SARS-CoV2 wild type and mutant specific humoral and T cell immunity is superior after vaccination than after natural infection

  1. Jennifer R. Richardson
  2. Ralph Götz
  3. Vanessa Mayr
  4. Martin J. Lohse
  5. Hans-Peter Holthoff
  6. Martin Ungerer

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Abstract

We investigated blood samples from fully SARS-CoV2-vaccinated subjects and from previously positive tested patients up to one year after infection with SARS-CoV2, and compared short- and long-term T cell and antibody responses, with a special focus on the recently emerged delta variant (B.1.617.2).

Methods and results

In 23 vaccinated subjects, we documented high anti-SARS-CoV2 spike protein receptor binding domain (RBD) antibody titers. Average virus neutralization by antibodies, assessed as inhibition of ACE2 binding to RBD, was 2.2-fold reduced for delta mutant vs. wild type (wt) RBD. The mean specific antibody titers were lower one year after natural infection than after vaccination; ACE2 binding to delta mutant vs. wt RBD was 1.65-fold reduced. In an additional group, omicron RBD binding was reduced compared to delta. Specific CD4+ T cell responses were measured after stimulation with peptides pools from wt, alpha, beta, gamma, or delta variant SARS-CoV2 spike proteins by flow cytometric intracellular cytokine staining. There was no significant difference in cytokine production of IFN-γ, TNF-α, or IL-2 between vaccinated subjects. T cell responses to wt or mutant SARS-CoV2 spike were significantly weaker after natural occurring infections compared to those in vaccinated individuals.

Conclusion

Antibody neutralisation of the delta mutant was reduced compared to wt, as assessed in a novel inhibition assay with a finger prick blood drop. Strong CD4 T cell responses were present against wt and mutant SARS-CoV2 variants, including the delta (B.1.617.2) strain, in fully vaccinated individuals, whereas they were partly weaker 1 year after natural infection. Hence, immune responses after vaccination are stronger compared to those after naturally occurring infection, pointing out the need of the vaccine to overcome the pandemic.

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  1. Version published to 10.1371/journal.pone.0266701
  2. Version published to 10.1101/2021.09.07.21262725v2 on medRxiv
  3. ScreenIT

    SciScore for 10.1101/2021.09.07.21262725: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    EthicsConsent: Research Subjects: Blood was collected from 67 volunteers with informed consent, who had been previously infected with SARS-CoV-2 corona virus or been vaccinated against the virus or been unexposed.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Qualitative Detection of anti-SARS-CoV2 Antibodies in Human Serum: Anti-SARS-CoV2 antibodies in human serum or in a drop of full blood were detected by a two-step incubation antigen sandwich ELISA using the RBD of the S1 protein of the SARS-CoV2 virus.
    anti-SARS-CoV2
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    The vectors were transfected into Flip-InTM-Chinese hamster ovary (CHO) cells (Life Technologies) using the Lipofectamine 2000 Reagent (Invitrogen, #11668-019), together with the plasmid pOG44, providing site-directed recombination.
    Flip-InTM-Chinese hamster ovary ( CHO )
    suggested: None
    Expression and purification of RBD-His and NP-Strep: The various CHO-spike-RBD-His cells and CHO-spike-NP-Strep cells were grown in suspension in ProCHO5, 4 mM L-glutamine and 600 µg/ml Hygromycin B in flasks to submaximal density at 37°C.
    CHO-spike-NP-Strep
    suggested: None
    Recombinant DNA
    SentencesResources
    Life Technologies) by adding the signal sequence METPAQLLFLLLLWLPDTTG at the beginning and cloned into the plasmid vector pcDNA5/FRT via BamHI and XhoI.
    pcDNA5/FRT
    suggested: RRID:Addgene_31983)
    The resulting vectors were called pcDNA5/CoV-RBD-His and pcDNA5/CoV-NP-Strep, respectively, and allow for expression and secretion of RBD-His or NP-Strep into the culture medium of mammalian cells under the control of the human cytomegalovirus (CMV) immediate-early enhancer/promoter.
    pcDNA5/CoV-RBD-His
    suggested: None
    pcDNA5/CoV-NP-Strep
    suggested: None
    The vectors were transfected into Flip-InTM-Chinese hamster ovary (CHO) cells (Life Technologies) using the Lipofectamine 2000 Reagent (Invitrogen, #11668-019), together with the plasmid pOG44, providing site-directed recombination.
    pOG44
    suggested: None
    Software and Algorithms
    SentencesResources
    Then, IC50 values minus blank values were calculated for each sample using results from all dilutions with SigmaPlot v14.5.
    SigmaPlot
    suggested: (SigmaPlot, RRID:SCR_003210)
    Statistical Analysis: Statistical analysis was performed using SPSS software version 26.
    SPSS
    suggested: (SPSS, RRID:SCR_002865)

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

    Results from JetFighter: We did not find any issues relating to colormaps.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    Results from scite Reference Check: We found no unreliable references.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  4. Version published to 10.1101/2021.09.07.21262725v1 on medRxiv