T.gondii p40
Description
Product Specs
For optimal stability, T.gondii p40 should be stored at -18°C. While it can remain stable at 4°C for up to one week, repeated freeze-thaw cycles should be avoided.
Q&A
FAQs for Researchers on T. gondii p40 (IL-12 p40)
What methodologies are used to detect IL-12 p40 production during T. gondii infection?
IL-12 p40 detection involves:
-
ELISA assays: Quantify cytokine levels in serum or supernatant (e.g., two-site ELISA with anti-IFN-γ antibodies) .
-
Flow cytometry: Identify IL-12 p40-producing cells using intracellular staining or reporter mice (e.g., YET40 mice with eYFP linked to IL-12 p40) .
-
Splenocyte cultures: Stimulate cells with T. gondii antigens (e.g., soluble tachyzoite antigen, STAg) and measure cytokine output .
How does IL-12 p40 contribute to the host immune response against T. gondii?
IL-12 p40 drives IFN-γ production by CD4+ Th1 cells, which is critical for controlling parasite replication. Key mechanisms include:
-
Activation of dendritic cells (DCs) and macrophages via CD40-CD40L interactions .
-
Promotion of autophagy in infected cells to limit parasite survival .
-
Maintenance of IFN-γ production in T cells during chronic infection .
What experimental models resolve contradictions in IL-12 p40 cellular sources during infection?
Conflicting reports on whether infected or bystander cells produce IL-12 p40 are addressed using:
-
Transgenic parasites: Toxoplasma-Cre strains combined with Ai6 Cre reporter mice to track parasite-host interactions .
-
Invasion-inhibited parasites: Pharmacologically treated parasites (e.g., 4-BPB) to distinguish phagocytosis from active invasion .
-
Flow cytometry with fixation protocols: Preserve intracellular cytokines while minimizing signal loss from phagocytosed parasites .
Key Finding: >90% of IL-12 p40 is produced by uninfected DCs and inflammatory monocytes, not infected cells .
How do researchers dissect IL-12 p40 regulation in genetically modified hosts?
-
IL-12 p40-deficient mice: Supplemented with recombinant IL-12 to study cytokine dynamics during acute/chronic infection .
-
Cell-specific knockouts: Use Cre-lox systems to delete IL-12 p40 in DCs, macrophages, or T cells.
-
TLR/MyD88 pathway analysis: Investigate IL-12 p40 induction via parasite profilin using TLR11/12-deficient models .
What statistical approaches address zero-inflation in T. gondii oocyst shedding data?
-
Zero-inflated beta regression: Models prevalence data with excess zeros (e.g., T. gondii oocyst shedding studies) .
-
Generalized linear mixed models (GLMMs): Account for random effects like study location or host species .
Table 1: Techniques for IL-12 p40 Research
How are conflicting roles of IL-12 p40 in ocular vs. systemic toxoplasmosis resolved?
-
Disease-specific models: Compare retinal (immunosuppressive TGF-β2/α-MSH microenvironment) and cerebral infections .
-
Cytokine blockade: Neutralize IL-12 p40 in ocular models to assess CD8+ T-cell cytotoxicity .
Data Interpretation and Contradictions
-
Paradox 1: IL-12 p40 is required for IFN-γ production but insufficient alone.
-
Paradox 2: Phagocytosis of T. gondii does not drive IL-12 p40 in vivo.
Product Science Overview
Toxoplasma gondii is an intracellular protozoan parasite responsible for toxoplasmosis, a widespread zoonotic infection. This parasite can infect nearly all warm-blooded animals, including humans. The infection is particularly concerning for immunocompromised individuals, pregnant women, and congenitally infected newborns .
Recombinant proteins are produced through genetic engineering techniques, where the gene encoding the protein of interest is inserted into an expression system, such as bacteria, yeast, or mammalian cells. For Toxoplasma gondii p40, the recombinant protein is typically produced in prokaryotic systems like Escherichia coli due to their efficiency and cost-effectiveness .
The production of recombinant Toxoplasma gondii p40 involves several steps:
- Gene Cloning: The gene encoding the p40 protein is cloned into an expression vector.
- Transformation: The vector is introduced into a host cell, commonly E. coli.
- Expression: The host cells are cultured under conditions that induce the expression of the p40 protein.
- Purification: The recombinant p40 protein is purified using techniques such as affinity chromatography to ensure high purity and yield .
The recombinant p40 protein has several applications:
- Serological Detection: It is used in diagnostic assays to detect antibodies against T. gondii in human and animal sera. Techniques like enzyme-linked immunosorbent assay (ELISA) utilize recombinant p40 to improve the sensitivity and specificity of toxoplasmosis diagnosis .
- Vaccine Development: The immunogenic properties of p40 make it a potential candidate for vaccine development. Research is ongoing to evaluate its efficacy in inducing protective immunity against T. gondii .
