SARS-CoV-2 serological analysis of COVID-19 hospitalized patients, pauci-symptomatic individuals and blood donors

Grzelak, Ludivine; Temmam, Sarah; Planchais, Cyril; Demeret, Caroline; Huon, Christèle; Guivel, Florence; Staropoli, Isabelle; Chazal, Maxime; Dufloo, Jérémy; Planas, Delphine; Buchrieser, Julian; Rajah, Maaran Michael; Robinot, Rémy; Porrot, Françoise; Albert, Mélanie; Chen, Kuang-Yu; Crescenzo, Bernadette; Donati, Flora; Anna, François; Souque, Philippe; Gransagne, Marion; Bellalou, Jacques; Nowakowski, Mireille; Backović, Marija; Bouadma, Lila; Fèvre, Lucie Le; Hingrat, Quentin Le; Descamps, Diane; Pourbaix, Anabelle; Yazdanpanah, Yazdan; Tondeur, Laura; Besombes, Camille; Ungeheuer, Marie‐Noëlle; Mellon, Guillaume; Morel, Pascal; Rolland, Sophie; Rey, F.A.; Behillil, Sylvie; Enouf, Vincent; Lemaître, A.; Créach, M.A.; Pêtres, Stéphane; Escriou, Nicolas; Charneau, Pierre; Fontanet, Arnaud; Hoën, Bruno; Bruel, Timothée; Éloit, Marc; Mouquet, Hugo; Schwartz, Olivier; Werf, Sylvie van der

doi:10.1101/2020.04.21.20068858

Cited by 76 publications

(110 citation statements)

References 35 publications

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“…The flow cytometry-based method we employed demonstrated the highest degree of specificity and sensitivity. A similar method was also recently used in an independent study, reaching comparable results, except no cross-reactivity with HCoV was suggested 9 . Although not specifically investigated in the latter study, detection of cross-reactivity with HCoV was deliberately avoided by higher dilution of serum samples.…”

Section: Discussionmentioning

confidence: 80%

“…Moreover, infection with SARS-CoV-2 has recently been shown to increase IgG seroreactivity to HCoVs 21 , supporting the existence of shared epitopes. Numerous serology assays that are being developed have also detected SARS-CoV-2reactive antibodies in a considerable proportion of SARS-CoV-2-uninfected individuals, which has often been considered as unspecific binding, rather than HCoV-elicited specific antibodies [9][10][11][12][13][14][15][16][17][18][19][20]22 . Based on our data, we would argue that this is the result of cross-reactivity between HCoVs.…”

mentioning

confidence: 99%

See 1 more Smart Citation

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

Ng¹,

Faulkner²,

Cornish³

et al. 2020

Preprint

126

170

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Several related human coronaviruses (HCoVs) are endemic in the human population, causing mild respiratory infections 1 . Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the etiologic agent of Coronavirus disease 2019 , is a recent zoonotic infection that has quickly reached pandemic spread 2,3 . Zoonotic introduction of novel coronaviruses is thought to occur in the absence of pre-existing immunity in the target human population. Using diverse assays for detection of antibodies reactive with the SARS-CoV-2 Spike (S) glycoprotein, we demonstrate the presence of pre-existing immunity in uninfected and unexposed humans to the new coronavirus. SARS-CoV-2 S-reactive antibodies, exclusively of the IgG class, were readily detectable by a sensitive flow cytometry-based method in SARS-CoV-2-uninfected individuals with recent HCoV infection and targeted the S2 subunit. In contrast, SARS-CoV-2 infection induced higher titres of SARS-CoV-2 Sreactive IgG antibodies, as well as concomitant IgM and IgA antibodies throughout the observation period of 6 weeks since symptoms onset. HCoV patient sera also variably reacted with SARS-CoV-2 S and nucleocapsid (N), but not with the S1 subunit or the receptor binding domain (RBD) of S on standard enzyme immunoassays. Notably, HCoV patient sera exhibited specific neutralising activity against SARS-CoV-2 S pseudotypes, according to levels of SARS-CoV-2 S-binding IgG and with efficiencies comparable to those of COVID-19 patient sera. Distinguishing pre-existing and de novo antibody responses to SARS-CoV-2 will be critical for serology, seroprevalence and vaccine studies, as well as for our understanding of susceptibility to and natural course of SARS-CoV-2 infection. ResultsImmune cross-reactivity among seasonally spreading human coronaviruses (HCoVs) has long been hypothesised to provide cross-protection, albeit transient, against infection with distinct HCoV types 1,4,5 . To determine the degree of cross-reactivity between HCoVs and the recently introduced zoonotic coronavirus SARS-CoV-2, we developed a sensitive flow cytometry-based assay for detection of SARS-CoV-2-binding antibodies. Sera from COVID-19 patients at University College London Hospitals (UCLH) (Table S1), contained high levels of IgG, IgM and IgA antibodies recognising the wild-type Spike (S) glycoprotein of SARS-CoV-2 expressed on the surface of HEK293T cells, whereas control sera did not (Fig. 1a). Notably, sera from a proportion patients with confirmed HCoV infection collected before or during the early spread of SARS-CoV-2 in the UK (Table S1), also contained SARS-CoV-2 S-specific antibodies (Fig. 1a). However, the latter sera contained only lower levels of S-specific IgG and no IgM or IgA antibodies, which clearly distinguished them from COVID-19 patient sera (Fig. 1a). The SARS-CoV-2 S protein is proteolytically processed into the S1 and S2 subunits that mediate target cell attachment and entry, respectively 6,7 . S2 exhibits a higher degree of homology among coronaviruses than S1 (Extended data Fig. 1...

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Section: Discussionmentioning

confidence: 80%

mentioning

confidence: 99%

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

Ng¹,

Faulkner²,

Cornish³

et al. 2020

Preprint

126

170

View full text Add to dashboard Cite

show abstract

“…There are a large number of immunological assays capable of measuring the antibody response to SARS-CoV-2 including neutrilization assays, ELISA, Luminex, Luciferase Immunoprecipitation System (LIPS), peptide microarrays and more [34].…”

Section: Discussionmentioning

confidence: 99%

Serological signatures of SARS-CoV-2 infection: Implications for antibody-based diagnostics

Rosado

Pelleau

Cockram

et al. 2020

Preprint

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BackgroundThe antibody response generated following infection with SARS-CoV-2 is expected to decline over time. This may cause individuals with confirmed SARS-CoV-2 infection to test negative according to serological diagnostic tests in the months and years following symptom onset. MethodsA multiplex serological assay was developed to measure IgG and IgM antibody responses to four SARS-CoV-2 Spike (S) antigens: spike trimeric ectodomain (S tri ), its receptor-binding domain (RBD), spike subunit 1 (S1), and spike subunit 2 (S2).Antibody responses were measured in serum samples from patients in French hospitals with RT-qPCR confirmed infection (n = 259), and negative control serum samples collected before the start of the SARS-CoV-2 epidemic in 2019 (n = 335).The multiplex antibody data was used to train a random forests algorithm for classifying individuals with previous SARS-CoV-2 infection. A mathematical model of antibody kinetics informed by prior information from other coronaviruses was used to estimate time-varying antibody responses and assess the potential sensitivity and classification performance of serological diagnostics during the first year following symptom onset. ResultsIgG antibody responses to one S antigen identified individuals with previous RT-qPCR confirmed SARS-CoV-2 infection with 90.3% sensitivity (95% confidence interval (CI); 86.1%, 93.4%) and 99.1% specificity (95% CI; 97.4%, 99.7%). Using a serological signature of IgG to four antigens, it was possible to identify infected individuals with 96.1% sensitivity (95% CI; 93.0%, 97.9%) and 99.1% specificity (95% CI; 97.4%, 99.7%). Antibody responses to SARS-CoV-2 increase rapidly 1-2 weeks after symptom onset, with antibody responses predicted to peak within 2-4 weeks. Informed by prior data from other coronaviruses, one year following symptom onset antibody responses are predicted to decay by approximately 60% from the peak response. Depending on the selection of sero-positivity cutoff, we estimate that the sensitivity of serological diagnostics may reduce to 56% -97% after six months, and to 49% -93% after one year. ConclusionSerological signatures based on antibody responses to multiple antigens can provide more accurate and robust serological classification of individuals with previous SARS-CoV-2 infection. Changes in antibody levels over time may cause reductions in the sensitivity of serological diagnostics leading to an underestimation of sero-prevalence. It is essential that data continue to be collected to evaluate this potential risk.

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“…Briefly, hACE2 was PCR amplified adding SpeI, XhoI, and attB sites using forward primer (5'-TCC CTC ACT AGT ACA AGT TTG TAC AAA AAA GCA GGC TGC CAC CAT GTC AAG CTC TTC CTG GCT C -3') and reverse primer (5'-AAA AAA CTC GAG ACC ACT TTG TAC AAG AAA GCT GGG TTT AAG CGG GCG CCA CCT -3') and cloned into pLenti6-H2B-mCherry using XhoI and SpeI sites. phCMV-SARS-CoV2-S was previously described 55 . pLV-EF1α-TMPRSS2-IRES-Hygro was generated by gateway cloning of pCSDest-TMPRSS2 into pLV-EF1α-IRES-Hygro-DEST.…”

Section: Methodsmentioning

confidence: 99%

Syncytia formation by SARS-CoV-2 infected cells

Buchrieser

Dufloo

Hubert

et al. 2020

Preprint

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113

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Severe cases of COVID-19 are associated with extensive lung damage and the presence of infected multinucleated syncytial pneumocytes. The viral and cellular mechanisms regulating the formation of these syncytia are not well understood. Here, we show that SARS-CoV-2 infected cells express the viral Spike protein (S) at their surface and fuse with ACE2-positive neighbouring cells. Expression of S without any other viral proteins triggers syncytia formation. Type-I interferon (IFN)-induced transmembrane proteins (IFITMs), a family of restriction factors that block the entry of many viruses, inhibit S-mediated fusion, with IFITM1 being more active than IFITM2 and IFITM3. On the contrary, the TMPRSS2 serine protease, which is known to enhance infectivity of cell-free virions, processes both S and ACE2 and increases syncytia formation by accelerating the fusion process. TMPRSS2 thwarts the antiviral effect of IFITMs. Our results show that the pathological effects of SARS-CoV-2 are modulated by cellular proteins that either inhibit or facilitate syncytia formation.

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SARS-CoV-2 serological analysis of COVID-19 hospitalized patients, pauci-symptomatic individuals and blood donors

Cited by 76 publications

References 35 publications

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

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

Serological signatures of SARS-CoV-2 infection: Implications for antibody-based diagnostics

Syncytia formation by SARS-CoV-2 infected cells

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