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Evaluation of recombinant Mycoplasma hyopneumoniae P97/P102 paralogs formulated with selected adjuvants as vaccines against mycoplasmal pneumonia in pigs.

Vaccine (2014-06-17)
Lauren K Woolley, Shayne A Fell, Jocelyn R Gonsalves, Benjamin B A Raymond, Damian Collins, Tracey A Kuit, Mark J Walker, Steven P Djordjevic, Graeme J Eamens, Cheryl Jenkins
ABSTRACT

Pig responses to recombinant subunit vaccines containing fragments of eight multifunctional adhesins of the Mycoplasma hyopneumoniae (Mhp) P97/P102 paralog family formulated with Alhydrogel(®) or Montanide™ Gel01 were compared with a commercial bacterin following experimental challenge. Pigs, vaccinated intramuscularly at 9, 12 and 15 weeks of age with either of the recombinant formulations (n=10 per group) or Suvaxyn(®) M. hyo (n=12), were challenged with Mhp strain Hillcrest at 17 weeks of age. Unvaccinated, challenged pigs (n=12) served as a control group. Coughing was assessed daily. Antigen-specific antibody responses were monitored by ELISA in serum and tracheobronchial lavage fluid (TBLF), while TBLF was also assayed for cytokine responses (ELISA) and bacterial load (qPCR). At slaughter, gross and histopathology of lungs were quantified and damage to epithelial cilia in the porcine trachea was evaluated by scanning electron microscopy. Suvaxyn(®) M. hyo administration induced significant serological responses against Mhp strain 232 whole cell lysates (wcl) and recombinant antigen F3P216, but not against the remaining vaccine subunit antigens. Alhydrogel(®) and Montanide™ Gel01-adjuvanted antigen induced significant antigen-specific IgG responses, with the latter adjuvant eliciting comparable Mhp strain 232 wcl specific IgG responses to Suvaxyn(®) M. hyo. No significant post-vaccination antigen-specific mucosal responses were detected with the recombinant vaccinates. Suvaxyn(®) M. hyo was superior in reducing clinical signs, lung lesion severity and bacterial load but the recombinant formulations offered comparable protection against cilial damage. Lower IL-1β, TNF-α and IL-6 responses after challenge were associated with reduced lung lesion severity in Suvaxyn(®) M. hyo vaccinates, while elevated pathology scores in recombinant vaccinates corresponded to cytokine levels that were similarly elevated as in unvaccinated pigs. This study highlights the need for continued research into protective antigens and vaccination strategies that will prevent Mhp colonisation and establishment of infection.

MATERIALS
Product Number
Brand
Product Description

Supelco
Hexamethyldisilazane, for GC derivatization, LiChropur, ≥99.0% (GC)
Sigma-Aldrich
Hexamethyldisilazane, produced by Wacker Chemie AG, Burghausen, Germany, ≥97.0% (GC)
Sigma-Aldrich
Carbon nanofibers, graphitized (iron-free), composed of conical platelets, D × L 100 nm × 20-200 μm
Sigma-Aldrich
Carbon nanofibers, graphitized, platelets(conical), >98% carbon basis, D × L 100 nm × 20-200 μm
Supelco
N,O-Bis(trimethylsilyl)acetamide, for GC derivatization, LiChropur, ≥98.5% (GC)
Carbon - Vitreous, rod, 200mm, diameter 1.0mm, glassy carbon
Carbon - Vitreous, foil, 100x100mm, thickness 2.0mm, glassy carbon
Carbon - Vitreous, foil, 8x8mm, thickness 0.5mm, glassy carbon
Carbon - Vitreous, foil, 100x100mm, thickness 1.0mm, glassy carbon
Carbon - Vitreous, rod, 200mm, diameter 10mm, glassy carbon
Carbon - Vitreous, foil, 100x100mm, thickness 6.0mm, glassy carbon
Carbon - Vitreous, foil, 10mm disks, thickness 0.2mm, glassy carbon
Carbon - Vitreous, foam, 150x150mm, thickness 2.5mm, bulk density 0.05g/cm3, porosity 96.5%
Carbon - Vitreous, rod, 200mm, diameter 3.0mm, glassy carbon
Carbon - Vitreous, rod, 200mm, diameter 5.0mm, glassy carbon
Carbon - Vitreous, foil, 8mm disks, thickness 1.0mm, glassy carbon
Carbon - Vitreous, rod, 200mm, diameter 7.0mm, glassy carbon
Carbon - Vitreous, foil, 10x10mm, thickness 4.0mm, glassy carbon
Carbon - Vitreous, foil, 50x50mm, thickness 4.0mm, glassy carbon
Carbon - Vitreous, rod, 100mm, diameter 1.0mm, glassy carbon
Carbon - Vitreous, foam, 275x330mm, thickness 3.2mm, bulk density 0.05g/cm3, porosity 96.5%
Carbon - Vitreous, rod, 50mm, diameter 1.0mm, glassy carbon
Carbon - Vitreous, foam, 275x330mm, 0.05g.cmué, porosity 96.5%, 24 pores/cm
Carbon - Vitreous, foam, 150x150mm, 0.05g.cmué, porosity 96.5%, 24 pores/cm
Carbon - Vitreous, rod, 5mm, diameter 3.0mm, glassy carbon
Carbon - Vitreous, foil, 25x25mm, thickness 0.5mm, glassy carbon
Carbon - Vitreous, foam, 300x300mm, thickness 20mm, bulk density 0.05g/cm3, porosity 96.5%
Carbon - Vitreous, foil, 10x10mm, thickness 1.0mm, glassy carbon
Carbon - Vitreous, tube, 100mm, outside diameter 10mm, inside diameter 3mm, wall thickness 3.5mm, glassy carbon
Carbon - Vitreous, foil, 25x25mm, thickness 4.0mm, glassy carbon