Document text
Principal Investigator: Leonid B. Margolis
Organization: EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT
Fiscal Year: 2023
Award: $1,895,506
Funding agency: Eunice Kennedy Shriver National Institute of Child Health and Human Development
1. Cytokines in HIV disease
We followed longitudinally 90 people living with HIV (PWH) (59 women and 31 men) already virally suppressed and on antiretroviral therapy (ART) on average for 6 years. We measured the concentrations of 41 cytokines in their blood plasma and followed their trajectory overtime. We found that long ART had a significant effect on the trajectory of 9 cytokines. In particular, the concentration of 8 cytokines continued to decrease over time (CD163; IP-10; CXCL13; I-TAC; MCP-1; MIG; MIP-3 and TNF-), while LBP concentration slightly increased over time. Our results suggest that long ART continues to modulate pro-inflammatory cytokines past the initial decrease in concentrations observed during ART suppressive therapy when viral load becomes undetectable.
2. Extracellular vesicle-associated cytokines in non-progressors among PWH
We evaluated the cytokine profile associated with extracellular vesicles (EVs) in plasma samples from well-characterized cohorts of PWH with different virological control status. PWH were divided into 5 groups: ART-nave; ART-treated with undetectable viremia; elite controllers (EC) who controlled viremia (both transient controllers (TC) and persistent controllers (PC)); and uninfected individuals. Levels of 39 cytokines within and on the surface of EVs isolated from their stored plasma were quantified. Random forest, principal component analysis, and decision tree analyses were performed to identify specific cytokines as potential signatures of each study group. EC showed the highest levels of EV-associated cytokines, especially when PC were compared to TC. Higher levels of EV IL-18 were identified as a biomarker for EC population in the context of suppressed viremia, and higher levels of EV IL-3 and TRAIL were best at discriminating the PC phenotype from TC. In conclusion, higher levels of EV-associated cytokines in plasma may represent a new signature for the EC population.
3. EVs in immune activation of HIV-infected tissues under ART
We investigated the mechanisms by which EVs from HIV-1-infected/ART-suppressed lymphoid tissues lead to immune activation. Using fluorescently labeled EVs and flow cytometry, we identified B cells as a prime target cell population of EVs from these tissues. Over time, the amount of pre-germinal center B cells increased followed by increased germinal center B cells, and B cell activation markers (CD40, CD80, CD86). By day 9, effector memory T cells increased, and natural killer cells increased activation markers (CD38, CD69, CD80, CD86, HLA-DR). The increase in cytokines was traced to multiple cell types including CD4 T cells (MIP-1a, MIP-1b, RANTES), CD8 T cells (IFN-g, MIP-1a, MIP-1b, RANTES, TNFa), B cells (MIP-1a, MIP-1b, RANTES), NK cells (RANTES) and monocytes (RANTES). Understanding the cascade of events triggered by EVs from may lead to potential therapeutic targets for suppression of improper immune activation in HIV-infected individuals under ART.
4. Localization of an HIV-1 protein on EVs
We used several techniques to determine the localization of Nef in EVs. All approaches, including vesicle permeabilization, stripping of surface proteins, and remixing of EVs with Nef, led to the same conclusion: over 90% of the Nef molecules in the EV particles were located at the surface of the EVs. This result is consistent with Nef incorporation in the EVs via accession of the maturing late endosomes and/or the multivesicular bodies, and represent molecules associated with EVs during MVB fusion with the plasma membrane. Although only about 0.5% of total secreted Nef was associated with EVs, the Nef EVs were more biologically potent than soluble Nef. The distribution of Nef in EVs isolated from Nef-transfected cells was confirmed in EVs isolated from lymphocytes from PWH. Our study suggests that Nef can be released from cells in two forms: the dominant form is free protein, and the minor form is Nef associated with EVs with high bioactivity. Our results suggest that Nef EVs can be targeted by immunotherapy to treat HIV-associated co-morbidities.
5. EV-associated cytokines in complicated pregnancies leading to fetal death
We found five proteins (CXCL6, endoglin, MIP-3, PlGF, and TFPI) to be different in concentrations in cases of fetal death complicated by preeclampsia compared to those without preeclampsia. Our data suggest that mechanisms of fetal death complicated with preeclampsia include the dysregulation of angiogenic, coagulation and inflammatory pathways.
Among cases of fetal death, unsupervised clustering based on the proteins differentially expressed in either the EV or soluble fractions of patients with fetal death relative to controls revealed three major clusters of patients with distinct clinical and placental histopathological characteristics. Cluster 1 included 19 cases of fetal death that had 33 differentially expressed proteins in either the EV or the soluble fraction. Many of these cases of fetal death had two or more features consistent with maternal vascular malperfusion (MVM) and preeclampsia. Cluster 2 contained 12 cases of fetal death, and only PDL1 in the soluble fraction had different concentrations in this subgroup compared to controls. Fewer cases involved MVM lesions or preeclampsia but included the highest percentage of unexplained fetal death cases. Cluster 3 included 16 cases of fetal death with 21 proteins differentially expressed in either the EV or the soluble fraction of patients compared to the controls and fewer features of MVM and preeclampsia than cluster 1. Overall, our results demonstrate the value of including EV-associated biomarkers to improve the prediction of fetal death and understand the pathophysiology of this obstetrical syndrome.
6. Blocking SARS-CoV-2 transmission
Nanocellulose particles were obtained from ivory nuts from a palm tree from the Arecaceae family that grows in tropical regions of South America and modified with dense sulfate charges. Due to its derivation from the endosperm of the palms seed, it was designated endospermic nanocellulose (ENC). The unique primary cell walls yielded more homogeneous and thinner cellulose nanoparticles compared to cell walls from other vegetal tissues. We evaluated the feasibility of sulfonated ENC as an inhibitor of viral transmission first by ELISA, which demonstrated effective blocking of SARS-CoV-2 S protein interaction with ACE2 in the presence of ENC, and allowed us to identify optimal working concentrations of ENC. We then used two different methods of blocking transmission of virus to cell cultures using ENC: a non-specific method of ENC encapsulation of SARS-CoV-2 pseudovirus particles or 293T-ACE2 target cells, and a highly specific method immobilizing molecular lures, anti-SARS-CoV-2 spike (S) protein antibodies and soluble ACE2 receptors, in ENC to bind virus. Both methods demonstrated that ENC effectively blocked cell infection by SARS-CoV-2 pseudoviruses, demonstrating the universal binding property of ENC and its usefulness for development of new strategies to prevent viral transmission. This material has potential applications as a film for masks and filters, to bind viral particles and increasing the effectiveness of these filtration devices, or to create adsorbent surfaces to capture or inactivate viruses. The concentrations of ENC used in these experiments will lend themselves easily to aerosolization making them ideal for such purposes. Nanoparticles that are deemed safe for in vivo use also have potential to enhance bioavailability of natural compounds, or for drug delivery.
Terms: <(TNF)-α><2019 novel corona virus><2019 novel coronavirus><2019-nCoV><2019-nCoV S protein><2019-nCoV spike glycoprotein><2019-nCoV spike protein><ACE2><AIDS><AIDS Virus><APO2L><Acquired Immune Deficiency><Acquired Immune Deficiency Syndrome><Acquired Immune Deficiency Syndrome Virus><Acquired Immuno-Deficiency Syndrome><Acquired Immunodeficiency Syndrome><Acquired Immunodeficiency Syndrome Virus><Acquired Immunologic Deficiency Syndrome><Affect><Alkanesulfonates><Alkyl Sulfonates><Alkylsulfonate Compound><Antibodies><Apo-2L><Arecaceae><Attention><B blood cells><B cell><B cells><B-Cell Activation><B-Cell Attracting Chemokine 1><B-Cells><B-Lymphocyte Chemoattractant><B-Lymphocytes><B-cell><B7-1><B7-H1><B7H1><BB1><BCA1><BLC gene><BLC protein><Binding><Bioavailability><Biological Availability><Biological Markers><Blood Plasma><Blood Vessels><Blood monocyte><Body Tissues><Bp50><CCL2><CCL2 gene><CCL23><CCL23 gene><CCL3><CCL3 gene><CCL5><CD105 Antigen><CD183><CD274><CD28LG><CD28LG1><CD4 Cells><CD4 Positive T Lymphocytes><CD4 T cells><CD4 helper T cell><CD4 lymphocyte><CD4+ T-Lymphocyte><CD4-Positive Lymphocytes><CD40><CD8 Cell><CD8 T cells><CD8 lymphocyte><CD8+ T cell><CD8+ T-Lymphocyte><CD8-Positive Lymphocytes><CD8-Positive T-Lymphocytes><CD80><CD80 gene><CD86><CD86 gene><CDW40><CK-BETA-8><CKA-3><CKR-L2><CKb8><CMKAR3><COVID-19 S protein><COVID-19 infection><COVID-19 spike glycoprotein><COVID-19 spike protein><COVID-19 transmission><COVID-19 virus><COVID-19 virus infection><COVID-19 virus transmission><COVID19 S protein><COVID19 infection><COVID19 spike glycoprotein><COVID19 spike protein><COVID19 virus><CRG-2><CXCL10><CXCL10 gene><CXCL11><CXCL11 gene><CXCL13><CXCL13 gene><CXCL6><CXCL6 gene><CXCR3><CXCR3 gene><Cachectin><Cell Body><Cell Communication><Cell Culture Techniques><Cell Interaction><Cell Wall><Cell membrane><Cell-to-Cell Interaction><Cells><Cellulose><Characteristics><Charge><Chemokine (C-C Motif) Ligand 5><Chemokine (C-C motif) Ligand 3><Chemokine (C-X-C Motif) Receptor 3><Chemokine, CC Motif, Ligand 2><Chemokine, CXC Motif, Ligand 13><Ckb-8><Ckb-8-1><Clinical><Clotting><CoV-2><CoV2><Coagulation><Coagulation Process><Collaborations><Cross Sectional Analysis><Cross-Sectional Analyses><Cross-Sectional Studies><Cross-Sectional Survey><Cytokine Activation><Cytoplasmic Membrane><Cytotoxic cell><D17S136E><Data><Decision Trees><Derivation><Derivation procedure><Development><Devices><Disease><Disease Frequency Surveys><Disorder><Drug Delivery><Drug Delivery Systems><Drug Targeting><Dysfunction><ELISA><EPH Gestosis><Early treatment><Effectiveness><Encapsulated><Endoglin><Enzyme-Linked Immunosorbent Assay><Eosinophil-Mast Cell Growth-Factor><Erythrocyte Burst-Promoting Factor><Event><Family><Fetal Death><Film><Filtration><Filtration Fractionation><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><Functional disorder><G Protein-Coupled Receptor 9><G0S19-1><GCP-2><GPR9><Germinal Center><Goals><H174><HIV><HIV Infections><HIV-1><HIV-I><HIV1><HLA-BR><HLA-BR Antigens><HLA-D-Related Antigens><HLA-DR><HLA-DR Antigens><HLA-MT><HLA-MT Antigens><HTLV-III Infections><HTLV-III-LAV Infections><Hematopoietic Cytokine><Human><Human Immunodeficiency Virus Type 1><Human Immunodeficiency Viruses><Human T-Lymphotropic Virus Type III Infections><Human immunodeficiency virus 1><I-TAC><IFI10><IFN-Gamma><IFN-Gamma-Inducing Factor Gene><IFN-g><IFN-gamma-Inducing Factor><IFN-γ><IFNG><IFNγ><IGIF><IGIF Gene><IL-1 Gamma><IL-1 Gamma Gene><IL-18><IL-18 Gene><IL-1g><IL-1g Gene><IL-3><IL-3 Gene><IL18><IL18 Protein><IL18 gene><IL1F4><IL1F4 Gene><IL3><IL3 Gene><IL3 Protein><INP10><IP-10><IP-9><IP10><IP10 Receptor><IP10-Mig receptor><IP10-R><IP9><Immune Cell Activation><Immune Interferon><Immune mediated therapy><Immunologically Directed Therapy><Immunotherapy><Individual><Infection><Inflammatory><Interferon Gamma><Interferon Type II><Interferon-Gamma-Inducing Factor Gene><Interferon-gamma-Inducing Factor><Interleukin 18 (Interferon-Gamma-Inducing Factor)><Interleukin 18 (Interferon-Gamma-Inducing Factor) Gene><Interleukin 18 Proprotein><Interleukin 18 Proprotein Gene><Interleukin-1 Gamma><Interleukin-1 Gamma Gene><Interleukin-18><Interleukin-18 Precursor><Interleukin-18 Precursor Gene><K lymphocyte><LAB7><LAV-HTLV-III><LD78ALPHA><Label><Lead><Leanness><Lesion><Link><Lymphadenopathy-Associated Virus><Lymphatic Tissue><Lymphatic cell><Lymphocyte><Lymphocytic><Lymphoid Tissue><MCAF><MCP-1><MCP1><MGC12320><MGC12320 Gene><MGC17164><MGC9013><MIP 1alpha><MIP-1-alpha><MIP-1a><MIP-3><MIP1A><MIP3><MOB-1><MPIF-1><Macrophage Activation><Macrophage-Derived TNF><Marrow monocyte><Masks><Mast-Cell Colony-Stimulating Factor><Measures><Mediating><Membrane Protein Gene><Membrane Proteins><Membrane-Associated Proteins><Methods><Mig Receptor><Mig-R><MigR><Minor><Modern Man><Molecular><Molecular Interaction><Monocyte Chemoattractant Protein-1><Monocyte Chemotactic Protein-1><Monocyte Chemotactic and Activating Factor><Monocyte Chemotactic and Activating Protein><Monocyte Chemotactive and Activating Factor><Monocyte Secretory Protein JE><Monocyte-Derived TNF><Multivesicular Body><NK Cells><Natural Compound><Natural Killer Cells><Nuts><Outcome><P-CSF><P-Cell Stimulating Factor><PD-L1><PDL-1><PDL1><PGF gene><PLGF-2><Palmae><Pathogenesis><Pathology><Pathway interactions><Patients><Pb element><Permeability><Persons><Phenotype><Physiologic Availability><Physiopathology><PlGF><PlGF protein><Placental Growth Factor><Plasma><Plasma Membrane><Plasma Serum><Polyanhydroglucuronic Acid><Population><Pre-Eclampsia><Preeclampsia><Pregnancy Complications><Pregnancy Toxemias><Prevent viral transmission><Principal Component Analyses><Principal Component Analysis><Programmed Cell Death 1 Ligand 1><Programmed Death Ligand 1><Property><Proteins><Proteinuria-Edema-Hypertension Gestosis><Proteomics><Public Health><RANTES><Receptor Protein><Reporting><Research><Reticuloendothelial System, Serum, Plasma><Role><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV-2 S protein><SARS-CoV-2 infection><SARS-CoV-2 inhibitor><SARS-CoV-2 spike glycoprotein><SARS-CoV-2 spike protein><SARS-CoV-2 transmission><SARS-CoV2><SARS-CoV2 S protein><SARS-CoV2 infection><SARS-CoV2 spike glycoprotein><SARS-CoV2 spike protein><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><SCYA2><SCYA23><SCYA3><SCYA5><SCYB10><SCYB11><SCYB13><SCYB6><SCYB9B><SIS delta><SIS-delta><SISd><Sampling><Severe Acute Respiratory Coronavirus 2><Severe Acute Respiratory Distress Syndrome CoV 2><Severe Acute Respiratory Distress Syndrome Corona Virus 2><Severe Acute Respiratory Distress Syndrome Coronavirus 2><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome coronavirus 2 S protein><Severe acute respiratory syndrome coronavirus 2 infection><Severe acute respiratory syndrome coronavirus 2 inhibitor><Severe acute respiratory syndrome coronavirus 2 spike glycoprotein><Severe acute respiratory syndrome coronavirus 2 spike protein><Severe acute respiratory syndrome related corona virus 2><Small Inducible Cytokine A2><Small Inducible Cytokine A3><Small Inducible Cytokine A5><Small Inducible Cytokine Subfamily B, Member 13><South America><Stem Cell Inhibitor><Structure of germinal center of lymph node><Subgroup><Sulfate><Surface><Surface Proteins><T memory cell><T-Cell RANTES Protein><T-Cell Specific Protein p288><T-Cells><T-Lymphocyte><T4 Cells><T4 Lymphocytes><T8 Cells><T8 Lymphocytes><TCP228><TFPI><TL2><TNF><TNF A><TNF Alpha><TNF gene><TNF-α><TNFA><TNFRSF5><TNFRSF5 gene><TNFSF10><TNFSF10 gene><TNFα><TRAIL><Techniques><Thinness><Time><Tissues><Transfection><Transforming Growth Factor P Receptor III><Transmission><Trees><Tumor Necrosis Factor><Tumor Necrosis Factor Receptor Superfamily Member 5 Gene><Tumor Necrosis Factor-alpha><Unexplained fetal death><Up-Regulation><Upregulation><Vesicle><Viral><Viral Burden><Viral Diseases><Viral Load><Viral Load result><Viral Pathogenesis><Viremia><Virus><Virus Diseases><Virus Inhibitors><Virus-HIV><Woman><Wuhan coronavirus><activated B cells><aerosolized><alpha-Cellulose><angiotensin converting enzyme 2><angiotensin converting enzyme II><anti-microbial><anti-retroviral therapy><anti-retroviral treatment><antimicrobial><antiretroviral therapy><antiretroviral treatment><b-R1><bio-markers><biologic marker><biomarker><biomarker identification><block SARS-CoV-2><block severe acute respiratory syndrome coronavirus 2><cell culture><cell cultures><cell type><co-morbid><co-morbidity><cohort><comorbidity><compare to control><comparison control><complications during pregnancy><coronavirus disease 2019 S protein><coronavirus disease 2019 infection><coronavirus disease 2019 spike glycoprotein><coronavirus disease 2019 spike protein><coronavirus disease 2019 transmission><coronavirus disease 2019 virus><coronavirus disease 2019 virus transmission><coronavirus disease-19 virus><cytokine><developmental><differential expression><differentially expressed><early therapy><enzyme linked immunoassay><experience><experiment><experimental research><experimental study><experiments><extracellular vesicles><fetus death><flow cytophotometry><gIP-10><hCoV19><heavy metal Pb><heavy metal lead><human tissue><identification of biomarkers><immune activation><immune therapeutic approach><immune therapeutic interventions><immune therapeutic regimens><immune therapeutic strategy><immune therapy><immune-based therapies><immune-based treatments><immuno therapy><improved><in vivo><infected with COVID-19><infected with COVID19><infected with SARS-CoV-2><infected with SARS-CoV2><infected with coronavirus disease 2019><infected with severe acute respiratory syndrome coronavirus 2><inhibit SARS-CoV-2><inhibit severe acute respiratory syndrome coronavirus 2><lFN-Gamma><late endosome><lymph cell><marker identification><memory T lymphocyte><men><monocyte><nCoV2><nano><nano particle><nano-sized particle><nanoparticle><nanosized particle><naturally occurring compound><nervous system development><obstetric care><obstetrical syndromes><p50><particle><pathophysiology><pathway><placenta growth factor><plasmalemma><pre-eclamptic><pregnancy toxemia/hypertension><pregnancy-related complications><prevent><preventing><product placenta growth factor><programmed cell death ligand 1><programmed cell death protein ligand 1><protein death-ligand 1><random forest><receptor><respiratory><severe acute respiratory syndrome coronavirus 2 transmission><social role><sulfonate><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><therapeutic target><thymus derived lymphocyte><tissue factor pathway inhibitor><transcriptional differences><transmission blocking><transmission process><transmitted COVID-19><transmitted SARS-CoV-2><transmitted coronavirus disease 2019><transmitted severe acute respiratory syndrome coronavirus 2><vascular><vesicle release><vesicular release><viraemia><viral infection><viral inhibitor><viral sepsis><viral transmission><virus infection><virus pathogenesis><virus transmission><virus-induced disease><virusemia>