Efficacy of two adjuvant systems to promote humoral immunity to the pre-proghrelin peptide obestatin in pigs: consequences for the growth of piglets to weaning

Abstract

The poor antigenicity of peptide antigens demands the selection of effective adjuvants to induce humoral immunity. The peptides obestatin and ghrelin from the pro-hormone pre-proghrelin were initially identified as antagonistic in regulating feeding behaviour, with obestatin being suppressive. The efficacy of two adjuvant systems, DEAE with the oil polysorbate emulsion of BP85:Span80 and the surfactant-oil system Montanide (ISA 50v) were therefore assessed with an obestatin-ovalbumin conjugate injected into late pregnant sows. This enabled the supply of antibodies directed against obestatin to newborn piglets through colostrum with the objective of promoting ghrelin secretion and therefore increasing feeding behaviour. Pregnant Landrace × Large White sows (n = 28) were immunised with 0.5 mg obestatin-ovalbumin in 2 mL DEAE:BP85:Span80 (DEAE; n = 14) or with 2 mL Montanide (ISA 50v: n = 14) as adjuvants at days 91 and 105 of gestation. After farrowing, piglets remained with their mothers during the lactation period and were weighed after weaning at Day 28. Antibody titres (unitless) in colostrum were assessed by ELISA as 5543 ± 2388 and 3139 ± 1151 for the DEAE and Montanide adjuvants respectively. These were associated with total IgG of 67.7 ± 3 and 82.3 ± 4.8 mg/mL respectively (P = 0.018). Piglet plasma titres were 5100 ± 1576 and 5762 ± 1688 for DEAE and Montanide respectively at Day 5 postpartum. These titres were still detectable through to Day 28 (titres of 1213 ± 389 and 665 ± 203 respectively (P = 0.176). However, sow colostral antibody titres were not related to piglet antibody concentrations on D5 (r = –0.225, P = 0.341). Sow plasma antibody titres were not related to titres at Day 28 in piglets across treatments (r = 0.198, P = 0.402). The concentration of ghrelin in colostrum was 672 ± 78 and 666 ± 39 pg/mL for the DEAE and Montanide groups, respectively, leading to piglet plasma concentrations on Day 5 of 1105 ± 164 and 530 ± 84 pg/mL (P = 0.002). Animals grew from birthweights of 1.7 ± 0.1 and 1.8 ± 0.1 (P = 0.993) to 7.7 ± 1.2 and 7.8 ± 1.0 kg (P = 0.295) at weaning, representing growth rates of 200.5 ± 52.9 and 225.5 ± 53.4 g/day (P = 0.181). There was a significant negative correlation between piglet D28 antibody titre and growth rate to weaning with the Montanide adjuvant (r = 0.116, P = 0.035) but not for the DEAE (r = –0.118, P = 0.411). Although both adjuvants were capable of generating high antibody titres, the DEAE dextran was likely to be the most effective adjuvant to induce a humoral immune response to develop further with a commercial vaccine.