Journal: medRxiv
Article Title: SARS-CoV-2 mRNA vaccination exposes progressive adaptive immune dysfunction in patients with chronic lymphocytic leukemia
doi: 10.1101/2022.12.19.22283645
Figure Lengend Snippet: (A and B) Comparison of SARS-CoV-2 vaccine-elicited spike (A) and RBD (B) ELISA IgG antibody titers in HC (n=30) and CLL (n=95) patients expressed as half-maximal effective concentrations (EC 50 ). (C and D) SARS-CoV-2 vaccine-elicited spike (C) and RBD (D) ELISA IgG antibody titers stratified by CLL disease status: treatment-naïve (Naïve) (n=45), on-therapy (On Tx) (n=34), off-therapy in clinical remission (CR) (n=9), and off therapy and relapsed or refractory (R/R) (n=7). (E-G) Comparison of SARS-CoV-2 vaccine-elicited neutralizing antibody titers for HC (n=30) and CLL patients against (E) D614G (CLL; n=95) and (F) delta (CLL; n=93) spike variants expressed as the reciprocal half-maximal inhibitory dilution (ID 50 ) as determined in an HIV-based pseudovirus neutralization assay or by (G) ACE2/RBD (Wuhan) binding inhibition (CLL; n=95) at a 1:25 dilution. (H-J) SARS-CoV-2 vaccine elicited NAb titers for D614G (H) and delta (I) S variants and ACE2/RBD binding frequencies (J) stratified by CLL disease status. Bars indicate the median with 95% CI. Dotted black lines indicate assay sensitivity cutoffs (EC 50 values of <100 by ELISA, ID 50 values of <20 in the neutralization assay, and >90% ACE2 binding in the RBD-inhibition assay). P values were determined by the Mann-Whitney test (A-B, E-G) or Dunn’s test of multiple comparisons following a Kruskal-Wallis test (C-D, H-J).
Article Snippet: Plates were incubated at room temperature for 1 h, washed 4 times with PBST, and 50 µl of biotinylated goat anti-human ACE2 (R&D) diluted at 0.1 µg/ml in PBST was added to the wells.
Techniques: Enzyme-linked Immunosorbent Assay, Neutralization, Binding Assay, Inhibition, MANN-WHITNEY