Development of anELISAto detect circulating anti‐asparaginase antibodies in dogs with lymphoid neoplasia treated withEscherichia colil‐asparaginase

Abstract

Resistance to E. coli L-asparaginase in canine lymphoma occurs frequently with repeated administration, a phenomenon often attributed, without substantiation, to the induction of neutralizing antibodies. To test the hypothesis that treated dogs develop antibodies against the drug, we created an ELISA to measure plasma anti-asparaginase IgG responses. Using samples from dogs that had received multiple doses, specific reactivity against L-asparaginase was demonstrated, while naïve patients’ samples were negative. The optimized ELISA appeared sensitive, with endpoint titers >1,600,000 in positive control dogs. Intra- and inter-assay coefficients of variation were 3.6 and 14.5%. The assay was supported by the observation that ELISA-positive plasma could immunoprecipitate asparaginase activity. When clinical patients were evaluated, 3/10 dogs developed titers after a single injection; with repeated administration, 4/7 dogs were positive. L-asparaginase antibodies showed reduced binding to the PEGylated drug formulation. The ELISA should prove useful in investigating the potential correlation of antibody responses with resistance.

Efficacy and adverse events of continuous l‐asparaginase administration for canine large cell lymphoma of presumed gastrointestinal origin

“…This is called silent inactivation and is considered to be an asymptomatic hypersensitivity reaction 25,30–32 . The presence of an anti‐asparaginase antibody contributes to the development of hypersensitivity reactions and reduces the therapeutic effect of l ‐asp 18,25,31,32 . In addition, in human medicine, it is recommended to switch to PEG‐asparaginase or Erwinia asparaginase in cases of allergic reaction or silent inactivation 25 .…”

“…The antibody against L-asp, a heterologous protein derived from bacteria, is involved in L-asp hypersensitivity reactions. 18 Therefore, hypersensitivity reactions are more likely to occur in the second and subsequent doses than in the first dose. 16 However, Blake et al recently reported that the incidence of hypersensitivity reactions was 1.6% even when L-asp was administered at a median of three times per dog and a maximum of 11 times.…”

L-Asparaginase, Doxorubicin, Vincristine, and Prednisolone (LHOP) Chemotherapy as a First-Line Treatment for Dogs with Multicentric Lymphoma

“…The other major adverse concern about l-asparaginase administration is the formation of antibodies and the development of drug resistance. Antibodies potentially could reduce or neutralize asparaginase activity in different ways, including the increased clearance by opsonization and the binding of Ig-drug complexes to Fc receptors and the direct inhibition by steric hindrance or by the binding to the proper catalytic site [ 30 ]. In a previous study, the average antibody prevalence was 30% (3/10) in dogs treated with single l-asparaginase, and the prevalence increased to 57% (4/7) after two l-asparaginase injections [ 30 ].…”

“…In a previous study, the average antibody prevalence was 30% (3/10) in dogs treated with single l-asparaginase, and the prevalence increased to 57% (4/7) after two l-asparaginase injections [ 30 ]. However, the route of administration could affect the frequency of antibody formation [ 30 ]. The dogs received subcutaneous injections of l-asparaginase in that study, which was different from our intramuscular route.…”

“…To produce a better outcome, we used intramuscular injection in all l-asparaginase treatments. Identifying the route of administration that is less immunogenic is important not only for preventing clinical hypersensitivity but also for preserving tumour responsiveness to the drug by diminishing antibody formation [ 30 ]. Further study is needed to clarify the outcome and antibody formation between different administration routes.…”