key: cord-293826-2p7dqacd
authors: Lee, Cheryl Yi-Pin; Amrun, Siti Naqiah; Chee, Rhonda Sin-Ling; Goh, Yun Shan; Mak, Tze-Minn; Octavia, Sophie; Yeo, Nicholas Kim-Wah; Chang, Zi Wei; Tay, Matthew Zirui; Torres-Ruesta, Anthony; Carissimo, Guillaume; Poh, Chek Meng; Fong, Siew-Wai; Bei, Wang; Lee, Sandy; Young, Barnaby Edward; Tan, Seow-Yen; Leo, Yee-Sin; Lye, David C.; Lin, Raymond T. P.; Maurer-Stroh, Sebastien; Lee, Bernett; Cheng-I, Wang; Renia, Laurent; Ng, Lisa F.P.
title: Neutralizing antibodies from early cases of SARS-CoV-2 infection offer cross-protection against the SARS-CoV-2 D614G variant
date: 2020-10-09
journal: bioRxiv
DOI: 10.1101/2020.10.08.332544
sha: 
doc_id: 293826
cord_uid: 2p7dqacd

The emergence of a SARS-CoV-2 variant with a point mutation in the spike (S) protein, D614G, has taken precedence over the original Wuhan isolate by May 2020. With an increased infection and transmission rate, it is imperative to determine whether antibodies induced against the D614 isolate may cross-neutralize against the G614 variant. In this report, profiling of the anti-SARS-CoV-2 humoral immunity reveals similar neutralization profiles against both S protein variants, albeit waning neutralizing antibody capacity at the later phase of infection. These findings provide further insights towards the validity of current immune-based interventions. IMPORTANCE Random mutations in the viral genome is a naturally occurring event that may lead to enhanced viral fitness and immunological resistance, while heavily impacting the validity of licensed therapeutics. A single point mutation from aspartic acid (D) to glycine (G) at position 614 of the SARS-CoV-2 spike (S) protein, termed D614G, has garnered global attention due to the observed increase in transmissibility and infection rate. Given that a majority of the developing antibody-mediated therapies and serological assays are based on the S antigen of the original Wuhan reference sequence, it is crucial to determine if humoral immunity acquired from the original SARS-CoV-2 isolate is able to induce cross-detection and cross-protection against the novel prevailing D614G variant.

Laurent Renia: Infectious Diseases Horizontal Technology Centre, A*STAR, 8A 50

Biomedical Grove, Immunos #04-06, Singapore 138648. Phone: (+65)-64070007. 51

Email: renia_laurent@immunol.a-star.edu.sg 52

The emergence of a SARS-CoV-2 variant with a point mutation in the spike (S) 54 protein, D614G, has taken precedence over the original Wuhan isolate by May 2020. 55

With an increased infection and transmission rate, it is imperative to determine 56 whether antibodies induced against the D614 isolate may cross-neutralize against 57 the G614 variant. In this report, profiling of the anti-SARS-CoV-2 humoral immunity 58 reveals similar neutralization profiles against both S protein variants, albeit waning 59 neutralizing antibody capacity at the later phase of infection. These findings provide 60 further insights towards the validity of current immune-based interventions. 61

Random mutations in the viral genome is a naturally occurring event that may lead to 63 enhanced viral fitness and immunological resistance, while heavily impacting the 64 validity of licensed therapeutics. A single point mutation from aspartic acid (D) to 65 glycine (G) at position 614 of the SARS-CoV-2 spike (S) protein, termed D614G, has 66 garnered global attention due to the observed increase in transmissibility and 67 infection rate. Given that a majority of the developing antibody-mediated therapies 68 and serological assays are based on the S antigen of the original Wuhan reference 69 sequence, it is crucial to determine if humoral immunity acquired from the original 70 SARS-CoV-2 isolate is able to induce cross-detection and cross-protection against 71 the novel prevailing D614G variant. 72

Coronavirus disease 2019 is the consequence of an infection by severe 74 acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which emerged in Wuhan, 75 China, in December 2019 (1). The rapid expansion of the COVID-19 pandemic has 76 affected 213 countries and territories, with a global count of more than 36 million 77 laboratory-confirmed human infection cases to date (2). An inevitable impact of this 78 pandemic is the accumulation of immunologically relevant mutations among the viral 79 populations due to natural selection or random genetic drift, resulting in enhanced 80 viral fitness and immunological resistance (3, 4). For instance, antigenic drift was 81 previously reported in other common cold coronaviruses, OC43 and 229E, as well as 82 in SARS-CoV (5-7). 83

In early March 2020, a non-synonymous mutation from aspartic acid (D) to 84 glycine (G) at position 614 of SARS-CoV-2 spike (S) protein was identified (8) Figure 1A and 1B). All patients showed a decrease in IgM response 102 ( Figure 1A) , and a prolonged IgG response over time ( Figure 1B) . Although one 103 recent study has demonstrated similar neutralization profiles against both D614 and 104 G614 SARS-CoV-2 pseudoviruses, the virus clade by which the six individuals were 105 infected with was not identified (9). According to Singapore's SARS-CoV-2 clade 106 pattern from December 2019 till July 2020, the D614G mutation only appeared in 107

February 2020 ( Figure 1C ). Hence, with knowledge on the D614G status of a subset 108 of COVID-19 patients (n=44 infected with D614, n=6 infected with G614, n=7 109 containing all other clades: O, S, L, V, G, GH or GR; Table 1, Figure 1C infections. In addition, determining the level of cross-reactivity is essential for 139 immunosurveillance, as well as to identify broadly neutralizing antibodies or epitopes 140 (23). Here, we confirm that cross-reactivity occurs at the functional level of the 141 humoral response on both the S protein variants. Our results, together with the 142 recent serological evaluation (22), strongly suggest that existing serological assays 143 will be able to detect both D614 and G614 clades of SARS-CoV-2 with a similar 144 sensitivity. However, it is of clinical relevance to assess if cross-reactivity between 145 the variants may enhance viral infection when neutralizing antibodies are present at 146 suboptimal concentrations (24). More importantly, further studies using monoclonal 147 antibodies are necessary to validate the cross-reactivity profiles between both 148 SARS-CoV-2 S variants. 149

Overall, our study shows that the D614G mutation on the S protein does not 150 impact SARS-CoV-2 neutralization by the host antibody response, nor confer viral 151 resistance against the humoral immunity. Hence, there should be negligible impact 152 towards the efficacy of antibody-based therapies and vaccines that are currently 153 being developed. 154

The authors would like to thank the study participants who donated their blood 156 samples to this project, and the healthcare workers caring for the COVID-19 157 patients. The authors also wish to thank Ding Ying and the Singapore Infectious 

New SARS-like virus in China triggers alarm

Insights into RNA 194 synthesis, capping, and proofreading mechanisms of SARS-coronavirus

Making Sense of Mutation: What 197 D614G Means for the COVID-19 Pandemic Remains Unclear

Genetic drift of human 199 coronavirus OC43 spike gene during adaptive evolution

Analysis of human coronavirus 229E spike and 202 nucleoprotein genes demonstrates genetic drift between chronologically distinct 203 strains

Cross-205 host evolution of severe acute respiratory syndrome coronavirus in palm civet and 206 human

The 209 D614G mutation in the SARS-CoV-2 spike protein reduces S1 shedding and 17

Structures of Human Antibodies Bound to SARS-CoV-2 Spike 239 Reveal Common Epitopes and Recurrent Features of Antibodies

Epidemiologic Perspective on Surveillance and Control. Frontiers in immunology

Safety and efficiency 245 of endoscopic resection versus laparoscopic resection in gastric gastrointestinal 246 stromal tumours: A systematic review and meta-analysis

Viral Mutation 249 Rates

Models of RNA virus evolution and their roles 251 in vaccine design

D614G Spike Variant Does Not Alter IgG, IgM, or IgA Spike Seroassay Performance

Serologic cross-reactivity of SARS-CoV-2 with endemic and seasonal 257 Betacoronaviruses. medRxiv : the preprint server for health sciences

A perspective on potential antibody-dependent enhancement of SARS-CoV-2. 261