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Single-cell mapping of peripheral immune dynamics in pregnant women after Moderna mRNA COVID-19 vaccination
This study was approved by the Institutional Review Board of Taichung Veterans General Hospital (IRB Number CF21270A). Three pregnant women receiving two doses of Moderna mRNA-1273 (Spikevax) were enrolled after informed consent. PBMCs were isolated from 8 mL of blood using density-gradient separation. Anti-SARS-CoV2-IgG detection IgG antibodies in serum against the receptor-binding domain of the spike protein S1 subunit of SARS-CoV-2 were measured using an electrochemiluminescence immunoassay following the manufacturer’s instructions (Elecsys® Anti-SARS-CoV-2 S assay, Roche Diagnostics, Basel, Switzerland). The measurement range was 0.40–250 U/mL, and < 0.8 and ≥ 0.80 U/mL were recorded as negative and positive, respectively. Sample preparation and scRNA-seq library construction This study followed the Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 protocol (10x Genomics CG000388, Rev A, Pleasanton, CA, USA). Cryopreserved PBMCs were thawed at 37 °C for 1 min and then cultured in RPMI with 10% FBS and 10 ng/mL DNase I to prevent aggregation. Each sample was labelled with Cell Multiplex oligos (PN-1000261, 10x Genomics), and up to 14 samples were pooled for loading onto the Chromium Single Cell Instrument. Each channel processed ~49,500 cells and targeted a recovery of 30,000 cells at 1600 cells/μg. Libraries were prepared using Dual Index...
