Roberto Da Costa R P, Carvalho H, Agrícola R, Alpoim-Moreira J, Martins C, Ferreira-Dias G
Escola Superior Agrária de Coimbra, 3040 Bencanta, Coimbra, Portugal.
Reprod Domest Anim. 2003 Dec;38(6):464-9. doi: 10.1046/j.0936-6768.2003.00464.x.
The purpose of this study was to evaluate different parameters of the immune status in the mare, during the follicular and the luteal phases of the oestrous cycle, in two consecutive years. Functional competence of peripheral blood neutrophils, such as chemotaxis, phagocytosis and oxidative burst was assessed under physiological cyclic conditions (Exp. I). In the second year of this study (Exp. II), besides peripheral blood neutrophil phagocytosis and oxidative burst analysis, circulating lymphocyte subsets were also characterized. The reproductive status in a total of 17 adult cycling mares was evaluated by ultrasonography and further confirmed by plasma progesterone levels. Chemotaxis tests were performed using porous membranes inserted in transwell chambers. Lipopolysaccharide (LPS) from Escherichia coli and N-formyl-Met-Leu-Phe (fMLP) were used as chemoattractants. Measurement of phagocytosis and oxidative burst in blood neutrophils were assessed by flow cytometry using commercially available kits. Quantification of T-lymphocyte subsets was assessed by indirect immunofluorescence staining after incubation with monoclonal antibodies specific for CD2, CD3, CD4 and CD8 cell markers by flow cytometry. Natural killer cells and B cells were estimated mathematically. No significant difference was found in migration, phagocytosis and oxidative burst at either phase of the oestrous cycle. Statistical analysis of total white blood cell counts also showed no significant difference between either phase of the oestrous cycle, although there was a tendency for blood neutrophils to increase in number under the progesterone influence (p = 0.09). Lymphocytic subpopulations did not differ throughout the oestrous cycle. Overall, our results suggest that luteal and follicular phases in cycling mares may not influence the immune status of the mare.
本研究的目的是连续两年评估发情周期中卵泡期和黄体期母马免疫状态的不同参数。在生理周期条件下评估外周血中性粒细胞的功能能力,如趋化性、吞噬作用和氧化爆发(实验一)。在本研究的第二年(实验二),除了分析外周血中性粒细胞的吞噬作用和氧化爆发外,还对循环淋巴细胞亚群进行了表征。通过超声检查评估了总共17匹成年发情母马的生殖状态,并通过血浆孕酮水平进一步确认。趋化性测试使用插入Transwell小室的多孔膜进行。来自大肠杆菌的脂多糖(LPS)和N-甲酰甲硫氨酸亮氨酸苯丙氨酸(fMLP)用作趋化剂。使用市售试剂盒通过流式细胞术评估血液中性粒细胞的吞噬作用和氧化爆发。通过与针对CD2、CD3、CD4和CD8细胞标志物的单克隆抗体孵育后,通过间接免疫荧光染色和流式细胞术评估T淋巴细胞亚群的定量。通过数学方法估算自然杀伤细胞和B细胞。在发情周期的任何一个阶段,迁移、吞噬作用和氧化爆发均未发现显著差异。对白细胞总数的统计分析也表明,发情周期的任何一个阶段之间均无显著差异,尽管在孕酮影响下血液中性粒细胞数量有增加的趋势(p = 0.09)。淋巴细胞亚群在整个发情周期中没有差异。总体而言,我们的结果表明,发情母马的黄体期和卵泡期可能不会影响母马的免疫状态。