The Effects of Polyhexamethylene Biguanide (PHMB) and TLR Agonists Alone or as Polyplex Nanoparticles against <em>Leishmania infantum</em> Promastigotes and Amastigotes
Dogs are the main reservoir for <i>Leishmania infantum,</i> manifesting from a subclinical to a fatal disease. Limited treatments are available, although new antiparasitics and immunomodulators are pursued. Polyhexamethylene biguanide (PHMB) has a broad antimicrobial spectrum, including...
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MDPI AG
2020-11-01
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| Series: | Veterinary Sciences |
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| Online Access: | https://www.mdpi.com/2306-7381/7/4/179 |
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| author | Pamela Martínez-Orellana Marta Baxarias Liam Good Laia Solano-Gallego |
| author_facet | Pamela Martínez-Orellana Marta Baxarias Liam Good Laia Solano-Gallego |
| author_sort | Pamela Martínez-Orellana |
| collection | DOAJ |
| description | Dogs are the main reservoir for <i>Leishmania infantum,</i> manifesting from a subclinical to a fatal disease. Limited treatments are available, although new antiparasitics and immunomodulators are pursued. Polyhexamethylene biguanide (PHMB) has a broad antimicrobial spectrum, including antiparasitic activity. Here, we evaluated the potential for Toll-like receptor agonists (TLRa) and PHMB alone, and as polyplex nanoparticles containing PHMB and TLR4 or TLR9 agonists, to selectively kill <i>L. infantum</i>. Susceptibility of <i>L. infantum</i> promastigotes to PHMB, miltefosine, and allopurinol was performed, and the half-maximum inhibitory concentrations (IC<sub>50</sub>) were determined. Then, DH-82 cells were infected and treated with PHMB alone or combined with TLR4a (MPLA-SM) or TLR9a (CpG ODNs) and allopurinol alone. The IC<sub>50</sub> values of <i>L. infantum</i> promastigotes were PHMB (1.495 µM), miltefosine (9.455 µM), and allopurinol (0.124 µM). After infection, treated DH-82 cells displayed a lower percentage (<i>p =</i> 0.0316), intensity (<i>p =</i> 0.0002), and index of infection (<i>p =</i> 0.0022) when compared to non-treated cells. PHMB induced lower percentage of infection alone (<i>p =</i> 0.043), in combination with TLR9a (<i>p =</i> 0.043), and with TLR4a (<i>p =</i> 0.0213). Supernatants were collected and used to measure TNF-α and IL-6 levels. Increased TNF-α was observed after PHMB <i>plus</i> TLR4a, relative to uninfected and infected untreated macrophages (<i>p =</i> 0.043). PHMB combined with TLR4a shows promise as a potential anti-<i>L. infantum</i> drug combination, as well as inducer of proinflammatory response, as demonstrated by decreased infection and increased TNF-α production. |
| first_indexed | 2024-03-10T14:51:42Z |
| format | Article |
| id | doaj.art-4efca8acd94248d1a64c4fe75a7cb07f |
| institution | Directory Open Access Journal |
| issn | 2306-7381 |
| language | English |
| last_indexed | 2024-03-10T14:51:42Z |
| publishDate | 2020-11-01 |
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| series | Veterinary Sciences |
| spelling | doaj.art-4efca8acd94248d1a64c4fe75a7cb07f2023-11-20T20:54:42ZengMDPI AGVeterinary Sciences2306-73812020-11-017417910.3390/vetsci7040179The Effects of Polyhexamethylene Biguanide (PHMB) and TLR Agonists Alone or as Polyplex Nanoparticles against <em>Leishmania infantum</em> Promastigotes and AmastigotesPamela Martínez-Orellana0Marta Baxarias1Liam Good2Laia Solano-Gallego3Departament de Medicina i Cirurgia Animals, Facultat de Veterinària, Universitat Autònoma de Barcelona, Bellaterra, 08193 Barcelona, SpainDepartament de Medicina i Cirurgia Animals, Facultat de Veterinària, Universitat Autònoma de Barcelona, Bellaterra, 08193 Barcelona, SpainDepartment of Pathobiology and Population Sciences, The Royal Veterinary College, London NW1 0NH, UKDepartament de Medicina i Cirurgia Animals, Facultat de Veterinària, Universitat Autònoma de Barcelona, Bellaterra, 08193 Barcelona, SpainDogs are the main reservoir for <i>Leishmania infantum,</i> manifesting from a subclinical to a fatal disease. Limited treatments are available, although new antiparasitics and immunomodulators are pursued. Polyhexamethylene biguanide (PHMB) has a broad antimicrobial spectrum, including antiparasitic activity. Here, we evaluated the potential for Toll-like receptor agonists (TLRa) and PHMB alone, and as polyplex nanoparticles containing PHMB and TLR4 or TLR9 agonists, to selectively kill <i>L. infantum</i>. Susceptibility of <i>L. infantum</i> promastigotes to PHMB, miltefosine, and allopurinol was performed, and the half-maximum inhibitory concentrations (IC<sub>50</sub>) were determined. Then, DH-82 cells were infected and treated with PHMB alone or combined with TLR4a (MPLA-SM) or TLR9a (CpG ODNs) and allopurinol alone. The IC<sub>50</sub> values of <i>L. infantum</i> promastigotes were PHMB (1.495 µM), miltefosine (9.455 µM), and allopurinol (0.124 µM). After infection, treated DH-82 cells displayed a lower percentage (<i>p =</i> 0.0316), intensity (<i>p =</i> 0.0002), and index of infection (<i>p =</i> 0.0022) when compared to non-treated cells. PHMB induced lower percentage of infection alone (<i>p =</i> 0.043), in combination with TLR9a (<i>p =</i> 0.043), and with TLR4a (<i>p =</i> 0.0213). Supernatants were collected and used to measure TNF-α and IL-6 levels. Increased TNF-α was observed after PHMB <i>plus</i> TLR4a, relative to uninfected and infected untreated macrophages (<i>p =</i> 0.043). PHMB combined with TLR4a shows promise as a potential anti-<i>L. infantum</i> drug combination, as well as inducer of proinflammatory response, as demonstrated by decreased infection and increased TNF-α production.https://www.mdpi.com/2306-7381/7/4/179TLR4 agonistTLR9 agonistPHMB<i>Leishmania infantum</i>caninemacrophages |
| spellingShingle | Pamela Martínez-Orellana Marta Baxarias Liam Good Laia Solano-Gallego The Effects of Polyhexamethylene Biguanide (PHMB) and TLR Agonists Alone or as Polyplex Nanoparticles against <em>Leishmania infantum</em> Promastigotes and Amastigotes Veterinary Sciences TLR4 agonist TLR9 agonist PHMB <i>Leishmania infantum</i> canine macrophages |
| title | The Effects of Polyhexamethylene Biguanide (PHMB) and TLR Agonists Alone or as Polyplex Nanoparticles against <em>Leishmania infantum</em> Promastigotes and Amastigotes |
| title_full | The Effects of Polyhexamethylene Biguanide (PHMB) and TLR Agonists Alone or as Polyplex Nanoparticles against <em>Leishmania infantum</em> Promastigotes and Amastigotes |
| title_fullStr | The Effects of Polyhexamethylene Biguanide (PHMB) and TLR Agonists Alone or as Polyplex Nanoparticles against <em>Leishmania infantum</em> Promastigotes and Amastigotes |
| title_full_unstemmed | The Effects of Polyhexamethylene Biguanide (PHMB) and TLR Agonists Alone or as Polyplex Nanoparticles against <em>Leishmania infantum</em> Promastigotes and Amastigotes |
| title_short | The Effects of Polyhexamethylene Biguanide (PHMB) and TLR Agonists Alone or as Polyplex Nanoparticles against <em>Leishmania infantum</em> Promastigotes and Amastigotes |
| title_sort | effects of polyhexamethylene biguanide phmb and tlr agonists alone or as polyplex nanoparticles against em leishmania infantum em promastigotes and amastigotes |
| topic | TLR4 agonist TLR9 agonist PHMB <i>Leishmania infantum</i> canine macrophages |
| url | https://www.mdpi.com/2306-7381/7/4/179 |
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