Toxoplasma gondii activates NLRP12 inflammasome pathway in the BALB/c murine model.

The host resistance against Toxoplasma gondii (T. gondii) infection is related to the initiation of the immune response. The study aimed to investigate the role of the leucine-rich repeat family, pyrin domain -containing protein 12 (NLRP12), and cytoplasmic nucleotide-binding domain in the inflammasome-mediated cell death during murine toxoplasmosis. Groups of BALB/c mice (n = 10) were inoculated intraperitoneally with live tachyzoites, excretory-secretory antigens (ESAs) of T. gondii RH strain, and RPMI. The gene expression levels of NLRP12, caspase-3, caspase-1, IL-1ß, IL-18, ASC, and Bcl-2 were measured in the peritoneal cells using quantitative real-time PCR, while the determination of NLRP12 protein level was measured by Western blot. Also, the intracellular reactive oxygen species (ROS) production was investigated. Quantitative and comparative analyses showed that injection of tachyzoites significantly increased NLRP12, caspase-3, caspase-1, IL-1ß, IL-18, and ASC genes mRNA expression levels (p<0.01). Contrary to the acute infection, the Bcl-2 gene was significantly expressed in the ESAs group (p<0.0001). The level of NLRP12 protein was significantly higher in the mice that received tachyzoites and ESAs in comparison to the control group (p<0.0001). These findings provide an inside into the host-T. gondii interaction and NLRP12 regulation, which is important for the modulation of the immunological response.

Protective effect of a DNA vaccine cocktail encoding ROP13 and GRA14 with Alum nano-adjuvant against Toxoplasma gondii infection in mice.

Toxoplasma gondii is an obligate intracellular protozoan parasite that can cause serious public health problems. The development of a safe and effective vaccine against T. gondii is urgently needed to prevent and control the spread of toxoplasmosis. The aim of this study was to evaluate the immune responses induced by a pcGRA14 + pcROP13 vaccine cocktail in BALB/c mice. All groups were immunized intramuscularly three times at two-week intervals. The production of anti-Toxoplasma gondii lysate antigen (TLA) antibodies, lymphocyte proliferation, serum levels of IFN-γ and IL-4 cytokines and the survival time were monitored after vaccination and challenged with the virulent RH strain of T. gondii. The results showed that immunization with the pcGRA14 + pcROP13 DNA vaccine significantly increased the production of specific IgG antibodies and cytokines against toxoplasmosis. Interestingly, high levels of IgG2a and IFN-γ were found in animals vaccinated with DNA vaccine cocktail. Furthermore, immunized mice challenged with the RH strain of T. gondii showed prolonged survival time when compared to control groups (P <0.05). The present study demonstrates the potential of a DNA cocktail vaccine expressing pcGRA14 and pcROP13 in developing specific immune responses and providing effective protection against T. gondii infection.

IL-17 and IL-22 elicited by a DNA vaccine encoding ROP13 associated with protection against Toxoplasma gondii in BALB/c mice.

Toxoplasma gondii, an intracellular parasitic protozoan, is capable of infecting man and all warm-blooded animals. Cell-mediated immunity is vital in mounting protective responses against T. gondii infection. Recent studies have shown that T-helper (Th) 17 responses may play a key role in parasite control. In this current study, we constructed a DNA vaccine encoding T. gondii ROP13 in a pcDNA vector. Groups of BALB/c mice were immunized intramuscularly with pcROP13 or controls and challenged with the RH strain of T. gondii. The results showed that immunization with pcROP13 could elicit an antibody response against T. gondii. The expression of the canonical Th17 cytokines, interleukin (IL)-17 and IL-22, were significantly increased after immunization with pcROP13 compared with control groups ( p < 0.05). Furthermore, vaccination resulted in a significant decrease in parasite load ( p < 0.05). The induction of Th17 related cytokines, using a ROP13 DNA vaccine, against T. gondii should be considered as a potential vaccine approach for the control of toxoplasmosis.

Quantification of Toxoplasma gondii in the tissues of BALB/c mice after immunization with nanoliposomal excretory-secretory antigens using Real-Time PCR.

BACKGROUND: Toxoplasmosis is an infectious disease caused by the intracellular parasite Toxoplasma gondii. Although almost 1/3 of the world's population are seropositive, there is no effective vaccine against toxoplasmosis. Therefore, the development of an effective vaccine for control of toxoplasmosis is one of major concerns in parasitology. The aim of this study was to evaluate the efficacy of nano-liposomal excretory-secretory antigens (NLESA) in BALB/c mice. MATERIALS AND METHODS: Excretory-secretory antigens (ESA) was obtained from tachyzoites, encapsulated in the liposome and studied by scanning electron microscope. BALB/c mice were immunized with NLESA and ESA, sterile phosphate-buffered saline (PBS). Immunization was performed three times at 14-day intervals and challenged with 1 × 104 tachyzoites of T. gondii RH strain four weeks later. The parasite load of mice blood, brain and spleen tissues were determined using quantitative PCR targeted at the repeated element (RE) gene. RESULTS: The immunization with NLESA and ESA induced a significant increase of anti-Toxoplasma IgG antibody compared with PBS group (P < 0.05). After challenge with tachyzoites, qPCR analyses showed significant reduction of parasite load in NLESA and ESA immunized mice compared with control group (P < 0.05). Also, NLESAs were more effective than ESAs and showed significantly reduced parasite load in blood (P = 0.001) and brain tissue (P = 0.01). DISCUSSION: The vaccination with NLESA showed more promising results comparing to ESA. Further studies are recommended in order to achieve effectiveness of the vaccine against T. gondii.