000063036 001__ 63036 000063036 005__ 20190709135519.0 000063036 0247_ $$2doi$$a10.3389/fcimb.2017.00266 000063036 0248_ $$2sideral$$a101714 000063036 037__ $$aART-2017-101714 000063036 041__ $$aeng 000063036 100__ $$aRodríguez-Arce, I. 000063036 245__ $$aInactivation of the thymidylate synthase thyA in non-typeable Haemophilus influenzae modulates antibiotic resistance and has a strong impact on its interplay with the host airways 000063036 260__ $$c2017 000063036 5060_ $$aAccess copy available to the general public$$fUnrestricted 000063036 5203_ $$aAntibacterial treatment with cotrimoxazol (TxS), a combination of trimethoprim and sulfamethoxazole, generates resistance by, among others, acquisition of thymidine auxotrophy associated with mutations in the thymidylate synthase gene thyA, which can modify the biology of infection. The opportunistic pathogen non-typeable Haemophilus influenzae (NTHi) is frequently encountered in the lower airways of chronic obstructive pulmonary disease (COPD) patients, and associated with acute exacerbation of COPD symptoms. Increasing resistance of NTHi to TxS limits its suitability as initial antibacterial against COPD exacerbation, although its relationship with thymidine auxotrophy is unknown. In this study, the analysis of 2, 542 NTHi isolates recovered at Bellvitge University Hospital (Spain) in the period 2010–2014 revealed 119 strains forming slow-growing colonies on the thymidine low concentration medium Mueller Hinton Fastidious, including one strain isolated from a COPD patient undergoing TxS therapy that was a reversible thymidine auxotroph. To assess the impact of thymidine auxotrophy in the NTHi-host interplay during respiratory infection, thyA mutants were generated in both the clinical isolate NTHi375 and the reference strain RdKW20. Inactivation of the thyA gene increased TxS resistance, but also promoted morphological changes consistent with elongation and impaired bacterial division, which altered H. influenzae self-aggregation, phosphorylcholine level, C3b deposition, and airway epithelial infection patterns. Availability of external thymidine contributed to overcome such auxotrophy and TxS effect, potentially facilitated by the nucleoside transporter nupC. Although, thyA inactivation resulted in bacterial attenuation in a lung infection mouse model, it also rendered a lower clearance upon a TxS challenge in vivo. Thus, our results show that thymidine auxotrophy modulates both the NTHi host airway interplay and antibiotic resistance, which should be considered at the clinical setting for the consequences of TxS administration. 000063036 536__ $$9info:eu-repo/grantAgreement/ES/MINECO/SAF2012-31166$$9info:eu-repo/grantAgreement/ES/MINECO/SAF2015-66520-R 000063036 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/ 000063036 590__ $$a3.52$$b2017 000063036 591__ $$aMICROBIOLOGY$$b40 / 125 = 0.32$$c2017$$dQ2$$eT1 000063036 591__ $$aIMMUNOLOGY$$b64 / 155 = 0.413$$c2017$$dQ2$$eT2 000063036 592__ $$a1.703$$b2017 000063036 593__ $$aMedicine (miscellaneous)$$c2017$$dQ1 000063036 593__ $$aInfectious Diseases$$c2017$$dQ1 000063036 593__ $$aMicrobiology (medical)$$c2017$$dQ1 000063036 593__ $$aMicrobiology$$c2017$$dQ1 000063036 593__ $$aImmunology$$c2017$$dQ2 000063036 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion 000063036 700__ $$aMartí, S. 000063036 700__ $$aEuba, B. 000063036 700__ $$aFernández-Calvet, A. 000063036 700__ $$aMoleres, J. 000063036 700__ $$aLópez-López, N. 000063036 700__ $$0(orcid)0000-0002-5225-661X$$aBarberán, M.$$uUniversidad de Zaragoza 000063036 700__ $$aRamos-Vivas, J. 000063036 700__ $$aTubau, F. 000063036 700__ $$aLosa, C. 000063036 700__ $$aArdanuy, C. 000063036 700__ $$aLeiva, J. 000063036 700__ $$aYuste, J.E. 000063036 700__ $$aGarmendia, J. 000063036 7102_ $$11009$$2773$$aUniversidad de Zaragoza$$bDpto. Patología Animal$$cÁrea Sanidad Animal 000063036 773__ $$g7 (2017), 266 [19 pp]$$tFrontiers in cellular and infection microbiology$$x2235-2988 000063036 8564_ $$s2657305$$uhttps://zaguan.unizar.es/record/63036/files/texto_completo.pdf$$yVersión publicada 000063036 8564_ $$s12097$$uhttps://zaguan.unizar.es/record/63036/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada 000063036 909CO $$ooai:zaguan.unizar.es:63036$$particulos$$pdriver 000063036 951__ $$a2019-07-09-11:56:18 000063036 980__ $$aARTICLE