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\begin{document}

\title{Cross-reactive antibody responses to coronaviruses elicited by
SARS-CoV-2 infection or vaccination}



\author[1]{Richard SH Lee}%
\author[1]{Samuel Cheng}%
\author[1]{Jin  Zhao}%
\author[1]{Annie YS Tsoi}%
\author[1]{Kaman KM Lau}%
\author[1]{Coco HC Chan}%
\author[1]{John KC Li}%
\author[2]{David Hui}%
\author[1]{Malik Peiris}%
\author[1]{Hui-Ling Yen}%
\affil[1]{The University of Hong Kong School of Public Health}%
\affil[2]{The Chinese University of Hong Kong}%


\vspace{-1em}



  
  \date{May 22, 2024}


\begingroup
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\selectlanguage{english}
\begin{abstract}
Background The newly emerged SARS-CoV-2 possesses shared antigenic
epitopes with other human coronaviruses. We investigated if COVID-19
vaccination or SARS-CoV-2 infection may boost cross-reactive antibodies
to other human coronaviruses. Methods Pre- and post-vaccination sera
from SARS-CoV-2 na\selectlanguage{ngerman}ïve healthy subjects who received three doses of the
mRNA vaccine (BioNTech, BNT) or the inactivated vaccine (CoronaVac, CV)
were used to monitor the level of cross-reactive antibodies raised
against other human coronaviruses by enzyme-linked immunosorbent assay.
In comparison, convalescent sera from COVID-19 patients with or without
prior vaccination history were also tested. Pseudoparticle
neutralization assay was performed to detect neutralization antibody
against MERS-CoV. Results Among SARS-CoV-2 infection naïve subjects, BNT
or CV significantly increased the anti-S2 antibodies against
Betacoronaviruses (OC43 and MERS-CoV) but not Alphacoronaviruses (229E).
The pre-vaccination antibody response to the common cold human
coronaviruses did not negatively impact the post-vaccination antibody
response to SARS-CoV-2. Cross-reactive antibodies that binds to the S2
protein of MERS-CoV were similarly detected from the convalescent sera
of COVID-19 patients with or without vaccination history. However, these
anti-S2 antibodies do not possess neutralizing activity in MERS-CoV
pseudoparticle neutralisation tests. Conclusions Our results suggest
that SARS-CoV-2 infection or vaccination may potentially modulate
population immune landscape against previously exposed or novel human
coronaviruses. The findings have implications for future
sero-epidemiological studies on MERS-CoV.%
\end{abstract}\selectlanguage{ngerman}%



\sloppy


Original Article

\textbf{Cross-reactive antibody responses to coronaviruses elicited by
SARS-CoV-2 infection or vaccination}

Richard SH Lee\textsuperscript{1}, Samuel MS Cheng\textsuperscript{1},
Jin Zhao\textsuperscript{1}, Annie YS Tsoi\textsuperscript{1}, Kaman KM
Lau\textsuperscript{1}, CoCo HC Chan\textsuperscript{1}, John KC
Li\textsuperscript{1}, David SC Hui\textsuperscript{2}, Malik
Peiris\textsuperscript{1,3*}, Hui-Ling Yen\textsuperscript{1*}

\textsuperscript{1}School of Public Health, LKS Faculty of Medicine, The
University of Hong Kong

\textsuperscript{2}Department of Medicine and Therapeutics, CUHK, Prince
of Wales Hospital, HKSAR, China

\textsuperscript{3}Centre for Immunology \& Infection, Hong Kong Science
Park, Hong Kong, China

Running title: SARS-CoV-2 and cross-reactive antibodies

Key words: SARS-CoV-2, human coronaviruses, cross-reactive antibody,
COVID-19 vaccines, convalescent sera

Word count: 214 words (abstract), 2211 words (text); Figures: 3;
References: 22.

Notes:

\begin{enumerate}
\tightlist
\item
  The authors declare no conflict of interest.
\item
  Data is available upon request from the corresponding author.
\item
  This study was supported by the HKU URC Seed Funding for Strategic
  Interdisciplinary Research Scheme 2021/22, Contract (U01AI151810) of
  NIAID, National Institutes of Health, USA, and the Theme-based
  Research Scheme (\#T11-705/21-N) of the Research Grants Council of the
  Hong Kong SAR Government, China.
\item
  The study was approved by the institutional review board of the Hong
  Kong West Cluster of the Hospital Authority of Hong Kong (Reference
  No.: UW20-169) and the Joint Chinese University of Hong Kong-New
  Territories East Cluster Clinical Research Ethics Committee (Reference
  No.: 2020.229).
\item
  Correspondence: Hui-Ling Yen (hyen@hku.hk) and Malik Peiris
  (malik@hku.hk).
\end{enumerate}

\textbf{Background}

The newly emerged SARS-CoV-2 possesses shared antigenic epitopes with
other human coronaviruses. We investigated if COVID-19 vaccination or
SARS-CoV-2 infection may boost cross-reactive antibodies to other human
coronaviruses.

\textbf{Methods}

Pre- and post-vaccination sera from SARS-CoV-2 na\selectlanguage{ngerman}ïve healthy subjects
who received three doses of the mRNA vaccine (BioNTech, BNT) or the
inactivated vaccine (CoronaVac, CV) were used to monitor the level of
cross-reactive antibodies raised against other human coronaviruses by
enzyme-linked immunosorbent assay. In comparison, convalescent sera from
COVID-19 patients with or without prior vaccination history were also
tested. Pseudoparticle neutralization assay was performed to detect
neutralization antibody against MERS-CoV.

\textbf{Results}

Among SARS-CoV-2 infection naïve subjects, BNT or CV significantly
increased the anti-S2 antibodies against Betacoronaviruses (OC43 and
MERS-CoV) but not Alphacoronaviruses (229E). The pre-vaccination
antibody response to the common cold human coronaviruses did not
negatively impact the post-vaccination antibody response to SARS-CoV-2.
Cross-reactive antibodies that binds to the S2 protein of MERS-CoV were
similarly detected from the convalescent sera of COVID-19 patients with
or without vaccination history. However, these anti-S2 antibodies do not
possess neutralizing activity in MERS-CoV pseudoparticle neutralisation
tests.

\textbf{Conclusions}

Our results suggest that SARS-CoV-2 infection or vaccination may
potentially modulate population immune landscape against previously
exposed or novel human coronaviruses. The findings have implications for
future sero-epidemiological studies on MERS-CoV.

\textbf{INTRODUCTION}

SARS-CoV-2 is a newly emerged human coronavirus (HCoV) that has rapidly
swept through the globe and resulted in significant public health and
socioeconomic loss. As a member of the genus Betacoronavirus, SARS-CoV-2
possesses shared epitopes with other HCoVs including the common cold
Alphacoronaviruses (229E, NL63) and Betacoronaviruses (OC43, HKU1), as
well as two newly emerged Betacoronaviruses, SARS-CoV-1 in 2002 and
MERS-CoV in 2012 (1, 2). Prior exposures to common cold HCoVs may
provide cross-protective humoral or cell-mediated immunity. However,
previous studies showed conflicting results on whether pre-existing
antibodies towards common cold HCoVs provide cross-protection against
SARS-CoV-2 infection or severe disease outcomes (3, 4, 5, 6). On the
other hand, pre-existing memory T cells that are likely elicited in
response to common cold CoVs have been consistently associated with
cross-protection among SARS-CoV-2-exposed healthcare workers or
household contacts (7, 8).

Since 2020, 7.7 billion SARS-CoV-2 infected and re-infected cases have
been reported to WHO and 13.5 billion doses of COVID-19 vaccines have
been administered globally. The extensive exposure to SARS-CoV-2 through
infection and immunization may substantially affect the population
immune landscape and susceptibility to other HCoVs. The pre-existing
immunity to common cold HCoVs may potentially result in back-boosting of
antibodies against the conserved S2 epitopes upon SARS-CoV-2 infection
or vaccination as reported previously (9, 10, 11, 12, 13). In addition,
de novo antibody response against SARS-CoV-2 may also cross-react with
other HCoVs (10, 14). A better understanding on the cross-reactive
antibody responses may provide guidance on the development of
pan-coronavirus vaccines.

While the spike-encoding mRNA and the whole-virion inactivated vaccines
have been the main COVID-19 vaccines administered globally to date, most
studies so far have focused on the effect of mRNA vaccines. The mRNA
vaccines adopted the pre-fusion conformation of the spike protein while
the pre-fusion and post-fusion conformations have been reported for the
inactivated vaccines (15). Inactivated vaccines also contain additional
SARS-CoV-2 structural proteins that may stimulate humoral or
cell-mediated immunity. Using pre- and post- vaccination sera, we
firstly compared the cross-reactive antibody responses against different
HCoVs elicited by mRNA and inactivated COVID-19 vaccines in individuals
previously infection-naïve for SARS-CoV-2. We also separately
investigated cross-reactive antibody response in convalescent sera of
SARS-CoV-2 patients with or without prior vaccination history.

\textbf{METHODS}

\textbf{Study design} . Sera were collected from healthy subjects
enrolled in a longitudinal study for monitoring population immunity to
SARS-CoV-2 infection and vaccination in Hong Kong. The study was
approved by the institutional review board of the Hong Kong West Cluster
of the Hospital Authority of Hong Kong (Reference No.: UW20-169) and the
Joint Chinese University of Hong Kong-New Territories East Cluster
Clinical Research Ethics Committee (Reference No.: 2020.229). Enrolled
participants were followed up every 6 months for sera collection and
self-reported SARS-CoV-2 infections that have been RT-PCR confirmed.
Pre- and post-vaccination sera were collected from age-matched (27-73
years old) individuals who received 3-doses of BioNTech (BNT) (n=20) or
CoronaVac (CV) (n = 21) in 2021-2022. The pre-vaccination sera were
collected on the day of receiving the first dose of the BNT or CV
vaccine, and the post-vaccination sera were collected 4-8 week after
receiving the third dose of the vaccine. The convalescent sera from
SARS-CoV-2 infected subjects without (n=20) or with vaccination history
(n=20 each for BNT and CV) were collected from study participants after
self-reported SARS-CoV-2 infections. The archived pre-pandemic sera in
2019 from healthy blood donors (n=20) were used as controls.

\textbf{Enzyme-linked immunosorbent assay (ELISA).} MaxiSorp 96-well
plates (Thermo Fisher) were coated with 0.1 \selectlanguage{greek}µ\selectlanguage{english}g recombinant spike {[}S1,
S2, or full-length S (S1+S2), as indicated{]} or nucleocapsid proteins
of OC43, 229E, SARS-CoV-2, or MERS-CoV (Sino Biological) per well
overnight at 4\selectlanguage{ngerman}°C. The plates were washed with PBST (PBS containing
0.05\% Tween 20) and blocked with blocking buffer (5\% non-fat milk in
PBST) for 2 hours. Human sera were heat treated at 56°C for 30 minutes
and were serially 3-fold diluted from 1:100 to 1:2700 with blocking
buffer. Diluted sera (100 \selectlanguage{greek}µ\selectlanguage{english}L/ well) were added in duplicate to the plate
and incubated for 1 hour, followed by detection using 1:10000 diluted
HRP-conjugated goat anti-human IgG secondary antibody (100 \selectlanguage{greek}µ\selectlanguage{english}L/ well).
TMB substrate (100 \selectlanguage{greek}µ\selectlanguage{english}L/ well) (Thermo Scientific) was added to the plate
for colorimetric signal formation for 10 minutes and stopped by adding
50 \selectlanguage{greek}µ\selectlanguage{english}l/well of 2M sulphuric acid. Plates were read at wavelength of 450
nm for absorbance (OD 450nm). In each ELISA plate, the mean OD 450nm
from wells without human sera (n=8 per plate) was calculated as the
background. The area under the curve (AUC) were calculated for each
serially diluted sera after subtracting the background.

\textbf{MERS-CoV~spike pseudoparticle neutralisation
tests~(ppNT).}Luciferase expressing HlV/MERS-RBD pseudoparticles (5 ng
of p24) were pre-incubated with serially diluted sera at 4\selectlanguage{ngerman}°C for 30
minutes and added to Vero E6 cells in triplicates. Infection was
determined by quantifying the firefly luciferase activity at 2 days
post-infection (Promega Corporation,) using the Microbeta luminometer
(PerkinElmer). The highest serum dilution that gave \selectlanguage{english}[?]90\% reduction of
the maximal luciferase activity (eg. in the absence of antibody) was
regarded as the ppNT antibody titre (16).

\textbf{Statistical analysis} . The difference of grouped AUC of pre-
and post-vaccination against each antigen within vaccination group was
analysed with the Wilcoxon test. The individual AUC difference between
pre- and post-vaccination of the same individual was calculated and
compared with the AUC difference between two vaccination groups with the
Mann-Whitney~test. Correlation between AUC ratio of samples against
SARS-CoV-2 versus other Human Coronaviruses were analysed using
Spearman's rank correlation. The statistical significance of all
statistical tests was set at p \textless{} 0.05.

\textbf{RESULTS}

\textbf{COVID-19 vaccination increased cross-reactive antibody responses
against other Betacoronaviruses.} We first compared the pre- and post-
COVID-19 vaccination antibody responses towards various HCoVs among
individuals who have not been previously infected with SARS-CoV-2. Both
BNT (Figure 1A) and CV (Figure 1B) vaccinees showed significant increase
of antibody AUC against the spike protein of Betacoronaviruses after
vaccination, including SARS-CoV-2 (S1+S2), MERS-CoV (S2), OC43 (S2). In
contrast, the post-vaccination sera showed a modest, but significant
decrease in median antibody AUC against Alphacoronavirus 229E (S2),
possible suggesting a waning antibody response over time. Only CV
vaccinees showed significantly increased antibody AUC against the
nucleocapsid (N) protein of SARS-CoV-2, MERS-CoV and 229E (Figure 1B).
Comparing the BNT and CV vaccination responses (Figure 1C), BNT induced
significantly greater S-binding antibody for SARS-CoV-2 (S1+S2) and
MERS-CoV (S2), while CV induced significantly greater N-binding
antibodies for SARS-CoV-2, MERS-CoV, OC43, and 229E.

Pre-vaccination sera showed high baseline binding antibodies for OC43
and 229E (Figure 1A \& 1B). As the baseline antibody titres towards
common cold HCoVs may affect the post-exposure antibody response against
SARS-CoV-2 (5), we analysed the correlation between the pre-vaccination
antibody levels against other HCoVs versus the post-vaccination antibody
response against SARS-CoV-2. For BNT vaccinees, their post-vaccination
antibody AUC against S of SARS-CoV-2 was not affected by the
pre-vaccination antibody against OC43 or 229E (Figure 2A). For CV
vaccinees, their post-vaccination antibody AUC against S of SARS-CoV-2
was marginally correlated with the pre-vaccination antibody against
229E-S1+S2 (Spearman's \selectlanguage{greek}ρ\selectlanguage{english}=0.46, p\textless{}0.05) and 229E-S2 (Spearman's
\selectlanguage{greek}ρ\selectlanguage{english}=0.44, p\textless{}0.05). The post-vaccination antibody AUC against the
N protein of SARS-CoV-2 did not correlate with the pre-vaccination
antibody responses against the N proteins of OC43 or 229E. These results
suggest that the de novo antibody response against SARS-CoV-2 after
vaccination was not negatively affected by the baseline antibody
responses against previously exposed HCoV.

The BNT vaccinees' post-vaccination antibody AUC against S of SARS-CoV-2
was positively correlated with the post-vaccination anti-MERS-CoV-S2
(Spearman's \selectlanguage{greek}ρ\selectlanguage{english}=0.63, p\textless{}0.01) and anti-OC43-S1+S2 (Spearman's
\selectlanguage{greek}ρ\selectlanguage{english}=0.57, p\textless{}0.01) responses (Figure 2B). For CV vaccinees, the
post-vaccination antibody AUC against S of SARS-CoV-2 was positively
correlated with anti-OC43-S1+S2 response (Spearman's \selectlanguage{greek}ρ\selectlanguage{english}=0.47,
p\textless{}0.05), and their post-vaccination antibody AUC against N of
SARS-CoV-2 was positively correlated with anti-MERS-N response
(Spearman's \selectlanguage{greek}ρ\selectlanguage{english}=0.73, p\textless{}0.001) (Figure 2B). Taken together, both
the mRNA or inactivated COVID-19 vaccines may boost antibody responses
against conserved epitopes shared with previously exposed (OC43) or
novel (MERS-CoV) Betacoronaviruses.

\textbf{SARS-CoV-2 infection increased cross-reactive antibody responses
against MERS-CoV.} We further investigated if SARS-CoV-2 infection may
similarly boost cross-reactive antibody response against MERS-CoV S and
N proteins (Figure 3A). Convalescent sera from SARS-CoV-2 patients
without a history of COVID-19 vaccination (n=20), those who have been
infected and vaccinated (n=20), and those who were vaccinated and had a
breakthrough infection (n=20) were compared with pre-pandemic sera
collected from healthy adults in 2019 (n=20). Compared to the
pre-pandemic sera (using the mean+3SD AUC value as threshold), 41 out of
60 (71.7\%) convalescent sera showed increased antibody against MERS-CoV
S2, while 6 of 60 (10\%) showed increased antibody against MERS-CoV S1.
In regard to the antibody response to the S2 protein of MERS-CoV-2,
those who were vaccinated followed by a breakthrough infection generally
showed higher AUC than those who were infected without vaccination
history, although the differences were not significant. In addition,
those who were infected followed by BNT vaccination showed greater
anti-S2 antibody response than those who were infected followed by CV
vaccination, suggesting that BNT vaccination may better expand the
breath of antibody response than CV vaccination among those who were
infected. Compared to the pre-pandemic sera, increase in the anti-N
protein antibody AUC was also observed from those who were infected
followed by CV vaccination or those who were CV vaccinated followed by
breakthrough infection.

\textbf{Cross-reactive antibody against MERS-CoV S2 were
non-neutralizing.} Pseudoparticle neutralization test (ppNT) was used to
evaluate if the cross-reactive anti-S2 antibodies possess neutralizing
activity against MERS-CoV (Figure 3B). None, except one subject who was
vaccinated with BNT followed by SARS-CoV-2 infection, showed
neutralizing antibody at 1:10 dilution using ppNT assay. Despite of
non-neutralizing, sera from subjects with CV vaccination followed by
infection (mean \selectlanguage{ngerman}± SD \% inhibition = 44.6 ± 27.8) showed greater
inhibition against MERS-CoV than the sera of pre-pandemic controls (15.3
± 16.9) (Kruskal-Wallis test, p=0.0435). Taken together, these results
suggest that the majority of the cross-reactive anti-MERS-CoV-S2
antibodies detected after vaccination or infection were
non-neutralizing.

\textbf{DISCUSSION}

Majority of the global population have been exposed to SARS-CoV-2
through infection or vaccination to date. As SARS-CoV-2 share common
epitopes with other HCoVs, it is anticipated that SARS-CoV-2 exposure
may boost cross-reactive antibodies towards other HCoV. Using the pre-
and post- vaccination sera of un-infected subjects, we show that both
the mRNA vaccine (BNT) and the inactivated vaccine (CV) increased
cross-reactive antibodies against the S2 protein of the two
Betacoronaviruses, OC43 and MERS-CoV, but not Alphacoronavirus 229E. CV
vaccination further boosted anti-N protein antibodies against MERS-CoV
and 229E. The antibody response against S2 protein of MERS-CoV were also
detected from 41 out of 60 (71.7\%) convalescent sera of SARS-CoV-2
patients with or without COVID-19 vaccination history. Our results are
in line with a recent study that detected high prevalence of
cross-reactive antibodies to spike proteins of viruses in
the\emph{Orthocoronavirinae} among the post-COVID-19 population (17).
Taken together, these results suggest that SARS-CoV-2 exposure may
modulate population antibody response towards other human coronaviruses.

The high level pre-existing antibodies against OC43 and 229E generated
from prior infection or vaccination have been shown to impact on the de
novo humoral responses against SARS-CoV-2 (5). Among our study subjects,
no negative impact was observed in the correlation analysis between
pre-vaccination antibody levels against OC43 or 229E and the
post-vaccination antibody levels against SARS-CoV-2. Due to various
public health and social measures implemented in Hong Kong during
COVID-19 pandemic, the activity of various respiratory viruses have been
reduced, which may limit recent exposure of our study population to
common cold HCoVs. As co-circulation of SARS-CoV-2 and other HCoVs is
anticipated, follow up studies are needed to understand how pre-existing
immunity may shape the antibody landscapes of various HCoVs.

Both COVID-19 vaccines back-boosted antibodies against the S2 domain of
OC43, which aligned with the results reported from previous studies (9,
10, 11, 12, 13). Furthermore, the de novo antibody response to
SARS-CoV-2 generated after vaccination or infection were cross-reactive
with S2 of MERS-CoV. The conserved region on S2 stem-helix domains
across Betacoronaviruses may explain the high cross reactivity of
anti-S2 antibodies (18), while other studies identified cross-reactive
neutralizing antibody that targets S2 region (19, 20, 21, 22). By
comparing the anti-MERS-CoV antibodies from infected subjects with or
without vaccination history, we noted that those who have been
vaccinated, followed by a SARS-CoV-2 breakthrough infection generally
showed higher anti-MERS S2 AUC than those who were infected without
vaccination history. These study subjects were vaccinated with the
prototype virus followed by infection with the BA.2 Omicron variant in
2022. In addition, higher anti-MERS S2 AUC was detected from those
infected followed by BNT vaccination compared to those who were infected
without vaccination or those infected followed by CV vaccination. These
results suggest that BNT vaccine may stimulate broader antibody response
than CV among those who were previously infected. Taken together, these
findings have implications for future sero-epidemiological studies on
MERS-CoV. While binding antibody responses to MERS-CoV S1 is still
likely to be specific for MERS-CoV infection, binding antibody to
MERS-CoV S2 should no longer be considered as a specific marker for
MERS-CoV infection.

Using the ppNT assay that specifically detect neutralizing antibodies
targeting the receptor binding domain (RBD) of MERS-CoV, we showed that
the anti-MERS-CoV antibodies were non-neutralizing. However, other
neutralising mechanisms such as inhibition of fusion peptide cannot be
ruled out (19). Further studies are needed to evaluate if these
cross-reactive antibodies possess Fc-mediated effector functions and if
they confer protection in vivo.

\textbf{ACKNOWLEDGMENTS}

This study was supported by the HKU URC Seed Funding for Strategic
Interdisciplinary Research Scheme 2021/22, Contract (U01AI151810) of
NIAID, National Institutes of Health, USA, and the Theme-based Research
Scheme (\#T11-705/21-N) of the Research Grants Council of the Hong Kong
SAR Government, China.

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\textbf{FIGURE LEGENDS}

\textbf{Figure 1. COVID-19 vaccination increased cross-reactive antibody
responses against other Betacoronaviruses.} (A) Pre- and post-
vaccination antibody responses of BNT vaccinees (n=20) to the S and N
proteins of different human coronaviruses, Wilcoxon test was used to
compare the pre and post-vaccination antibody response. (B) Pre- and
post- vaccination antibody responses of CV vaccinees (n=21) to the S and
N proteins of different human coronaviruses, Wilcoxon test was used to
compare the pre and post-vaccination antibody response. (C) Differences
between BNT and CV vaccinees in the post-vaccination antibody responses
against the S and N proteins of different human coronaviruses,
Mann-Whitney~test was used to compare antibody responses of BNT and CV
vaccinees.

\textbf{Figure 2. Correlation between post-vaccination antibody response
against SARS-CoV-2 versus antibody response against other human
coronaviruses.} (A) Post-vaccination antibody response against
SARS-CoV-2 versus pre-vaccination antibody responses against other
HCoVs. (B) Post-vaccination antibody response against SARS-CoV-2 versus
post- vaccination antibody responses against other HCoVs. The Spearman
coefficient were shown, * p\textless{}0.05, ** p\textless{}0.01.

\textbf{Figure 3. SARS-CoV-2 infection increased cross-reactive antibody
responses against MERS-CoV.} (A) Antibody responses against MERS-CoV S1,
S2, or N proteins detected from sera of healthy donors collected in 2019
prior to the COVID-19 pandemic (control sera, n=20), subjects infected
with SARS-CoV-2 without vaccination history (infected, n=20), subjects
infected with SARS-CoV-2 followed by 2-doses CV vaccination (infected +
CV, n=10), subjects infected with SARS-CoV-2 followed by 2-doses BNT
vaccination (infected + BNT, n=10), subjects vaccinated with 2-doses of
CV followed by SARS-CoV-2 infection (CV + infected, n=10), subjects
vaccinated with 2-doses of BNT followed by SARS-CoV-2 infection (BNT +
infected, n=10). The threshold AUC values (mean+3SD) determined from the
pre-pandemic sera were shown in dotted lines. (B) Neutralizing antibody
response against MERS-CoV were determined using the ppNT assay. In
addition to the pre-pandemic sera and convalescent sera tested above, we
also determined neutralizing antibody responses against MERS-CoV among
those who have been vaccinated with 3-doses of BNT (n=20) or CV (n=20)
without a history of SARS-CoV-2 infection. Sera were diluted at 1:10
dilution and any sample with \selectlanguage{english}[?] 90\% inhibition was considered positive.



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