key: cord-1038203-azk7hxhe
authors: Lozano-Rodríguez, Roberto; Valentín-Quiroga, Jaime; Avendaño-Ortiz, José; Martín-Quirós, Alejandro; Pascual-Iglesias, Alejandro; Terrón, Verónica; Montalbán-Hernández, Karla; Casalvilla-Dueñas, José Carlos; Bergón-Gutiérrez, Marta; Alcamí, José; Garcia-Pérez, Javier; Cascajero, Almudena; García-Garrido, Miguel Ángel; Balzo-Castillo, Álvaro del; Peinado, María; Gómez, Laura; Llorente-Fernández, Irene; Martín-Miguel, Gema; Herrero-Benito, Carmen; Benito, José Miguel; Rallón, Norma; Vela-Olmo, Carmen; López-Morejón, Lissette; Cubillos-Zapata, Carolina; Aguirre, Luis A.; Fresno, Carlos del; López-Collazo, Eduardo
title: Cellular and humoral functional responses after BNT162b2 mRNA vaccination differ longitudinally between naïve and subjects recovered from COVID-19
date: 2021-12-21
journal: Cell Rep
DOI: 10.1016/j.celrep.2021.110235
sha: 87981d942dfcedad8865703df0c52f44af2c10b4
doc_id: 1038203
cord_uid: azk7hxhe

We have analyzed BNT162b2 vaccine-induced immune responses in naïve and individuals recovered from COVID-19, both early (fourteen days) and late (almost eight months) after vaccination. Plasma S-specific immunoglobulins peak after one vaccine shot in individuals recovered from COVID-19, while a second dose is needed in naïve subjects, although the latter group shows reduced levels all-along the analyzed period. Despite the neutralization capacity against SARS-CoV-2 mirrors this behavior early after vaccination, both groups show comparable neutralizing antibodies and S-specific B cells levels late post-vaccination. When studying cellular responses, naïve individuals exhibit higher SARS-CoV-2-specific cytokines production, CD4+ T cells activation and proliferation than individuals recovered from COVID-19, with patent inverse correlations between humoral and cellular variables early post-vaccination. However, almost eight months post-vaccination, SARS-CoV-2-specific responses are comparable between both groups. Our data indicate that previous history of COVID-19 differentially determines the functional T and B cell-mediated responses to BNT162b2 vaccination over time.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and its associated Figure 2B) . However, only naïve individuals showed a robust induction of most of the 156 cytokines analyzed (IL-2, IL-4, IL-6, IL-10 and TNF), although this S-peptide-specific 157 response in naïve subjects was exclusive of sample 2 ( Figure 2C) . Interestingly, IFN- 158 expression showed a specific pattern, mirroring the CCL2 production ( Figure 2B and C) . 159 To further examine this differential outcome, cytokine production was analyzed by intracellular 160 FACS staining. The expression of IL-2, TNF, IFN- and Granzyme B were consistently 161 induced in CD4 + T cells after ex vivo stimulation with the S-peptide pool in both naïve and 162 individuals recovered from COVID-19, early after vaccination (sample 2) ( Figure 2D and S4) , 163 with less robust responses in CD8 + T cells. However, more than seven months after vaccination 164 (sample 3), this SARS-CoV-2-specific response was negligible in both groups ( Figure 2D and 165 S4). 166 Interestingly, the analysis of the intracellular cytokine production increment induced by SARS- CoV-2 Spike antigen-stimulated and non-stimulated PBMCs confirmed a more powerful CD4 + 176 lymphoproliferative activity in naïve than individuals recovered from COVID-19, specifically, 177 early after vaccination (sample 2) ( Figure 2G ). 178 These data suggest a strong SARS-CoV-2 Spike-specific T cell response early after vaccination 179 in naïve subjects (but not in COVID-19-recovered), that declines over time. 

182 decline along the time. 183 Next, we dissected the SARS-CoV-2 Spike-specific T cell responses. First, we analyzed the 184 phenotype of both proliferative (CFSE dim ) and non-proliferative (CFSE bright ) CD4 + T cells after Figure 3C ). Although in a less robust way, similar behaviors were observed for CD8 + T cells ( Figure S5A-C) . 194 We next analyzed intracellular cytokines production in CD4 + T cell subpopulations induced by Humoral and cellular activation features inversely correlated early after vaccination. 211 Based on the differential behavior of SARS-CoV-2 Spike-specific humoral and cellular 212 responses between naïve and individuals recovered from COVID-19, we explored whether 213 these features could identify subjects belonging to these two groups, in an unsupervised generated a clearer discrimination between naïve and individuals recovered from COVID-19 217 in sample 2 than in sample 3 ( Figure 4A ). 218 Next, we depicted the correlation between the analyzed variables once they were classified 219 based on their functionality. Along these lines, humoral parameters quantified in plasma were 220 confronted to the cellular response after ex vivo cellular stimulation with SARS-CoV-2 Spike 221 peptide pool ( Figure 4B ). This representation suggested inverse correlations early after 222 vaccination (sample 2) between humoral and cellular responses, particularly IgG production 223 and neutralization capacity with SARS-CoV-2 Spike-specific pro-inflammatory cytokines 224 (CCL2, CXCL10, IFN and IL-2) production and CD4 + T cells proliferation ( Figure 4B ). 225 However, these correlations were attenuated more than seven months after vaccination (sample 

Of note, our data on early humoral responses support these findings. We also observed that 274 antibody levels and the neutralizing capacity of plasma from individuals recovered from 275 COVID-19 was higher than that of naïve subjects early after vaccination (samples 1 and 2), an 276 effect suggested but not fully analyzed in a previous study (Gobbi et al., 2021) . However, more 277 than seven months after vaccination, individuals recovered from COVID-19 still showed 278 higher antibody titers but comparable neutralizing antibodies to naïve subjects. These data 279 highlight the need to discriminate between antibody titers and neutralizing capacity. In In our study, we have analyzed SARS-CoV-2 Spike-specific responses in T cells after 306 restimulation with a peptide pool covering this antigen. This assay showed a differential 307 response between naïve and individuals recovered from COVID-19 early after vaccination, 308 with a more pronounced activation of CD4 + T cells in naïve subjects. This was revealed by a 309 higher induction of cytokine production and proliferation after restimulation, particularly in 339 Sample size is a limitation of this study. Considering the high number of immune variables 340 analysed and their complexity, we decided to perform our study with a not so large but well-341 controlled cohort of participants. We believe that this approach has allowed us to reach clear 342 conclusions, but a multicentre cohort with a larger number of patients would be desiderable. Table S1 . Table S3 . 

Efficacy and Safety of the mRNA-629 1273 SARS-CoV-2 Vaccine

Waning of IgG, Total and Neutralizing 632

Antibodies 6 Months Post-Vaccination with BNT162b2 in Healthcare Workers

Phenotypical and functional specialization of 635 FOXP3+ regulatory T cells

Evidence for 638 retained spike-binding and neutralizing activity against emerging SARS-CoV-2 variants in 639 serum of COVID-19 mRNA vaccine recipients

Waning of BNT162b2

Vaccine Protection against SARS-CoV-2 Infection in Qatar

Evidence of SARS-CoV-2-Specific 645

Six Months After Vaccination With the BNT162b2 mRNA Vaccine. Front 646 Immunol 12

SARS-CoV-2 649 mRNA vaccine design enabled by prototype pathogen preparedness

Antibody Persistence 652 through 6 Months after the Second Dose of mRNA-1273 Vaccine for Covid-19

Antibody responses to the BNT162b2 mRNA 656 vaccine in individuals previously infected with SARS-CoV-2

Antibody response to the bnt162b2 mrna covid-19 659 vaccine in subjects with prior sars-cov-2 infection

Waning Immunity after the BNT162b2 666 Vaccine in Israel

Targets of T Cell 669 Responses to SARS-CoV-2 Coronavirus in Humans with COVID-19 Disease and Unexposed 670 individuals

BNT162b2 vaccination induces 673 durable SARS-CoV-2 specific T cells with a stem cell memory phenotype

Delayed Second Dose versus Standard 676 Regimen for Covid-19 Vaccination

Antibody Responses in Seropositive Persons after a Single Dose of SARS-CoV-2 mRNA 680

One dose of SARS-CoV-2 vaccine exponentially increases antibodies in individuals 683 who have recovered from symptomatic COVID-19

Covid-19: Third vaccine dose boosts immune response but may not be 685 needed, say researchers

UMAP: Uniform Manifold Approximation 687 and Projection for Dimension Reduction

Correlates of protection against SARS-CoV-2 in 690 rhesus macaques

Phase I/II study of COVID-19 RNA 693 vaccine BNT162b1 in adults

Dynamics of antibody response to 696

BNT162b2 vaccine after six months: a longitudinal prospective study

Detection of SARS-CoV-2-Specific Humoral and Cellular Immunity in 700 COVID-19 Convalescent individuals. Immunity. antibodies against severe acute respiratory syndrome-CoV-2 variants of concern: A 704 systematic review of in vitro studies

Safety and Efficacy of the 707

BNT162b2 mRNA Covid-19 Vaccine

Effect of previous SARS-CoV-2 infection on 710 humoral and T-cell responses to single-dose BNT162b2 vaccine

Effect of previous SARS-CoV-2 infection on 714 humoral and T-cell responses to single-dose BNT162b2 vaccine

Immunological memory and neutralizing activity to a 718 single dose of COVID-19 vaccine in previously infected individuals

SARS-CoV-2-specific T cell responses and 722 correlations with COVID-19 patient predisposition

Maturation and persistence of 726 the anti-SARS-CoV-2 memory B cell response

Safety and Efficacy of the BNT162b2 729 mRNA Covid-19 Vaccine through 6 Months

SARS-CoV-2 mRNA vaccines induce 732 persistent human germinal centre responses

ChAdOx1 nCoV-19 vaccine (AZD1222) against SARS-CoV-2: an interim analysis of four 736 randomised controlled trials in Brazil, South Africa, and the UK

HIGHLIGHTS  History of SARS-CoV-2 infection affects longitudinal responses to BNT162b2 vaccine  Lower humoral but enhanced cellular responses early after vaccine in naïve subjects  Comparable humoral and cellular responses almost eight months after vaccination  Similar S-specific B cells late after vaccine in naïve and recovered from COVID-19 eTOC Lozano-Rodríguez et al. show that naïve subjects have enhanced SARS-CoV-2 Spike-specific T reactions, but reduced humoral specific responses compared to individuals recovered from COVID-19. However, almost eight months after vaccination, comparable specific responses are observed, with equivalent levels of SARS-CoV-2specific B cells and neutralizing antibodies.