Serological and Cytokine Profiling of Toxoplasmosis

T. gondii-specific antibody and cytokine levels were determined by ELISA as reported previously [18 (link),26 (link)]. Diluted immune sera (1:100), urine (1:3), vaginal samples (1:3), fecal supernatants (1:3), intestine samples (1:100), and brain supernatants (undiluted) were used as primary antibodies. Briefly, 96-well plates were coated with T. gondii RH (2 μg/mL) and blocked prior to incubation with the primary antibodies listed above. Anti-mouse IgG and IgA secondary antibodies conjugated with horseradish peroxidase (HRP) were used to detect the antigen-specific responses. The production of pro-inflammatory cytokines interferon-gamma (IFN-γ) and interleukin-6 (IL-6) were measured from the brain supernatants using the cytokine ELISA kits (BD Biosciences, San Jose, CA, USA).