Document text
Principal Investigator: Heather Kalish
Organization: NATIONAL INSTITUTE OF BIOMEDICAL IMAGING AND BIOENGINEERING
Fiscal Year: 2024
Award: $892,557
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 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.
In collaboration with NEI, we are developing protocols to image mouse eyes that have been dosed with small molecule therapy agents that address inherited retinal disease. The protocols will help identify the spatial distribution of these agents within the eye. Additionally the protocols will help to identify novel lipids and peptides that are observed in eyes with this disease.
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 saliva samples from patients being treated with AAV2hAQP1 therapy
- Analysis of serum samples to quantify the amount of Mn, Co present in pediatric patients with PPGL tumors
- 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
Sub Project #5
Automation of Aedes aegypti sera assay for the analysis of IgG in serum samples from people exposed to mosquitos
Our unit is automating the IgG ELISa developed in Dr Luiz Oliveira's lab in order to process the 4900 samples provided by his unit and additionally process samples provided by other labs under MTA. The unit has successfully reproduced the results of the manual assay using automation. Additionally, the assay has been altered to provide for more dynamic range between positive and negative samples. The automated assay will be used to process 4900 samples from Dr Oliveira's lab and then process samples from several other studies through MTA.
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><Address><Aedes><After Care><After-Treatment><Aftercare><Antibodies><Asialia><Assay><Automation><Autopsy><B blood cells><B cell><B cells><B-Cells><B-Lymphocytes><B-cell><Binding><Bioassay><Biological><Biological Assay><Blood Plasma><Blood Sample><Blood Serum><Blood specimen><Body Tissues><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-2><CoV2><Collaborations><Communities><Contrast Agent><Contrast Drugs><Contrast Media><Coupled><Culicidae><Detection><Development><Diagnosis><Disease><Disorder><Dose><Drugs><ELISA><Enrollment><Enzyme-Linked Immunosorbent Assay><Evaluation><Exhibits><Exposure to><Eye><Eyeball><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><Hereditary><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><Immune><Immune response><Immunes><Immunochemistry><Immunoglobulin A><Immunoglobulin G><Immunoglobulin M><Immunological response><Immunology><Immunomodulation><Individual><Inductively Coupled Plasma Mass Spectrometry><Infection><Inflammatory Response><Inherited><Interleukin 7 Precursor><Interleukin 7 Precursor Gene><Interleukin-7><Interleukin-7 Gene><Investigators><K element><LAV-HTLV-III><Label><Laboratories><Lipids><Lymphadenopathy-Associated Virus><Lymphopoietin-1><MALD-MS><MALDI><MALDI-MS><Manganese><Manuals><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><Mosquitoes><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><Persons><Pharmaceutical Preparations><Plasma><Plasma Serum><Potassium><Procedures><Process><Proteins><Protocol><Protocols documentation><Radiopaque Media><Reaction><Regulation><Research><Research Personnel><Research Specimen><Researchers><Resolution><Reticuloendothelial System, Serum, Plasma><Retinal Diseases><Retinal Disorder><Role><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><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 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><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 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 exposure><Severe acute respiratory syndrome related corona virus 2><Sodium><Spatial Distribution><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 coronavirus disease 2019><antibody against severe acute respiratory syndrome coronavirus 2><antibody to COVID-19><antibody to SARS-CoV-2><antibody to coronavirus disease 2019><antibody to severe acute respiratory syndrome coronavirus 2><aptyalism><biologic><child patients><community spread><community transmission><community-level spread><community-level transmission><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><immune engineering><immune modulation><immune regulation><immune response to vaccination><immune response to vaccines><immune system response><immunodeficiency><immunoengineering><immunologic reactivity control><immunomodulatory><immunoregulation><immunoregulatory><immunoresponse><infection by hepatitis c virus><instrumentation><interest><manganese oxide><manganous-manganic oxide><matrix assisted laser desorption ionization><medical personnel><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><murine model><nCoV2><nano particle><nano-sized particle><nanoparticle><nanosized particle><necropsy><non A, non B hepatitis><non-A, non-B hepatitis><novel><participant enrollment><particle><patient enrollment><pediatric patients><post treatment><postmortem><resolutions><retina disease><retina disorder><retinopathy><saliva analysis><saliva sample><salivary sample><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><small molecule><social role><spectrophotometer><tool><treatment provider><tumor><ultrasound><uptake><vaccine associated immune response><vaccine immune response><vaccine immunogenicity><vaccine induced immune response><vaccine response><vaccine responsiveness><vaccine-induced response>