Lian Xihua, Scott-Thomas Amy, Lewis John G, Bhatia Madhav, Chambers Stephen T
Department of Pathology and Biomedical Science, University of Otago, Christchurch 8140, New Zealand.
Department of Ultrasound Medicine, The Second Clinical Medical School, Fujian Medical University, Quanzhou 362000, China.
J Fungi (Basel). 2024 Dec 2;10(12):832. doi: 10.3390/jof10120832.
Due to the high morbidity and mortality rates of invasive aspergillosis (IA) and the importance of early IA detection for successful treatment and subsequent outcome, this study aimed to determine a time course of detectable antigen in a mouse model of IA and correlate it with tissue invasion by using two novel monoclonal antibodies, 1D2 and 4E4, that can be used to detect the -derived glycoproteins. Immunocompromised mice were randomly divided into five groups: uninfected control, and inoculation with conidia from , , and . Conidia (2 × 10 cells/mL) were administered intravenously via tail vein injection. Three mice from each group were euthanised at each time point (6 h, 12 h, 18 h, 24 h, and 48 h) after inoculation. Urine and blood were collected for analysis using a double-sandwich ELISA using 1D2 and 4E4. Liver, spleen, and kidney tissues were harvested for tissue staining. The levels of liver injury in the IA mice progressively increased with time after inoculation with conidia. Following inoculation with , swollen conidia were identified in the spleen, as well as antigens in blood after 18 h. Hyphae were detected in the spleen, liver, and kidney after 48 h. For the antibodies detected hyphae in the liver and spleen as well as circulating antigens in blood samples 48 h after inoculation. Tissue injury was observed in the mice inoculated with and . , but there was no evidence of fungal invasion or antigens in the blood. Antigens were not detectable in mouse urine but could be detected in glomeruli of the kidney by immunofluorescence. In conclusion, the mAb-based antigen detection double-sandwich ELISA results were consistent with the IHC results in this study. Novel monoclonal antibodies 1D2 and 4E4 can serve as tools for the early identification of IA in mice infected by and . This study also suggests the potential usefulness of this approach in human disease.
由于侵袭性曲霉病（IA）的高发病率和高死亡率，以及早期检测IA对于成功治疗及后续预后的重要性，本研究旨在通过使用两种可用于检测特定来源糖蛋白的新型单克隆抗体1D2和4E4，确定IA小鼠模型中可检测抗原的时间进程，并将其与组织侵袭相关联。免疫受损小鼠被随机分为五组：未感染对照组，以及接种来自不同菌株分生孢子的组。分生孢子（2×10个细胞/毫升）通过尾静脉注射进行静脉给药。接种后在每个时间点（6小时、12小时、18小时、24小时和48小时）对每组三只小鼠实施安乐死。收集尿液和血液，使用基于1D2和4E4的双夹心ELISA进行分析。采集肝脏、脾脏和肾脏组织进行组织染色。接种特定菌株分生孢子后，IA小鼠的肝损伤水平随时间逐渐升高。接种另一菌株后，在脾脏中发现肿胀的分生孢子，接种18小时后血液中也出现抗原。48小时后在脾脏、肝脏和肾脏中检测到菌丝。接种又一菌株后，48小时后抗体在肝脏和脾脏中检测到菌丝以及血液样本中的循环抗原。接种另外两菌株的小鼠出现组织损伤，但没有真菌侵袭或血液中抗原的证据。在小鼠尿液中未检测到抗原，但通过免疫荧光可在肾脏肾小球中检测到。总之，本研究中基于单克隆抗体的抗原检测双夹心ELISA结果与免疫组化结果一致。新型单克隆抗体1D2和4E4可作为早期鉴定受特定菌株感染小鼠中IA的工具。本研究还表明这种方法在人类疾病中的潜在用途。
