key: cord-0270165-uifh4ax9 authors: Faustini, S. E.; Shields, A. M.; Banham, G.; Wall, N.; Al-Taei, S.; Tanner, C.; Ahmed, Z.; Efstathiou, E.; Townsend, N.; Price, R.; Curry, G.; Robertson, L.; Nesbit, A.; Black, A.; MOORE, J.; McLaughlin, J.; Farnan, J.; COVID-HD Birmingham Study Group,; PITCH consortium,; Cunningham, A. F.; Harper, L.; Moore, T.; Drayson, M. T.; Richter, A. G. title: Cross reactivity of spike glycoprotein induced antibody against delta and omicron variants before and after third SARS-CoV-2 vaccine dose date: 2022-01-01 journal: nan DOI: 10.1101/2021.12.30.21268308 sha: 5f11650edd20564a5ab5a592c01661c60227a4c5 doc_id: 270165 cord_uid: uifh4ax9 Variants of SARS-CoV-2 may evade natural and vaccine induced immunity and monoclonal antibody immunotherapeutics. There is an urgent need to know how well antibodies, induced by healthy and Clinically Extremely Vulnerable (CEV) patients, will bind and thus help reduce transmission and severity of infection from variants of concern (VOC). This study determines the cross-reactive binding of serum antibodies obtained prior to and 28 days after a third vaccination in three cohorts; a health care worker cohort who received three doses of Pfizer-BioNtech (PPP), a cohort of CEV patients received two doses of the AstraZeneca-ChAdOx1-nCoV-19 (AAP) vaccine, followed by a third PFZ vaccine and a haemodialysis cohort that had a mixture of two AZ or PFZ vaccines followed by a PFZ booster. Six months post second vaccine there was evidence of antibody waning with 58.9% of individuals in the HD cohort seropositive against Wuhan, 34.4% Delta and 62.2% Omicron strains. For the AAP cohort, equivalent figures were 62.5%, 45.8% and 91.7% and the PPP cohort 92.2%, 90% and 91.1%. Post third dose vaccination there were universal increases in seropositivity and median optical density. For the HD cohort, 98.8% were seropositive to the Wuhan strain, 97.6% against Delta and 100% against Omicron strains. For the PPP and AAP cohorts, 100% were seropositive against all 3 strains. Lastly, we examined the WHO NIBSC 20/136 standard and there was no loss of antibody binding to either VOC. Similarly, a dilution series of Sotrovimab (GSK) found this therapeutic monoclonal antibody bound similarly to all VOC. The Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) pandemic continues to affect almost every country in the world. Variants of SARS-CoV-2 may evade natural and vaccine-induced immunity and thus continuously reset our progress in mitigating the disastrous impacts of the virus on our health and socioeconomic well-being. In the UK, two main vaccines have formed the basis of the national immunisation strategy: the AstraZeneca ChAdOx1 nCoV-19 vaccine (AZ) (1) and the Pfizer-BioNtech COVID-19 vaccine (PFZ) (2) . Both elicit immune responses targeting the original, wildtype SARS-CoV-2 (Wuhan) spike glycoprotein and both have been shown to reduce the incidence of severe disease in clinical trials (1, 2) . With the emergence of novel variants of concern (VOC) that demonstrate increased transmissibility (3, 4) , it is necessary to understand whether current vaccines will provide protection against further novel VOC. In mid-2021 the Delta VOC became the dominant strain worldwide and currently the Omicron VOC is likely to becoming the dominant SARS-CoV2 strain in the UK and across the world (5) . Indeed, both strains may even continue to co-exist within the same population. These rapid shifts in the pre-dominance of VOC outpaces and impairs the development and testing in clinical trials of new VOC-tailored vaccines. We do not yet know how well the different vaccine strategies applied in the UK will reduce the transmission of and severity of disease arising from rapidly emerging VOC in the general population and immunological vulnerable subgroups. After vaccination, detection of serum IgG against the SARS-CoV-2 spike glycoprotein correlates with future protection against COVID-19 infection (6) . Nevertheless, it is not fully understood whether IgG binds as similarly to the spike glycoprotein from currently circulating VOC as to the vaccine encoded spike glycoprotein and therefore likely to offer protection against infection. This knowledge gap is especially pronounced for immunocompromised or other clinically extremely vulnerable (CEV) patients such as haemodialysis (HD) patients. In this study, we have designed and utilised three ELISA tests to measure IgG levels against SARS-CoV-2 spike glycoprotein; identical except for the use of spike glycoprotein from the original Wuhan strain or from the B.1.617.2 (Delta) or B.1.1.529 (Omicron) VOC. We have used these ELISAs to determine cross reactivity of spike glycoprotein induced antibody against Delta and Omicron variants before and after third SARS-CoV-2 vaccine dose. These results show the value of the additional vaccination on enhancing IgG responses to the spike glycoprotein from both VOCs, even in HD and CEV patients. . CC-BY-ND 4.0 International license It is made available under a perpetuity. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted January 1, 2022. Secondly, individuals attending general practice for vaccination in Ulster who were considered Clinically Extremely Vulnerable (CEV) as part of The Pandemic Study, who received two doses of the AZ vaccine, followed by a third immunization of the PFZ vaccine. Lastly, individuals on haemodialysis under renal care at the University Hospitals Birmingham that had a mixture of two AZ vaccines or two PFZ vaccines followed by a PFZ booster (9) . Samples were taken at 6 months following their 2 nd vaccination of their primary course, prior to the 3 rd booster vaccination with PFZ and also 28 days In addition we ran serial dilutions at 1 in 2 (250 -7.8 IU/mL) of the WHO standard NIBSC 20/136 (13) . We also ran the therapeutic monoclonal antibody therapy Sotrovimab (Glaxo Smith Kline) in serial dilutions. A therapeutic dose of 500mg is given to patients at a concentration of which is 62.5 mg/ml. The COCO study was ethically approved for this work by the London -Camden and Kings Cross Research Ethics Committee on behalf of the United Kingdom Health Research Authority -reference . CC-BY-ND 4.0 International license It is made available under a perpetuity. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted January 1, 2022. All analyses were undertaken on GraphPad Prism 9 (San Diego, California). Normality testing was performed by Kolmogorov -Smirnov testing and data was non parametric. Groups were compared using a Kruskal-Wallis test with Dunn's multiple comparison test. Statistical significance was accepted when p is less than 0.05. The HD cohort had an average age of 66 years (range 55-75) with male gender predominance (61%). Of this cohort 70.3% had AZ as their primary course and the remaining 29.7% PFZ. There were 90 pre 3 rd dose samples tested and 85 samples from those receiving a third vaccination. For the COCO health care worker cohort that had 3 PFZ vaccine courses (PPP) the median age was 47 (range 25-64). Male gender accounted for 21% of this cohort. Ninety pre 3 rd dose samples were tested and 60 from individuals vaccinated a third time. The CEV cohort from general practice had a median age of 51 years (range 28-89). Forty four percent of the cohort were male gender. This cohort had two AZ vaccines as their primary course and then a third vaccination with PFZ (termed the AAP cohort). is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted January 1, 2022. We compared the antibody concentration, defined as median optical density (OD), between the PPP and AAP groups and found the median ODs were consistently higher in the AAP cohort for the Understanding whether antibodies induced to existing vaccines or strains of SARS-CoV-2 can bind and function against novel SARS-CoV-2 VOC and their enhancement or otherwise after additional immunisations, is of critical importance in guiding public health policy during the ongoing SARS-CoV-2 pandemic. Coupled with this is the necessity to understand how well these responses are sustained over time, whilst de novo vaccines are developed. This knowledge is of particular relevance for clinically vulnerable groups who are typically immunocompromised and either do not make robust antibody responses to vaccination or fail to retain antibody responses over time. Consistent with early data for healthy adult populations, studies already available show that sera from individuals vaccinated twice binds and neutralises omicron less well than sera from individuals immunized three times (14) (15) (16) . A distinct feature of the current study is the demonstration that in a . CC-BY-ND 4.0 International license It is made available under a perpetuity. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted January 1, 2022. ; https://doi.org/10.1101/2021.12.30.21268308 doi: medRxiv preprint cohort of haemodialysis (HD) patients that there is significant waning, 6 months post second dose, of vaccine-induced antibodies against the original vaccine-strain spike glycoprotein (Wuhan) and the Delta and Omicron VOC in comparison to healthy controls. This population of patients are at greater risk of severe SARS-CoV-2 infection, are difficult to shield from infection as they need to attend hospital routinely and also can show diminished protection from vaccination, despite being capable of maintaining their antibody responses to this virus (9, 17) . In HD patients, seropositivity ranged from 34.4% to 62.2% six-months after their initial two-dose immunisation schedule, consistently lower than healthy-controls regardless of whether their primary vaccination schedule employed the PFZ or the AZ vaccine. Encouragingly, following a third vaccination to HD and other CEV patients, antibody levels against all three spike proteins significantly increased and overall seroprevalence against each spike protein exceeded 97%. These data provide reassurance that booster immunisations with currently available vaccines induce relevant humoral immunogenicity against the highly transmissible Omicron VOC (18) . We have previously demonstrated that an extended dosing interval improved the immunogenicity of an initial two-dose Pfizer immunisation in healthy individuals (19) . We demonstrate that, in healthy controls, a heterologous vaccination strategy (i.e. AZ prime, followed by PFZ booster), induced antibody responses of greater magnitude against the Wuhan and Delta, but not the Omicron VOC. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted January 1, 2022. ; https://doi.org/10.1101/2021.12.30.21268308 doi: medRxiv preprint is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted January 1, 2022. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted January 1, 2022. Pre-2019 controls -health controls samples from pre-2019. Pre -pre 3 rd dose of vaccine and 6 months post 2 nd dose. Post -28 days post 3 rd dose of vaccine. PPP -3 Pfizer-BioNtech vaccines given in this cohort. AAP -two AstraZeneca ChAdOx1 nCoV-19 vaccines and the one Pfizer-BioNtech vaccine given in this cohort Safety and efficacy of the ChAdOx1 nCoV-19 vaccine (AZD1222) against SARS-CoV-2: an interim analysis of four randomised controlled trials in Brazil, South Africa, and the UK Safety and Efficacy of the BNT162b2 mRNA Covid-19 Vaccine Estimated transmissibility and impact of SARS-CoV-2 lineage B.1.1.7 in England. Science (80-) WHO World Health Organisation. Tracking SARS-CoV-2 variants Omicron (B.1.1.529) Variant. CDC COVID-19 Sci Briefs Hemodialysis patients make long-lived antibodies against sarscov-2 that may be associated with reduced reinfection Development of a high-sensitivity ELISA detecting IgG, IgA and IgM antibodies to the SARS-CoV-2 spike glycoprotein in serum and saliva Validation of a combined ELISA to detect IgG, IgA and IgM antibody responses to SARS-CoV-2 in mild or moderate non-hospitalised patients Site-specific glycan analysis of the SARS-CoV-2 spike WHO International Standard First WHO International Standard for anti-SARS-CoV-2 immunoglobulin (human) NIBSC code: 20/136 Instructions for use (Version 2.0, Dated 17/12/2020) Plasma neutralization properties of the SARS-CoV-2 Omicron variant mRNA-based COVID-19 vaccine boosters induce neutralizing immunity against SARS-CoV-2 Omicron variant Omicron has extensive but incomplete escape of Pfizer BNT162b2 elicited 10 The Electrostatic Potential of the Omicron Variant Spike is Higher than in Delta and Delta-plus Variants: a Hint to Higher Transmissibility? Safety and immunogenicity of seven COVID-19 vaccines as a third dose (booster) following two doses of ChAdOx1 nCov-19 or BNT162b2 in the UK (COV-BOOST): a blinded, multicentre, randomised, controlled, phase 2 trial Delayed production of neutralizing antibodies correlates with fatal COVID-19 We thank the staff and patients that have kindly volunteered for this study. Thanks also to Abingdon Health for the Wuhan and Delta antigen.