Impact of orally administered Antibodies on the Outcome of Experimental Paratuberculosis Infection in Rabbits
Description
This dataset contains the experimental data generated in a rabbit model of Mycobacterium avium subsp. paratuberculosis (MAP) infection designed to investigate whether anti-MAP antibodies present during oral challenge influence infection outcome, bacterial dissemination and immune responses. Juvenile female New Zealand White rabbits (approximately 8 weeks old) were assigned to four experimental groups: non-infected control (NIC, n = 1), infected control receiving MAP pre-incubated with phosphate-buffered saline (IC, n = 2), infected with MAP pre-incubated with homologous anti-MAP antibodies (IAbMAP, n = 4), and infected with MAP pre-incubated with irrelevant antibodies obtained from healthy animals (IAbHealthy, n = 4). Animals received three consecutive oral inoculations (days 0–2) and were monitored for 145 days post-infection. The information is organized in two excel files: 1- entitled Setting MAP_AbTreatment before challenge, containing data for sera selection to Ab pools preparation (AbMAP, from MAP-immunized rabbits; AbHealthy, from healthy rabbits) and to set the experimental conditions for MAP:Ab pool incubation before challenge of rabbits. Sera for AbHealthy pool were selected out of 10 candidates based on the lowest reactivity against MAP by ELISA (worksheet entitled MAP reactivity_AbHealthy pool); while for AbMAP pool sera were selected considering also the avidity levels assessed by ELISA after urea treatment of the antigen-binded antibodies (worksheet entitled MAP reactivity and avidity_AbMAP pool). To establish the amount of antibodies required for MAP–antibody pre-incubation, a preparative titration experiment was performed. Aliquots containing 10⁹ CFU of MAP were incubated with 100, 10, or 1 µg of γ-globulins from each Ab pool, in a final volume of 2 mL. Samples were centrifuged and the supernatants recovered. The level of reactivity against MAP of Ab pools prior and after incubation (to assess antibody consumption) with MAP were evaluated. These data are presented in the worksheet entitled MAP_AbTreatment before challenge. 2- entitled Variables measured in MAP challenged Rabbits, containing data about antibody levels and MAP counts recovered from different tissues (ileum, ileocecal valve, cecum, sacculus rotundus, vermiform appendix, mesenteric lymph node, spleen, liver, erector spinae and diaphragmatic muscles) and rectal feces, at necropsy (CFU/g). Antibody levels were measured by ELISA: IgG at the seric compartment at the time course of infection (0, 45, 100 and 145 days post-infection -dpi-) against different antigens (live, inactivated or sonicated MAP, Paratuberculosis Protoplasmic Antigen -PPA- and liporarabinomannan -LAM-); and IgA in cecal content at final time point. Sera were examined at 1:100 and cecal contents at 1:2.
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Steps to reproduce
References: NRS: negative reference sera PRS: positive reference sera ELISA Units (EU): ELISA units were calculated as the average OD of the sample/average OD of 3 MAP-immunized rabbits as positive reference sera x 100. When appropriate, cut-off was defined as the mean EU of NRS + 2 standard deviation (SD).
Institutions
- Universidad de Buenos AiresBuenos Aires F.D., Buenos Aires