2004;78:8312C8321. isolated in the 1940s, whereas both West African and Asian strains were discovered in the 1960s. Identification and diagnosis of ZIKV has been and continues to be confounded by its overlap in geographic range, vector space, symptomology and serological cross-reactivity with other flaviviruses such as dengue computer virus (DENV) (Ioos et al., 2014; Zammarchi et al., 2015). A large body of literature has provided evidence for a potential dual role for CD8+ T cells in protection and pathogenesis during DENV contamination (Screaton et al., 2015; Tang et al., 2015; Weiskopf and Sette, 2014; Zellweger and Shresta, 2014). Epidemiologic studies indicate that Severe Dengue is most often seen in individuals experiencing a heterotypic DENV infection after prior seroconversion to at least one of the other three serotypes (Guzman et al., 2000; Sangkawibha et al., 1984). Some studies showed cross-reactive CD8 T cells are more activated during secondary infection (Mongkolsapaya et al., 2003) with a suboptimal T cell phenotype (Mongkolsapaya et al., 2006) (Imrie et al., 2007; Mangada and Rothman, 2005) suggesting a possible pathogenic role for cross-reactive T cells. However, recently emerging literature points to a protective role for T cells in DENV infection (Weiskopf et al., 2013; Weiskopf et al., 2015), and our previous work on DENV using mouse models (Prestwood et al., 2012b; Yauch et al., 2010; Yauch et al., 2009; Zellweger et al., 2014; Zellweger et al., 2013; Zellweger et al., 2015) in C57BL/6 and 129/Sv mice lacking type I IFN receptor (IFNAR) alone or both type Cannabichromene I and II IFN receptors (AB6, A129, and AG129) has provided multiple lines of evidence indicating a protective role for CD8+ T cells. H-2b mouse models of ZIKV infection recently have been established in WT C57BL/6 mice treated with blocking anti-IFNAR monoclonal antibody and in gene-deficient mice that globally lack IFNAR or both IFNAR and type II IFN receptors (Dowall et al., 2016; Govero et al., 2016; Lazear et al., 2016; Rossi et al., 2016). To investigate IFN receptor-competent CD8+ T cell responses in H-2b mice, in the present study we established a model of ZIKV infection in LysMCre+IFNARfl/fl C57BL/6 mice, which lack IFNAR in a subset of myeloid cells but express normal IFNAR levels on T cells, B cells, and most dendritic cells (Clausen et al., 1999; Diamond et al., 2011). We infected both LysMCre+IFNARfl/fl C7BL/6 mice and anti-IFNAR antibody-treated wild-type (WT) C57BL/6 mice with ZIKV MR766 and FSS13025 strains and mapped the H-2b-restricted CD8+ T cell responses. Additionally, we demonstrated a protective role for CD8+ T cells in controlling ZIKV infection in LysMCre+IFNARfl/fl mice. Our work provides an immunocompetent LATS1 and well-characterized H-2b mouse model for investigating protective gene deletion is efficient in Cannabichromene mature macrophages (83C98%) and granulocytes (100%) but partial for CD11C+ splenic dendritic cells (16%) (Clausen et al., 1999; Diamond et al., 2011). LysMCre+IFNARfl/fl and WT C57BL/6 mice were infected intravenously with MR766 or FSS13025, and levels of infectious virus in serum, liver, spleen, and brain at 1 and 3 Cannabichromene days after infection were determined. At day 1 post-infection, the infectious virus was detectable in all of the tissues tested in LysMCre+IFNARfl/fl mice infected with MR766 (Figure 2A) and FSS13025 (Figure 2B), whereas virus was undetectable in WT mice. At day Cannabichromene 3 post-infection, infectious ZIKV were still detectable in tissues of LysMCre+IFNARfl/fl mice. Based on these results, LysMCre+IFNAR1fl/fl mice, unlike WT mice, are susceptible to ZIKV infection. Open in a separate.
Further supporting this possibility, cell biological studies link NIMA to both cell tip growth and the modulation of interphase microtubule functions
Further supporting this possibility, cell biological studies link NIMA to both cell tip growth and the modulation of interphase microtubule functions. suppressor colonies were isolated and spread on plates and allowed to grow either at permissive or semi-permissive temperatures (35C). The data shows that although are unable to form colonies at this temperature (A) colonies that also carry suppressor mutations are able to GDC-0834 Racemate do so (B and C).(PDF) pgen.1004248.s003.pdf (311K) GUID:?0949F642-A3E6-4B7A-90DD-4FF7884FF381 Figure S4: (A) The cell tip location of NIMA is unchanged in the absence of ESCRT complex function. NIMA-GFP is detectable at 28% of WT cell tips (n?=?117; strain KF005) and a comparable 31% of (n?=?129; strain MGH61) cell tips at 35C. (B) NIMA-GFP levels at the cell tip decrease in mitosis when NIMA displays its characteristic nuclear location. Bar, 5 m.(PDF) pgen.1004248.s004.pdf (37K) GUID:?B4151E74-0B65-47C7-970D-33B80159A43A Figure S5: Colony growth of strains expressing ectopic NIMA constructs. (A) Growth of the indicated strains carrying driven NIMA constructs under conditions when ectopic NIMA is not expressed (lactose) or is expressed (threonine) compared to WT. (B) Growth of a strain carrying cell at 35C. Delay ?=? 0.81 s. Play rate ?=? 30 fps. Length of movie ?=? 7 min.(AVI) pgen.1004248.s011.avi (2.0M) GUID:?9BE1D983-1C19-4245-8A2E-5ADF4903C3EE Table S1: Genotypes of strains used in the study.(PDF) pgen.1004248.s012.pdf (58K) GUID:?F4C7F2EA-D875-4CBC-B83B-D826774863DC Abstract The Never in GDC-0834 Racemate Mitosis A (NIMA) kinase (the founding member of the Nek family of kinases) has been considered a mitotic specific kinase with nuclear restricted roles in the model fungus the results of a synthetic lethal screen performed in using the NIMA ortholog and genes encoding proteins of the Endosomal Sorting Complex Required for Transport (ESCRT) pathway. Absence of ESCRT pathway functions in combination with partial GDC-0834 Racemate NIMA ICAM1 function causes enhanced cell growth defects, including an inability to maintain a single polarized dominant cell tip. These genetic insights suggest NIMA potentially has interphase functions in addition to its established mitotic functions at nuclei. We therefore generated endogenously GFP-tagged NIMA (NIMA-GFP) which was fully functional to follow its interphase locations using live cell spinning disc 4D confocal microscopy. During interphase some NIMA-GFP locates to the tips of rapidly growing cells and, when expressed ectopically, also locates to the tips of cytoplasmic microtubules, suggestive of non-nuclear interphase functions. In support of this, perturbation of NIMA function either by ectopic overexpression or through partial inactivation results in marked cell tip growth defects with excess NIMA-GFP promoting multiple growing cell tips. Ectopic NIMA-GFP was found to locate to the plus ends of microtubules in an EB1 dependent manner, while impairing NIMA function altered the dynamic localization of EB1 and the cytoplasmic microtubule network. Together, our genetic and cell biological analyses reveal novel nonnuclear interphase functions for NIMA involving microtubules and the ESCRT pathway for normal polarized fungal cell tip growth. These insights extend the roles of NIMA both spatially and temporally and indicate that this conserved protein kinase could help integrate cell cycle progression with polarized cell growth. Author Summary All organisms have to integrate cell growth, and often the polarization of cell growth, with the rate of progression through the cell cycle. One of the most highly polarized modes of growth found in nature is displayed by the ubiquitous filamentous fungi. How the regulation of mitotic divisions is linked to polarized growth remains a mystery, but might involve mitotic regulators. One key mitotic regulator identified in the model filamentous fungus is the NIMA kinase, the founding member of the Nek family of protein kinases. This kinase is known to play mitotic specific roles within nuclei. Our genetic studies reported here reveal unexpected interactions between NIMA and six components of a pathway required for the turnover of cell.