Molnár Anna Sára, Murillo-Picco Andrea, Jiménez-Fortunato Clara, Solano-Gallego Laia
Department of Immunology, Faculty of Science, Eötvös Loránd University, 1117 Budapest, Hungary.
Departament de Medicina i Cirurgia Animals, Facultat de Veterinària, Universitat Autònoma de Barcelona (UAB), 08193 Bellaterra, Spain.
Animals (Basel). 2024 Nov 30;14(23):3464. doi: 10.3390/ani14233464.
Canine leishmaniosis (CanL) is a zoonotic disease caused by , where increased interferon-gamma (IFN-γ) levels are associated with controlling the infection and mild to moderate disease. Therefore, monitoring IFN-γ concentrations is essential for monitoring the immune responses in CanL. This study compared a faster, cost-effective IFN-γ release whole blood assay in tubes (WBA-T) with a standardized version (WBA-S) in 41 dogs at different states of infection. WBA-T was performed at 24, 48, and 72 h of incubation with three conditions: blood, blood with -soluble antigen (LSA), and blood with mitogen ConA. WBA-S was performed in plates, with blood diluted and incubated for five days using the same conditions. Supernatants (WBA-S) or plasma (WBA-T) were harvested for IFN-γ measurement by ELISA. No significant differences were observed in terms of IFN-γ concentration between WBA-T and WBA-S under LSA conditions. However, the 48 h incubation period during WBA-T showed the highest median of IFN-γ concentration compared to other incubation periods and WBA-S. The IFN-γ concentrations under ConA stimulation in WBA-S were significantly higher than in WBA-T at all incubation times studied. In conclusion, WBA-T stimulated with LSA at 48 h incubation time was shown to be the most appropriate for assessing IFN-γ production.
犬利什曼病（CanL）是一种由……引起的人畜共患病，其中干扰素-γ（IFN-γ）水平升高与控制感染以及轻度至中度疾病相关。因此，监测IFN-γ浓度对于监测CanL中的免疫反应至关重要。本研究在41只处于不同感染状态的犬中，将一种更快、更具成本效益的管内IFN-γ释放全血检测法（WBA-T）与一种标准化方法（WBA-S）进行了比较。WBA-T在孵育24、48和72小时时进行，有三种条件：血液、含可溶性抗原（LSA）的血液和含丝裂原刀豆蛋白A（ConA）的血液。WBA-S在平板中进行，血液稀释后在相同条件下孵育五天。通过酶联免疫吸附测定（ELISA）收集上清液（WBA-S）或血浆（WBA-T）用于测量IFN-γ。在LSA条件下，WBA-T和WBA-S之间的IFN-γ浓度未观察到显著差异。然而，与其他孵育时间和WBA-S相比，WBA-T的48小时孵育期显示出IFN-γ浓度的中位数最高。在所有研究的孵育时间，WBA-S中ConA刺激下的IFN-γ浓度均显著高于WBA-T。总之，在48小时孵育时间用LSA刺激的WBA-T被证明最适合评估IFN-γ的产生。
