Anamnestic anti-pZP CD4+ and CD8+ T-cell responses in PBMC from pony mares immunized with either native pZP, a recombinant zona pellucida vaccine or adjuvanted saline (controls)

Mares were randomly assigned to pZP, reZP and control groups (n=7 per group). Treatments consisted of a primary vaccination or saline (V1; Day 0) incorporating Freund’s modified complete adjuvant, followed by a single booster (V2; Day 35) incorporating Freund’s incomplete adjuvant.

PBMC were isolated and cryopreserved on Day 0 (immediately prior to V1) and Day 70 (five weeks post V2). Relative proliferation of T-lymphocytes in response to pZP antigen was assessed using carboxyfluorescein diacetate succinimidyl ester dilution with immunophenotyping, analysed via flow cytometry (Accuri C6).