To use proteomic analysis to identify qualitatively and quantitatively mammalian protein components of commercial veterinary vaccines against canine distemper, leptospirosis, borreliosis, and rabies.
25 licensed veterinary vaccines (from 4 different manufacturers) against canine distemper and leptospirosis, borreliosis, and rabies (3-year and 1-year durations of immunity).
Duplicate samples from a single-lot vial of each vaccine were prepared by acetone precipitation and proteolysis with trypsin and Lys-C protease mix. Peptides mixtures (1 μg) were analyzed by liquid chromatography-tandem mass spectrometry using an Orbitrap Fusion Lumos mass spectrometer. Liquid chromatography-tandem mass spectroscopy data were searched against a Bos taurus protein database using MaxQuant to identify and quantify mammalian proteins in the vaccines. Identified proteins were classified by function and network analysis to visualize interactions.
The largest number of mammalian proteins was identified in 3-year rabies vaccines (median, 243 proteins; range, 184 to 339 proteins) and 1-year rabies vaccines (median, 193 proteins; range, 169 to 350 proteins). Borrelia and leptospirosis-distemper (L&D) vaccines had the lowest number of proteins. Rabies vaccines had the highest number of identified proteins in common (n = 316); 33 were unique to 1-year products and 44 were found in 3-year products. Borrelia and L&D vaccines had 16 and 22 uniquely identified proteins, respectively. The protein classifications were primarily modulators of protein-binding activity, enzymes, transfer-carrier proteins, cytoskeletal proteins, defense-immunity proteins, calcium-binding proteins, and extracellular matrix proteins.
This study demonstrates proteomics application to evaluate quality differences among different vaccines, identifying potential stimulants of desirable and undesirable immune responses.
Objective—To determine whether routine vaccination
induces antibodies against bovine thyroglobulin
and autoantibodies against canine thyroglobulin in
Animals—20 healthy research Beagles and 16 healthy
Procedure—For the research Beagles, 5 dogs were
vaccinated with a multivalent vaccine and a rabies
vaccine, 5 dogs received only the multivalent vaccine,
5 dogs received only the rabies vaccine, and 5 dogs
were unvaccinated controls. The multivalent vaccine
was administered at 8, 10, 12, 16, 20, 26, and 52
weeks of age and every 6 months thereafter. The
rabies vaccine was administered at 16 and 52 weeks
of age and then once per year. Blood was collected
from all dogs at 8, 16, and 26 weeks of age and then
4 times yearly. Assays for antibodies directed against
bovine and canine thyroglobulin were performed prior
to and 2 weeks after each yearly vaccination. For the
pet dogs, blood was collected prior to and 2 weeks
after 1 vaccination.
Results—In the research Beagles, there was a significant
increase in anti-bovine thyroglobulin antibodies in
all vaccinated dogs, compared with control dogs.
There was a significant increase in anti-canine thyroglobulin
antibodies in the 2 groups of dogs that
received the rabies vaccine but not in the group that
received the multivalent vaccine alone. In the pet
dogs, there was a significant increase in anti-canine
thyroglobulin antibodies after vaccination but no significant
change in anti-bovine thyroglobulin antibodies.
Conclusions and Clinical Relevance—Recent vaccination
may result in increased anti-canine thyroglobulin
antibodies. Whether these antibodies have a deleterious
effect on canine thyroid function is unknown.
(J Am Vet Med Assoc 2002;221:515–521)
Objective—To determine the effect of vaccination on
serum concentrations of total and antigen-specific IgE
Animals—20 female Beagles.
Procedure—Groups of 5 dogs each were vaccinated
repeatedly between 8 weeks and 4 years of age with
a multivalent and rabies vaccine, a multivalent vaccine
only, or a rabies vaccine only. A fourth group of 5 dogs
served as unvaccinated controls. Serum concentrations
of total immunoglobulins and antigen-specific
IgE were determined following vaccination.
Results—The multivalent vaccine had little effect
on serum total IgE concentrations. The concentration
of IgE increased slightly following vaccination
for rabies at 16 weeks and 1 year of age and
increased greatly after vaccination at 2 and 3 years
of age in most dogs, with a distinct variation
between individual dogs. Vaccination had no effect
on serum concentrations of IgA, IgG, and IgM as
measured at 2 and 3 years of age. The rabies vaccine
contained aluminum adjuvant in contrast to the
multivalent vaccine. An increase of IgE that was
reactive with vaccine antigens, including bovine
serum albumin and bovine fibronectin, was detected
in some of the dogs vaccinated for rabies. There
was no significant correlation between serum concentrations
of total IgE and antigen-specific IgE following
vaccination. Serum total IgE concentration
rapidly returned to preimmunization concentrations
in most dogs, but high concentrations of antigenspecific
Conclusions and Clinical Relevance—Vaccination of
dogs for rabies increases serum concentrations of total
IgE and induces IgE specific for vaccine antigens, including
tissue culture residues. Vaccination history should be
considered in the interpretation of serum total IgE concentrations.
(Am J Vet Res 2002;63:611–616)