Buy It Now. Add to cart. Weiss and Kami Kim , Hardcover. Be the first to write a review About this product. About this product Product Information This second edition of Toxoplasma gondii reflects the significant advances in the field in the last 5 years including new information on the genomics, epigenomics and proteomics of T. T gondii remains the best model system for studying the entire Apicomplexa group of protozoans, which includes Malaria, making this new edition essential for a broad group of researchers and scientists.
Toxoplasmosis is caused by a one-celled protozoan parasite known as Toxoplasma gondii..
The infection produces a wide range of clinical syndromes in humans, land and sea mammals, and various bird species. Approx 1 in 4 more than 60 million people in the USA are infected with the parasite, and in the UK between 0. Most humans contract toxoplasmosis by eating contaminated raw or undercooked meat particularly pork , vegetables, or milk products, by coming into contact with the T. The parasite damages the ocular and central nervous systems causing behavioral and personality alterations as well as fatal necrotizing encephalitis.
It is especially dangerous for the fetus of an infected pregnant woman and for individuals with compromised immune systems, such as HIV infected patients. Completely updated, the 2nd edition presents recent advances driven by new information on the genetics and genomics of the pathogen since Toxoplasma gondii is the third most common cause of lethal waterborne illness in the US Presents a complete review of molecular and cellar biology and immunology of Toxoplasma gondii combined with methods and resources for working with this pathogen.
Provides the latest information concerning the epidemiology diagnosis, treatment and prevention of toxoplasmosis. Offers a single source reference for a wide range of scientists and physicians working with this pathogen including parasitologists, cell and molecular biologists, veterinarians, neuroscientists, physicians, and food scientists.
Additional Product Features Dewey Edition. A one source resource for these molecular techniques is essential to the continuing progress in this research field. Cushion, Ph. Show More Show Less. No ratings or reviews yet. Be the first to write a review. Combined with the results with genes encoding known proteins, these findings are concordant with the similarities in host responses to sporozoites and tachyzoites reported here i.
Inner-membrane complex IMC proteins and functionally related proteins play a crucial role in parasite replication, motility, and host cell invasion. As might be expected, therefore, tachyzoites, which are the rapidly dividing form of T. There are 68 annotated AP2 domain-containing transcription factors in the T. Some of these are cell-cycle-regulated [ 65 ] and have been implicated in the transcriptional regulation of interconversion of tachyzoites to bradyzoites [ 66 , 67 ] and of virulence determinants, including ROP18 [ 68 ].
The metabolic state of intracellular sporozoites has not previously been investigated and so we analyzed our transcriptomic data for clues to how these parasites might compare to tachyzoites in this regard. On the other hand, the genes with higher expression in sporozoites than in tachyzoites show some degree of significant enrichment in riboflavin, nicotinate and nicotinamide, purine, and pyrimidine metabolisms Table Of the three developmental forms capable of infection, Toxoplasma sporozoites have been the least studied.
Comparing these data to those from infection with tachyzoites and previous studies on day 10 oocysts allowed us to identify host and parasite genes that may be specifically involved in these different stages of a Toxoplasma infection. Our findings indicate that the primary response of rat IECs to Toxoplasma gondii , whether as sporozoite or tachyzoite, involves an NF-kB-like inflammatory response.
This response is marked by a significant increase in expression of genes encoding proteins associated with NF-kB signaling, including chemokines and inflammatory cytokines.
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For instance, Ccl20 , Cxcl1 , and Ccl2 , which were among the highly expressed genes during Toxoplasma infection, are involved in recruitment and activation of immune cells, such as neutrophils and lymphocytes [ 75 , 76 ]. The absence of alterations in the host transcriptome in the presence of frozen-thawed sporozoites indicates that inactive sporozoites do not provide pathogen-associated molecular patterns PAMPs , at least not ones recognized by these IECs; instead, it appears that active parasite processes lead to the observed host gene activation.
Toxoplasma Gondii: The Model Apicomplexan - Perspectives and Methods
The slightly stronger host response measured in the TZ vs. SPZ cultures could be due to either an inherent property of tachyzoites or the slightly higher MOI for the tachyzoite infections 0. Achieving a higher MOI may be difficult with the available amounts of infectious sporozoites but the advent of single-cell transcriptomic analyses will help circumvent this problem [ 77 ]. To identify sporozoite-specific effectors that may contribute to the transcriptomic changes observed in infected IECs, we also profiled the transcriptomes of the infecting sporozoites and tachyzoites.
Transcripts encoding other well-characterized tachyzoite effectors [ 51 , 53 ], namely ROP16, ROP5, and ROP18 were also all expressed at similar levels in both sporozoites and tachyzoites. GRA24, GRA16, and the newly identified GRA28, have all been shown to localize to the host nucleus [ 33 , 79 ] and all three had a significantly higher expression in infecting sporozoites compared to tachyzoites. Whether these effector proteins localize and function in a similar fashion during infection with sporozoites remains to be studied.
Of perhaps greater importance is that our results show 85 genes encoding putatively secreted, uncharacterized proteins that have higher expression in the sporozoites compared to tachyzoites. It is likely like that some of these proteins are effectors that contribute to the ability of sporozoites to initiate an infection in the intestine of a new host.
The need for differential expression of these genes could reflect the different cell type being infected by a sporozoite, almost exclusively an intestinal endothelial cell, vs. The effectors needed for these different stages and locations of an infection could be very different. Analysis of the infecting sporozoite transcriptome revealed a significant increase in the expression of several AP2 transcription factors, suggesting that these play a role in the regulation of the differences in gene expression seen between sporozoites and tachyzoites.
Further support for this inference comes from the recent findings that AP2 transcription factors, such as TgAP2XI-4, which was found here to be increased in tachyzoites, are involved in the transcriptional regulation of tachyzoite-to-bradyzoite interconversion [ 67 ]. The different transcript levels for a large number of genes from extracellular D10 sporulated oocysts vs. It could be that after sporulation is complete, there are major changes in the transcriptome if the sporozoites do not quickly invade a new host. Alternatively, the differences observed could be due to the fact that we are looking 8 hours post-infection.
As previously reported, invaded sporozoites transition to the tachyzoite form about 12 hours after infection [ 34 ] and so our finding few if any transcripts for several genes that were high in D10 oocysts could indicate that these transcripts are rapidly degraded as part of the differentiation process. What fraction of the differences reflects this vs.
Overall, our results show that, at least within the constraints of the in vitro model used here, and although the response of IECs to sporozoite infection is qualitatively similar to that seen with tachyzoites, many transcriptomic differences are seen, especially on the parasite side. Future studies will seek to characterize these genes and determine the role they play in enabling sporozoites to initiate a successful infection. Table D: Final list of rat genes with average exon reads included in analysis pipeline with RPKM values and fold changes.
Table E: Final list of Toxoplasma genes with average adjusted exon reads that were included in analysis pipeline with RPKM values and fold-changes. Table G: Rat genes with increased expression during infection with tachyzoites. Table H: Toxoplasma genes with higher expression in intracellular sporozoites compared to tachyzoites. Table I: Toxoplasma genes with higher expression in intracellular tachyzoites compared to sporozoites.
Only genes with at least 20 reads in TZ are included in the table. Table J: Significantly higher genes encoding hypothetical proteins with predicted signal peptides in SPZ vs. Data curation: PSG. Formal analysis: PSG. Software: PSG. Browse Subject Areas? Click through the PLOS taxonomy to find articles in your field. Introduction Toxoplasma gondii is one of the most successful eukaryotic pathogens of medical and veterinary importance, as it can infect humans and a very large number of warm-blooded animals worldwide [ 1 ].
In vitro model of infection of intestinal epithelium Two independent experiments were performed with two technical replicates per experiment. Uninfected IEC UN : 25, IEC cells were seeded in each well of well tissue culture-treated plates in IEC medium and incubated for 48 hours to reach confluence approximately one million cells. Frozen-thawed sporozoites control fzSPZ : Two million frozen-thawed sporozoites were added to a well of confluent IEC obtained as described in 1.
Active sporozoites infection SPZ : Two million freshly excysted sporozoites, estimated from hemocytometer counts, were used to infect a well with confluent IEC The target nominal multiplicity of infection MOI was 2. MA filtered lysate was added to confluent IEC in a volume matching the tachyzoite inoculum. Download: PPT. Fig 1. Experimental layout for in vitro infection of the intestine. Results In vitro infection of intestinal epithelial cells and parameters for RNAseq analysis Following ingestion by an intermediate host, sporozoites are excysted and rapidly invade intestinal epithelial cells IECs [ 12 ].
History and Life Cycle of Toxoplasma gondii
Fig 2. Quantification of infection in IEC cells exposed to sporozoites or tachyzoites. Toxoplasma sporozoites trigger an NF-kB-like signature response in IEC cells Given that the oocyst wall includes complex polysaccharides, proteins, and acid-fast lipids [ 37 ], the first pairwise analysis we performed was to check for possible pathogen-associated molecular patterns PAMPs in the oocyst-derived preparations of sporozoites.
Table 1. Rat genes with significantly higher expression in IECs upon infection with sporozoites. Table 2. Host pathways significantly enriched in IECs infected with sporozoites or tachyzoites. Table 3. Rat genes with significantly higher expression in IECs infected with tachyzoites but not with sporozoites. Intracellular sporozoites are transcriptionally different from maturing sporozoites in day 10 sporulated oocysts In addition to data on differences in the host transcriptome, which was the primary objective of this study, our dataset allowed us to assess transcriptomic differences between tachyzoites and the sporozoites at the time point used here, 8 hpi.
Table 4. Expression dynamics of oocyst-associated genes from unsporulated oocysts to intracellular sporozoites. The transcriptomic profile of infecting sporozoites is distinct from that of tachyzoites Next, we compared the transcriptomic data on the intracellular sporozoites with the data for the intracellular tachyzoites. Table 5. Top 50 genes with significantly higher expression in sporozoites compared to tachyzoites. Table 6. Top 50 genes with significantly higher expression in tachyzoites compared to sporozoites.
Shared and distinct sets of genes encoding secreted proteins Toxoplasma proteins derived from the specialized secretory organelles, namely micronemes, rhoptries, and dense granules, are critical for invasion, intracellular growth and modulation of host responses. Table 7. Selected known secreted proteins with similar expression in sporozoites and tachyzoites. Table 8. Differentially expressed genes encoding microneme proteins. Table 9. Differentially expressed genes encoding rhoptry proteins.
Table Differentially expressed genes encoding dense granule or dense granule-like proteins. Genes involved in gene expression and cell division Inner-membrane complex IMC proteins and functionally related proteins play a crucial role in parasite replication, motility, and host cell invasion.
Differentially expressed genes encoding IMC proteins. Differentially expressed genes encoding AP2 transcription factors. Genes encoding metabolic enzymes The metabolic state of intracellular sporozoites has not previously been investigated and so we analyzed our transcriptomic data for clues to how these parasites might compare to tachyzoites in this regard. Metabolic pathways differentially enriched in intracellular sporozoites vs.
Discussion Of the three developmental forms capable of infection, Toxoplasma sporozoites have been the least studied.
Supporting information. S1 File. Supplementary tables described below. References 1. Toxoplasmosis in Wild and Domestic Animals. Elsevier Academic Press; Toxoplasmosis snapshots: Global status of Toxoplasma gondii seroprevalence and implications for pregnancy and congenital toxoplasmosis. Int J Parasitol. Human Toxoplasma infection.
Structures of Toxoplasma gondii tachyzoites, bradyzoites, and sporozoites and biology and development of tissue cysts. Clinical Microbiology Reviews. Toxoplasma oocysts as a public health problem. Trends in Parasitology. Soil survival of Toxoplasma oocysts in Kansas and Costa Rica. Am J Trop Med Hyg.
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Strategies to discover the structural components of cyst and oocyst walls. Eukaryotic Cell. Proton-sensing G-protein-coupled receptors. Inflamm Bowel Dis. A Rabdependent pathway contributes to Salmonella typhi host restriction. Science Tang BL. Microbes Infect. Cell Host Microbe. Immunol Rev. Production of IL by macrophages infected with Toxoplasma gondii depends on the parasite genotype. J Immunol. Tumor necrosis factor-alpha triggers anti-toxoplasmal activity of IFN-gamma primed macrophages.
Molecular characterisation of a novel family of cysteine-rich proteins of Toxoplasma gondii and ultrastructural evidence of oocyst wall localisation.