Veterinary Physiology, Vetsuisse Faculty University of Bern, Bremgartenstrasse 109a, CH-3001 Bern, Switzerland.
J Dairy Sci. 2013 Feb;96(2):889-96. doi: 10.3168/jds.2012-5410. Epub 2012 Dec 6.
During a mammary immune response, the integrity of the blood-milk barrier is negatively affected and becomes leaky. The aim of the present study was to demonstrate the blood origin, and to investigate changes in the concentration, of various constituents including immunoglobulins in blood and milk during the early phase of lipopolysaccharide (LPS)-induced mastitis. Five lactating dairy cows received continuous β-hydroxybutyrate (BHBA) clamp infusions to maintain elevated BHBA blood concentrations (1.5 to 2.0 mmol/L) from 48 h before and 8h after LPS administration. One udder quarter was infused with 200 μg of Escherichia coli LPS. A second quarter served as control. Milk and blood samples were taken hourly for 8h postchallenge (PC). The somatic cell count in LPS-challenged quarters was increased from 4h PC to the end of the experiment compared with control quarters. In LPS-challenged quarters, l-lactate, BHBA, lactate dehydrogenase (LDH), IgG(1), and IgG(2) were increased at 3h PC and remained elevated until the end of experiment (8h PC) compared with control quarters. In addition, the optical density values in milk in a nonquantitative ELISA for antibodies directed against bluetongue virus (used as a measure of nonspecific antibody transfer; all animals were vaccinated) increased and, thus, indicates an increase in these antibodies in response to LPS treatment. l-Lactate concentration also increased in blood 2h PC and in the milk of control quarters during the experiment from 3h PC. A second experiment was conducted in vitro to investigate a possible contribution from destructed milk cells to l-lactate concentration and activity of LDH in milk. Aliquots of milk samples (n=8) were frozen (-20°C) or disrupted with ultrasound, respectively. Freeze thawing and ultrasound treatment increased LDH in milk samples, but had no effect on l-lactate concentrations. Results suggest that intramammary infusion of LPS induces a systemic response, as evidenced by an elevation of blood l-lactate concentration. The concomitant changes of all investigated components suggest that they were blood derived. However, the increase in blood components in the milk is not necessarily supportive of the mammary immune system, and likely a side effect of reduced blood-milk barrier integrity.
在乳腺免疫反应过程中,血乳屏障的完整性受到负面影响,变得易渗透。本研究的目的是证明血液来源,并研究脂多糖(LPS)诱导乳腺炎早期阶段各种成分(包括免疫球蛋白)在血液和牛奶中的浓度变化。 给五只泌乳奶牛连续输注β-羟基丁酸(BHBA),以在 LPS 给药前 48 小时至给药后 8 小时内维持升高的 BHBA 血液浓度(1.5 至 2.0 mmol/L)。一个乳房象限用 200 μg 大肠杆菌 LPS 输注。第二个象限作为对照。攻毒后每小时采集一次牛奶和血液样本,共 8 小时。与对照象限相比,LPS 攻毒象限中的体细胞计数从攻毒后 4 小时开始增加,直至实验结束。在 LPS 攻毒象限中,l-乳酸、BHBA、乳酸脱氢酶(LDH)、IgG(1)和 IgG(2)在 3 小时攻毒时增加,并保持升高直到实验结束(8 小时攻毒)与对照象限相比。此外,用于测量非特异性抗体转移的抗蓝舌病毒非定量 ELISA 中的牛奶光密度值增加,表明这些抗体因 LPS 处理而增加。在攻毒后 2 小时,血液中 l-乳酸浓度也增加,并且在实验过程中,从 3 小时攻毒开始,对照象限的牛奶中 l-乳酸浓度也增加。进行了第二项体外实验,以研究受损乳细胞对牛奶中 l-乳酸浓度和 LDH 活性的可能贡献。分别将牛奶样本等分试样(n=8)冷冻(-20°C)或用超声破坏。冷冻解冻和超声处理增加了牛奶样品中的 LDH,但对 l-乳酸浓度没有影响。结果表明,乳内输注 LPS 会引起全身反应,这表现为血液 l-乳酸浓度升高。所有研究成分的伴随变化表明它们是血液来源的。然而,血液成分在牛奶中的增加不一定支持乳腺免疫系统,很可能是血乳屏障完整性降低的副作用。