Free Standard AU & NZ Shipping For All Book Orders Over $80!
Register      Login
Animal Production Science Animal Production Science Society
Food, fibre and pharmaceuticals from animals
RESEARCH ARTICLE

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

P. F. Geale A , P. A. Sheehy A , C. Giles A D , P. C. Thomson B C and P. C. Wynn A C E
+ Author Affiliations
- Author Affiliations

A Sydney School of Veterinary Science, University of Sydney, Werombi Road Camden, NSW 2570, Australia.

B School of Life and Environmental Sciences, University of Sydney, Werombi Road Camden, NSW 2570, Australia.

C E.H. Graham Centre for Agricultural Innovation, Charles Sturt University, Wagga Wagga, NSW 2650, Australia.

D Future Industries Institute, University of South Australia, Mawson Lakes, SA 5095, Australia.

E Corresponding author. Email: pwynn@csu.edu.au

Animal Production Science 60(3) 356-362 https://doi.org/10.1071/AN18404
Submitted: 8 July 2018  Accepted: 29 May 2019   Published: 3 December 2019

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.

Additional keywords: colostrum, lactation, obestatin antibodies, piglet growth.


References

Alloatti G, Arnoletti E, Bassino E, Penna C, Perrelli MG, Ghé C, Muccioli G (2010) Obestatin affords cardioprotection to the ischemic-reperfused isolated rat heart and inhibits apoptosis in cultures of similarly stressed cardiomyocytes. American Journal of Physiology. Heart and Circulatory Physiology 299, H470–H481.
Obestatin affords cardioprotection to the ischemic-reperfused isolated rat heart and inhibits apoptosis in cultures of similarly stressed cardiomyocytes.Crossref | GoogleScholarGoogle Scholar | 20525876PubMed |

Andrade S, Pinho F, Ribeiro AM, Carriera M, Casanueva FF, Roy P, Monteiro MP (2013) Immunization against active ghrelin using virus-like particles for obesity treatment. Current Pharmaceutical Design 19, 6551–6558.
Immunization against active ghrelin using virus-like particles for obesity treatment.Crossref | GoogleScholarGoogle Scholar | 23859551PubMed |

Annunziato F, Romagnani C, Romagnani S (2015) The 3 major types of innate and adpative cell-mediated effector immunity. The Journal of Allergy and Clinical Immunology 135, 626–635.
The 3 major types of innate and adpative cell-mediated effector immunity.Crossref | GoogleScholarGoogle Scholar | 25528359PubMed |

AOAC (1960) ‘Method (8) 15.029 Fat analysis. Official methods of analysis of AOAC International.’ (AOAC International: Rockville, MD, USA)

Ascarateil S, Puget A, Koziol M (2015) Safety data of Montanide ISA 51 VG and Montanide ISA 720 VG, two adjuvants dedicated to human therapeutic vaccines. Journal for Immunotherapy of Cancer 3, 428

Butler JE, Lager KM, Splichal I, Francis D, Kacskovics I, Sinkora M, Wertz N, Sun J, Zhao Y, Brown W, DeWald R, Dierks S, Muyldermans S, Lunney J, McCray P, Rogers CS, Welsh MJ, Navarro P, Klobasa F, Habe F, Ramsoondar J (2009) The piglet as a model for B cell and immune system development. Veterinary Immunology and Immunopathology 128, 147–170.
The piglet as a model for B cell and immune system development.Crossref | GoogleScholarGoogle Scholar | 19056129PubMed |

Chartrel N, Alvear-Perez AR, Leprince J, Iturrioz X, Reaux-Le Goazingo A, Audinot V, Chomarat P, Coge F, Nosjean O, Rodriguez M, Galizzi JP, Boutin JA, Vaudry H, Llorens-Cortes C (2007) Comment on ‘Obestatin, a peptide encoded by the ghrelin gene, opposes ghrelin’s effects on feed intake’. Science 315, 766
Comment on ‘Obestatin, a peptide encoded by the ghrelin gene, opposes ghrelin’s effects on feed intake’.Crossref | GoogleScholarGoogle Scholar | 17289961PubMed |

Coffman RL, Sher A, Seder RA (2010) Vaccine adjuvants: putting innate immunity to work. Immunity 33, 492–503.
Vaccine adjuvants: putting innate immunity to work.Crossref | GoogleScholarGoogle Scholar | 21029960PubMed |

Dunshea FR, Colantoni C, Howard K, McCauley I, Jackson PKA, Long K, Lopaticki S, Nugent EA, Simons JA, Walker J, Hennessy DP (2001) Vaccination of boars with a GnRH vaccine (Improvac) eliminates boar taint and increases growth performance. Journal of Animal Science 79, 2524–2535.
Vaccination of boars with a GnRH vaccine (Improvac) eliminates boar taint and increases growth performance.Crossref | GoogleScholarGoogle Scholar | 11721830PubMed |

Erşahin M, Özsavcı D, Şener A, Özakpınar ÖB, Toklu HZ, Akakin D, Şener G, Yeğen BÇ (2013) Obestatin alleviates subarachnoid haemorrhage-induced oxidative injury in rats via its anti-apoptotic and antioxidant effects. Brain Injury 27, 1181–1189.
Obestatin alleviates subarachnoid haemorrhage-induced oxidative injury in rats via its anti-apoptotic and antioxidant effects.Crossref | GoogleScholarGoogle Scholar | 23895491PubMed |

Fromme B, Eftkhari P, Van Reganmortel M, Hoebeke J, Katz A, Millar R (2003) A novel retro-inverse gonadotropin releasing hormone (GnRH)immunogen elicits Abs that neutralise the activity of native GnRH. Endocrinology 144, 3262–3269.

Gesmundo I, Gallo D, Favaro E, Ghigo E, Granarta R (2013) Obestatin: a new metabolic player in the pancreas and white adipose tissue. IUMB Life 65, 976–982.
Obestatin: a new metabolic player in the pancreas and white adipose tissue.Crossref | GoogleScholarGoogle Scholar |

Granata R, Baragli A, Settanni F, Scarlatti F, Ghigo E (2010) Unravelling the role of the ghrelin gene peptides in the endocrine pancreas. Journal of Molecular Endocrinology 45, 107–118.
Unravelling the role of the ghrelin gene peptides in the endocrine pancreas.Crossref | GoogleScholarGoogle Scholar | 20595321PubMed |

Granata R, Gallo D, Luque RM, Baragli A, Scarlatti F, Grande C, Gesmundo I, Córdoba-Chacón J, Bergandi L, Settanni F, Togliatto G, Volante M, Garetto S, Annunziata M, Chanclón B, Gargantini E, Rocchietto S, Matera L, Datta G, Morino M, Brizzi MF, Huy Ong H, Camussi G, Castaño JP, Papotti M, Ghigo E (2012) Obestatin regulates adipocyte function and protects against diet-induced insulin resistance and inflammation. The FASEB Journal 26, 3393–3411.
Obestatin regulates adipocyte function and protects against diet-induced insulin resistance and inflammation.Crossref | GoogleScholarGoogle Scholar | 22601779PubMed |

Gurriarán-Rodríguez U, Al-Massadi O, Roca-Rivada A, Crujeiras AB, Gallego R, Pardo M, Seoane LM, Pazos Y, Casanueva FF, Camiña JP (2011) Obestatin as a regulator of adipocyte metabolism and adipogenesis. Journal of Cellular and Molecular Medicine 15, 1927–1940.
Obestatin as a regulator of adipocyte metabolism and adipogenesis.Crossref | GoogleScholarGoogle Scholar | 21029370PubMed |

Hampe CS (2012) Protective role of anti-idiotypic antibodies in autoimmunity – lessons for type 1 diabetes. Autoimunity 45, 320–331.
Protective role of anti-idiotypic antibodies in autoimmunity – lessons for type 1 diabetes.Crossref | GoogleScholarGoogle Scholar |

Hardy CM, Braid AL (2007) Vaccines for immunological control of fertility in animals. Revue Scientifique et Technique (International Office of Epizootics) 26, 461–470.
Vaccines for immunological control of fertility in animals.Crossref | GoogleScholarGoogle Scholar |

Irvine DJ, Swartz MA, Szeto GL (2013) Engineering synthetic vaccines using cues from natural immunity. Nature Materials 12, 978–990.
Engineering synthetic vaccines using cues from natural immunity.Crossref | GoogleScholarGoogle Scholar | 24150416PubMed |

Klobasa F, Werhahn E, Butler JE (1987) Composition of sow milk during lactation. Journal of Animal Science 64, 1458–1466.
Composition of sow milk during lactation.Crossref | GoogleScholarGoogle Scholar | 3583950PubMed |

Kojima M, Hosoda H, Date Y, Nakazato M, Matsuo H, Kangawa K (1999) Ghrelin is a growth-hormone-releasing acylated peptide from stomach. Nature 402, 656–660.
Ghrelin is a growth-hormone-releasing acylated peptide from stomach.Crossref | GoogleScholarGoogle Scholar | 10604470PubMed |

Langer P (2009) Differences in the composition of colostrum and milk in eutherians reflect differences in immunoglobulin transfer. Journal of Mammalogy 90, 332–339.
Differences in the composition of colostrum and milk in eutherians reflect differences in immunoglobulin transfer.Crossref | GoogleScholarGoogle Scholar |

Li P, Wang F (2015) Polysaccharides: candidates of promising vaccine adjuvants. Drug Discoveries & Therapeutics 9, 88–93.
Polysaccharides: candidates of promising vaccine adjuvants.Crossref | GoogleScholarGoogle Scholar |

Loizou S, McCrea JD, Rudge AC, Reynolds R, Boyles CC, Harris EN (1985) Measurement of anti-cardiolipin antibodies by an enzyme linked immunosorbent assay (ELISA). Journal of clinical Experimental Immunology 62, 738–745.

Lu SC, Xu J, Chinookoswong N, Liu S, Steavenson S, Gegg C, Brankow D, Lindberg R, Veniant M, Gu W (2009) An acyl-ghrelin-specific neutralizing antibody inhibits the acute ghrelin-mediated orexigenic effects in mice. Molecular Pharmacology 75, 901–907.
An acyl-ghrelin-specific neutralizing antibody inhibits the acute ghrelin-mediated orexigenic effects in mice.Crossref | GoogleScholarGoogle Scholar | 19129426PubMed |

Meeusen ENT, Walker J, Peters A, Pastoret P-P, Jungersen G (2007) Current status of veterinary vaccines. Clinical Microbiology Reviews 20, 489–510.
Current status of veterinary vaccines.Crossref | GoogleScholarGoogle Scholar |

Monteiro M (2014) Obesity vaccines. Human Vaccines and Therapeutics 10, 1–9.

Nakazato M, Murakami N, Date Y, Kojima M, Matsuo H, Kangawa K, Matsukura S (2001) A role for ghrelin in the central regulation of feeding. Nature 409, 194–198.
A role for ghrelin in the central regulation of feeding.Crossref | GoogleScholarGoogle Scholar | 11196643PubMed |

Nogueiras R, Pfluger P, Tovar S, Arnold M, Mitchell S, Morris A, Perez-Tilve D, Vazguez MJ, Wiedmer P, Castenada TR, Dimarchi R, Tschop M, Schurmann A, Joost H-G, Williams LM, Langhans W, Dieguez C (2007) Effects of obestatin on energy balance and growth hormone secretion in rodents. Endocrinology 148, 21–26.
Effects of obestatin on energy balance and growth hormone secretion in rodents.Crossref | GoogleScholarGoogle Scholar | 17008393PubMed |

Paige CJ, Wu GE (1989) The B cell repertoire. The FASEB Journal 3, 1818–1824.
The B cell repertoire.Crossref | GoogleScholarGoogle Scholar | 2497040PubMed |

Pattison S (1994) ‘Immunomodulation of ovine adrenocortical cell function.’ (University of Sydney: Sydney)

Piedrafita D, Preston S, Kemp J, de Veer M, Sherrard J, Kraska T, Elhay M, Meeusen E (2013) The effect of different adjuvants on immune parameters and protection following vaccination of sheep with a larval-specific antigen of the gastrointestinal nematode, Haemonchus contortus. PLoS One 8, e78357
The effect of different adjuvants on immune parameters and protection following vaccination of sheep with a larval-specific antigen of the gastrointestinal nematode, Haemonchus contortus.Crossref | GoogleScholarGoogle Scholar | 24205209PubMed |

Purcell AW, McCluskey J, Rossjohn J (2007) More than one reason to rethink the use of peptides in vaccine design. Nature Reviews. Drug Discovery 6, 404–414.
More than one reason to rethink the use of peptides in vaccine design.Crossref | GoogleScholarGoogle Scholar | 17473845PubMed |

Qi X, Li L, Yang G, Liu J, Li K, Tang Y, Liou HGB (2007) Circulating obestatin levels in normal subjects and in patients with impaired glucose regulation and type 2 diabetes mellitus. Horumon To Rinsho 66, 593–597.

Ren AJ, Guo ZF, Wang YK, Lin L, Zheng X (2009) Obestatin, obesity and diabetes. Peptides 30, 439–444.
Obestatin, obesity and diabetes.Crossref | GoogleScholarGoogle Scholar | 18992781PubMed |

Rice JM, Madison RM (1972) Subcutaneous injections of vaccine adjuvant DEAE-dextran induce local sarcomas in mice. Nature: New Biology 236, 28
Subcutaneous injections of vaccine adjuvant DEAE-dextran induce local sarcomas in mice.Crossref | GoogleScholarGoogle Scholar |

Rooke JA, Bland IM (2002) The acquisition of passive immunity in the new-born piglet. Livestock Production Science 78, 13–23.
The acquisition of passive immunity in the new-born piglet.Crossref | GoogleScholarGoogle Scholar |

Spencer GS (1984) New approach to regulation of growth using immunization against somatostatin: discussion paper. Journal of the Royal Society of Medicine 77, 496–500.

Vizcarra JA, Kirby JD, Kim SK, Galyean ML (2007) Active immunization against ghrelin decreases weight gain and alters plasma concentrations of growth hormone in growing pigs. Domestic Animal Endocrinology 33, 176–189.
Active immunization against ghrelin decreases weight gain and alters plasma concentrations of growth hormone in growing pigs.Crossref | GoogleScholarGoogle Scholar | 16793235PubMed |

Vizcarra JA, Karges SL, Wetteman RP (2012) Immunization of beef heifers against gonadotropin-releasing hormone prevents luteal activity and pregnancy: effect of conjugation to different proteins and effectiveness of adjuvants. Journal of Animal Science 90, 1479–1488.
Immunization of beef heifers against gonadotropin-releasing hormone prevents luteal activity and pregnancy: effect of conjugation to different proteins and effectiveness of adjuvants.Crossref | GoogleScholarGoogle Scholar | 22147471PubMed |

Wen Y, Collier JH (2015) Supramolecular peptide vaccines: tuning adaptive immunity. Current Opinion in Immunology 35, 73–79.
Supramolecular peptide vaccines: tuning adaptive immunity.Crossref | GoogleScholarGoogle Scholar | 26163376PubMed |

Westerink MA, Smithson SL, Hutchins WA, Widera G (2001) Development and characterization of anti-idiotype based peptide and DNA vaccines which mimic the capsular polysaccharide of Neisseria meningitidis serogroup C. International Reviews of Immunology 20, 251–261.
Development and characterization of anti-idiotype based peptide and DNA vaccines which mimic the capsular polysaccharide of Neisseria meningitidis serogroup C.Crossref | GoogleScholarGoogle Scholar | 11878768PubMed |

Wortley KE, Anderson KE, Garcia K, Murray JD, Malinova I, Liu R, Moncrieffe M, Thabet K, Cox HJ, Yancopoulos GD, Wiegand SJ, Sleeman MW (2004) Genetic deletion of ghrelin does not decrease food intake but influences metabolic fuel preference Proceedings of the National Academy of Sciences of the United States of America 101, 8227–8232.
Genetic deletion of ghrelin does not decrease food intake but influences metabolic fuel preferenceCrossref | GoogleScholarGoogle Scholar | 15148384PubMed |

Wren AM, Small CJ, Ward HL, Murphy KG, Dakin CL, Taheri S, Kennedy AR, Roberts GH, Morgan DG, Ghatei MA, Bloom SR (2000) The novel hypothalamic peptide ghrelin stimulates food intake and growth hormone secretion. Endocrinology 141, 4325–4328.
The novel hypothalamic peptide ghrelin stimulates food intake and growth hormone secretion.Crossref | GoogleScholarGoogle Scholar | 11089570PubMed |

Wynn PC, Shahneh AZ, Rigby RDG, Behrendt R, Giles LR, Gooden JM, Jones MR (1995) Physiological consequences of the induction of auto-immunity to adrenocorticotropin (ACTH). Livestock Production Science 42, 247–254.
Physiological consequences of the induction of auto-immunity to adrenocorticotropin (ACTH).Crossref | GoogleScholarGoogle Scholar |

Yang Y, Heo S, Zheng J, Yun J, Shinde P, Choi J, Yang B, Chae B (2008) Effects of dietary energy and lysine intake during late gestation and lactation on blood metabolites, hormones, milk composition and reproductive performance in multiparous sows. Archives of Animal Nutrition 62, 10–21.
Effects of dietary energy and lysine intake during late gestation and lactation on blood metabolites, hormones, milk composition and reproductive performance in multiparous sows.Crossref | GoogleScholarGoogle Scholar |

Zhang JV, Ren PG, Avsian-Ketchmer O, Luo CW, Raunch R, Klein C, Hsueh AJ (2005) Obestatin, a peptide encoded by the ghrelin gene, opposes ghrelin’s effects on food intake. Science 310, 996–999.
Obestatin, a peptide encoded by the ghrelin gene, opposes ghrelin’s effects on food intake.Crossref | GoogleScholarGoogle Scholar | 16284174PubMed |

Zhu M, Wang R, Nie G (2014) Applications of nanomaterials as vaccine adjuvants. Human Vaccines & Immunotherapeutics 10, 2761–2774.
Applications of nanomaterials as vaccine adjuvants.Crossref | GoogleScholarGoogle Scholar |

Zorrilla EP, Iwasaki S, Moss JA, Chang J, Orsuji J, Inoue K, Meijler MM, Janda KD (2006) Vaccination against weight gain. Proceedings of the National Academy of Sciences of the United States of America 103, 13226–13231.
Vaccination against weight gain.Crossref | GoogleScholarGoogle Scholar | 16891413PubMed |