Dynamics of humoral and cellular response to three doses of anti-SARS-CoV-2 BNT162b2 vaccine in patients with hematological malignancies and older subjects

Valentina Laquintana¹ Carla Mottini² Francesco Marchesi² Benedetta Marcozzi² Irene Terrenato³ Eleonora Sperandio² Luisa De Latouliere² Francesca Carrieri² FULVIA PIMPINELLI² Martina Pontone² Raul Pellini² Flaminia Campo² Laura Conti² Celeste Accetta² Chiara Mandoj² Fabrizio Petrone² Ornella Di Bella² Branka Vujovic⁴ Aldo Morrone⁵ Mirco Compagnone⁶ Eugenia Principato⁷ Eleonora Pinto⁸ Elena Papa² Paolo Falcucci² Antonia L. Malfa² Matteo Pallocca² Federico De Marco² Giulia Piaggio^9* Gennaro Ciliberto¹⁰ Andrea Mengarelli² Simona Di Martino^11*

• ¹Pathological Anatomy Unit, Research and Advanced Technologies Department, Regina Elena National Cancer Institute (IRCCS), Italy
• ²IRCCS National Cancer Institute Regina Elena, Roma, Italy
• ³Biostatistical Unit, Regina Elena National Cancer Institute, Hospital Physiotherapy Institutes (IRCCS), Italy
• ⁴Direzione Sanitaria Istituti Fisioterapici Ospitalieri, Italy
• ⁵IRCCS San Gallicano Dermatological Institute, Roma, Italy
• ⁶NeoMatrix, Italy
• ⁷Magna Græcia University, Italy
• ⁸Takis s.r.l., Italy
• ⁹Regina Elena National Cancer Institute, Hospital Physiotherapy Institutes (IRCCS), Italy
• ¹⁰Scientific Direction, IRCCS Regina Elena National Cancer Institute, Istituti Fisioterapici Ospitalieri (IFO), Italy
• ¹¹Department of Pathology, Regina Elena National Cancer Institute, Hospital Physiotherapy Institutes (IRCCS), Italy

Background: Few data are available about the durability of the response, the induction of neutralizing antibodies, and the cellular response upon the third dose SARS-CoV-2 vaccine in hemato-oncological patients. Objective: To investigate the antibody and cellular response to the BNT162b2 vaccine in patients with hematological malignancy. Methods: We measured SARS-CoV-2 anti-spike antibodies, anti-Omicron neutralizing antibodies, and T-cell responses 1 month after the third dose of vaccine in 93 fragile patients with hematological malignancy (FHM), 51 fragile not oncological subjects (FNO) aged 80-92, and 47 employees of the hospital (healthcare workers, (HW), aged 23-66 years. Blood samples were collected at day 0 (T0), 21 (T1), 35 (T2), 84 (T3), 168 (T4), 351 (T pre-3D), and 381 (T post-3D) after the first dose of vaccine. Serum IgG antibodies against S1/S2 antigens of SARS-CoV-2 spike protein were measured at every time point. Neutralizing antibodies were measured at T2, T3 (anti-Alpha), T4 (anti-Delta), and T post-3D (anti-Omicron). T cell response was assessed at T post-3D. Results: An increase in anti-S1/S2 antigen antibodies compared to T0 was observed in the three groups at T post-3D. After the third vaccine dose, the median antibody level of FHM subjects was higher than after the second dose and above the putative protection threshold, although lower than in the other groups. The neutralizing activity of antibodies against the Omicron variant of the virus was tested at T2 and T post-3D. 42.3% of FHM, 80,0% of FNO and 90,0% of HW had anti-Omicron neutralizing antibodies at T post-3D. To get more insight into the breadth of antibody responses, we analyzed neutralizing capacity against BA.4/BA.5, BF.7, BQ.1, XBB.1.5 Omicron variants,. The memory T-cell response was lower in FHM than in FNO and HW cohorts. Data on breakthrough infections and deaths suggested that the positivity threshold of the test is protective after the third dose of the vaccine in all cohorts. Conclusions: FHM have a relevant response to the BNT162b2 vaccine, with increasing antibody levels after the third dose coupled with, although low, a T-cell response. FHM need repeated vaccine doses to attain a protective immunological response.