Preexisting and de novo humoral immunity to SARS-CoV-2 in humans

doi:10.1126/science.abe1107

. 2020 Dec 11;370(6522):1339-1343.

doi: 10.1126/science.abe1107. Epub 2020 Nov 6.

Preexisting and de novo humoral immunity to SARS-CoV-2 in humans

Kevin W Ng¹, Nikhil Faulkner¹, Georgina H Cornish¹, Annachiara Rosa², Ruth Harvey³, Saira Hussain³, Rachel Ulferts⁴, Christopher Earl⁵, Antoni G Wrobel⁶, Donald J Benton⁶, Chloe Roustan⁷, William Bolland¹, Rachael Thompson¹, Ana Agua-Doce⁸, Philip Hobson⁸, Judith Heaney⁹, Hannah Rickman⁹, Stavroula Paraskevopoulou⁹, Catherine F Houlihan^{9

10}, Kirsty Thomson⁹, Emilie Sanchez⁹, Gee Yen Shin⁹, Moira J Spyer^{9

11}, Dhira Joshi¹², Nicola O'Reilly¹², Philip A Walker⁷, Svend Kjaer⁷, Andrew Riddell⁸, Catherine Moore¹³, Bethany R Jebson^{14

15}, Meredyth Wilkinson^{14

15}, Lucy R Marshall^{14

15}, Elizabeth C Rosser^{14

16}, Anna Radziszewska^{14

16}, Hannah Peckham^{14

16}, Coziana Ciurtin^{14

16}, Lucy R Wedderburn^{14

15}, Rupert Beale⁴, Charles Swanton¹⁷, Sonia Gandhi¹⁸, Brigitta Stockinger¹⁹, John McCauley³, Steve J Gamblin⁶, Laura E McCoy²⁰, Peter Cherepanov²¹, Eleni Nastouli^{22

11}, George Kassiotis^{23

24}

Affiliations

¹ Retroviral Immunology, The Francis Crick Institute, London NW1 1AT, UK.
² Chromatin Structure and Mobile DNA Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
³ Worldwide Influenza Centre, The Francis Crick Institute, London NW1 1AT, UK.
⁴ Cell Biology of Infection Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
⁵ Signalling and Structural Biology Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
⁶ Structural Biology of Disease Processes Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
⁷ Structural Biology STP, The Francis Crick Institute, London NW1 1AT, UK.
⁸ Flow Cytometry STP, The Francis Crick Institute, London NW1 1AT, UK.
⁹ University College London Hospitals (UCLH) NHS Trust, London NW1 2BU, UK.
¹⁰ Division of Infection and Immunity, University College London (UCL), London WC1E 6BT, UK.
¹¹ Department of Population, Policy and Practice, Great Ormond Street Institute for Child Health (ICH), UCL, London WC1N 1EH, UK.
¹² Peptide Chemistry, The Francis Crick Institute, London NW1 1AT, UK.
¹³ Public Health Wales, University Hospital of Wales, Cardiff CF14 4XW, UK.
¹⁴ Centre for Adolescent Rheumatology Versus Arthritis at UCL, UCLH, Great Ormond Street Hospital (GOSH), London WC1N 3JH, UK.
¹⁵ UCL Great Ormond Street Institute for Child Health (ICH), UCL, London WC1N 1EH, UK.
¹⁶ Centre for Rheumatology Research, Division of Medicine, UCL, London, WC1E 6BT, UK.
¹⁷ Cancer Evolution and Genome Instability Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
¹⁸ Neurodegeneration Biology Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
¹⁹ AhRimmunity Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
²⁰ Division of Infection and Immunity, University College London (UCL), London WC1E 6BT, UK. george.kassiotis@crick.ac.uk e.nastouli@ucl.ac.uk peter.cherepanov@crick.ac.uk l.mccoy@ucl.ac.uk.
²¹ Chromatin Structure and Mobile DNA Laboratory, The Francis Crick Institute, London NW1 1AT, UK. george.kassiotis@crick.ac.uk e.nastouli@ucl.ac.uk peter.cherepanov@crick.ac.uk l.mccoy@ucl.ac.uk.
²² University College London Hospitals (UCLH) NHS Trust, London NW1 2BU, UK. george.kassiotis@crick.ac.uk e.nastouli@ucl.ac.uk peter.cherepanov@crick.ac.uk l.mccoy@ucl.ac.uk.
²³ Retroviral Immunology, The Francis Crick Institute, London NW1 1AT, UK. george.kassiotis@crick.ac.uk e.nastouli@ucl.ac.uk peter.cherepanov@crick.ac.uk l.mccoy@ucl.ac.uk.
²⁴ Department of Medicine, Faculty of Medicine, Imperial College London, London W2 1PG, UK.

PMID: 33159009
PMCID: PMC7857411
DOI: 10.1126/science.abe1107

Preexisting and de novo humoral immunity to SARS-CoV-2 in humans

Kevin W Ng et al. Science. 2020.

. 2020 Dec 11;370(6522):1339-1343.

doi: 10.1126/science.abe1107. Epub 2020 Nov 6.

Authors

Affiliations

¹ Retroviral Immunology, The Francis Crick Institute, London NW1 1AT, UK.
² Chromatin Structure and Mobile DNA Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
³ Worldwide Influenza Centre, The Francis Crick Institute, London NW1 1AT, UK.
⁴ Cell Biology of Infection Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
⁵ Signalling and Structural Biology Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
⁶ Structural Biology of Disease Processes Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
⁷ Structural Biology STP, The Francis Crick Institute, London NW1 1AT, UK.
⁸ Flow Cytometry STP, The Francis Crick Institute, London NW1 1AT, UK.
⁹ University College London Hospitals (UCLH) NHS Trust, London NW1 2BU, UK.
¹⁰ Division of Infection and Immunity, University College London (UCL), London WC1E 6BT, UK.
¹¹ Department of Population, Policy and Practice, Great Ormond Street Institute for Child Health (ICH), UCL, London WC1N 1EH, UK.
¹² Peptide Chemistry, The Francis Crick Institute, London NW1 1AT, UK.
¹³ Public Health Wales, University Hospital of Wales, Cardiff CF14 4XW, UK.
¹⁴ Centre for Adolescent Rheumatology Versus Arthritis at UCL, UCLH, Great Ormond Street Hospital (GOSH), London WC1N 3JH, UK.
¹⁵ UCL Great Ormond Street Institute for Child Health (ICH), UCL, London WC1N 1EH, UK.
¹⁶ Centre for Rheumatology Research, Division of Medicine, UCL, London, WC1E 6BT, UK.
¹⁷ Cancer Evolution and Genome Instability Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
¹⁸ Neurodegeneration Biology Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
¹⁹ AhRimmunity Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
²⁰ Division of Infection and Immunity, University College London (UCL), London WC1E 6BT, UK. george.kassiotis@crick.ac.uk e.nastouli@ucl.ac.uk peter.cherepanov@crick.ac.uk l.mccoy@ucl.ac.uk.
²¹ Chromatin Structure and Mobile DNA Laboratory, The Francis Crick Institute, London NW1 1AT, UK. george.kassiotis@crick.ac.uk e.nastouli@ucl.ac.uk peter.cherepanov@crick.ac.uk l.mccoy@ucl.ac.uk.
²² University College London Hospitals (UCLH) NHS Trust, London NW1 2BU, UK. george.kassiotis@crick.ac.uk e.nastouli@ucl.ac.uk peter.cherepanov@crick.ac.uk l.mccoy@ucl.ac.uk.
²³ Retroviral Immunology, The Francis Crick Institute, London NW1 1AT, UK. george.kassiotis@crick.ac.uk e.nastouli@ucl.ac.uk peter.cherepanov@crick.ac.uk l.mccoy@ucl.ac.uk.
²⁴ Department of Medicine, Faculty of Medicine, Imperial College London, London W2 1PG, UK.

PMID: 33159009
PMCID: PMC7857411
DOI: 10.1126/science.abe1107

Abstract

Zoonotic introduction of novel coronaviruses may encounter preexisting immunity in humans. Using diverse assays for antibodies recognizing SARS-CoV-2 proteins, we detected preexisting humoral immunity. SARS-CoV-2 spike glycoprotein (S)-reactive antibodies were detectable using a flow cytometry-based method in SARS-CoV-2-uninfected individuals and were particularly prevalent in children and adolescents. They were predominantly of the immunoglobulin G (IgG) class and targeted the S2 subunit. By contrast, SARS-CoV-2 infection induced higher titers of SARS-CoV-2 S-reactive IgG antibodies targeting both the S1 and S2 subunits, and concomitant IgM and IgA antibodies, lasting throughout the observation period. SARS-CoV-2-uninfected donor sera exhibited specific neutralizing activity against SARS-CoV-2 and SARS-CoV-2 S pseudotypes. Distinguishing preexisting and de novo immunity will be critical for our understanding of susceptibility to and the natural course of SARS-CoV-2 infection.

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Figures

**Fig. 1. Flow cytometric detection and specificity of antibodies reactive with SARS-CoV-2 S.**
(A) Detection of IgG, IgA, and IgM in five individuals from each indicated group. IgM levels are indicated by a heatmap. (B to D) Inhibition of SARS-CoV-2 S binding of sera from SARS-CoV-2–infected (SARS-CoV-2⁺, n = 10) or SARS-CoV-2–uninfected (SARS-CoV-2⁻ HCoV⁺, n = 6) patients by soluble S1 or S2. (B) Flow cytometry profile of one representative patient per group. (C) Mean frequency of positive cells. *P = 0.015; **P = 0.006, one-way analysis of variance (ANOVA) on ranks. (D) Mean staining intensity [mean fluorescence intensity (MFI) of sample as a percentage of negative control MFI]. In (C) and (D), dots represent individual samples from one of three similar experiments.

**Fig. 2. Prevalence of SARS-CoV-2 S–cross-reactive antibodies detected by different methods.**
(A) Flow cytometry and ELISA results for each sample in cohorts A and C to E listed in table S1. (B) Flow cytometry and ELISA results for serum samples from SARS-CoV-2–uninfected pregnant women. (C to E) SARS-CoV-2 S–cross-reactive antibodies in healthy children and adolescents. (C) Representative flow cytometry profiles of seronegative donors (Negative) or COVID-19 patients (Positive) and of SARS-CoV-2–uninfected adolescents with SARS-CoV-2 cross-reactive antibodies. (D) Frequency of cells stained with all three antibody classes (IgG⁺IgM⁺IgA⁺) or only with IgG (IgG⁺) ranked by their IgG⁺IgM⁺IgA⁺ frequency. The dashed line denotes the assay sensitivity cutoff. (E) Flow cytometry and ELISA results for each sample. (F) Prevalence of SARS-CoV-2 S–cross-reactive antibodies in the indicated age groups (line) and frequency of cells that stained only with IgG (dots) in all samples for which the date of birth was known. The heatmaps in (A), (B), and (E) represent the quartile values above each assay’s technical cutoff.

**Fig. 3. Neutralization of SARS-CoV-2 S pseudotypes and authentic SARS-CoV-2 by SARS-CoV-2–infected and –uninfected patient sera.**
(A) Inhibition of transduction efficiency of SARS-CoV-2 S or VSVg pseudotypes by adult COVID-19 patients who seroconverted (SARS-CoV-2⁺ Adults Ab⁺) or not (SARS-CoV-2⁺ Adults Ab⁻) and SARS-CoV-2–uninfected adult donors (SARS-CoV-2⁻ Adults Ab⁺) or children and adolescent donors (SARS-CoV-2⁻ Children/Adolescents Ab⁺) with SARS-CoV-2 S–binding antibodies. Each line is an individual serum sample. (B) Authentic SARS-CoV-2 neutralization titers of sera from the same donors as in (A), as well as SARS-CoV-2–uninfected donors without SARS-CoV-2 S–binding antibodies (Ab⁻). Dots represent individual samples. *P = 0.037; **P = 0.014; ns, not significant by one-way ANOVA on ranks.

**Fig. 4. Mapping of cross-reactive epitopes in SARS-CoV-2 S.**
(A) Signal intensity for each overlapping peptide along the length of SARS-CoV-2 S covered in the peptide arrays using pooled sera with (Ab⁺) or without (Ab⁻) flow cytometry–detectable SARS-CoV-2 S–reactive antibodies. Differentially recognized peaks are boxed. (B) Alignment of the amino acid sequences of SARS-CoV-2 and HCoV S glycoproteins. Boxes indicate predicted core epitopes. (C) Mapping of predicted epitopes targeted on the trimeric SARS-CoV-2 spike. The S1 (blue) and S2 (pink) subunits of one monomer are colored. Epitopes are shown for one monomer; the circled dashed line represent the membrane proximal region not present in the structure. (D) Left: Reactivity with the S glycoproteins of each HCoV of the indicated sera with (Ab⁺) or without (Ab⁻) flow cytometry–detectable SARS-CoV-2 S–reactive antibodies as determined by flow cytometry. Each column is an individual sample. Rows depict the staining for each antibody class. Right: Correlation coefficients between percentages of IgG staining for SARS-CoV-2 S and IgG, IgM, and IgA staining for each HCoV S glycoprotein.

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Comment in

Remembering seasonal coronaviruses.
Guthmiller JJ, Wilson PC. Guthmiller JJ, et al. Science. 2020 Dec 11;370(6522):1272-1273. doi: 10.1126/science.abf4860. Science. 2020. PMID: 33303605 No abstract available.
Do childhood colds help the body respond to COVID?
Brazil R. Brazil R. Nature. 2021 Nov;599(7886):540-541. doi: 10.1038/d41586-021-03087-0. Nature. 2021. PMID: 34795432 No abstract available.

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References

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1. Callow K. A., Parry H. F., Sergeant M., Tyrrell D. A., The time course of the immune response to experimental coronavirus infection of man. Epidemiol. Infect. 105, 435–446 (1990). 10.1017/S0950268800048019 - DOI - PMC - PubMed
1. Severance E. G., Bossis I., Dickerson F. B., Stallings C. R., Origoni A. E., Sullens A., Yolken R. H., Viscidi R. P., Development of a nucleocapsid-based human coronavirus immunoassay and estimates of individuals exposed to coronavirus in a U.S. metropolitan population. Clin. Vaccine Immunol. 15, 1805–1810 (2008). 10.1128/CVI.00124-08 - DOI - PMC - PubMed
1. Dijkman R., Jebbink M. F., El Idrissi N. B., Pyrc K., Müller M. A., Kuijpers T. W., Zaaijer H. L., van der Hoek L., Human coronavirus NL63 and 229E seroconversion in children. J. Clin. Microbiol. 46, 2368–2373 (2008). 10.1128/JCM.00533-08 - DOI - PMC - PubMed
1. Huang A. T., Garcia-Carreras B., Hitchings M. D. T., Yang B., Katzelnick L. C., Rattigan S. M., Borgert B. A., Moreno C. A., Solomon B. D., Trimmer-Smith L., Etienne V., Rodriguez-Barraquer I., Lessler J., Salje H., Burke D. S., Wesolowski A., Cummings D. A. T., A systematic review of antibody mediated immunity to coronaviruses: Kinetics, correlates of protection, and association with severity. Nat. Commun. 11, 4704 (2020). 10.1038/s41467-020-18450-4 - DOI - PMC - PubMed

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[1] Aldridge R. W., Lewer D., Beale S., Johnson A. M., Zambon M., Hayward A. C., Fragaszy E. B., Seasonality and immunity to laboratory-confirmed seasonal coronaviruses (HCoV-NL63, HCoV-OC43, and HCoV-229E): Results from the Flu Watch cohort study. Wellcome Open Res. 5, 52 (2020). 10.12688/wellcomeopenres.15812.1 - DOI - PMC - PubMed

[2] Aldridge R. W., Lewer D., Beale S., Johnson A. M., Zambon M., Hayward A. C., Fragaszy E. B., Seasonality and immunity to laboratory-confirmed seasonal coronaviruses (HCoV-NL63, HCoV-OC43, and HCoV-229E): Results from the Flu Watch cohort study. Wellcome Open Res. 5, 52 (2020). 10.12688/wellcomeopenres.15812.1 - DOI - PMC - PubMed

[3] Callow K. A., Parry H. F., Sergeant M., Tyrrell D. A., The time course of the immune response to experimental coronavirus infection of man. Epidemiol. Infect. 105, 435–446 (1990). 10.1017/S0950268800048019 - DOI - PMC - PubMed

[4] Callow K. A., Parry H. F., Sergeant M., Tyrrell D. A., The time course of the immune response to experimental coronavirus infection of man. Epidemiol. Infect. 105, 435–446 (1990). 10.1017/S0950268800048019 - DOI - PMC - PubMed

[5] Severance E. G., Bossis I., Dickerson F. B., Stallings C. R., Origoni A. E., Sullens A., Yolken R. H., Viscidi R. P., Development of a nucleocapsid-based human coronavirus immunoassay and estimates of individuals exposed to coronavirus in a U.S. metropolitan population. Clin. Vaccine Immunol. 15, 1805–1810 (2008). 10.1128/CVI.00124-08 - DOI - PMC - PubMed

[6] Severance E. G., Bossis I., Dickerson F. B., Stallings C. R., Origoni A. E., Sullens A., Yolken R. H., Viscidi R. P., Development of a nucleocapsid-based human coronavirus immunoassay and estimates of individuals exposed to coronavirus in a U.S. metropolitan population. Clin. Vaccine Immunol. 15, 1805–1810 (2008). 10.1128/CVI.00124-08 - DOI - PMC - PubMed

[7] Dijkman R., Jebbink M. F., El Idrissi N. B., Pyrc K., Müller M. A., Kuijpers T. W., Zaaijer H. L., van der Hoek L., Human coronavirus NL63 and 229E seroconversion in children. J. Clin. Microbiol. 46, 2368–2373 (2008). 10.1128/JCM.00533-08 - DOI - PMC - PubMed

[8] Dijkman R., Jebbink M. F., El Idrissi N. B., Pyrc K., Müller M. A., Kuijpers T. W., Zaaijer H. L., van der Hoek L., Human coronavirus NL63 and 229E seroconversion in children. J. Clin. Microbiol. 46, 2368–2373 (2008). 10.1128/JCM.00533-08 - DOI - PMC - PubMed

[9] Huang A. T., Garcia-Carreras B., Hitchings M. D. T., Yang B., Katzelnick L. C., Rattigan S. M., Borgert B. A., Moreno C. A., Solomon B. D., Trimmer-Smith L., Etienne V., Rodriguez-Barraquer I., Lessler J., Salje H., Burke D. S., Wesolowski A., Cummings D. A. T., A systematic review of antibody mediated immunity to coronaviruses: Kinetics, correlates of protection, and association with severity. Nat. Commun. 11, 4704 (2020). 10.1038/s41467-020-18450-4 - DOI - PMC - PubMed

[10] Huang A. T., Garcia-Carreras B., Hitchings M. D. T., Yang B., Katzelnick L. C., Rattigan S. M., Borgert B. A., Moreno C. A., Solomon B. D., Trimmer-Smith L., Etienne V., Rodriguez-Barraquer I., Lessler J., Salje H., Burke D. S., Wesolowski A., Cummings D. A. T., A systematic review of antibody mediated immunity to coronaviruses: Kinetics, correlates of protection, and association with severity. Nat. Commun. 11, 4704 (2020). 10.1038/s41467-020-18450-4 - DOI - PMC - PubMed

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Preexisting and de novo humoral immunity to SARS-CoV-2 in humans

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Preexisting and de novo humoral immunity to SARS-CoV-2 in humans

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