Association study of HLA with the kinetics of SARS-CoV-2 spike specific IgG antibody responses to BNT162b2 mRNA vaccine

BNT162b2, an mRNA-based SARS-CoV-2 vaccine (Pfizer-BioNTech), is one of the most effective COVID-19 vaccines and has been approved by more than 130 countries worldwide. However, several studies have reported that the COVID-19 vaccine shows high interpersonal variability in terms of humoral and cellular responses, such as those with respect to SARS-CoV-2 spike protein immunoglobulin (Ig)G, IgA, IgM, neutralizing antibodies, and CD4+ & CD8+ T cells. The objective of this study is to investigate the kinetic changes in anti-SARS-CoV-2 spike IgG (IgG-S) profiles and adverse reactions and their associations with HLA profiles among 100 hospital workers from the Center Hospital of the National Center for Global Health and Medicine (NCGM), Tokyo, Japan. DQA1*03:03:01 (P = 0.017; Odd ratio (OR) 2.80, 95%Confidence interval (CI) 1.05-7.25) was significantly associated with higher IgG-S production after two doses of BNT162b2 while DQB1*06:01:01:01 (P = 0.028, OR 0.27, 95%CI 0.05-0.94) was significantly associated with IgG-S declines after two doses of BNT162b2. No HLA alleles were significantly associated with either local symptoms or fever. However, C*12:02:02 (P = 0.058; OR 0.42, 95%CI 0.15-1.16), B*52:01:01 (P = 0.031; OR 0.38, 95%CI 0.14-1.03), DQA1*03:02:01 (P = 0.028; OR 0.39, 95%CI 0.15-1.00) and DPB1*02:01:02 (P = 0.024; OR 0.45, 95%CI 0.21-0.97) appeared significantly associated with protection against systemic symptoms after two doses of BNT162b2 vaccination. Further studies with larger sample sizes are clearly warranted to determine HLA allele associations with the production and long-term sustainability of IgG-S after COVID-19 vaccination.


Introduction
The concept of messenger RNA (mRNA) vaccination stems from 1987, when Robert 2 Malone confirmed that human cells can absorb cationic liposomes containing mRNA and can 3 create proteins from those mRNA sequences (1). However, over the years, academic 4 laboratories and companies working on mRNA had come to the consensus that mRNA is too 5 prone to degradation to be used effectively as a drug or a vaccine (2). From the end of the 20 th 6 century, research into mRNA vaccines mainly focused on influenza disease (3) and cancer 7 (4), with testing in animal models yielding satisfactory results. Such results inspired CureVac, 8 BioNTech, and Moderna to focus on transforming mRNA into a drug platform (5)(6)(7). In 2005, 9 the Karikó and Weissman team performed a landmark experiment in which an mRNA 10 vaccine was used to successfully suppress RNA recognition by Toll-like receptors (TLRs) 11 using modified pseudouridine, a uridine analog (8). All this previous work contributed to the 12 rapid development of coronavirus disease 2019 (COVID-19) mRNA vaccines within days of 13 the genome for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) being 14 released online (9). 15 BNT162b2, an mRNA-based SARS-CoV-2 vaccine (Pfizer-BioNTech), is one of the 16 most effective COVID-19 vaccines (10) and has been approved by more than 130 countries 17 worldwide. However, several studies have reported that the COVID-19 vaccine shows high 18 interpersonal variability in terms of humoral and cellular responses, such as those with 19 respect to anti-SARS-CoV-2 spike immunoglobulin (Ig)G (11-17), IgA (15, 16), IgM (16), 2 The National Center for Global Health and Medicine (NCGM) is a Japanese 3 government-designated medical center for the treatment of COVID-19 patients. We recruited 4 and monitored a total of 100 hospital workers 20 years of age from the period between 5 March and June 2021. Details of the study design are explained in previous studies(26, 27). 6 All participants were vaccinated with two doses of the BNT162b2 mRNA-based SARS-CoV- 7 2 vaccine (Pfizer-BioNTech) according to the standard protocol (two doses of 30 µg, 8 administered 3 weeks apart) (18,28). All participants received the first dose of the 9 BNT162b2 vaccine in March 2021 and the second dose was administered 21 days after the 10 first dose. Blood was drawn from volunteers on day 1 (immediately after the first dose).

11
Samples were then collected on day 15, day 29 (7 days after the second dose) and day 61.  Self-reported questionnaire 18 Participants were requested to complete a questionnaire regarding the adverse  Next-generation sequencing (NGS)-based HLA genotyping 5 NGS HLA genotyping was performed using AllType™ NGS Assays (One Lambda, 6 West Hills, CA) on an Ion GeneStudio S5 sequencing system (Thermo Fisher Scientific, 7 Waltham, MA). HLA-A, -C, -B, DRB1, DQA1, DQB1, DPA1, DPB1 targeted gene 8 amplification, HLA library preparation, HLA template preparation, and HLA library loading 9 onto an ion 530v1 chip in an Ion Chef library preparation robot (Thermo Fisher Scientific) 10 and final sequencing in an Ion GeneStudio S5 sequencer (Thermo Fisher Scientific) were 11 performed following the instructions from the vendor.

6
Basic characteristics of participants 7 A total of 100 hospital workers at the NCGM were recruited, comprising 32 men and 8 68 women. Participants were further classified into 5 age strata to observe the changes of 9 IgG-S in relation to age: 20-29 years old; 30-39 years old; 40-49 years old; 50-59 years old; 10 and 60 years old ( Table 1). The BMI of participants ranged from 17 kg/m 2 to 38 kg/m 2 , 11 with the highest BMI observed in the group 60-years old (mean standard deviation, 26    Figure 2c) were apparent for participants. However, although it is 36 not statistically significant, strong decliners (∆ 20,000 AU/mL) were observed to be mainly 37 All rights reserved. No reuse allowed without permission. perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in The copyright holder for this this version posted February 2, 2022.
concentrated among the female group (n=15, 23%), compared with the male group (n=3, 1 9.4%). 2 3 IgG-N 4 None of the participants were seropositive for IgG-N across the five survey time   9 We classified the self-reported body temperature into 4 categories: T1, < 37.5 C; T2,  Table 3).  (Table 4).  Table 5). Due to the limited sample size, none of the 32 abovementioned HLA alleles remained significant associated with IgG-S levels after multiple 33 correction, but a potential correlation between individual HLA alleles and responsiveness to 34 BNT162b2 vaccination was still suggested. perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in  perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in Research into the associations between HLA alleles and IgG-S antibody levels after 5 vaccination remain limited, but a small Italian cohort (n=56) found no associations between 6 HLA alleles and IgG-S levels after BNT162b2 vaccination. In the present study, we identified 7 several HLA alleles showing potential associations with the kinetics of IgG-S antibody levels. 8 Further studies with larger sample sizes are clearly warranted to determine HLA allele 9 associations with the production and long-term sustainability of IgG-S antibody levels after 10 COVID-19 vaccination. perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in    perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in   perpetuity. preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in

Otani J, Ohta R, Sano C. Association between Immunoglobulin G Levels and Adverse Effects
Following Vaccination with the BNT162b2 Vaccine among Japanese Healthcare Workers. Vaccines (Basel). 2021;9(10).
All rights reserved. No reuse allowed without permission. perpetuity.
preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in The copyright holder for this this version posted February 2, 2022. ;