Department of Medical Microbiology, Radboudumc Center for Infectious Diseases, Radboud University Medical Center , Nijmegen, The Netherlands.
Department of Pharmacy, Radboudumc Center for Infectious Diseases, Radboud University Medical Center , Nijmegen, The Netherlands.
Antimicrob Agents Chemother. 2023 Nov 15;67(11):e0087423. doi: 10.1128/aac.00874-23. Epub 2023 Oct 25.
Rifampicin is recommended for the treatment of complex pulmonary disease alongside azithromycin and ethambutol. We evaluated the azithromycin-ethambutol backbone with and without rifampicin in an intracellular hollow fiber model and performed RNA sequencing to study the differences in adaptation. In an hollow fiber experiment, we simulated epithelial lining fluid pharmacokinetic profiles of the recommended 3-drug (rifampicin, ethambutol, and azithromycin) or a 2-drug (ethambutol and azithromycin) treatment. THP-1 cells infected with ATCC700898 were exposed to these regimens for 21 days. We determined intra- and extra-cellular bacterial load- and THP-1 cell densities on days 0, 3, 7, 14, and 21, alongside RNA sequencing. The emergence of macrolide resistance was studied by inoculating intra- and extra-cellular fractions of azithromycin-containing Middlebrook 7H10 agar plates. Complete pharmacokinetic profiles were determined at days 0 and 21. Both therapies maintained stasis of both intra- and extra-cellular bacterial populations for 3 days, whilst regrowth coinciding with the emergence of a macrolide-resistant subpopulation was seen after 7 days. THP-1 cell density remained static. Similar transcriptional profiles were observed for both therapies that were minimally influenced by exposure duration. Transcriptional response was slightly larger during 2-drug treatment. Rifampicin did not add to the antimycobacterial effect to the 2-drug therapy or suppression of emergence resistance. RNA transcription was not greatly altered by the addition of rifampicin, which may be due to strong transcriptional influence of azithromycin and host cells. This questions the role of rifampicin in the currently recommended therapy. These findings should be confirmed in clinical trials.
利福平联合阿奇霉素和乙胺丁醇推荐用于治疗复杂肺部疾病。我们在细胞内中空纤维模型中评估了阿奇霉素-乙胺丁醇骨干方案联合和不联合利福平的效果，并进行 RNA 测序以研究适应性差异。在中空纤维实验中，我们模拟了推荐的三联药物（利福平、乙胺丁醇和阿奇霉素）或二联药物（乙胺丁醇和阿奇霉素）治疗的上皮衬液药代动力学特征。将 THP-1 细胞感染 ATCC700898 后，用这些方案处理 21 天。我们在第 0、3、7、14 和 21 天测定细胞内外细菌负荷和 THP-1 细胞密度，并进行 RNA 测序。通过在含有阿奇霉素的 Middlebrook7H10 琼脂平板的细胞内外部分接种来研究大环内酯类耐药的出现。在第 0 和 21 天测定完整的药代动力学特征。两种治疗方案均在第 3 天维持细胞内外细菌种群的稳定，而在第 7 天出现耐大环内酯亚群时则出现了再生长。THP-1 细胞密度保持稳定。两种治疗方案的转录谱相似，受暴露时间的影响最小。二联治疗时转录反应稍大。利福平对二联疗法的抗分枝杆菌作用或对耐药性出现的抑制作用没有帮助。添加利福平并没有大大改变 RNA 转录，这可能是由于阿奇霉素和宿主细胞的转录强烈影响。这就提出了利福平在目前推荐的治疗方案中的作用问题。这些发现应在临床试验中得到证实。
