PMID- 34368811
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20231107
VI  - 2
DP  - 2021
TI  - Isolation and characterization of extracellular vesicles in saliva of children 
      with asthma.
PG  - 29-48
LID - 10.20517/evcna.2020.09 [doi]
AB  - AIM: To confirm the presence of extracellular vesicles (EVs) in cell-free saliva 
      (CFS) of children with asthma and describe the isolated EV population. METHODS: A 
      pooled sample of CFS EVs isolated from 180 participants using ExoQuick-TC was 
      examined in downstream analyses. Transmission electron microscopy (TEM) was used 
      to confirm the presence of EVs. Nanoparticle tracking analysis (NTA) and single 
      particle interferometric reflectance imaging sensing (SP-IRIS) with fluorescence 
      were used for sizing, counting, and phenotyping of EVs. Capillary immunoassays 
      were used for protein quantitation. RESULTS: TEM confirmed the presence of EVs of 
      diverse sizes, indicating the prep contained a heterogeneous population of EVs. 
      Capillary immunoassays confirmed the presence of EV-associated proteins (CD9, 
      CD63, CD81, ICAM-1, and ANXA5) and indicated limited cellular contamination. As 
      others have also reported, there were discrepancies in the EV sizing and 
      enumeration across platforms. Fluorescent NTA detected particles with a mode 
      diameter of ~90 nm, whereas SP-IRIS reported sizes of ~55-60 nm that more closely 
      approximated the TEM results. Consistent with protein immunoassay results, 
      SP-IRIS with fluorescence showed that the majority of these EVs were CD9- and 
      CD63-positive, with little expression of CD81. CONCLUSION: EVs from CFS can be 
      isolated using a high-throughput method that can be scaled to large 
      epidemiological studies. To our knowledge, we are the first to characterize CFS 
      EVs from patients with asthma. The use of CFS EVs as potential novel biomarkers 
      in asthma warrants further investigation and opens a new avenue of research for 
      future studies.
FAU - Comfort, Nicole
AU  - Comfort N
AD  - Department of Environmental Health Sciences, Columbia University Mailman School 
      of Public Health, New York, NY 10032, USA.
FAU - Bloomquist, Tessa R
AU  - Bloomquist TR
AD  - Department of Environmental Health Sciences, Columbia University Mailman School 
      of Public Health, New York, NY 10032, USA.
FAU - Shephard, Alex P
AU  - Shephard AP
AD  - NanoView Biosciences, Malvern Hills Science Park, Malvern, Worcestershire WR14 
      3SZ, UK.
FAU - Petty, Carter R
AU  - Petty CR
AD  - Brigham and Women's Hospital, Boston, MA; Boston Children's Hospital, Clinical 
      Research Center Boston, MA 02115, USA.
FAU - Cunningham, Amparito
AU  - Cunningham A
AD  - Harvard Medical School, Boston, MA 02115, USA.
FAU - Hauptman, Marissa
AU  - Hauptman M
AD  - Harvard Medical School, Boston, MA 02115, USA.
AD  - Division of Allergy and Immunology, Boston Children's Hospital, Boston, MA 02115, 
      USA.
FAU - Phipatanakul, Wanda
AU  - Phipatanakul W
AD  - Harvard Medical School, Boston, MA 02115, USA.
AD  - Division of Allergy and Immunology, Boston Children's Hospital, Boston, MA 02115, 
      USA.
FAU - Baccarelli, Andrea
AU  - Baccarelli A
AD  - Department of Environmental Health Sciences, Columbia University Mailman School 
      of Public Health, New York, NY 10032, USA.
LA  - eng
GR  - F31 ES030973/ES/NIEHS NIH HHS/United States
GR  - R01 ES025225/ES/NIEHS NIH HHS/United States
GR  - S10 RR027699/RR/NCRR NIH HHS/United States
GR  - U01 AI110397/AI/NIAID NIH HHS/United States
PT  - Journal Article
DEP - 20210330
PL  - United States
TA  - Extracell Vesicles Circ Nucl Acids
JT  - Extracellular vesicles and circulating nucleic acids
JID - 101775065
PMC - PMC8340923
MID - NIHMS1691208
OTO - NOTNLM
OT  - Extracellular vesicles
OT  - NTA
OT  - SP-IRIS
OT  - TEM
OT  - asthma
OT  - biomarker
OT  - exosome
OT  - saliva
COIS- Conflicts of interest Alex P. Shephard is employed by NanoView Biosciences. All 
      other authors declared that there are no conflicts of interest.
EDAT- 2021/08/10 06:00
MHDA- 2021/08/10 06:01
PMCR- 2021/08/05
CRDT- 2021/08/09 06:51
PHST- 2021/08/09 06:51 [entrez]
PHST- 2021/08/10 06:00 [pubmed]
PHST- 2021/08/10 06:01 [medline]
PHST- 2021/08/05 00:00 [pmc-release]
AID - 10.20517/evcna.2020.09 [doi]
PST - ppublish
SO  - Extracell Vesicles Circ Nucl Acids. 2021;2:29-48. doi: 10.20517/evcna.2020.09. 
      Epub 2021 Mar 30.