Secretory microneme proteins induce T-cell recall responses in mice chronically infected with Toxoplasma gondii

Microneme (MIC) proteins play important roles in the recognition, adhesion, and invasion of host cells by Toxoplasma gondii. Previous studies have shown that MIC proteins are highly immunogenic in the mouse and recognized by human serum antibodies. Here we report that T. gondii antigens MIC1, MIC3, MIC4, and MIC6 were capable of inducing memory responses leading to production of gamma interferon (IFN-γ) by T cells from T. gondii-infected mice. Production of IFN-γ was demonstrated using enzyme-linked immunosorbent spot (ELISPOT) assay and also intracellular cytokine staining. All four MIC antigens displayed very high sensitivity (100%) and specificity (86 to 100%) for detecting chronic infection. Interestingly, IFN-γ was produced by both CD4 ⁺ and CD8 ⁺ T cells in BALB/c mice but primarily by CD4 ⁺ T cells in C57BL/6 mice. Phenotypic characterization of IFN-γ-producing CD4 ⁺ and CD8 ⁺ T cells in BALB/c mice and CD4 ⁺ T cells in C57BL/6 mice revealed effector memory T cells (CD44 ^hi CD62L ^lo ) as the predominant cells that contributed to IFN-γ production in response to MIC antigens. Effector memory responses were seen in mice of different major histocompatibility complex class II (MHC-II) haplotypes, suggesting that MIC antigens contain epitopes that are broadly recognized.