The transcription factor BATF operates as an essential differentiation checkpoint in early effector CD8+ T cells

The transcription factor BATF is required for interleukin 17 (IL-17)-producing helper T cell (TH17) and follicular helper T cell (TFH) differentiation. Here, we show that BATF also has a fundamental role in regulating effector CD8+ T cell differentiation. BATF-deficient CD8+ T cells show profound defects in effector expansion and undergo proliferative and metabolic catastrophe early after antigen encounter. BATF, together with IRF4 and Jun proteins, binds to and promotes early expression of genes encoding lineage-specific transcription-factors (T-bet and Blimp-1) and cytokine receptors, while paradoxically repressing genes encoding effector molecules (IFN-γ and granzyme B). Thus, BATF amplifies TCR-dependent transcription factor expression and augments inflammatory signal propagation but restrains effector gene expression. This checkpoint prevents irreversible commitment to an effector fate until a critical threshold of downstream transcriptional activity has been achieved

Turfgrass research report 1996

Preemergence herbicide efficacy on crabgrass-1996 / J. Street and R. Stewart --Postemergence herbicide efficacy on crabgrass -1996 / J. Street and R. Stewart -- General turfgrass broadleaf weed control evaluation / W. Pound -- Nonselective herbicide evaluation / W. Pound -- Postemergent yellow nutsedge evaluation / W. Pound and R. Stewart -- Preemergent broadleaf weed control / W. Pound -- Control of ant mounds in turfgrass-1996 / D. J. Shetlar, H. D. Niemczyk and K. T. Power -- Control of hairy chinch bugs in turfgrass -1996 / D. J. Shetlar, H. D. Niemczyk and K. T. Power -- Preventive and curative insecticide applications for control of hairy chinch bugs in turfgrass - 1996 / D. J. Shetlar, H. D. Niemczyk and K. T. Power -- Quick kill of black cutworm larvae in bentgrass - 1996 / D. J. Shetlar, H. D. Niemczyk and K. T. Power -- Control of black cutworm, Agrotis ipsilon (Hufnagel), and sod webworms (Pyralidae: Crambinae) in creeping bentgrass, Agrostis palustris Hudson, with spinosad formulations, Columbus, Ohio, 1996 / D. J. Shetlar, H. D. Niemczyk and M. Belcher -- Control of black cutworm, Agrotis ipsilon (Hufnagel), and sod webworms (Pyralidae: Crambinae) in creeping bentgrass, Agrostis palustris Hudson, with observations on black turfgrass Ataenius adults, Ataenius spretulus (Haldeman), Columbus, Ohio, 1996 / D. J. Shetlar, H. D. Niemczyk and M. Belcher -- Control of black cutworm, Agrotis ipsilon (Hufnagel), sod webworms (Pyralidae: Crambinae), and black turfgrass Ataenius adults, Ataenius spretulus (Haldeman), in creeping bentgrass, Agrostis palustris Hudson, Columbus, Ohio, 1996 / D. J. Shetlar, H. D. Niemczyk and M. Belcher -- Control of bluegrass billbug, Sphenophorus parvulus Gyllenhal, larvae in lawn turf, Wooster, Ohio, 1996 / D. J. Shetlar, H. D. Niemczyk and M. Belcher -- Efficacy of spinosad applied prior to oviposition for control of bluegrass billbug larvae in turfgrass - 1996 / D. J. Shetlar, H. D. Niemczyk and K. T. Power -- Efficacy of insecticides applied prior to oviposition for control of bluegrass billbug larvae in turfgrass - 1996 / D. J. Shetlar, H. D. Niemczyk and K. T. Power -- Application of insecticides to kill black turfgrass Ataenius adults prior to egg laying on golf course fairways-1996 / D. J. Shetlar, H. D. Niemczyk and K. T. Power -- Preventive applications for control of Japanese beetle larvae in turfgrass - 1996 / D. J. Shetlar, H. D. Niemczyk and K. T. Power -- Influence of application date on the efficacy of insecticides applied for control of Japanese beetle larvae in turfgrass - 1996 / D. J. Shetlar, H. D. Niemczyk and K. T. Power -- Application of insecticides for preventive control of Japanese beetle larval populations in turfgrass - 1996 / D. J. Shetlar, H. D. Niemczyk and K. T. Power -- Application of Bacillus thuringiensis strain 'buibui' for control of white grubs in turfgrass -1996 / D. J. Shetlar, H. D. Niemczyk and K. T. Power -- Curative control of masked chafer larvae in turfgrass -1996 / D. J. Shetlar, H. D. Niemczyk and K. T. Power -- A field test of RH-0345 2SC and 2.5G at 2.0 lb.ai/ acre and Merit 75WP at 0.3 lb. ai/ acre for control of black cutworm larvae on golf course greens / H. D. Niemczyk -- Dollar spot control study - 1996 / K. Danneberger, J. Rimelspach, M. Boehm, and J. Taylor -- quality ratings for various fungicide treatments on creeping bentgrass putting green turf / K. Danneberger, J. Rimelspach, M. Boehm, and J. Taylor -- Brown patch control study on creeping bentgrass turf - 1996 / K. Danneberger, J. Rimelspach, M. Boehm, and J. Taylor -- Brown patch control on tall fescue / K. Danneberger, J. Rimelspach, M. Boehm, and J. Taylor -- Yellow tuft study-1996 / K. Danneberger, J. Rimelspach, M. Boehm, and J. Taylor -- Creeping bentgrass melting-out study / K. Danneberger, J. Taylor, R. Golembiewski, G. Bell, J. Rimelspach, and M. Boehm -- Kentucky bluegrass melting-out study / K. Danneberger, J. Rimelspach, M. Boehm and J. Taylor -- Red thread control study on perennial ryegrass / J. Rimelspach, K. Danneberger, and M. Boehm -- Red thread control study on Kentucky bluegrass / J. Rimelspach, K. Danneberger and M. Boehm -- Evaluation of fungicides for the control of red thread in Kentucky bluegrass, 1996 / J. Rimelspach, M. Boehm, K. Danneberger and J. Taylor -- Evaluation of fungicides for the control of red thread in perennial ryegrass, 1996 / J. Rimelspach, M. Boehm, K. Danneberger and J. Taylor -- Pink snow mold control study-1995-1996 / J. Rimelspach and K. Danneberger -- Primo & Sentinel applications on Poa annua quality / K. Danneberger, J. Taylor, R. Golembiewski, and G. Bell -- Nitrogen source, rate, and timing effect on Kentucky bluegrass -1996 / J. R. Street and R. M. Stewart -- Natural organic source evaluation on a Kentucky bluegrass-perennial ryegrass mixture-1996 / J. R. Street and R. M. Stewart -- Polymer-coated urea source and rate effect on Kentucky bluegrass / J. R. Street and R. M. Stewart -- Polymer-coated urea and IBDU fertilizer performance on Kentucky bluegrass / J. R. Street and R. M. Stewart -- 1993 NTEP bentgrass (fairway/tee) cultivar evaluation / J. A. Taylor -- 1993 NTEP fineleaf fescue cultivar evaluation / J. Taylor -- 1994 NTEP perennial ryegrass cultivar evaluation / J. Taylor -- 1995 Kentucky bluegrass (medium/high input) cultivar evaluation / J. Taylor -- 1995 NTEP Kentucky bluegrass (low input) cultivar evaluation / J. Taylor -- Controlling annual bluegrass and rough bluegrass in creeping bentgrass fairways: a nutritional approach / G. E. Bell, E. Odorizzi and T. K. Danneberger -- "Primo" growth regulator evaluation on creeping bentgrass / W. Pound and R. Stewart -- Influence of dollar spot on a blend of two creeping bentgrass cultivars / R. C. Golembiewski, T. K. Danneberger and P. M. Sweeney -- Dollar spot severity as influenced by Primo, creeping bentgrass cultivars, and nitrogen fertility / R. C. Golembiewski and T. K. Danneberger -- Identification of bulk samples of perennial ryegrass cultivars with RAPD Markers / P. M. Sweeney and T. K. Danneberge

Polyfunctional T cell responses in children in early stages of chronic Trypanosoma cruzi infection contrast with monofunctional responses of long-term infected adults

Background: Adults with chronic Trypanosoma cruzi exhibit a poorly functional T cell compartment, characterized by monofunctional (IFN-γ-only secreting) parasite-specific T cells and increased levels of terminally differentiated T cells. It is possible that persistent infection and/or sustained exposure to parasites antigens may lead to a progressive loss of function of the immune T cells. Methodology/Principal Findings: To test this hypothesis, the quality and magnitude of T. cruzi-specific T cell responses were evaluated in T. cruzi-infected children and compared with long-term T. cruzi-infected adults with no evidence of heart failure. The phenotype of CD4+ T cells was also assessed in T. cruzi-infected children and uninfected controls. Simultaneous secretion of IFN-γ and IL-2 measured by ELISPOT assays in response to T. cruzi antigens was prevalent among T. cruzi-infected children. Flow cytometric analysis of co-expression profiles of CD4+ T cells with the ability to produce IFN-γ, TNF-α, or to express the co-stimulatory molecule CD154 in response to T. cruzi showed polyfunctional T cell responses in most T. cruzi-infected children. Monofunctional T cell responses and an absence of CD4+TNF-α+-secreting T cells were observed in T. cruzi-infected adults. A relatively high degree of activation and differentiation of CD4+ T cells was evident in T. cruzi-infected children. Conclusions/Significance: Our observations are compatible with our initial hypothesis that persistent T. cruzi infection promotes eventual exhaustion of immune system, which might contribute to disease progression in long-term infected subjects.Fil: Albareda, María Cecilia. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud. Instituto Nacional de Parasitología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Provincia de Buenos Aires. Ministerio de Salud. Hospital Interzonal de Agudos "Eva Perón"; ArgentinaFil: de Rissio, Ana María. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud. Instituto Nacional de Parasitología; ArgentinaFil: Tomas, Gonzalo. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud. Instituto Nacional de Parasitología; ArgentinaFil: Serjan, Alicia. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos "Juan A. Fernández"; ArgentinaFil: Alvarez, María Gabriela. Provincia de Buenos Aires. Ministerio de Salud. Hospital Interzonal de Agudos "Eva Perón"; ArgentinaFil: Viotti, Rodolfo Jorge. Provincia de Buenos Aires. Ministerio de Salud. Hospital Interzonal de Agudos "Eva Perón"; ArgentinaFil: Fichera, Laura Edith. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud. Instituto Nacional de Parasitología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Esteva, Mónica Inés. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud. Instituto Nacional de Parasitología; ArgentinaFil: Potente, Daniel Fernando. Provincia de Buenos Aires. Ministerio de Salud. Hospital Interzonal de Agudos "Eva Perón"; ArgentinaFil: Armenti, Alejandro. Provincia de Buenos Aires. Ministerio de Salud. Hospital Interzonal de Agudos "Eva Perón"; ArgentinaFil: Tarleton, Rick L.. University of Georgia; Estados UnidosFil: Laucella, Susana Adriana. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud. Instituto Nacional de Parasitología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Hrs and SNX3 Functions in Sorting and Membrane Invagination within Multivesicular Bodies

After internalization, ubiquitinated signaling receptors are delivered to early endosomes. There, they are sorted and incorporated into the intralumenal invaginations of nascent multivesicular bodies, which function as transport intermediates to late endosomes. Receptor sorting is achieved by Hrs—an adaptor-like protein that binds membrane PtdIns3P via a FYVE motif—and then by ESCRT complexes, which presumably also mediate the invagination process. Eventually, intralumenal vesicles are delivered to lysosomes, leading to the notion that EGF receptor sorting into multivesicular bodies mediates lysosomal targeting. Here, we report that Hrs is essential for lysosomal targeting but dispensable for multivesicular body biogenesis and transport to late endosomes. By contrast, we find that the PtdIns3P-binding protein SNX3 is required for multivesicular body formation, but not for EGF receptor degradation. PtdIns3P thus controls the complementary functions of Hrs and SNX3 in sorting and multivesicular body biogenesis

The Nucleocapsid Region of HIV-1 Gag Cooperates with the PTAP and LYPXnL Late Domains to Recruit the Cellular Machinery Necessary for Viral Budding

HIV-1 release is mediated through two motifs in the p6 region of Gag, PTAP and LYPXnL, which recruit cellular proteins Tsg101 and Alix, respectively. The Nucleocapsid region of Gag (NC), which binds the Bro1 domain of Alix, also plays an important role in HIV-1 release, but the underlying mechanism remains unclear. Here we show that the first 202 residues of the Bro1 domain (Broi) are sufficient to bind Gag. Broi interferes with HIV-1 release in an NC–dependent manner and arrests viral budding at the plasma membrane. Similar interrupted budding structures are seen following over-expression of a fragment containing Bro1 with the adjacent V domain (Bro1-V). Although only Bro1-V contains binding determinants for CHMP4, both Broi and Bro1-V inhibited release via both the PTAP/Tsg101 and the LYPXnL/Alix pathways, suggesting that they interfere with a key step in HIV-1 release. Remarkably, we found that over-expression of Bro1 rescued the release of HIV-1 lacking both L domains. This rescue required the N-terminal region of the NC domain in Gag and the CHMP4 binding site in Bro1. Interestingly, release defects due to mutations in NC that prevented Bro1 mediated rescue of virus egress were rescued by providing a link to the ESCRT machinery via Nedd4.2s over-expression. Our data support a model in which NC cooperates with PTAP in the recruitment of cellular proteins necessary for its L domain activity and binds the Bro1–CHMP4 complex required for LYPXnL–mediated budding

The amyloid precursor protein controls PIKfyve function

While the Amyloid Precursor Protein (APP) plays a central role in Alzheimer's disease, its cellular function still remains largely unclear. It was our goal to establish APP function which will provide insights into APP's implication in Alzheimer's disease. Using our recently developed proteo-liposome assay we established the interactome of APP's intracellular domain (known as AICD), thereby identifying novel APP interactors that provide mechanistic insights into APP function. By combining biochemical, cell biological and genetic approaches we validated the functional significance of one of these novel interactors. Here we show that APP binds the PIKfyve complex, an essential kinase for the synthesis of the endosomal phosphoinositide phosphatidylinositol-3,5-bisphosphate. This signalling lipid plays a crucial role in endosomal homeostasis and receptor sorting. Loss of PIKfyve function by mutation causes profound neurodegeneration in mammals. Using C. elegans genetics we demonstrate that APP functionally cooperates with PIKfyve in vivo. This regulation is required for maintaining endosomal and neuronal function. Our findings establish an unexpected role for APP in the regulation of endosomal phosphoinositide metabolism with dramatic consequences for endosomal biology and important implications for our understanding of Alzheimer's disease

The Sodium/Proton Exchanger NHE8 Regulates Late Endosomal Morphology and Function

The pH and lumenal environment of intracellular organelles is considered essential for protein sorting and trafficking through the cell. We provide the first evidence that a mammalian sodium (potassium)/proton exchanger, NHE8, plays a key role in the control of protein trafficking and endosome morphology

Enhanced Transferrin Receptor Expression by Proinflammatory Cytokines in Enterocytes as a Means for Local Delivery of Drugs to Inflamed Gut Mucosa

Therapeutic intervention in inflammatory bowel diseases (IBDs) is often associated with adverse effects related to drug distribution into non-diseased tissues, a situation which attracts a rational design of a targeted treatment confined to the inflamed mucosa. Upon activation of immune cells, transferrin receptor (TfR) expression increases at their surface. Because TfR is expressed in all cell types we hypothesized that its cell surface levels are regulated also in enterocytes. We, therefore, compared TfR expression in healthy and inflamed human colonic mucosa, as well as healthy and inflamed colonic mucosa of the DNBS-induced rat model. TfR expression was elevated in the colonic mucosa of IBD patients in both the basolateral and apical membranes of the enterocytes. Increased TfR expression was also observed in colonocytes of the induced colitis rats. To explore the underlying mechanism CaCo-2 cells were treated with various proinflammatory cytokines, which increased both TfR expression and transferrin cellular uptake in a mechanism that did not involve hyper proliferation. These findings were then exploited for the design of targetable carrier towards inflamed regions of the colon. Anti-TfR antibodies were conjugated to nano-liposomes. As expected, iron-starved Caco-2 cells internalized anti-TfR immunoliposomes better than controls. Ex vivo binding studies to inflamed mucosa showed that the anti-TfR immunoliposomes accumulated significantly better in the mucosa of DNBS-induced rats than the accumulation of non-specific immunoliposomes. It is concluded that targeting mucosal inflammation can be accomplished by nano-liposomes decorated with anti-TfR due to inflammation-dependent, apical, elevated expression of the receptor