Objective—To develop mouse monoclonal and rabbit
polyclonal antibodies against immunoglobulin of
Argentine boa constrictors and to demonstrate the
ability of these reagents to detect antibody responses
in boa constrictors by use of an ELISA and western
Procedure—Boa constrictors were immunized with
2,4-dinitrophenylated bovine serum albumin (DNP-BSA).
Each snake received biweekly inoculations of
250 µg of DNP-BSA (half SC, half IP) for a total of 6
inoculations followed by monthly inoculations for 3
months. Preimmune blood samples were collected.
Subsequently, blood was collected immediately prior
to each booster inoculation. Anti-DNP antibodies
were isolated from immune plasma samples by affinity
chromatography. Affinity-purified boa anti-DNP
immunoglobulin was used for production of polyclonal
and monoclonal antibodies. An ELISA and western
blot analysis were used to monitor immune responses,
for purification of boa anti-DNP immunoglobulin,
and for assessment of polyclonal and monoclonal
Results—A 6-fold increase in optical density (OD405)
of immune boa plasma, compared with preimmune
plasma, was detected by the polyclonal antibody, and
a 12- and 15-fold increase was detected by monoclonal
antibodies HL1787 and HL1785, respectively,
between weeks 4 and 8. Results of western blot
analysis confirmed anti-DNP antibody activity in
immunized boa plasma and in affinity column eluates.
Polyclonal and monoclonal antibodies detected specific
anti-DNP antibody responses in immunized boas.
Conclusions and Clinical Relevance—Polyclonal
and monoclonal antibodies recognized boa constrictor
immunoglobulin. These antibodies may be useful in
serologic tests to determine exposure of snakes to
pathogens. (Am J Vet Res 2003;64:388–395)
Objective—To evaluate the safety and efficacy of a vaccine containing plasmid DNA with an insert encoding human tyrosinase (ie, huTyr vaccine) as adjunctive treatment for oral malignant melanoma (MM) in dogs.
Animals—111 dogs (58 prospectively enrolled in a multicenter clinical trial and 53 historical controls) with stage II or III oral MM (modified World Health Organization staging scale, I to IV) in which locoregional disease control was achieved.
Procedures—58 dogs received an initial series of 4 injections of huTyr vaccine (102 μg of DNA/injection) administered transdermally by use of a needle-free IM vaccination device. Dogs were monitored for adverse reactions. Surviving dogs received booster injections at 6-month intervals thereafter. Survival time for vaccinates was compared with that of historical control dogs via Kaplan-Meier survival analysis for the outcome of death.
Results—Kaplan-Meier analysis of survival time until death attributable to MM was determined to be significantly improved for dogs that received the huTyr vaccine, compared with that of historical controls. However, median survival time could not be determined for vaccinates because < 50% died of MM before the end of the observation period. No systemic reactions requiring veterinary intervention were associated with vaccination. Local reactions were primarily limited to acute wheal or hematoma formation, mild signs of pain at the injection site, and postvaccination bruising.
Conclusions and Clinical Relevance—Results support the safety and efficacy of the huTyr DNA vaccine in dogs as adjunctive treatment for oral MM.
Impact for Human Medicine—Response to DNA vaccination in dogs with oral MM may be useful in development of plasmid DNA vaccination protocols for human patients with similar disease.