key: cord-271544-i20105lq authors: Poston, Daniel; Weisblum, Yiska; Wise, Helen; Templeton, Kate; Jenks, Sara; Hatziioannou, Theodora; Bieniasz, Paul title: Absence of SARS-CoV-2 neutralizing activity in pre-pandemic sera from individuals with recent seasonal coronavirus infection date: 2020-10-11 journal: medRxiv DOI: 10.1101/2020.10.08.20209650 sha: doc_id: 271544 cord_uid: i20105lq Cross-reactive immune responses elicited by seasonal coronaviruses might impact SARS-CoV-2 susceptibility and disease outcomes. We measured neutralizing activity against SARS-CoV-2 in pre-pandemic sera from patients with prior PCR-confirmed seasonal coronavirus infection. While neutralizing activity against seasonal coronaviruses was detected in nearly all sera, cross-reactive neutralizing activity against SARS-CoV-2 was undetectable. Since the initial description in December 2019 of a novel human coronavirus SARS-CoV-2, 43 there has been a global effort to identify underlying the underlying causes for the great range of 44 disease severity observed, from mild or even asymptomatic infection to severe respiratory 45 distress and death. One hypothesis is that cross reactive immune responses, elicited by prior 46 infection with seasonal coronaviruses impacts the course of SARS-CoV-2 infection, perhaps 47 providing a degree of protection against severe COVID-19 disease. 48 The endemic seasonal human coronaviruses (HCoVs)-HCoV-HKU1, HCoV-OC43, HCoV-50 NL63, and HCoV-229E-cause, mild or subclinical respiratory infections, with severe disease 51 being exceptionally rare [1] . Although there is low overall sequence homology between the 52 SARS-CoV-2 Spike (S) protein and those of the endemic HCoVs, overlapping T-cell epitopes 53 have been reported, particularly in the S2 subunit [2, 3] . It is possible that neutralizing antibodies 54 induced by seasonal HCoV infection could cross-react with similar epitopes in SARS-CoV-2 S. 55 Such antibodies could potentially afford some level of protection against and perhaps contribute 56 to the wide range of outcomes of SARS-CoV-2 infection. To investigate this possibility, we 57 analyzed sera that had been collected prior to the COVID-19 pandemic from patients with a 58 recent PCR-confirmed diagnosis of HCoV-OC43, HCoV-NL63, or HCoV-229E infection. Such 59 samples should contain neutralizing antibodies against the respective seasonal HCoV, without 60 the possibility of prior SARS-CoV-2 infection, allowing us to specifically test whether 61 antibodies elicited by seasonal HCoV infection can neutralize SARS-CoV-2. Our results indicate 62 a lack of SARS-CoV-2 cross-neutralization activity between the seasonal HCoVs and SARS-63 CoV-2. 64 . CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted October 11, 2020. is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted October 11, 2020. . https://doi.org/10.1101/2020.10.08.20209650 doi: medRxiv preprint dilutions and incubated at 37 o C for 1 hour. Virus serum mixtures were subsequently transferred 88 to 96-well plates containing 1x10 4 293T/ACE2cl.22 (for rVSV/SARS-2/GFP and HCoV-OC43) 89 or HT1080/ACE2cl.14 (for HCoV-NL63 and HCoV-229E) target cells/well. Infection was 90 allowed to proceed for 16 hours (rVSV/SARS-2) or 24 hours (HCoV-OC43, HCoV-NL63, 91 HCoV-229E). The numbers of rVSV/SARS-2/GFP was assessed by flow cytometric detection of 92 GFP expression as described previously To assess whether prior infection by seasonal coronaviruses could elicit antibodies with 104 neutralization activity against SARS-CoV-2, we identified 37 serum samples collected prior to 105 the COVID19 pandemic from patients who were diagnosed using PCR with a seasonal 106 coronavirus 11-291 days (median 80 = days) prior to serum sample collection. Of these 20 were 107 diagnosed with HCoV-OC43 infection, 10 were diagnosed with HCoV-NL63 infection and 7 108 were diagnosed with HCoV-229E infection. We also developed flow cytometry-based 109 coronavirus neutralization assays based on the detection of nucleocapsid expression in HCoV-110 . CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted October 11, 2020. is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted October 11, 2020. . https://doi.org/10.1101/2020.10.08.20209650 doi: medRxiv preprint neutralize rVSV/SARS-CoV-2/GFP with NT50 values ranging from 96 to 5400. Overall these 134 data strongly suggest that only pandemic sera, and not pre-pandemic sera have neutralizing 135 activity against SARS-CoV-2, and further suggest that pre-existing serological immunity to 136 seasonal coronaviruses is not a major driver of the diverse outcome of SARS-CoV-2 infection. with this virus is more common, which is in line with previous observations suggesting that 148 reinfection with HCoV-OC43 and HCoV-229E occurs at a greater frequency than HCoV-149 NL63[5-8] and that infection with HCoV-OC43 is common this geographic locale. 150 While we cannot exclude the possibility that that seasonal coronavirus elicit cross-neutralizing 151 antibodies, the divergence between seasonal coronaviruses S proteins would suggest limited 152 cross reactivity (HCoV-OC43 S shares 22.6 and 24.5% identical amino acids with and HCoV-153 229E and HCoV-NL63 S respectively, while HCoV-NL63 and HCoV-229E S share 55% 154 identical amino acids) Accordingly, none of the samples tested had any neutralization activity 155 against SARS-CoV-2 whose spike protein shares 24%-29% amino acid identity with the seasonal 156 . CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted October 11, 2020. . https://doi.org/10.1101/2020.10.08.20209650 doi: medRxiv preprint coronaviruses. In agreement with the notion that there is little cross reactivity between seasonal 157 HCoV neutralizing antibodies and SARS-CoV-2, many of the monoclonal antibodies cloned 158 from SARS-CoV-2 infected individuals contain very low levels of somatic hypermutation [9] , 159 suggesting that they arise from de novo rather than recall B-cell responses. However, instances of Infectious Diseases R37AI640003 (to PDB) and R01AI078788 (to TH). DP was supported by a 179 . CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint this version posted October 11, 2020. CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted October 11, 2020. . https://doi.org/10.1101/2020.10.08.20209650 doi: medRxiv preprint Epidemiology, genetic recombination, and pathogenesis of 187 coronaviruses SARS-CoV-2-reactive T cells in healthy donors and 189 patients with COVID-19 Viral epitope profiling of COVID-19 patients reveals 191 cross-reactivity and correlates of severity Measuring SARS-CoV-2 neutralizing 193 antibody activity using pseudotyped and chimeric viruses Seasonal coronavirus protective 195 immunity is short-lasting Coronavirus and Other 197 Respiratory Illnesses Comparing Older with Young Adults The dominance of human coronavirus OC43 and 200 NL63 infections in infants Epidemiology and clinical 202 presentations of the four human coronaviruses 229E, HKU1, NL63, and OC43 detected 203 over 3 years using a novel multiplex real-time PCR method Convergent antibody responses CoV-2 in convalescent individuals Broad neutralization of SARS-related viruses by 208 human monoclonal antibodies Cross-reactive serum and memory B cell responses to 210 spike protein in SARS-CoV-2 and endemic coronavirus infection Pre-existing and de novo humoral immunity to SARS-212 CoV-2 in humans CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review)The copyright holder for this preprint this version posted October 11, 2020. . https://doi.org/10.1101/2020.10.08.20209650 doi: medRxiv preprint . CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review)