Immunization with excreted/secreted proteins in AS/n mice activating cellular and humoral response against Toxoplasma gondii infection.

This study investigated how Toxoplasma gondii excretory-secretory antigens (ESA) stimulate the humoral and cellular response in infected hosts. We evaluated IFN-γ, IL-4 TNF-α, and IL-10 levels as well as humoral response of ESA-immunized AS/n mice. T. gondii lysate antigen (TLA), a crude antigen, was used in all experiments to evaluate the immune response. Chronic infected and naive mice were used as control groups, since the immune response is well known. The challenge experiments showed the parasitemia levels, determined by real time PCR and survival index. The naive group had early mortality and higher parasitemia than the ESA-immunized mouse group. In addition the chronic infected group had no parasitemia and mortality. Both ESA-immunized and chronic infected mice produced a similar level of IFN-γ and TNF-α. ESA, also, activated cells from immunized mice to produce IL-4 and IL-10 in lower levels compared to those cells collected from chronic mice but sufficient to modulate IFN-γ and TNF-α synthesis, preventing an excessive immune response that could cause extensive inflammation and host tissue damage. After 6 weeks, ESA-immunized mice had low IgM and IgG2a levels and high IgG1 levels. Purified anti-ESA IgG were able to opsonize tachyzoites (RH strain), and mice that received these parasites had lower parasitemia, and mortality was delayed 48 h, compared with the same results from those receiving parasites opsonized with IgG purified from naive mice. The protective immune response in the chronic infection was efficient in protecting the host against infection caused by other T. gondii strain and ESA participate in stimulating the host humoral and cellular responses. The immunization assays showed that ESA can elicit high IgG1, IFN-γ and TNF-α production and, a lower amount of IgM, IgG2, IL-10 and IL-4, suggesting a mixed Th1/Th2 profile.

The spider acylpolyamine Mygalin is a potent modulator of innate immune responses.

Mygalin is an antibacterial molecule isolated from the hemocytes of the spider Acanthoscurria gomesiana. It was identified as bis-acylpolyamine spermidine. We evaluated the modulator effects of synthetic Mygalin in the innate immune response. We demonstrate that Mygalin induces IFN-γ synthesis by splenocytes increasing the nitrite secretion by splenocytes and macrophages. A specific inhibitor of iNOS abrogated Mygalin-induced nitrite production in macrophages independent of IFN-γ activation. In addition, Mygalin-activated macrophages produced TNF-α but not IL-1ß, demonstrating that Mygalin does not act directly on the inflammasome. Furthermore, this compound did not affect spontaneous or Concanavalin A-induced proliferative responses by murine splenocytes and did not induce IL-5 or apoptosis of splenocytes or bone marrow-derived macrophages. These data provide evidence that Mygalin modulates the innate immune response by inducing IFN-γ and NO synthesis. The combined immune regulatory and antibacterial qualities of Mygalin should be explored as a strategy to enhance immune responses in infection.