Cooperative action of SP-A and its trimeric recombinant fragment with polymyxins against Gram-negative respiratory bacteria
The exploration of therapies combining antimicrobial lung proteins and conventional antibiotics is important due to the growing problem of multidrug-resistant bacteria. The aim of this study was to investigate whether human SP-A and a recombinant trimeric fragment (rfhSP-A) have cooperative antimicr...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-09-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2022.927017/full |
| _version_ | 1798036510899961856 |
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| author | Juan Manuel Coya Víctor Fraile-Ágreda Lidia de Tapia Belén García-Fojeda Alejandra Sáenz José A. Bengoechea Nina Kronqvist Jan Johansson Cristina Casals |
| author_facet | Juan Manuel Coya Víctor Fraile-Ágreda Lidia de Tapia Belén García-Fojeda Alejandra Sáenz José A. Bengoechea Nina Kronqvist Jan Johansson Cristina Casals |
| author_sort | Juan Manuel Coya |
| collection | DOAJ |
| description | The exploration of therapies combining antimicrobial lung proteins and conventional antibiotics is important due to the growing problem of multidrug-resistant bacteria. The aim of this study was to investigate whether human SP-A and a recombinant trimeric fragment (rfhSP-A) have cooperative antimicrobial activity with antibiotics against pathogenic Gram-negative bacteria. We found that SP-A bound the cationic peptide polymyxin B (PMB) with an apparent dissociation constant (KD) of 0.32 ± 0.04 µM. SP-A showed synergistic microbicidal activity with polymyxin B and E, but not with other antibiotics, against three SP-A-resistant pathogenic bacteria: Klebsiella pneumoniae, non-typable Haemophilus influenzae (NTHi), and Pseudomonas aeruginosa. SP-A was not able to bind to K. pneumoniae, NTHi, or to mutant strains thereof expressing long-chain lipopolysaccharides (or lipooligosaccharides) and/or polysaccharide capsules. In the presence of PMB, SP-A induced the formation of SP-A/PMB aggregates that enhance PMB-induced bacterial membrane permeabilization. Furthermore, SP-A bound to a molecular derivative of PMB lacking the acyl chain (PMBN) with a KD of 0.26 ± 0.02 μM, forming SP-A/PMBN aggregates. PMBN has no bactericidal activity but can bind to the outer membrane of Gram-negative bacteria. Surprisingly, SP-A and PMBN showed synergistic bactericidal activity against Gram-negative bacteria. Unlike native supratrimeric SP-A, the trimeric rfhSP-A fragment had small but significant direct bactericidal activity against K. pneumoniae, NTHi, and P. aeruginosa. rfhSP-A did not bind to PMB under physiological conditions but acted additively with PMB and other antibiotics against these pathogenic bacteria. In summary, our results significantly improve our understanding of the antimicrobial actions of SP-A and its synergistic action with PMB. A peptide based on SP-A may aid the therapeutic use of PMB, a relatively cytotoxic antibiotic that is currently being reintroduced into clinics due to the global problem of antibiotic resistance. |
| first_indexed | 2024-04-11T21:13:54Z |
| format | Article |
| id | doaj.art-7efda82791254f3697a49e40591c3553 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-04-11T21:13:54Z |
| publishDate | 2022-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-7efda82791254f3697a49e40591c35532022-12-22T04:02:53ZengFrontiers Media S.A.Frontiers in Immunology1664-32242022-09-011310.3389/fimmu.2022.927017927017Cooperative action of SP-A and its trimeric recombinant fragment with polymyxins against Gram-negative respiratory bacteriaJuan Manuel Coya0Víctor Fraile-Ágreda1Lidia de Tapia2Belén García-Fojeda3Alejandra Sáenz4José A. Bengoechea5Nina Kronqvist6Jan Johansson7Cristina Casals8Department of Biochemistry and Molecular Biology, Complutense University of Madrid, Madrid, SpainDepartment of Biochemistry and Molecular Biology, Complutense University of Madrid, Madrid, SpainDepartment of Biochemistry and Molecular Biology, Complutense University of Madrid, Madrid, SpainDepartment of Biochemistry and Molecular Biology, Complutense University of Madrid, Madrid, SpainDepartment of Biochemistry and Molecular Biology, Complutense University of Madrid, Madrid, SpainWellcome-Wolfson Institute for Experimental Medicine, Queen’s University Belfast, Belfast, United KingdomDepartment of Biosciences and Nutrition, Neo, Karolinska Institutet, Huddinge, SwedenDepartment of Biosciences and Nutrition, Neo, Karolinska Institutet, Huddinge, SwedenDepartment of Biochemistry and Molecular Biology, Complutense University of Madrid, Madrid, SpainThe exploration of therapies combining antimicrobial lung proteins and conventional antibiotics is important due to the growing problem of multidrug-resistant bacteria. The aim of this study was to investigate whether human SP-A and a recombinant trimeric fragment (rfhSP-A) have cooperative antimicrobial activity with antibiotics against pathogenic Gram-negative bacteria. We found that SP-A bound the cationic peptide polymyxin B (PMB) with an apparent dissociation constant (KD) of 0.32 ± 0.04 µM. SP-A showed synergistic microbicidal activity with polymyxin B and E, but not with other antibiotics, against three SP-A-resistant pathogenic bacteria: Klebsiella pneumoniae, non-typable Haemophilus influenzae (NTHi), and Pseudomonas aeruginosa. SP-A was not able to bind to K. pneumoniae, NTHi, or to mutant strains thereof expressing long-chain lipopolysaccharides (or lipooligosaccharides) and/or polysaccharide capsules. In the presence of PMB, SP-A induced the formation of SP-A/PMB aggregates that enhance PMB-induced bacterial membrane permeabilization. Furthermore, SP-A bound to a molecular derivative of PMB lacking the acyl chain (PMBN) with a KD of 0.26 ± 0.02 μM, forming SP-A/PMBN aggregates. PMBN has no bactericidal activity but can bind to the outer membrane of Gram-negative bacteria. Surprisingly, SP-A and PMBN showed synergistic bactericidal activity against Gram-negative bacteria. Unlike native supratrimeric SP-A, the trimeric rfhSP-A fragment had small but significant direct bactericidal activity against K. pneumoniae, NTHi, and P. aeruginosa. rfhSP-A did not bind to PMB under physiological conditions but acted additively with PMB and other antibiotics against these pathogenic bacteria. In summary, our results significantly improve our understanding of the antimicrobial actions of SP-A and its synergistic action with PMB. A peptide based on SP-A may aid the therapeutic use of PMB, a relatively cytotoxic antibiotic that is currently being reintroduced into clinics due to the global problem of antibiotic resistance.https://www.frontiersin.org/articles/10.3389/fimmu.2022.927017/fullcollectin SP-Arecombinant trimeric fragmentmultidrug-resistant bacteriamicrobial infectionlungpolymyxin B |
| spellingShingle | Juan Manuel Coya Víctor Fraile-Ágreda Lidia de Tapia Belén García-Fojeda Alejandra Sáenz José A. Bengoechea Nina Kronqvist Jan Johansson Cristina Casals Cooperative action of SP-A and its trimeric recombinant fragment with polymyxins against Gram-negative respiratory bacteria Frontiers in Immunology collectin SP-A recombinant trimeric fragment multidrug-resistant bacteria microbial infection lung polymyxin B |
| title | Cooperative action of SP-A and its trimeric recombinant fragment with polymyxins against Gram-negative respiratory bacteria |
| title_full | Cooperative action of SP-A and its trimeric recombinant fragment with polymyxins against Gram-negative respiratory bacteria |
| title_fullStr | Cooperative action of SP-A and its trimeric recombinant fragment with polymyxins against Gram-negative respiratory bacteria |
| title_full_unstemmed | Cooperative action of SP-A and its trimeric recombinant fragment with polymyxins against Gram-negative respiratory bacteria |
| title_short | Cooperative action of SP-A and its trimeric recombinant fragment with polymyxins against Gram-negative respiratory bacteria |
| title_sort | cooperative action of sp a and its trimeric recombinant fragment with polymyxins against gram negative respiratory bacteria |
| topic | collectin SP-A recombinant trimeric fragment multidrug-resistant bacteria microbial infection lung polymyxin B |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2022.927017/full |
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