https://orcid.org/0000-0003-0630-9175
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Sandhya Bansal, Timothy Fleming, Thalachallour Mohanakumar, Response to Comment on “Cutting Edge: Circulating Exosomes with COVID Spike Protein Are Induced by BNT162b2 (Pfizer-BioNTech) Vaccination prior to Development of Antibodies: A Novel Mechanism for Immune Activation by mRNA Vaccines”, The Journal of Immunology, Volume 208, Issue 8, April 2022, Pages 1833–1834, https://doi.org/10.4049/jimmunol.2101067
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We used minor modifications for the isolation of circulating exosomes using a commercial precipitation kit followed by 0.22 u filtration, the quality of exosomes isolated had a mean size of 158.7 + 5.4 nm, and the NanoSight data were provided in Fig. 1A. For the Western blot, we have used a commercial Ab specific to SARS-CoV-2 (mAb 1A9 from Thermo Fisher Scientific). We found exosomes with S2 on day 14 after the first and second dose, and finally 4 mo after the second dose and no further time points were taken.
The article by Ogata et al. (1) is very different from our article and an entirely different study, so comparing the two results is not appropriate. Ogata et al. (1) did not deal with circulating exosomes and they analyzed S1, whereas we detected S2 on exosomes. We developed an in-house ELISA for quantifying Abs specific to the spike and nucleocapsid proteins. The details of commercial reagents used (Abs from Thermo Fisher Scientific and proteins from Sino Biological) were provided in our article. Because we did not perform serial analysis starting from day 14 following the first and second dose of vaccine for circulating exosomes with S2, we cannot state how long these Ags persisted in the exosomes. As stated above, we analyzed Ab development and exosomes on day 14 after the first vaccine dose, day 14 after the second dose, and finally at 4 mo.
