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一种新型体外培养系统的建立

The Establishment of a Novel In Vitro System for Culturing .

作者信息

Weerarathne Pabasara, Reichard Mason, Miller Craig, Scimeca Ruth C

机构信息

Department of Veterinary Pathobiology, College of Veterinary Medicine, Oklahoma State University, Stillwater, OK 74078, USA.

出版信息

Pathogens. 2024 Jul 4;13(7):565. doi: 10.3390/pathogens13070565.

DOI:10.3390/pathogens13070565
PMID:39057792
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11279574/
Abstract

Cytauxzoonosis, a highly fatal tick-borne disease in domestic cats caused by , poses diagnostic and therapeutic challenges due to the inability to culture the parasite in vitro. This study aimed to artificially replicate infection and characterize in vitro replication kinetics. Concanavalin A-activated feline embryonal macrophages (Fcwf-4) were plated at 3-5 × 10 cells/mL and incubated with -positive blood samples from either a (1) chronically infected bobcat (), (2) chronically infected domestic cat, or (3) acutely infected domestic cat with clinical signs of cytauxzoonosis. Temporal changes in parasite load were quantified by droplet digital PCR (ddPCR), and the inhibition of infection/replication was assessed using atovaquone, imidocarb dipropionate (ID), artemisinin, ponazuril, and neutralizing antibodies. Tick cell lines AAE2 and ISE6 were also tested for infection. In vitro inoculation with chronic infection led to transient replication, while acute infection resulted in sustained replication beyond 10 days post-inoculation. Atovaquone, ID, and artemisinin inhibited replication, and neutralizing antibodies prevented infection. The inoculation of tick cells in vitro indicated infection; however, parasite replication was not observed. The results of this study established an in vitro model for studying infection dynamics, assessing therapy efficacy, and testing vaccination strategies in cytauxzoonosis-infected cats.

摘要

嗜吞噬细胞无形体病是由[具体病原体未给出]引起的家猫中一种高度致命的蜱传疾病，由于无法在体外培养该寄生虫，给诊断和治疗带来了挑战。本研究旨在人工复制[具体病原体未给出]感染并表征其体外复制动力学。将伴刀豆球蛋白A激活的猫胚胎巨噬细胞（Fcwf-4）以3 - 5×10个细胞/毫升接种，并用来自以下样本的[具体病原体未给出]阳性血液样本进行孵育：（1）慢性感染的短尾猫；（2）慢性感染的家猫；或（3）具有嗜吞噬细胞无形体病临床症状的急性感染家猫。通过液滴数字PCR（ddPCR）定量寄生虫载量的时间变化，并使用阿托伐醌、双丙酸咪唑苯脲（ID）、青蒿素、硝唑尼特和中和抗体评估感染/复制的抑制情况。还对蜱细胞系AAE2和ISE6进行了感染测试。慢性感染的体外接种导致短暂复制，而急性感染导致接种后10天以上持续复制。阿托伐醌、ID和青蒿素抑制复制，中和抗体预防感染。体外接种蜱细胞表明发生了感染；然而，未观察到寄生虫复制。本研究结果建立了一个体外模型，用于研究嗜吞噬细胞无形体病感染猫的感染动态、评估治疗效果和测试疫苗接种策略。

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