p,0.05; p,0.01; p,0.001. Data are representative of two independent experiments. doi:10.1371/journal.pone.0031012.g003 for drug development and therapeutic intervention in CM. Studies in human clinical cases of CM and experiments in mouse models of P. berghei-induced CM have identified excessive TNF-a and IFNg-driven inflammatory response as a key determinant in CM pathogenesis; novel strategies to blunt this response have shown promise in the prevention and treatment of CM. We have implemented a genome-wide mutagenesis screen in mice to systematically identify genes and proteins that mediate pathological inflammatory responses during P. berghei infection in vivo, and whose pharmacological or genetic inactivation may protect from CM. We report on the first mutant identified in this screen. Linkage analyses, genomic DNA sequencing, and complementation studies in double heterozygotes have established that CM protection in this mutant is caused by a mutation in the amino-terminal FERM domain of Jak3. Jak3 is a cytosolic tyrosine kinase expressed primarily in the hematopoietic 80321-63-7 custom synthesis system that plays a critical role in: a) the ontogeny of 
different myeloid and lymphoid cells, and b) the response of these cells to stimulation by different cytokines. Jak3 interacts with the common gc chain of type 1 and 2 cytokine receptors, which includes IL-2, IL-4, IL-7, IL-9, IL-15 and IL-21. This interaction causes recruitment and phosphorylation of STAT family members to trigger downstream transcriptional responses in cells expressing such receptors. In humans, loss of either JAK3 or IL2RG causes autosomal and Xlinked T2B+ SCID, respectively. These patients lack T cells and NK cells, have normal numbers of immature and poorly functional B-lymphocytes, a clinical picture in agreement with the established roles of IL-7, IL-2, IL-15, and IL-4 in which associated signaling is impaired in JAK3 mutants. Jak3W81R homozygote mice showed a phenotype that overlaps T2B+ SCID in humans, and displayed an atrophied thymus, a low number of thymic and splenic CD8+ T cells and NK cells, as well as a near absence of B cells in the bone marrow. CD4+ T cells were present in normal numbers but appeared anergic and did not produce IFN-g in response to simulation with PMA or ionomycin Schematic representation of the Jak3 protein, showing the 7 PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/22181837 Jak Homology structural domains, and associated functional annotation. The position of the W81R mis-sense mutation discovered in pedigree 48 is shown. Multiple sequence alignment of the amino terminal portion of Jak3 surrounding W81 shows high conservation across Jak3 relatives. doi:10.1371/journal.pone.0031012.g004 7 A Jak3 Mutation Protects against Cerebral Malaria polarization). This immune phenotype closely resembled that of previously described Jak3 knockout mice. What is the mechanism underlying protection against P. bergheiinduced CM in Jak3W81R homozygotes IFN-g plays a critical role in initiating and amplifying pathological inflammatory response during CM, and mouse mutants lacking the IFN-g gene are protected against P. berghei-induced CM. Although the dominant cell type responsible for early production of IFN-g in vivo during P. berghei infection has been debated, NK cells, CD4+ and CD8+ T cells have all been implicated, and all three populations are affected in Jak3W81R homozygotes. In the case of NK cells, results either supporting or excluding a role for these cells in CM pathogenesis have been published. In one study, depletio
