Szczotka Maria, Kuźmak Jacek
Department of Biochemistry, National Veterinary Research Institute, 24-100 Puławy, Poland.
J Vet Res. 2024 Mar 23;68(1):19-33. doi: 10.2478/jvetres-2024-0012. eCollection 2024 Mar.
Bovine leukaemia virus (BLV) is a Deltaretrovirus responsible for enzootic bovine leukosis, the most common neoplastic disease of cattle. It deregulates the immune system, favouring secondary infections and changes in the blood and lymphatic tissues. Blood homeostasis depends on functional haematopoietic stem cells (HSCs). Bone marrow is populated by these cells, which express CD34 and CD35 surface antigens and produce and release cytokines involved in the maintenance of haematopoiesis. The aim of the study was determination of the profile of cytokine production by CD34 stem cells of cattle naturally infected with BLV.
The HSCs were generated from the blood and lymphoid organs of cows infected with BLV and healthy control cows with immunomagnetic separation and anti-CD34 monoclonal antibodies. Isolated CD34 cells were cultivated for two weeks with interleukin (IL)-4 and granulocyte-macrophage colony-stimulating factor. The levels of IL-6, IL-10, IL-12p40, IL-12p70, interferon gamma (IFN-γ) and tumour necrosis factor alpha (TNF-α) were determined in culture fluid by flow cytometry.
The expression of IL-6, IL-12p70 and TNF-α in blood HSCs was higher in BLV cows than in the control animals. In bone marrow HSCs of infected cows, IL-12, TNF-α and IFN-γ were more concentrated, but in these cows' spleen HSCs only expression of IL-10 was elevated. In HSCs isolated from the lymph nodes of leukaemic cows, only TNF-α secretion was lower than in control cows, the other cytokines being more potently secreted.
Infection with BLV caused statistically significant differences in cytokine expression by HSC CD34 cells.
牛白血病病毒(BLV)是一种δ逆转录病毒,可引发地方流行性牛白血病,这是牛最常见的肿瘤性疾病。它会破坏免疫系统,助长继发性感染以及血液和淋巴组织的变化。血液稳态依赖于功能性造血干细胞(HSC)。骨髓中存在这些细胞,它们表达CD34和CD35表面抗原,并产生和释放参与维持造血作用的细胞因子。本研究的目的是确定自然感染BLV的牛的CD34干细胞产生细胞因子的情况。
通过免疫磁珠分离法和抗CD34单克隆抗体,从感染BLV的奶牛和健康对照奶牛的血液及淋巴器官中分离出造血干细胞。分离出的CD34细胞用白细胞介素(IL)-4和粒细胞-巨噬细胞集落刺激因子培养两周。通过流式细胞术测定培养液中IL-6、IL-10、IL-12p40、IL-12p70、干扰素γ(IFN-γ)和肿瘤坏死因子α(TNF-α)的水平。
BLV感染奶牛血液造血干细胞中IL-6、IL-12p70和TNF-α的表达高于对照动物。在感染奶牛的骨髓造血干细胞中,IL-12、TNF-α和IFN-γ更为集中,但在这些奶牛的脾脏造血干细胞中,只有IL-10的表达升高。在从白血病奶牛淋巴结分离出的造血干细胞中,只有TNF-α的分泌低于对照奶牛,其他细胞因子的分泌更为强烈。
感染BLV导致造血干细胞CD34细胞的细胞因子表达存在统计学上的显著差异。