ABSTRACT
Ebola virus disease (EVD) often leads to severe and fatal outcomes in humans with early supportive care increasing the chances of survival. Profiling the human plasma lipidome provides insight into critical illness as well as diseased states, as lipids have essential roles as membrane structural components, signaling molecules, and energy sources. Here we show that the plasma lipidomes of EVD survivors and fatalities from Sierra Leone, infected during the 2014-2016 Ebola virus outbreak, were profoundly altered. Focusing on how lipids are associated in human plasma, while factoring in the state of critical illness, we found that lipidome changes were related to EVD outcome and could identify states of disease and recovery. Specific changes in the lipidome suggested contributions from extracellular vesicles, viremia, liver dysfunction, apoptosis, autophagy, and general critical illness, and we identified possible targets for therapies enhancing EVD survival.
Subject(s)
Critical Illness/epidemiology , Hemorrhagic Fever, Ebola/genetics , Lipid Metabolism/genetics , Lipids/genetics , Adolescent , Adult , Child , Disease Outbreaks , Ebolavirus/genetics , Ebolavirus/pathogenicity , Female , Gene Expression Regulation/genetics , Hemorrhagic Fever, Ebola/blood , Hemorrhagic Fever, Ebola/pathology , Hemorrhagic Fever, Ebola/virology , Humans , Lipids/blood , Male , Sierra Leone/epidemiology , Young AdultABSTRACT
Autoradiography identified six 125I-labeled proteins, ranging in molecular weight (Mr) from 20,000 to approximately 110,000, on the plasmalemma of Eimeria bovis sporozoites. Immunoblotting with bovine antiserum generated by intravenous inoculations of sporozoites and with immune serum generated by per os inoculations of oocysts revealed that protein 20 (i.e., 20,000 Mr) was an immunodominant antigen on the surface of E. bovis sporozoites.
Subject(s)
Antigens, Protozoan/isolation & purification , Antigens, Surface/isolation & purification , Eimeria/immunology , Membrane Proteins/isolation & purification , Animals , Antigens, Protozoan/immunology , Antigens, Surface/immunology , Blotting, Western , Cattle , Eimeria/growth & development , Male , Membrane Proteins/immunology , Molecular WeightABSTRACT
Five monoclonal antibodies (MAbs) were partially characterized and tested for their ability to inhibit penetration of Madin-Darby bovine kidney (MDBK) cells by sporozoites of Eimeria bovis. By indirect fluorescent-antibody assays, all MAbs reacted with acetone-fixed sporozoites, but only two MAbs, EbS9 (immunoglobulin G1) and EbS11 (immunoglobulin G2a), localized specifically on the plasmalemma of live sporozoites. Two of the five MAbs also reacted with acetone-fixed first-generation merozoites of E. bovis; however, none of the MAbs reacted with live merozoites. Treatment of live sporozoites with EbS9 or EbS11 resulted in 79 and 73% decreases, respectively, in sporozoite penetration of MDBK cells. No significant differences in cell penetration occurred in MDBK cells inoculated with sporozoites that had been treated with the other three MAbs. Both EbS9 and EbS11 reacted in Western blots (immunoblots) of sporozoites with the same 20,000-relative-molecular-weight protein. The antigens against which these neutralizing MAbs react might be useful in immunizing against bovine coccidiosis.
Subject(s)
Antibodies, Monoclonal/immunology , Antibodies, Protozoan/immunology , Antigens, Protozoan/immunology , Antigens, Surface/immunology , Eimeria/immunology , Animals , Cell Line , Cells/parasitology , Dogs , Fluorescent Antibody Technique , Immunoglobulin G/immunology , Mice , Molecular WeightABSTRACT
Macrophage (M phi)-activating lymphokines present in concanavalin A-stimulated bovine T-lymphocyte cultures (ConAS) were studied by assessing their effects on Eimeria bovis and Toxoplasma gondii growth in cultured bovine monocytes (BM) and mouse M phi. The in vitro development of both parasites was assessed by incorporation of [3H]uracil and by microscopic examination of parallel cultures. Incorporation of [3H]uracil into infected cultures was an accurate indicator of growth of both E. bovis and T. gondii in BM and mouse M phi. Sporozoites of E. bovis underwent merogony in untreated BM but not in mouse M phi, whereas T. gondii developed in both cell types. Inhibition of T. gondii growth was greatest in ConAS-treated BM, whereas preincubation of mouse M phi with ConAS resulted in about 80% growth inhibition. There was no significant difference between the inhibition of either T. gondii sporozoite- or tachyzoite-induced growth in ConAS-treated cells, showing that activation pathways are equally effective against both stages. Treatment of ConAS with glycine-hydrochloride buffer (pH 2) resulted in a total loss of antiviral activity mediated by gamma interferon (IFN-gamma). When pH 2 dialyzed ConAS was used to activate BM, inhibition of T. gondii growth was only partially affected. Because bovine IFN-gamma does not activate mouse M phi and due to the partial effects of pH 2 on ConAS-induced growth inhibition, the major component(s) of ConAS responsible for T. gondii growth inhibition is distinct from IFN-gamma. Furthermore, IFN-gamma may act synergistically rather than being part of a priming sequence for M phi responsiveness to other lymphokines. Murine recombinant granulocyte-macrophage colony-stimulating factor (rGM-CSF) was tested for any microbistatic activity against T. gondii sporozoites and tachyzoites. There was no significant difference in either colony formation or [3H]uracil incorporation between rGM-CSF-treated and control cultures, regardless of host cell type. Thus, rGM-CSF does not induce adequate M phi activation to kill T. gondii and is not a major microbistatic component of ConAS. rGM-CSF also had no effect on T. gondii infection in vivo.