key: cord-0797288-npqbj123
authors: Takano, Tomohiro; Morikawa, Miwa; Adachi, Yu; Kabasawa, Kiyomi; Sax, Nicolas; Moriyama, Saya; Sun, Lin; Isogawa, Masanori; Nishiyama, Ayae; Onodera, Taishi; Terahara, Kazutaka; Tonouchi, Keisuke; Nishimura, Masashi; Tomii, Kentaro; Yamashita, Kazuo; Matsumura, Takayuki; Shinkai, Masaharu; Takahashi, Yoshimasa
title: Distinct Immune Cell Dynamics Correlate with the Immunogenicity and Reactogenicity of SARS-CoV-2 mRNA Vaccine
date: 2022-04-22
journal: Cell Rep Med
DOI: 10.1016/j.xcrm.2022.100631
sha: ac20ca5825a165479183be6143b9a7bc0749697f
doc_id: 797288
cord_uid: npqbj123

Two doses of Pfizer/BioNTech BNT162b2 mRNA vaccine elicit robust SARS-CoV-2-neutralizing antibodies with frequent adverse events. Here, by applying a high-dimensional immune profiling on 92 vaccinees, we identify six vaccine-induced immune dynamics that correlate with the amounts of neutralizing antibodies, the severity of adverse events, or both. The early dynamics of natural killer (NK)/monocyte subsets (CD16+ NK cells, CD56high NK cells, and non-classical monocytes), dendritic cell (DC) subsets (DC3s and CD11c- AS-DCs), and NKT-like cells are revealed as the distinct cell correlates for neutralizing antibody titers, severity of adverse events, and both, respectively. The cell correlates for neutralizing antibody or adverse events are consistently associated with elevation of IFN-γ-inducible chemokines but the chemokine receptors, CCR2 and CXCR3, are expressed in distinct manners between the two correlates; vaccine-induced expression on neutralizing antibody correlate and constitutive expression on adverse event correlate. The finding may guide vaccine strategies that balances immunogenicity and reactogenicity.

Monocytes are identified as a correlate for antibody responses following 165 SARS-CoV-2 mRNA vaccines 6 . In our study, the vaccinees with a more profound 166 reduction in non-classical monocytes mounted higher amounts of NT ( Figure 2F ). 

We evaluated the discriminative value of the identified cellular dynamics for 230 systemic symptoms and antibody titers following secondary vaccination. First, three cell 231 populations (NKT-like cells, DC3, and CD11c -AS-DCs) were selected ( Figure 5A ) and 232 combined into a Z-score to visualize the correlation with NT titers and systemic symptom scores ( Figure 5B ). The combined parameter showed a stronger correlation with 0.30 ~ -0.22 in Figures 4C-4E) , however failed to correlate with NT titers. Next, to assess 236 the utility of these three parameters for identifying a subgroup of vaccinees with severe 237 reactogenicity, the vaccinees were separated into matched groups that were positive 238 (i.e., post/pre ratios were below the median of the cohort) for all three parameters, and 239 unmatched groups that were negative (i.e., post/pre ratios were above the median of the 240 cohort) for three parameters. There was no significant difference in gender distribution 241 between the groups (matched group: male, n = 4; female, n = 15 and unmatched group: 242 male, n = 7; female, n = 11, p = 0.2953, Fisher's exact test). Classification by the three 243 parameters slightly, not significantly, increased the NT titers in the matched group, 244 whereas the matched group led to a significant 2.3-fold increase in systemic symptoms 245 ( Figure 5C ).

The two parameters (DC3s and CD11c -AS-DCs) linked to the AEs, but not to 247 NT titers, were combined ( Figure S6A ). The combined Z-score correlated with systemic 248 symptom scores but not with NT titers (Figures S6B). Also, the subgroups of the vaccinees that were matched or unmatched with two selected parameters mounted 250 equivalent NT titers, however, the matched group had higher systemic symptom scores 251 by 1.8-fold compared to those of the unmatched group ( Figure S6C ). Again, no 252 significant difference was observed in gender distribution between the classified groups subset were identified as specific correlates for AEs and those in NK/monocyte subsets 328 as correlates for NT titers. Importantly, based on the cellular parameters, we were able to classify the vaccinees who suffered from more severe AEs or those who mounted 330 more NT titers after the secondary vaccination.

subsets in the peripheral blood remains to be determined, but the available evidence suggest the following model ( Figure 7 ). The prompt IFN-γ production and the subsequent 334 activation of IFN-γ-inducible chemotaxis is a common trigger for the observed cell 468 Science. 374, abm0829. All data reported in this paper will be shared by the lead contact upon request. This paper 738 does not report the original code. Any additional information required to reanalyze the 739 data reported in this paper is available from the lead contact upon request. Table S1 ). The 

A recombinant reference monoclonal antibody (CR3022) was produced as 798 previously described 48, 49 . Briefly, the VH and VL genes of CR3022 were cloned into 799 expression vectors with human IgG1 heavy chain and kappa light chain, respectively.

The prepared expression vectors were transfected into Expi293F and the recombinant 801 monoclonal antibody was purified from the culture supernatant using a protein G column 802 (Thermo Fisher Scientific). 

and shown as dotted lines in Figure 1B .

RBD IgG titers of COVID-19 convalescent patients were partially referred from 819 data used in Figure 1C of a previous publication 7 . The criteria for data selection were titers. NT titers under the detection limit (IC50 = 20) were set to 20 and detection limit was 845 indicated as dotted line in Figure 1C .

NT titers of COVID-19 convalescent patients were partially referred from data 847 used in Figure 1D of a previous publication 7 . The criteria of data selection were the same 848 as described in ELISA section.

For MDSC analysis, after blocking non-specific antibody binding using Human TruStain with the following antibodies: CD45-APC-Alexa Fluor 700 (HI30, 1:300; BioLegend), 

FCS files were analyzed using FlowJo software (v.10.8.0, BD Biosciences).

Gating strategies for identifying immune cells are shown in Figure S1 . Figure S3 ).

In cases that the cell post/pre ratios inversely correlated with AEs or NT antibody 920 responses, the cell post/pre ratios below the median of the cohort were identified as 921 "positive", and vice versa for "negative". In Figure 5 , 6, and Figure S6 , participants whose 922 cellular parameters were all "positive" (all parameters were below the median of the 923 cohort) were defined as the matched group, and vice versa. 

Longitudinal analysis of local and systemic symptom scores ( Figure 1E ),

immune cell frequencies ( Figure S2 ) and plasma cytokine/chemokine concentrations 943 ( Figure S4A ) were performed using the Wilcoxon test. Fisher's exact test was performed 944 to examine the statistical difference in the incidence of symptoms between primary and 945 secondary vaccinations (Figures 1F and 1G ) and gender distributions in Ab response 3

R700 (BU15, 1:300, BD Biosciences)

CD1c-BUV737 (F10/21A3, 1:300, BD Biosciences)

CD141-BV605 (1A4, 1:300, BD Biosciences)

GHI/61, 1:300, BD Biosciences)

To examine the expression of CCR2

CD86, and CD69, and the binding of Annexin V, cells were stained with the following 878 antibodies: CD45-BUV805 (HI30, 1:150, BD Biosciences)

HLA-DR-APC-H7 (G46-6, 1:300, BD Biosciences)

BV421 (M5E2, 1:300, BD Biosciences), CD16-BUV395 (3G8, 1:300, BD Biosciences

CD11c-APC-R700 (BU15, 1:300, BD Biosciences), CD88-PE-Cyanine7 (S5/1, 1:300 (UCHT2, 1:300, BD Biosciences)

Siglec-6-BV650 (767329, 1:300, BD Biosciences)

CCR2-Brilliant Violet 785 (K036C2, 1:300, BioLegend), CXCR3-CD69-BV711 (FN50, 1:300, BD Biosciences), Annexin V-Alexa647

Scientific) Brilliant Violet 785 Mouse IgG2a

PE/Cyanine5 Mouse IgG1, κ Isotype control (MOPC-21

We thank Rutaro Iwabuchi, Akira Dosaka, Eriko Izumiyama, Emi Kohda, Rieko Iwaki,