![Time kinetics of humoral and cellular responses against spike and nonspike epitopes of SARS-CoV-2 in CLL patients after COVID-19 infection followed by 3 mRNA vaccination doses. (A) Serum and (B) salivary levels of S (spike)-receptor-binding domain (RBD) and N (nucleocapsid)-specific antibody responses in patients post–COVID-19 and pre- (pre vacc) and postvaccination (dose 2 [d2]) and pre- (dose 3) and postvaccination (dose 3) with indicated median values. (C) Spike-specific CD4+ (CD69+CD154+) and CD8+ (CD69+CD137+) T cells were detected by flow cytometry (activation-induced marker assay). Spike-specific CD4+ (D) and CD8+ (E) T-cell response against Wu-Hu1 (wild-type) and Omicron (BA.1) post–COVID-19 and prevaccination, prevaccination (dose 3), and postvaccination (dose 3). (F) ELISpot IFN-γ-specific T-cell responses to spike and membrane, envelope, and nucleocapsid (M + E + N) peptide pools post–COVID-19 and prevaccination, prevaccination (dose 3), and postvaccination (dose 3). White dots represent patients on BTKi treatment in the respective analysis. Number (n) of patients tested at each time point is indicated below graph. Assay upper limit of detection of 25 000 U/mL is shown as dotted line (A). Dashed line represents positive threshold for each assay: (A) 0.8 U/mL and 1 cutoff index (COI) respectively; (B) median fluorescence intensity (MFI) of 59 and 100 for S and N, respectively; (D-E) 0.05% and (F) 80 spot-forming units (SFU)/106 cells. Error bars represent the median (red line) and interquartile range. Kruskal-Wallis test with Dunn multiple comparison correction was used. ∗P < .05, ∗∗P < .01, ∗∗∗∗P < .0001. ns, not significant.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/140/22/10.1182_blood.2022016815/5/blood_bld-2022-016815-gr1.jpeg?Expires=1735800311&Signature=Q8sGfYC5VdOS1kuVEUADdle~EDaA35oEOud-xwZClB6xBMuHooZnTZ7uaaku5O1au-zq7gmPPfreRHn9Wl8s~jXoXa6kuZuYzS30wOVjSFucB0CO62UuzqdLd0TAixq5Ck1eE6a~0Xa5okAFybVmcMlXhAgxJP6gpV~ewi3U66012B~ZS~07-V2P9FPhDF-JQUm58S-Cgr6ntS9Or6qZg5-voO-UJuHT6DMh0-2g0HYhBrYPR4YtA239g325pbQ7aLFeU0WNeb3lEmCz46UjytRMw85BzAgUGeyoUnpPUeoaFdz4BXkrmfo2x5lZPvzsfjFNC~NX9KdtncKTRdpI9A__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Time kinetics of humoral and cellular responses against spike and nonspike epitopes of SARS-CoV-2 in CLL patients after COVID-19 infection followed by 3 mRNA vaccination doses. (A) Serum and (B) salivary levels of S (spike)-receptor-binding domain (RBD) and N (nucleocapsid)-specific antibody responses in patients post–COVID-19 and pre- (pre vacc) and postvaccination (dose 2 [d2]) and pre- (dose 3) and postvaccination (dose 3) with indicated median values. (C) Spike-specific CD4+ (CD69+CD154+) and CD8+ (CD69+CD137+) T cells were detected by flow cytometry (activation-induced marker assay). Spike-specific CD4+ (D) and CD8+ (E) T-cell response against Wu-Hu1 (wild-type) and Omicron (BA.1) post–COVID-19 and prevaccination, prevaccination (dose 3), and postvaccination (dose 3). (F) ELISpot IFN-γ-specific T-cell responses to spike and membrane, envelope, and nucleocapsid (M + E + N) peptide pools post–COVID-19 and prevaccination, prevaccination (dose 3), and postvaccination (dose 3). White dots represent patients on BTKi treatment in the respective analysis. Number (n) of patients tested at each time point is indicated below graph. Assay upper limit of detection of 25 000 U/mL is shown as dotted line (A). Dashed line represents positive threshold for each assay: (A) 0.8 U/mL and 1 cutoff index (COI) respectively; (B) median fluorescence intensity (MFI) of 59 and 100 for S and N, respectively; (D-E) 0.05% and (F) 80 spot-forming units (SFU)/106 cells. Error bars represent the median (red line) and interquartile range. Kruskal-Wallis test with Dunn multiple comparison correction was used. ∗P < .05, ∗∗P < .01, ∗∗∗∗P < .0001. ns, not significant.
