Journal: Vaccines

Article Title: Longitudinal Analyses after COVID-19 Recovery or Prolonged Infection Reveal Unique Immunological Signatures after Repeated Vaccinations

doi: 10.3390/vaccines10111815

Figure Lengend Snippet: Serum antibody features in COVID-19 patients during hospitalization: Graphs show serum anti-S-RBD IgM ( A ) and IgG ( B ) levels for each disease severity ( n = 9 patients/mild group; 10 patients/moderate I group; 9 patients/moderate II patients). Blue and red dots indicate admission and discharge, respectively; the duration of hospitalization is indicated at the bottom of each panel. Graphs show the correlation between anti-S-RBD IgM or IgG and neutralizing antibody titer ( C ) and age, as well as the change in the titer against the WT strain in patients infected with the WT and VOCs ( D ). Each dot denotes disease severity, and Spearman’s correlation coefficients are shown within the graph. Gray areas indicate a 95% confidence interval (CI) for the total. ( E ) Dot plot indicates the neutralizing potency against the WT strain by sex. Graphs show the serum-neutralizing antibody titer against WT ( F ), Kappa ( G ), and Delta ( H ) variants at each severity (moderate II is classified by the infected strains, n = 9 patients/mild group; 10 patients/moderate I group; 9 patients/WT-infected moderate II group; 8 patients/VOC-infected moderate II group). Statistical significances were determined using the Tukey–Kramer test and Welch’s t -test. All horizontal bars show mean values. Cross, deceased; #, long-term observed healthcare worker. * p < 0.05; *** p < 0.005; ns, not significant.

Article Snippet: Isolated PBMCs were incubated with His-tagged SARS-CoV-2 S-RBD recombinant protein (Cell Signaling Technology, Danvers, MA, USA) for 30 min on ice.

Techniques: Infection

Journal: Vaccines

Article Title: Longitudinal Analyses after COVID-19 Recovery or Prolonged Infection Reveal Unique Immunological Signatures after Repeated Vaccinations

doi: 10.3390/vaccines10111815

Figure Lengend Snippet: Longitudinal analyses of serum neutralizing antibody activities and PBMCs in a patient with high neutralizing potency: ( A ) Concentration of anti-S-RBD IgM and IgG in the serum of a healthcare worker (indicated by # in ) upon infection. ( B ) Fluctuation of each neutralizing antibody titer from onset to post-vaccination ( n = 1). ( C ) Representative FACS plots of cell population are shown in panels. The numbers indicate the positive rate of each cell population. Graphs show the absolute number of IgG + S-RBD + Bmems ( D ) and the frequency of lymphocyte ( E ), CD19 + CD27 + CD38 high , and IgG + Bmems (( F ); n = 1, 3–5 technical replicates). Black dots indicate technical replicates. Each number indicates the mean.

Article Snippet: Isolated PBMCs were incubated with His-tagged SARS-CoV-2 S-RBD recombinant protein (Cell Signaling Technology, Danvers, MA, USA) for 30 min on ice.

Techniques: Concentration Assay, Infection

Journal: Vaccines

Article Title: Longitudinal Analyses after COVID-19 Recovery or Prolonged Infection Reveal Unique Immunological Signatures after Repeated Vaccinations

doi: 10.3390/vaccines10111815

Figure Lengend Snippet: Affinity maturation of S-RBD-specific memory B-cells after vaccination: Pie charts show the distribution of heavy chain genes ( A ) and light chain genes ( B ) from onset to post-vaccination, comparing patients with the recovered individual ( n = 1) and a prolonged COVID-19 patient ( n = 1). The number in the inner circle indicates the number of sequenced clones. ( C ) Graphs show the number of somatic hypermutations in each antibody gene. Statistical significances were determined using Tukey–Kramer test. ( D ) Graphs show the distribution of CDR3 length in each antibody gene (heavy chains; n = 34–45 clones/recovered individual, 14 clones/prolonged COVID-19 patient; light chains; 18–66 clones/recovered individual, 15 clones/prolonged COVID-19 patient).

Article Snippet: Isolated PBMCs were incubated with His-tagged SARS-CoV-2 S-RBD recombinant protein (Cell Signaling Technology, Danvers, MA, USA) for 30 min on ice.

Techniques: Clone Assay