Immune response of the llama (Lama glama) to tetanus toxoid vaccination.

An ELISA was developed to measure serum concentrations of tetanus toxoid-specific immunoglobulins. The titers obtained with this assay were compatible with those obtained by the standard mouse toxin-neutralization test. Serum samples from 123 llamas were analyzed for ELISA titers to tetanus toxoid. Of the 82 vaccinated adults, 75 (91%) had titers greater than or equal to 1:50. The vaccination status and titers of weanlings and juveniles (3 to 12 months old) varied; of the 21 vaccinated, 17 (81%) had titers greater than or equal to 1:50 and 7 of 9 (78%) unvaccinated llamas had titers less than 1:50. The ELISA titers of unvaccinated llamas less than 8 weeks old (crias) were matched with the maternal titers. All crias with titers less than 1:50 had dams with titers greater than or equal to 1:50.

Levels of serum IgE in response to gastrointestinal nematodes in cattle.

Serum IgE concentration has been considered a valuable measurement in parasitic infections, yet little has been reported regarding cattle. This study examines the association of IgE levels of nematode-naïve Holstein steer calves and the level of gastrointestinal parasitism acquired by grazing irrigated pasture for 30 days. Total IgE levels were determined by enzyme-linked immunosorbent assay (ELISA) specific for bovine IgE on serum collected both before and after exposure to parasite-infected pastures. Following necropsy, parasite loads were determined by direct count from the contents of the abomasum, small intestine, cecum and large intestine; species of Ostertagia and Cooperia being the most common helminths found. Significant increases of IgE in the serum of calves with light infestations were seen, whereas calves with moderate to heavy infestations showed only mild IgE increases. With increased parasite burden, the frequency of increased IgE levels was reduced. Additionally, there appeared to be a seasonal correlation relating the level of serum IgE detected to the number of worms counted and to the course of parasite development.

Colostral transfer of bovine immunoglobulin E and dynamics of serum IgE in calves.

The role of IgE in protective immunity is becoming understood, therefore the colostral transfer of IgE and the age-dependent changes of IgE levels may be important for neonatal immunity. To investigate this question, serum samples were collected from range-fed Hereford cows and their calves from birth through 9 months of age. The sera were assayed for total IgE during the first week postpartum, indicating colostral transfer of IgE. Thereafter, serum levels declined rapidly within 3 weeks from birth. The IgE levels began to increase after 12 weeks of age, and in some cases reached adult levels. The passive transfer of maternal IgE through colostrum may be important in providing early protection from disease, especially against intestinal parasites.

Generation and characterization of murine monoclonal antibodies specific for bovine immunoglobulin E.

Monoclonal antibodies were produced against serum-derived bovine immunoglobulin E (IgE). Culture supernatants of hybridomas were initially screened by enzyme-linked immunosorbent assay (ELISA). Supernatant-derived antibodies were concentrated and further characterized using ELISA, reverse cutaneous anaphylaxis, immunohistochemical staining, and immunoblotting of IgE-containing samples separated by SDS-polyacrylamide gel electrophoresis (PAGE). Eight monoclonal antibodies showed specificity for bovine epsilon immunoglobulin heavy chain. Two antibodies (E2 and E32) reacted in immunoblots of SDS-PAGE of serum IgE under reducing conditions. Additionally, E2, E5, and E32 detected epsilon chain in serum separated by SDS-PAGE and then renatured. Antigen-specific IgE was detected in Western blots by E5 and E32. Immunoperoxidase staining of IgE-containing cells in mesenteric lymph node sections was detected with E5, E21 and E32. All eight antibodies produced positive reverse cutaneous anaphylaxis reactions in calf skin. All functioned well in ELISA as a plate-sensitizing reagent for quantitation of total IgE; E5 and E32 worked well as a primary antibody in antigen-specific IgE assays. These antibodies will be useful in research applications and in diagnostic assays.