Kirchner Manuela, Daugschies Arwid, Delling Cora
Institute of Parasitology, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Sachsen, Germany.
Parasit Vectors. 2025 Jul 12;18(1):281. doi: 10.1186/s13071-025-06926-5.
The two intestinal protozoan parasites Giardia duodenalis and Cryptosporidium parvum cause infections in a wide spectrum of vertebrates and have also been shown to infect suitable hosts simultaneously. To investigate potential effects between these parasites and on host cells, a co-infection model with IPEC-J2 cells was established.
Optimal infection conditions and several infection doses of both parasites were tested. The effect of Giardia growth medium on IPEC-J2 cells was analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay, while the effect of different infection doses of each parasite on host cell viability was investigated by CellTiter Blue cell viability assay. For co-infection, IPEC-J2 cells were first infected with C. parvum sporozoites, and 3.5 h later, G. duodenalis trophozoites were added. Parasite propagation during single infection and co-infection were analyzed by quantitative real-time polymerase chain reaction (qPCR) as well as immunofluorescent staining.
The infection with C. parvum sporozoites had no significant impact on cell viability, while G. duodenalis trophozoites affected cell culture in a dose dependent manner. The amount of gene copies of C. parvum in single and co-infected cells did not differ significantly, while statistically higher amounts of G. duodenalis gene copies in co-infected cell cultures were identified.
In this study, single infections and co-infections of IPEC-J2 cells with C. parvum and G. duodenalis were established and optimized over a period of 72 h.
两种肠道原生动物寄生虫，即十二指肠贾第虫和微小隐孢子虫，可感染多种脊椎动物，并且已证实它们也能同时感染合适的宿主。为了研究这些寄生虫之间以及对宿主细胞的潜在影响，建立了IPEC-J2细胞的共感染模型。
测试了两种寄生虫的最佳感染条件和几种感染剂量。使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）还原试验分析贾第虫生长培养基对IPEC-J2细胞的影响，同时通过CellTiter Blue细胞活力测定法研究每种寄生虫不同感染剂量对宿主细胞活力的影响。对于共感染，IPEC-J2细胞首先用微小隐孢子虫子孢子感染，3.5小时后加入十二指肠贾第虫滋养体。通过定量实时聚合酶链反应（qPCR）以及免疫荧光染色分析单感染和共感染期间的寄生虫增殖情况。
微小隐孢子虫子孢子感染对细胞活力没有显著影响，而十二指肠贾第虫滋养体以剂量依赖方式影响细胞培养。单感染和共感染细胞中微小隐孢子虫的基因拷贝数没有显著差异，而在共感染细胞培养物中，十二指肠贾第虫基因拷贝数在统计学上更高。
在本研究中，建立并优化了IPEC-J2细胞与微小隐孢子虫和十二指肠贾第虫的单感染和共感染模型，持续72小时。
