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\begin{document}

\title{COVID-19 vaccination with BNT162b2 and ChAdOx1 vaccines induces nasal
neutralizing antibodies}



\author[1]{Philippe Gevaert}%
\author[2]{Declercq Jozefien}%
\author[3]{Tobback Els}%
\author[2]{Stijn Vanhee}%
\author[1]{Natalie Deruyck}%
\author[4]{Gerlo Sarah}%
\author[4]{Vandekerckhove Linos}%
\affil[1]{Universiteit Gent}%
\affil[2]{Vlaams Instituut voor Biotechnologie}%
\affil[3]{Universitair Ziekenhuis Gent}%
\affil[4]{Universiteit Gent Vakgroep Inwendige Ziekten}%


\vspace{-1em}



  
  \date{June 17, 2024}


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\textbf{COVID-19 vaccination with BNT162b2 and ChAdOx1 vaccines induces
nasal neutralizing antibodies}

Declercq Jozefien\textsuperscript{(1,2,5)*}, Tobback
Els\textsuperscript{(3)*}, Vanhee Stijn\textsuperscript{(2,5)}, Natalie
De Ruyck\textsuperscript{(1)}, Gerlo Sarah\textsuperscript{(4)}, Gevaert
Philippe\textsuperscript{(1)**}, Vandekerckhove
Linos\textsuperscript{(3,4)**}

\textsuperscript{*}shared co-first authorship

** shared last authorship

\begin{enumerate}
\tightlist
\item
  Upper Airways Research Lab URL, Department of Otorhinolaryngology,
  Ghent University, Ghent, Belgium
\item
  Laboratory of immunoregulation, VIB Center for Inflammation Research,
  Ghent, Belgium
\item
  Department of General Internal Medicine, Ghent University Hospital,
  Ghent, Belgium
\item
  HIV Cure Research Centre, Department of Internal Medicine and
  Pediatrics, Ghent University, Ghent, Belgium
\item
  Department of Internal Medicine and Pediatrics, Ghent University,
  Ghent, Belgium
\end{enumerate}

Corresponding author:

Prof. Dr. Philippe Gevaert

Upper Airways Research Lab URL, Department of Otorhinolaryngology, Ghent
University,

C Heymanslaan 10, 1P1

Ghent, Belgium

+3293324922

Philippe.gevaert@ugent.be

Financial support: no funding

Word count: 591

\textbf{To the editor,}

to date, several vaccines against severe acute respiratory coronavirus 2
(SARS-CoV-2) have proven to effectively reduce severe illness (1,2). To
promote herd immunity and reduce virus circulation, vaccines need to
effectively reduce transmission risk. The nasal cavity is the first
entrance point for SARS-CoV-2 and it has been suggested that viral
replication is most efficient in the upper airways (3). Local
neutralizing antibodies (NAbs) in the nasal mucosa can play an important
role in preventing SARS-CoV-2 infection and transmission by limiting
viral replication and shedding. While induction of systemic neutralizing
humoral responses has been shown for both natural infection and upon
vaccination with BNT162b2 and ChAdOx1, the presence of NAbs in the nasal
mucosa upon vaccination remains unclear. A recent not peer reviewed
report described the potential of BNT162b2 to induce NAbs in the nasal
cavity, but did not consider prior COVID-19 as a potentiator of
vaccine-induced local immune responses (4). Local humoral responses
after vaccination with viral vector-based vaccines, another type of
frequently used SARS-CoV-2 vaccines, has not been investigated. As the
induction of nasal NAbs might be a key marker of vaccine efficacy, we
set out to determine whether SARS-CoV-2 mRNA (BNT162b2) and viral vector
based (ChAdOx1) vaccines can induce local neutralizing responses in the
nasal cavity.

To address this question, we collected serum and nasal secretions from
subjects visiting the COVID-19 vaccination center at the University
Hospital Ghent, Belgium. Sampling took place just before the first
SARS-CoV-2 vaccination, with either BNT162b2 or ChAdOx1, and at 13-40
days after the second dose with the same vaccine. All patients provided
written informed consent. Collection of nasal secretions was done as
described previously (5). In all samples, SARS-CoV-2 NAbs were
determined by using the Elabscience SARS-CoV-2 Neutralization Antibody
ELISA Kit. NAbs levels expressed as inhibition rates were determined,
with an inhibition [?]20\% considered positive and \textless{}20\%
considered negative. Forty-six subjects, mainly females, were included
in the study (Table 1). Half of these subjects had a history of prior
COVID-19. Prior to vaccination, 16 subjects had NAbs in serum and 4 in
nasal secretions. At second sampling, except for one, all subjects
showed NAbs in their serum, regardless of the vaccine received (Figure
1A+B). In nasal secretions, NAbs were observed in the majority of
subjects (n=23; 96\%) vaccinated with BNT162b2 and in about half of the
subjects (n=13; 59\%) vaccinated with ChAdOx1 at second sampling
(p=0.0032; Fisher's exact test) (Figure 1C). Moreover, the ACE2 binding
inhibition in nasal secretion was higher in subjects vaccinated with
BNT162b2 compared to those vaccinated with ChAdOx1 (p\textless{}0.0001;
2-way repeated-measures ANOVA with Sidak's multiple comparisons test)
(Figure 1D). Induction of NAbs occurred irrespective of prior SARS-CoV-2
infection or the presence of patient reported allergy to aeroallergens
(data not shown).

Taken together, our study shows that both BNT162b2 and ChAdOx1 vaccines
can effectively induce nasal NAbs, albeit variable in the ChAdOx1 arm.
Why only some subjects develop local NAbs after vaccination in the
former group is currently unclear and warrants follow up to fully
understand the underlying immunological mechanisms. As it is unclear
whether other local immunological mechanisms might induce protection
against infection and viral shedding, induction of mucosal NAbs might be
of key importance. Longitudinal follow-up of the described subjects is
needed to see whether vaccines can induce long lasting neutralizing
responses in the nasal mucosa. Failure to induce long-lasting NAbs
warrants rational booster design or other strategies, such as nasal
vaccination. Based on our findings, we advocate for the inclusion of
nasal mucosa NAbs measurements in vaccine efficacy trials and routine
testing procedures.\selectlanguage{english}
\begin{longtable}[]{@{}lll@{}}
\toprule
& \textbf{BNT162b2} & \textbf{ChAdOx1}\tabularnewline
& No (\%) & No (\%)\tabularnewline
\textbf{n=} & 24 & 22\tabularnewline
\textbf{Sex} & &\tabularnewline
Female & 18 (75\%) & 18 (82\%)\tabularnewline
Male & 6 (25\%) & 4 (18\%)\tabularnewline
\textbf{Age, median (IQR), y} & 42.5 (38.5-49.0) & 36.0
(25.0-42.0)\tabularnewline
\textbf{BMI, median (IQR), kg/m2} & 25.0 (23.0-27.1) & 23.7
(20.6-26.5)\tabularnewline
\textbf{Current smoking} & 1 (4\%) & 2 (9\%)\tabularnewline
\textbf{Prior COVID-19 infection} & 12 (50\%) & 11 (50\%)\tabularnewline
Confirmed by RT-PCR & 9 (75\%) & 10 (91\%)\tabularnewline
Confirmed by serology & 1 (8\%) & 1 (9\%)\tabularnewline
Self-reported & 2 (17\%) & 0 (0\%)\tabularnewline
\textbf{Patient reported allergy} & &\tabularnewline
Yes & 11 (46\%) & 8 (36\%)\tabularnewline
No & 13 (54\%) & 14 (64\%)\tabularnewline
\bottomrule
\end{longtable}

Table I. Subject baseline characteristics.

\textbf{Hosted file}

\verb`image1.emf`  available at \url{https://authorea.com/users/396618/articles/530709-covid-19-vaccination-with-bnt162b2-and-chadox1-vaccines-induces-nasal-neutralizing-antibodies}

\emph{Figure I. Effect of vaccination on the induction of systemic and
local neutralizing antibodies.} \textbf{A-B,} Percentage of patients
with SARS-CoV-2 neutralizing antibodies (a) and degree of ACE2 binding
inhibition (b) in serum prior and post vaccination with BNT162b2 and
ChAdOx1. \textbf{C-D,} Percentage of patients with SARS-CoV-2 NAbs (c)
and degree of ACE2 binding inhibition (d) in nasal secretions prior and
post vaccination with BNT162b2 and ChAdOx1.

Asterisks indicate statistical significance by two-way repeated-measures
ANOVA followed by Sidak's multiple comparisons test. *:
p\textless{}0.05, **: p\textless{}0.001, ***: p\textless{}0.0002, ****:
p\textless{}0,0001.

\textbf{References}

1. Polack FP, Thomas SJ, Kitchin N, et al. Safety and Efficacy of the
BNT162b2 mRNA Covid-19 Vaccine. NEJM. 2020; 383:2603-2615.

2. Voysey M, Costa Clemens SA, Madhi SA, et al. Safety and efficacy of
the ChAdOx1 nCoV-19 vaccine (AZD1222) against SARS-CoV-2: an interim
analysis of four randomized controlled trials in Brazil, South Africa,
and the UK. The Lancet. 2021; 397:99-111.

3. Sungnak W, Huang N, B\selectlanguage{ngerman}écavin C, et al. SARS-CoV-2 entry factors are
highly expressed in nasal epithelial cells together with innate immune
genes. Nat Med. 2020; 26: 681-687.

4. Chan RWY, Liu S, Cheung JY, et al. Study on the mucosal and
serological immune reponse to the novel coronavirus (SARS-CoV-2)
vaccines. MedRxiv preprint doi:
https://doi.org/10.1101/2021.06.15.21256661

5. Berings M, Arasi S, De Ruyck N, et al. Reliable mite-specific IgE
testing in nasal secretions by means of allergen microarray. JACI. 2017;
140:301-303.

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