal Epithelial Cells and Induce Moderate Inflammation. Int. J. Mol. Sci. 2023, 24, 8804. https://doi.org/10.3390/ijms24108804
Dentice Maidana S, Elean M, Fukuyama K, Imamura Y, Albarracín L, Saha S, Suda Y, Kurata S, Jure MÁ, Kitazawa H, et al. Hypermucoviscous Carbapenem-Resistant Klebsiella pneumoniae ST25 Infect Human Intestinal Epithelial Cells and Induce Moderate Inflammation. International Journal of Molecular Sciences. 2023; 24(10):8804. https://doi.org/10.3390/ijms24108804
Chicago/Turabian StyleDentice Maidana, Stefania, Mariano Elean, Kohtaro Fukuyama, Yoshiya Imamura, Leonardo Albarracín, Sudeb Saha, Yoshihito Suda, Shoichiro Kurata, María Ángela Jure, Haruki Kitazawa, and et al. 2023. "Hypermucoviscous Carbapenem-Resistant Klebsiella pneumoniae ST25 Infect Human Intestinal Epithelial Cells and Induce Moderate Inflammation" International Journal of Molecular Sciences 24, no. 10: 8804. https://doi.org/10.3390/ijms24108804