Focussing in the highly elevated IL-6 amounts in HP and LP offspring 3 h after LPS injection, these piglets seem to be more vunerable to bacterial endotoxin exposure with an elevated threat of an acute systemic inflammatory response syndrome [58]. replies of offspring to difficult circumstances or immune system problems afterwards in lifestyle. In the present study, we used our recently developed model of intrauterine growth restriction in which dietary protein to carbohydrate ratios in pregnant primiparous sows EPZ004777 are modulated [15]. The objective of this experiment was to investigate the effects of low (6.5%) and high (30%) protein:carbohydrate ratios in the diet of sows throughout pregnancy on the immune system of their offspring at different ages. To this end, the impact of the maternal diet on cortisol, protein and immunoglobulin levels in sow EPZ004777 blood over the course of pregnancy was determined, and the immune systems of piglets were evaluated by measuring total serum protein and immunoglobulin levels (IgG, IgM, IgA), lymphocyte proliferation, proportions of circulating lymphocyte subpopulations and cytokine levels at baseline and under challenging conditions (weaning and LPS administration). The intensity of the challenge was also measured by determining the response of the HPA axis. Methods All procedures including use and treatment of animals were in accordance with the German animal protection law and approved by the relevant authorities (Landesamt fr Landwirtschaft, Lebensmittelsicherheit und Fischerei Mecklenburg-Vorpommern, Germany; LVL M-V/TSD/7221.3-1.1-006/04; LALLF M-V/TSD/7221.3-1.2-05/06; LALLF M-V/TSD/7221.3-1.2-013/06). The present experimental investigation is part of a comprehensive study recently described by Rehfeldt et al. [15]. Animals and treatments A total of 42 primiparous German Landrace sows EPZ004777 and their litters, bred and raised in the experimental pig unit of our institute, were used for the experiment with 6 independent replicates. Housing and breeding management were as recently described EPZ004777 in detail [15]. The sows were fed an isoenergetic corn-barley and soybean meal diet (~13.7 MJ ME/kg) containing an adequate (AP, 12.1%; n?=?13), a low (LP, 6.5%; n?=?15) or a high (HP, 30%; n?=?14) protein level corresponding to protein:carbohydrate ratios of 1 1:5, 1:10.4, and 1:1.3, respectively, throughout gestation [15]. Diets were fed between 2.3 and 2.9 kg/d from early to late pregnancy to achieve an average target energy intake of ~34 MJ ME/d during gestation following the recommendations for primiparous sows [27]. The sows were fed twice daily, and water was provided for 15 min at 4C to separate plasma, which was analysed for cortisol. Whole blood samples were allowed to clot for 4 h at room temperature and centrifuged at 1000??for 15 min at 4C to obtain serum for analyses of total protein and immunoglobulins IgG, IgA and IgM. Plasma and serum samples were stored at ?20C until analysis. Litter size, piglets born alive and dead, individual birth weights and Rabbit Polyclonal to PDZD2 sex of piglets were recorded at birth. Runt piglets weighing less than 800 g were excluded from further experiments. From each experimental litter, usually three to four piglets (the lightest one, 1C2 of medium weight, the heaviest one) were sampled between 24 to 36 h after birth (day 1 (D1): AP, n?=?43; LP, n?=?51; HP, n?=?48). Three to four other piglets, in single cases only two piglets of each litter, were randomly assigned for sampling on D27 (AP, n?=?42; LP, n?=?46; HP, n?=?41), and EPZ004777 the remaining piglets were sampled on D80 (AP, n?=?24; LP, n?=?25; HP, n?=?20) or D180 (AP, n?=?23; LP, n?=?24; HP, n?=?21). Sex was almost equally distributed within diets. The timeline for blood sampling and distribution of animals in the different diet groups are summarised in Figure?1. Open in a separate window Figure 1 Timeline: summary of blood sampling and distribution of animals in different diet groups. Sows were fed adequate (AP, 12.1%), low (LP, 6.5%) and high (HP, 30%) dietary protein levels throughout gestation. Piglets were cross-fostered within 48 h after birth to multiparous sows fed a standard diet during pregnancy (Provital RF R.324.0; Trede & Pein, Dammfleth, Germany). The litters were standardised to 11 piglets with piglets from foster sows. After birth, experimental sows and foster sows were fed a single standard lactation diet (Provital LAC R.325.0; Trede & Pein, Dammfleth, Germany). Male piglets were castrated at four days of age. The piglets were weaned at D28 and housed in groups of four littermates per pen (2.5 m x 1.8 m) in a post-weaning room until D32. Thereafter, they were transferred to single-housing cages.