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Structural characteristics of circulating immune complexes in calves with bronchopneumonia: Impact on the quiescent leukocytes.
Research in Veterinary Science ( IF 2.4 ) Pub Date : 2020-09-09 , DOI: 10.1016/j.rvsc.2020.09.004
Marijana Kovačić 1 , Natalija Fratrić 2 , Aleksandra Arsić 1 , Slavko Mojsilović 1 , Ivana Drvenica 1 , Dragana Marković 1 , Irina Maslovarić 1 , Jelica Grujić Milanović 1 , Milan Ivanov 1 , Vesna Ilić 1
Affiliation  

Calf bronchopneumonia is accompanied by increased level of circulating immune complexes (CIC), and we analysed size, and protein and lipid constituents of these CIC with an attempt to elucidate the connection between the CIC structural properties and their capacity to modulate leukocyte function.

CIC of heathy calves (CICH) and calves with naturally occurring bronchopneumonia (CICD) were isolated by PEG precipitation and analysed by electrophoresis and chromatography. The predominant CIC proteins were IgG, albumin, and transferrin. Affinity isolated serum and CIC IgG coprecipitated several proteins, but only 75 and 80 kDa proteins bound CIC IgG, exclusively. 60 and 65 kDa proteins co-precipitated with CICD IgG, unlike CICH IgG. In both CICH and CICD, oleic acid-containing phospholipids predominated. In CICD, the content of oleic and vaccenic acid was higher than in CICH, while myristic, palmitic, stearic, linoleic and arachidonic acid showed lower content. Dynamic light scattering displayed difference in particle size distribution between CICH and CICD; 1280 nm large particles were present only in CICD.

The effect of CICH and CICD on mononuclear cells (MNC) and granulocytes was analysed in vitro. CICH and CICD, with slight difference in intensity, stimulate MNC apoptosis, promote cell cycle arrest of unstimulated MNC, and cell cycle progression of PHA stimulated MNC. Both CIC reduced granulocyte apoptosis after 24 h while after 48 h this effect was detected for CICD only.

These results indicate that structural differences of CICH and CICD might interfere with the CIC functional capacity, which we consider important for evaluation of CIC immunoregulatory function.



中文翻译:

犊牛支气管肺炎中循环免疫复合物的结构特征:对静态白细胞的影响。

小牛支气管肺炎伴随循环免疫复合物(CIC)水平升高,我们分析了这些CIC的大小,蛋白质和脂质成分,试图阐明CIC结构性质与其调节白细胞功能的能力之间的联系。

通过PEG沉淀分离出健康小牛的CIC(CIC H)和具有天然支气管肺炎的小牛(CIC D),并通过电泳和色谱法进行分析。主要的CIC蛋白是IgG,白蛋白和转铁蛋白。亲和分离的血清和CIC IgG共沉淀了几种蛋白质,但只有75和80 kDa的蛋白质仅与CIC IgG结合。与CIC H IgG不同,60和65 kDa的蛋白质与CIC D IgG共沉淀。在CIC H和CIC D中,含油酸的磷脂占主导地位。CIC D中油酸和痘苗酸的含量高于CIC H中,而肉豆蔻酸,棕榈酸,硬脂酸,亚油酸和花生四烯酸的含量较低。动态光散射显示了CIC H和CIC D之间的粒度分布差异; 仅CIC D中存在1280 nm大颗粒。

体外分析了CIC H和CIC D对单核细胞(MNC)和粒细胞的影响。CIC H和CIC D的强度略有不同,可刺激MNC凋亡,促进未受刺激的MNC的细胞周期停滞,以及PHA刺激的MNC的细胞周期进程。两种CIC均在24小时后降低了粒细胞凋亡,而48小时后仅对CIC D检测到了这种作用。

这些结果表明,CIC H和CIC D的结构差异可能会干扰CIC的功能,我们认为这对于CIC免疫调节功能的评估很重要。

更新日期:2020-09-15
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