Characterisation of a clinical isolated Aspergillus lentulus strain using a Galleria mellonella infection model.
Characterisation of a clinical isolated Aspergillus lentulus strain using a Galleria mellonella infection model.
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使用大蜡螟感染模型表征临床分离的迟缓曲霉菌株
DOI：
10.21037/jtd-20-961
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发表时间：
2021-03
影响因子：
2.5
通讯作者：
Abliz P
中科院分区：
文献类型：
--
作者：
Zhang LJ;Wang XD;Ji MS;Hasimu H;Abliz P
近年来，由香曲霉（aspergillus lentulus， A. lentulus）引起的侵袭性曲霉感染病例逐渐增多，这种真菌在形态上通常难以与烟曲霉（aspergillus fumigatus）区分。这些都对香菇侵袭性真菌感染的治疗提出了很大的挑战。本研究旨在探讨慢性阻塞性肺疾病（COPD）患者分离的香菇A. lentulus幼虫感染后抗真菌耐药性、毒力及炎症因子的变化，以反映宿主的免疫反应。采用形态学和分子生物学方法对慢阻肺患者分离的香菇进行鉴定，并测定其体外药敏。采用香菇侵染幼虫模型，测定香菇毒力因子及炎症反应相关因子。将其分为3组：香菇标准株、患者分离的香菇菌株；PBS的控制。将浓度为1×106 CFU的香菇菌悬液注射至幼虫体内形成感染模型，采用RT-PCR法测定(1,3)-β- d -葡聚糖和半乳甘露聚糖水平，并测定mellonella幼虫体内caspase-1和TNF-α浓度。结果表明，香菇对伊曲康唑、伏立康唑和米卡芬净均有良好的敏感性，而(1,3)-β- d -葡聚糖在两组均为阴性。两组患者半乳甘露聚糖水平均高于对照组，差异有统计学意义（P<0.05）。标准应变组与患者应变组比较，差异无统计学意义（P < 0.05）。幼虫感染后，两组大黄蜂幼虫体内caspase-1、TNF-α水平升高，且两组差异均有统计学意义（P<0.05）。但两组间差异无统计学意义（P < 0.05）。从慢性阻塞性肺病患者中分离出的香菇单胞菌与标准菌株在毒力因子和宿主炎症反应方面无显著差异。半乳甘露聚糖在早期检测香菇芽孢杆菌侵袭性感染方面更有优势。此外，caspase-1介导的炎性体途径可能参与了宿主对香菇的免疫反应。
In recent years, the number of invasive aspergillus infection cases caused by Aspergillus lentulus (A. lentulus) has gradually increased and this fungus is usually difficult to distinguish from Aspergillus fumigatus in morphology. All of these presents a great challenge to the treatment of invasive fungal infections caused by A. lentulus. The present study aims to discuss the antifungal resistance, virulence and inflammatory factors’ changes after the infection of larvae of A. lentulus separated from patients with chronic obstructive pulmonary disease (COPD) to reflect the host immune response. A. lentulus isolated from COPD patients was identified by morphology and molecular biology, and its drug sensitivity was determined in vitro. Then the virulence factors and inflammatory response related factors of A. lentulus were determined by the model of A. lentulus infecting larvae. These were divided into three groups: A. lentulus standard strain, A. lentulus strain isolated from patients; PBS control. The infection model was formed by injecting the suspension of A. lentulus at a concentration of 1×106 CFU into larvae, in order to determine the (1,3)-β-D-glucan and galactomannan levels, and determine the caspase-1 and TNF-α concentration in Galleria mellonella larvae by RT-PCR. The results revealed that A. lentulus had good sensitivity to itraconazole, voriconazole and micafungin, while (1,3)-β-D-glucan was negative in the two groups. The level of galactomannan in the two groups was higher than that in the control group, and the difference was statistically significant (P<0.05). However, there was no statistical difference between the standard strain group and patient strain group (P>0.05). After the infection of larvae, caspase-1 and TNF-α in the Galleria mellonella larvae increased in the two groups, and these elevated levels were statistically significant in both groups (P<0.05). However, there was no significant difference between the two groups (P>0.05). There is no significant difference in virulence factor and host inflammatory response between A. lentulus isolated from COPD patients and standard strains. Galactomannan has more advantages in the early detection of A. lentulus invasive infection. Furthermore, the caspase-1-mediated inflammasome pathway may be involved in the host immune response to A. lentulus.