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Licensed Unlicensed Requires Authentication Published by De Gruyter February 2, 2023

Assessment of humoral and cellular immunity after bivalent BNT162b2 vaccination and potential association with reactogenicity

  • Gian Luca Salvagno , Laura Pighi , Brandon M. Henry , Myriam Valentini , Beatrice Tonin , Damiano Bragantini , Gianluca Gianfilippi , Simone De Nitto , Mario Plebani ORCID logo and Giuseppe Lippi ORCID logo EMAIL logo



This study investigated the feasibility and clinical value of using a novel, automated and high-throughput SARS-CoV-2 Interferon Gamma Release Assay (IGRA), combined with total anti-SARS-CoV-2 antibodies assessment, for evaluating the immune response after bivalent BNT162b2 vaccination.


A cohort of healthcare workers, who already underwent primary vaccination and boosting with monovalent BNT162b2 vaccine, received a booster dose of the new BNT162b2 bivalent formulation. Blood samples were taken immediately before vaccination (T0) and 1 month afterwards (T1). Humoral and cellular immunity were assayed with Roche Elecsys Anti-SARS-CoV-2 and Roche Elecsys IGRA SARS-CoV-2, respectively.


The study population consisted of 51 subjects (median age: 43 years; 51% females). Total anti-SARS-CoV-2 antibodies and IGRA SARS-CoV-2 values increased at T1 from 9,050 to 25,000 BAU/mL (p<0.001), and from 0.44 to 0.78 IU/mL (p=0.385), accounting for median increase of 2.0 and 1.6 folds, respectively. Increased T1 values of total anti-SARS-CoV-2 antibodies and IGRA SARS-CoV-2 were recorded in 100% and 68.6% subjects, respectively. In those with baseline values below the median, post-vaccine levels displayed larger increases of 3.3 and 5.1 folds for anti-SARS-CoV-2 total antibodies and IGRA SARS-CoV-2, respectively. The variation of total anti-SARS-CoV-2 antibodies was inversely associated with their T0 values (r=−0.97; p<0.001), whilst that of IGRA SARS-CoV-2 was inversely associated with its T0 value (r=−0.58; p<0.001). No other signifcant associations were found with demographical or clinical variables, including side effects.


The bivalent BNT162b2 vaccine booster enhances humoral and cellular immunity against SARS-CoV-2, especially in recipients with lower baseline biological protection.

Corresponding author: Prof. Giuseppe Lippi, Section of Clinical Biochemistry, University Hospital of Verona, Piazzale L.A. Scuro, 10, 37134 Verona, Italy, Phone: 0039-045-8122970, Fax: 0039-045-8124308, E-mail:


The authors acknowledge the staff of the Service of Laboratory Medicine and the Medical Direction of the Pederzoli Hospital (Peschiera del Garda, Italy) for support and technical assistance.

  1. Research funding: None declared.

  2. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  3. Competing interests: Authors state no conflict of interest.

  4. Informed consent: Informed consent was obtained from all individuals included in this study.

  5. Ethical approval: All subjects recruited in this prospective observational study provided written informed consents for undergoing COVID-19 vaccination and for being included in the immunological prospective survey. The study was conducted in accordance with the Declaration of Helsinki and its protocol has been approved by the Ethics Committee of the Provinces of Verona and Rovigo (59COVIDCESC; November 8, 2021).


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Supplementary Material

This article contains supplementary material (

Received: 2023-01-14
Accepted: 2023-01-20
Published Online: 2023-02-02
Published in Print: 2023-06-27

© 2023 Walter de Gruyter GmbH, Berlin/Boston

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