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Comment
. 2021 Jan;35(1):289-292.
doi: 10.1038/s41375-020-01103-2. Epub 2020 Dec 9.

SARS-CoV-2 antibody responses in patients with acute leukaemia

J O'Nions  1 , L Muir  2 , J Zheng  3 , C Rees-Spear  2 , A Rosa  4 , C Roustan  4 , C Earl  4 , P Cherepanov  4 , R Gupta  3   5 , A Khwaja  3   5 , C Jolly  2 , L E McCoy  6
Affiliations
Comment

SARS-CoV-2 antibody responses in patients with acute leukaemia

J O'Nions et al. Leukemia. 2021 Jan.
No abstract available

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Conflict of interest statement

The authors declare that they have no conflict of interest.

Figures

Fig. 1
Fig. 1. Temporal dynamics and anti-viral function of SARS-CoV-2 antibody responses in acute leukaemia.
Serum samples from patients A–K were assayed on ELISA plates pre-coated with S1 (a) or N (b). Antigen production and assay conditions were as previously described [–6] except that all samples were treated with 0.5% NP40 before dilution in ELISA buffer. Absorbance was measured at 405 nm and data expressed as fold-change above blank background. The limit of detection (seropositivity) was determined as fold change >4 and is indicated by the dotted line marked *. The dotted line marked ** indicates the limit of quantification in the assay depicted in d–e, determined by the linear range of the standard curve. c ELISA plates were coated overnight at 4 °C with goat anti-human F(ab)’2. Serum samples from patients A–K, and commercial polyclonal IgG standard, were titrated in ELISA buffer and added to the ELISA plate. Binding was detected with goat anti-human IgG conjugated to peroxidase and absorbance read at 450 nm. IgG concentrations in serum were calculated based on interpolation from the IgG standard results using a four-parameter logistic (4PL) regression curve fitting model. Dotted lines marked with }* indicate the normal average range for IgG in human serum. d, e Sera supplemented with 0.5% NP40 from patients A–K were diluted in ELISA buffer and then added to a blocked ELISA plates pre-coated with the indicated antigen and three lanes of goat anti-human F(ab)’2 as per (c). Binding was detected with anti-IgG conjugated to alkaline phosphatase and absorbance measured at 405 nm. Antigen-specific IgG concentrations in serum were calculated based on interpolation from the IgG standard results using a four-parameter logistic (4PL) regression curve fitting model. The dotted line indicates the limit of quantification, which is determined by the linear range of the standard curve and higher than the limit of detection in a and b. f. Sequential serum samples from seropositive patients were titrated in duplicate and pre-incubated with luciferase-encoding HIV pseudotyped with the SARS-CoV-2 Spike for 1 h prior to the addition of HeLa cells expressing human ACE2 as previously described [7]. ID50 titres were only calculated in GraphPad Prism where at least two data points exhibited >50% neutralisation. ID50 values for each patient are plotted on the y-axis against days POS on the x-axis. g. ID50 values plotted against semi-quantitative S1 titres (μg/ml) for each sample. There is a trend to increasing ID50 with higher S1 titres but linear regression does not show a significant correlation (r = 0.06915, p = 0.0925). All data are from at least three independent experiments. h ID50 values and anti-S1 (μg/ml) values plotted against COVID-19 severity score for each patient as cited in Table 1. A trend line is shown but there were insufficient sample numbers to assess correlation and significance.
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References

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