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Principal Investigator: Heather Kalish
Organization: NATIONAL INSTITUTE OF BIOMEDICAL IMAGING AND BIOENGINEERING
Fiscal Year: 2023
Award: $789,255
Funding agency: National Institute of Biomedical Imaging and Bioengineering
Sub-project #1
MALDI imaging of tissue samples to identify novel proteins
MALDI-TOF Imaging is an emerging tool for the label free measurement of peptides, proteins, lipids, drugs and metabolites. The tissue can be imaged directly be the MALDI-TOF and combined with other modalities to study the molecular profiles as well as spatial placement of the tissue samples. Currently most tissue samples are imaged by MALDI and compared to histological stained tissues.
In collaboration with NIAID, we are developing protocols to image tissues and then compare them to confocal microscopy/histo-cytometry imaged sections. The confocal microscopy imaging provides cellular level of resolution and spatial mapping whereas MALDI-IMS provides more high-throughput proteomics and molecular mapping that can identify biomolecules associated with tissue inflammation and other processes. The goal is to use these techniques to image lymph nodes infected with HIV in hopes of identifying new biomarkers and providing spatial resolution of them.
In collaboration with LDRR, we are developing protocols to image mice tissue that has been irradiated with ultrasound. We are imaging tissues pre and post treatment to determine if any proteins, peptides or lipids are unregulated by this procedure and identify them if they are. The goal of is to determine if the mice exhibit an inflammatory response to the procedure.
Sub-Project #2
Trace metal analysis by Inductively Coupled Plasma (ICP)
ICP analysis with UV/Vis detection can be used to analyze samples for the presence of trace metals. The samples are introduced into a plasma flame and then detected by a continuous wavelength spectrophotometer to allow for quantitative measurements of the metal of interest. This technique allows researchers to determine yields for reactions, measure cellular uptake of metal compounds or determine metal levels in tissues in contrast agent experiments.
Studies using ICP-OES include:
- Analysis of manganese oxide nanoparticles for the amount of manganese in the particle and released in acidic sloutions
- Analysis of multimodal contrast agents to quantify the amount of iron and gadolinium
present
- Analysis of novel drug eluting beads to quantify the amount of boron present
- Analysis of poly(acrylic acid) gels to quantify the amount of sodium, calcium,
potassium and chlorine present.
- Analysis of saliva collected from patients diagnosed with Hepatitis C to quantify sodium and
potassium content
- Analysis of saliva collected from mice modeling dry mouth syndrome
Sub-project #3
High-throughput analysis of IgG glycosylation by matrix assisted laser desorption/ionization (MALDI) -time-of-flight (TOF)- mass spectrometry (MS).
The Laboratory of Immunoregulation at NIAID focuses its research efforts on the elucidation of cellular and molecular mechanisms of the regulation of the human immune response in health and disease. A major component of these research efforts involves understanding the immunopathogenesis of B cells in HIV infection. Dr. Susan Moir of the B-Cell Immunology Unit has recently described a role for IgG3 in regulating B cells in HIV-infected individuals enrolled in clinical research protocol 02-I-0202. The findings indicated that glycosylation of IgG3 and possibly other serum proteins played a role in the regulatory function described in the study; the findings were obtained from the analysis of specimens isolated from over 100 participants. The evaluation of glycosylation patterns in serum proteins must be performed using an approach that is compatible with processing a relatively large number of samples from a relatively small quantity of material. Preliminary assays will be performed on a few positive and negative controls, followed by a few longitudinal samples from individuals who either begin with a positive IgG3 profile and then lose it over time or acquire the profile over time.
Sub-project #4
Serosurvey to determine antibody presence to SARS-COV-2 protein
The Section for Immuno-Engineering in NIBIB has undertaken the development of and ELISA based serosurvey to detect the presence of IgG, IgM and IgA antibodies to SARS-COVID2 SPIKE and RBD proteins.
Studies using this serosurvey include:
- 9,300 healthy blood samples were collected at time point zero to determine the extent of
community spread of SARS-COVID-2 within the community.
- Approximately 7000 healthy blood samples were collected from the initial individuals at time
point six months to determine the extent of community spread and vaccination status.
- Approximately 5000 healthy blood samples were collected from the initial individuals at time
point one year to determine the extent of community spread and vaccination status.
- 2,542 samples from trauma patients to determine seropositivity for SARS-COV-2
- Autopsy samples from patients suspected of SARS-COV-2 death to determine seropositivity for
SARS-COV-2
- Approximately 15 samples from patients who experienced rebound infections after being
administered paxlovid to determine seropositivity
- Approximately 75 health care worker samples who have received boosters to determine
seropositivity
- Approximately 30 samples of patients enrolled in an IL-7 trial to determine seropositivity
- Approximately 4000 samples of individuals with select immunodeficiencies and immune
dysregulations compared to healthy volunteers to assess pre- and post- vaccine immune
responses
- Approximately 300 samples of individuals with Mitochondrial Disease to determine
seropositivity
Terms: <19S Gamma Globulin><2-propenoic acid><2019 novel corona virus><2019 novel coronavirus><2019-nCoV><7S Gamma Globulin><AIDS Virus><Acquired Immune Deficiency Syndrome Virus><Acquired Immunodeficiency Syndrome Virus><After Care><After-Treatment><Aftercare><Antibodies><Asialia><Assay><Autopsy><B blood cells><B cell><B cells><B-Cells><B-Lymphocytes><B-cell><Binding><Bioassay><Biologic Assays><Biological><Biological Assay><Biological Markers><Blood Plasma><Blood Sample><Blood specimen><Body Tissues><Boron><COVID><COVID-19 antibody><COVID-19 associated death><COVID-19 associated fatality><COVID-19 associated mortality><COVID-19 death><COVID-19 exposure><COVID-19 fatality><COVID-19 induced death><COVID-19 induced fatality><COVID-19 induced mortality><COVID-19 mortality><COVID-19 related death><COVID-19 related fatality><COVID-19 related mortality><COVID-19 virus><COVID19 associated death><COVID19 associated fatality><COVID19 associated mortality><COVID19 death><COVID19 fatality><COVID19 induced death><COVID19 induced fatality><COVID19 induced mortality><COVID19 mortality><COVID19 related death><COVID19 related fatality><COVID19 related mortality><COVID19 virus><Calcium><Chlorine><Cl element><Clinical Research Protocols><CoV disease><CoV-2><CoV2><Collaborations><Communities><Confocal Microscopy><Contrast Agent><Contrast Drugs><Contrast Media><Coupled><Detection><Development><Diagnosis><Disease><Disorder><Drugs><ELISA><Enrollment><Enzyme-Linked Immunosorbent Assay><Evaluation><Exhibits><Fe element><Gadolinium><Gd element><Gel><Goals><HCV infection><HIV><HIV Infections><HTLV-III Infections><HTLV-III-LAV Infections><Health><Health Care Providers><Health Personnel><Healthcare Providers><Healthcare worker><Hepatitis C><Hepatitis C virus infection><Hepatitis, Viral, Non-A, Non-B, Parenterally-Transmitted><Hepatitus C><Human><Human Immunodeficiency Viruses><Human T-Lymphotropic Virus Type III Infections><Hyposalivation><ICP-MS><IL-7><IL-7 Gene><IL7><IL7 Protein><IL7 gene><IgA><IgG><IgG3><IgM><Image><Image Cytometry><Immune><Immune response><Immunes><Immunochemistry><Immunodeficiency Disorder><Immunodeficiency Syndrome><Immunoglobulin A><Immunoglobulin G><Immunoglobulin M><Immunologic Deficiency Syndromes><Immunological Deficiency Syndromes><Immunological response><Immunology><Immunomodulation><Individual><Inductively Coupled Plasma Mass Spectrometry><Infection><Inflammation><Inflammatory Response><Interleukin 7 Precursor><Interleukin 7 Precursor Gene><Interleukin-7><Interleukin-7 Gene><Investigators><Iron><K element><LAV-HTLV-III><Label><Laboratories><Lipids><Lymph Node Reticuloendothelial System><Lymph node proper><Lymphadenopathy-Associated Virus><Lymphatic nodes><Lymphopoietin-1><MALD-MS><MALDI><MALDI-MS><Manganese><Maps><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Measurement><Measures><Medication><Metabolic Glycosylation><Metals><Mice><Mice Mammals><Mission><Mitochondrial Diseases><Mitochondrial Disorders><Mn element><Mn3O4><Modality><Modern Man><Molecular><Molecular Fingerprinting><Molecular Interaction><Molecular Profiling><Mouth Dryness><Murine><Mus><NIAID><NIBIB><Na element><National Institute of Allergy and Infectious Disease><National Institute of Biomedical Imaging and Bioengineering><Participant><Patients><Pattern><Paxlovid><Peptides><Pharmaceutic Preparations><Pharmaceutical Preparations><Plasma><Plasma Serum><Potassium><Procedures><Process><Proteins><Proteomics><Protocol><Protocols documentation><Radiopaque Media><Reaction><Regulation><Research><Research Personnel><Research Specimen><Researchers><Resolution><Reticuloendothelial System, Serum, Plasma><Role><SARS><SARS corona virus 2><SARS coronavirus disease><SARS-CO-V2><SARS-COVID-2><SARS-CoV disease><SARS-CoV-2><SARS-CoV-2 antibody><SARS-CoV-2 associated death><SARS-CoV-2 associated fatality><SARS-CoV-2 associated mortality><SARS-CoV-2 death><SARS-CoV-2 exposure><SARS-CoV-2 fatality><SARS-CoV-2 induced death><SARS-CoV-2 induced fatality><SARS-CoV-2 induced mortality><SARS-CoV-2 mortality><SARS-CoV-2 related death><SARS-CoV-2 related fatality><SARS-CoV-2 related mortality><SARS-CoV2><SARS-CoV2 antibody><SARS-CoV2 exposure><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Sampling><Serum Proteins><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><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome CoV disease><Severe Acute Respiratory Syndrome coronavirus disease><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 exposure><Severe acute respiratory syndrome related corona virus 2><Sodium><Specimen><Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization><Spectrophotometry><Spectroscopy, Mass, Matrix-Assisted Laser Desorption-Ionization><Syndrome><System><Techniques><Time><Tissue Sample><Tissue Stains><Tissue imaging><Tissues><Trace metal><Trauma patient><Vaccination><Virus-HIV><Work><Wuhan coronavirus><Xerostomia><acrylic acid><antibody against COVID-19><antibody against SARS-CoV-2><antibody against SARS-CoV2><antibody against coronavirus disease 2019><antibody against severe acute respiratory syndrome coronavirus 2><antibody to COVID-19><antibody to SARS-CoV-2><antibody to SARS-CoV2><antibody to coronavirus disease 2019><antibody to severe acute respiratory syndrome coronavirus 2><aptyalism><bio-markers><biologic><biologic marker><biomarker><community spread><community transmission><community-level spread><community-level transmission><corona virus disease><coronavirus disease><coronavirus disease 2019 antibody><coronavirus disease 2019 associated death><coronavirus disease 2019 associated fatality><coronavirus disease 2019 associated mortality><coronavirus disease 2019 death><coronavirus disease 2019 exposure><coronavirus disease 2019 fatality><coronavirus disease 2019 induced death><coronavirus disease 2019 induced fatality><coronavirus disease 2019 induced mortality><coronavirus disease 2019 mortality><coronavirus disease 2019 related death><coronavirus disease 2019 related fatality><coronavirus disease 2019 related mortality><coronavirus disease 2019 virus><coronavirus disease-19 mortality><coronavirus disease-19 virus><death due to COVID-19><death due to COVID19><death due to SARS-CoV-2><death due to coronavirus disease 2019><death due to severe acute respiratory syndrome coronavirus 2><death in COVID><death in COVID-19><death in SARS-CoV-2><death in coronavirus disease><death in coronavirus disease 2019><death in severe acute respiratory syndrome coronavirus 2><developmental><drug/agent><dry mouth><engineered immune system><enroll><enzyme linked immunoassay><experience><experiment><experimental research><experimental study><experiments><exposure to COVID-19><exposure to SARS-CoV-2><exposure to SARS-CoV2><exposure to Severe acute respiratory syndrome coronavirus 2><exposure to coronavirus disease 2019><fatality due to COVID-19><fatality due to COVID19><fatality due to SARS-CoV-2><fatality due to coronavirus disease 2019><fatality due to severe acute respiratory syndrome coronavirus 2><glycosylated IgG><glycosylated immunoglobulin G><glycosylation><hCoV19><hausmannite><health care personnel><health care worker><health provider><health workforce><healthcare personnel><healthy volunteer><hep C><hepatitis non A non B><high throughput analysis><histologic stains><histological stains><host response><hypoimmunity><imaging><immune deficiency disorder><immune engineering><immune modulation><immune regulation><immune system response><immunodeficiency><immunoengineering><immunologic reactivity control><immunomodulatory><immunoregulation><immunoregulatory><immunoresponse><infection by hepatitis c virus><instrumentation><interest><lymph gland><lymph nodes><lymphnodes><manganese oxide><manganous-manganic oxide><matrix assisted laser desorption ionization><medical personnel><microscope imaging><microscopic imaging><microscopy imaging><molecular profile><molecular signature><mortality due to COVID-19><mortality due to COVID19><mortality due to SARS-CoV-2><mortality due to coronavirus disease 2019><mortality due to severe acute respiratory syndrome coronavirus 2><mouse model><multi-modality><multimodality><murine model><nCoV2><nano particle><nano-sized particle><nanoparticle><nanosized particle><necropsy><new drug treatments><new drugs><new pharmacological therapeutic><new therapeutics><new therapy><next generation therapeutics><non A, non B hepatitis><non-A, non-B hepatitis><novel><novel drug treatments><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel therapeutics><novel therapy><participant enrollment><particle><patient enrollment><post treatment><postmortem><resolutions><saliva analysis><serology survey><seropositive><serosurvey><severe acute respiratory syndrome coronavirus 2 antibody><severe acute respiratory syndrome coronavirus 2 associated death><severe acute respiratory syndrome coronavirus 2 associated fatality><severe acute respiratory syndrome coronavirus 2 associated mortality><severe acute respiratory syndrome coronavirus 2 death><severe acute respiratory syndrome coronavirus 2 fatality><severe acute respiratory syndrome coronavirus 2 induced death><severe acute respiratory syndrome coronavirus 2 induced fatality><severe acute respiratory syndrome coronavirus 2 induced mortality><severe acute respiratory syndrome coronavirus 2 mortality><severe acute respiratory syndrome coronavirus 2 related death><severe acute respiratory syndrome coronavirus 2 related fatality><severe acute respiratory syndrome coronavirus 2 related mortality><social role><spectrophotometer><tool><treatment provider><ultrasound><uptake><vaccine immune response><vaccine immunogenicity><vaccine response><vaccine responsiveness><vaccine-induced response>