Document text
Principal Investigator: James E. Gern
Organization: UNIVERSITY OF WISCONSIN-MADISON
Fiscal Year: 2024
Award: $656,603
Funding agency: National Institute of Allergy and Infectious Diseases
Project Summary
Proteomics analysis of URECA nasal secretions.
Rationale. The URECA 14-17 year time point includes collection of nasal secretions by nasosorption to
complement information that will be gained by analyzing nasal cell RNA and DNA. Analysis of nasal specimens
collected at 11 year of age indicates that children with allergic asthma or chronic rhinitis have distinct
differences in nasal epithelial cell gene expression that include increased expression of T2 genes and reduced
expression of an array of other immune response genes. For example, children with a history of chronic rhinitis
beginning in early life have downregulation of a network of genes including canonical antiviral genes (STAT1,
IRF1) but also genes involved in other innate immune responses (e.g., GSMA, CCL5) and adaptive immune
responses (HLA-DRA, CD4, CD19). Notably, there is a strong negative correlation between the upregulated T2
cluster and the downregulation of other immune response genes. These findings suggest that T2 inflammation
can suppress a variety of mucosal immune responses, including antiviral, antibacterial, and innate and
adaptive responses.
Preliminary data. To further explore the hypothesis that airway
mucosal immunity is depressed in children with allergic rhinitis, we
have analyzed nasal secretions from children enrolled in the
Childhood Origins of Asthma (COAST study). Nasal secretions were
obtained by “nasal blow” from 9 y/o children who were sensitized to
aeroallergens and reported at least moderate nasal symptoms, and
an equal number of children with neither symptoms nor sensitization
(10 per group). The nasal samples were tested for immunoglobulin
content by multiplex ELISA (Milliplex, Millipore). IgE values were low
at the threshold of detection for this assay (40% undetectable), and
not significantly different between the groups (geomean 6 vs. 3
ng/mL, allergic vs. nonallergic respectively, ns). However, children with
allergic rhinitis had generally lower immunoglobulin levels than the
normal controls (Figure). However, there were significant differences
Figure _. Immunoglobulin levels in nasal
secretions from allergic and nonallergic
children. Geometric means +/- SD.
(p<0.05) in other immunoglobulins in nasal secretions including IgM (48 vs. 11 pg/mL), IgG2 (1365 vs. 196
pg/mL) and IgA (124 vs. 23 ng/mL). These findings suggest that allergic rhinitis is associated with reduced
immunoglobulins in nasal secretions, and that this could be related to greater susceptibility to both viral and
bacterial infections.
Experimental design. We proposal to conduct a pilot study with 12 samples of nasal secretions (nasosorption)
from URECA children with allergic rhinitis (aeroallergen sensitization and nasal symptoms) and 12 samples
from children with neither allergy nor allergic sensitization. We will use mass spectrometry (LC/MS/MS, UW
A.I.C. Mass Spectrometry Facility) to identify proteins in the secretions. Samples of nasal secretions are stored
in PBS, and we will extract with acetone and analyze protein content. Between-group comparisons will be used
to identify differentially expressed proteins. We will verify differences in protein content by analyzing the
remaining specimens (~300) with multiplex ELISA using custom panels targeting differentially expressed
proteins (Milliplex). Analyzing the entire group of specimens will enable differences in protein expression to be
compared across rhinitis and asthma phenotypes.
Anticipated results and interpretation. We anticipate that respiratory allergy will be associated with reduction in
critical proteins involved in innate and adaptive immune responses. These include IgA and other
immunoglobulins, granzyme A, and complement factors. If confirmed, these observations could help to explain
higher risk of infections (e.g., viral illnesses, sinusitis) in children with respiratory allergy, and higher rates of
colonization with bacterial pathogens such as Moraxella catarrhalis and Hemophilus influenzae.
Terms: <0-11 years old><19S Gamma Globulin><2-Propanone><Acetone><Active Follow-up><Address><Administrative Supplement><Affect><Age Years><Airway mucosa><Allergic><Allergic asthma><Allergic rhinitis><Allergic rhinitis due to allergen><Allergic rhinosinusitis><Allergy><Anti-Bacterial Agents><Asthma><Asthma in Children><Atopic rhinitis><B catarrhalis><B. catarrhalis><Bacterial Infections><Birth><Branhamella catarrhalis><Bronchial Asthma><CCL5><CD19><CD19 gene><COVID crisis><COVID epidemic><COVID pandemic><COVID-19 crisis><COVID-19 epidemic><COVID-19 era><COVID-19 global health crisis><COVID-19 global pandemic><COVID-19 health crisis><COVID-19 pandemic><COVID-19 period><COVID-19 public health crisis><COVID-19 years><Caring><Cell Body><Cells><Chemokine (C-C Motif) Ligand 5><Child><Child Youth><Childhood><Childhood Asthma><Children (0-21)><Chronic Rhinitis><Cohort Studies><Collection><Complement><Complement Proteins><Concurrent Studies><Custom><D17S136E><DNA><DNA Methylation><Data><Deoxyribonucleic Acid><Depressed mood><Dimethyl formaldehyde><Disease><Disorder><Down-Regulation><ELISA><Enrollment><Environmental Exposure><Enzyme-Linked Immunosorbent Assay><Epithelial Cells><Experimental Designs><Exposure to><Extrinsic asthma><Gene Chips><Gene Expression><Gene Expression Chip><GeneChip><Genes><Genetic><Granzyme><H influenzae><H. influenzae><Haemophilus influenzae><History><Hypersensitivity><IRF-1><IRF1><IRF1 gene><IgA><IgE><IgG2><IgM><Immune Globulins><Immune Response Genes><Immunoglobulin A><Immunoglobulin E><Immunoglobulin M><Immunoglobulins><Inflammation><Innate Immune Response><Interferon Regulatory Factor 1><Intervention><Intervention Strategies><Ir Gene><LC/MS><Leadership><Life><M catarrhalis><M. catarrhalis><MGC17164><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Mediating><Modification><Moraxella catarrhalis><Morbidity><Morbidity - disease rate><Mucosal Immune Responses><Mucosal Immunity><MyD32 protein><Nasal><Nasal Epithelium><Nasal Passages Nose><Non-Polyadenylated RNA><Nose><Parturition><Patients><Pediatric asthma><Phenotype><Pilot Projects><Predisposition><Proteins><Proteomics><RANTES><RNA><RNA Gene Products><Recording of previous events><Reporting><Research Specimen><Respiratory Mucosa><Respiratory System, Nose, Nasal Passages><Rhinitis><Rhinitis allergic atopic><Ribonucleic Acid><SARS-CoV-2 epidemic><SARS-CoV-2 global health crisis><SARS-CoV-2 global pandemic><SARS-CoV-2 pandemic><SARS-coronavirus-2 epidemic><SARS-coronavirus-2 pandemic><SCYA5><SIS delta><SIS-delta><SISd><STAT1><STAT1 gene><STAT91><Sampling><Self Care><Severe Acute Respiratory Syndrome CoV 2 epidemic><Severe Acute Respiratory Syndrome CoV 2 pandemic><Severe acute respiratory syndrome coronavirus 2 epidemic><Severe acute respiratory syndrome coronavirus 2 pandemic><Sinusitis><Small Inducible Cytokine A5><Specimen><Susceptibility><Symptoms><T-Cell RANTES Protein><T-Cell Specific Protein p288><TCP228><Testing><Time><United States><Urban Community><Viral><Viral Diseases><Viral Genes><Virus Diseases><active followup><adaptive immune response><aeroallergens><airborn allergen><airborne allergen><anti-bacterial><atopic asthma><bacteria infection><bacteria pathogen><bacterial disease><bacterial pathogen><cohort><comparator group><comparison group><complementation><coronavirus disease 2019 crisis><coronavirus disease 2019 epidemic><coronavirus disease 2019 global health crisis><coronavirus disease 2019 global pandemic><coronavirus disease 2019 health crisis><coronavirus disease 2019 pandemic><coronavirus disease 2019 public health crisis><coronavirus disease crisis><coronavirus disease epidemic><coronavirus disease pandemic><coronavirus disease-19 global pandemic><coronavirus disease-19 pandemic><customs><depressed><detection assay><differential expression><differentially expressed><disease control><disorder control><enroll><enzyme linked immunoassay><expression array><extrinsic allergic asthma><follow up><follow-up><followed up><followup><gene expression microarray><gene network><healthcare burden><high risk><histories><improved><infection risk><interferon-stimulated gene factor 1><interventional strategy><kids><liquid chromatography mass spectrometry><novel><pathogenic bacteria><pediatric><personal care><pilot study><prevent><preventing><protein expression><respiratory><response><sadness><severe acute respiratory syndrome coronavirus 2 global health crisis><severe acute respiratory syndrome coronavirus 2 global pandemic><transcriptional differences><urban children><urban environment><urban setting><viral infection><virus infection><virus-induced disease><youngster>