Heiser Axel, McCarthy Allison, Wedlock Neil, Meier Susanne, Kay Jane, Walker Caroline, Crookenden Mallory A, Mitchell Murray D, Morgan Stuart, Watkins Kate, Loor Juan J, Roche John R
AgResearch, Hopkirk Research Institute, Grasslands Research Centre, Palmerston North, New Zealand 4442.
AgResearch, Hopkirk Research Institute, Grasslands Research Centre, Palmerston North, New Zealand 4442.
J Dairy Sci. 2015 Feb;98(2):937-46. doi: 10.3168/jds.2014-8494. Epub 2014 Dec 5.
Peripartum, and especially during the transition period, dairy cows undergo dramatic physiological changes. These coincide with an increased risk of disease during the first 2 wk after calving and have been linked to dairy cows failing to achieve production as well as reproductive targets. Previous evidence suggests that these physiological changes affect the immune system and that transition dairy cows experience some form of reduced immunocompetence. However, almost all of these studies were undertaken in high-production, housed dairy cows. Grazing cows have much lower levels of production and this study aimed to provide clarity whether or not the dysfunctional attributes of the peripartum immune system reported in high production housed cows are evident in these animals. Therefore, cell culture techniques, flow cytometry, and quantitative PCR were applied to analyze the cellular composition of peripheral blood mononuclear cells from transition dairy cows as well as the performance of these cells in an in vitro assay. First, a combination of in vitro stimulation and quantitative PCR for cytokines was validated as a quantifiable immunocompetence assay in 29 cattle and a correlation of quantitative PCR and ELISA demonstrated. Second, the relative number of T helper cells, cytotoxic T cells, B cells, γδ T cells, natural killer cells, and monocytes in peripheral blood was measured, of which B cells and natural killer cells increased in number postcalving (n=29) compared with precalving. Third, following in vitro stimulation cytokine profiles indicated decreased expression of IFNγ, tumor necrosis factor, and IL-17 and increased expression of IL-10 wk 1 after calving, which later all returned to precalving values (n=39). Additionally, treatment of transition cows with a nonsteroidal anti-inflammatory drug (i.e., carprofen) administered on d 1, 3, and 5 postcalving (n=19; untreated control n=20) did not affect the cytokine expression at any time point. In conclusion, an immunocompetence assay has been developed that highlights a characteristic expression pattern for IFNγ, tumor necrosis factor, IL-17, and IL-10 that reflects a state of reduced immunocompetence in moderate-yielding pasture-based transition cows after calving, which is similar to that described for higher-yielding housed cows.
围产期,尤其是在过渡期,奶牛会经历巨大的生理变化。这些变化与产犊后前两周疾病风险增加同时出现,并与奶牛未能达到生产以及繁殖目标有关。先前的证据表明,这些生理变化会影响免疫系统,并且围产期奶牛会经历某种形式的免疫能力下降。然而,几乎所有这些研究都是在高产、圈养的奶牛身上进行的。放牧奶牛的产量要低得多,本研究旨在明确高产圈养奶牛中报道的围产期免疫系统功能失调特征在这些动物身上是否明显。因此,应用细胞培养技术、流式细胞术和定量PCR来分析围产期奶牛外周血单个核细胞的细胞组成以及这些细胞在体外试验中的表现。首先,在29头牛中验证了体外刺激和细胞因子定量PCR相结合作为一种可量化的免疫能力检测方法,并证明了定量PCR与ELISA之间的相关性。其次,测量外周血中辅助性T细胞、细胞毒性T细胞、B细胞、γδT细胞、自然杀伤细胞和单核细胞的相对数量,其中与产犊前相比,产犊后(n=29)B细胞和自然杀伤细胞数量增加。第三,体外刺激后,细胞因子谱显示产犊后第1周干扰素γ、肿瘤坏死因子和白细胞介素-17的表达降低,白细胞介素-10的表达增加,随后所有指标都恢复到产犊前水平(n=39)。此外,在产犊后第1、3和5天给围产期奶牛使用非甾体抗炎药(即卡洛芬)进行治疗(n=19;未治疗对照组n=20),在任何时间点都不影响细胞因子的表达。总之,已经开发出一种免疫能力检测方法,该方法突出了干扰素γ、肿瘤坏死因子、白细胞介素-17和白细胞介素-10的特征性表达模式,反映了中等产量的以牧场为基础的围产期奶牛产犊后免疫能力下降的状态,这与高产圈养奶牛中描述的情况相似。