Thrombocytopenia and splenic platelet-directed immune responses after IV ChAdOx1 nCov-19 administration
This article has been Reviewed by the following groups
Listed in
- Evaluated articles (ScreenIT)
Abstract
Vaccines against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are based on a range of novel platforms, with adenovirus-based approaches (like ChAdOx1 nCov-19) being one of them. Recently, a novel complication of SARS-CoV-2–targeted adenovirus vaccines has emerged: immune thrombocytopenia, either isolated, or accompanied by thrombosis (then termed VITT). This complication is characterized by low platelet counts, and in the case of VITT, also by platelet-activating platelet factor 4 antibodies reminiscent of heparin-induced thrombocytopenia, leading to a prothrombotic state with clot formation at unusual anatomic sites. Here, we detected antiplatelet antibodies targeting platelet glycoprotein receptors in 30% of patients with proven VITT (n = 27) and 42% of patients with isolated thrombocytopenia after ChAdOx1 nCov-19 vaccination (n = 26), indicating broad antiplatelet autoimmunity in these clinical entities. We use in vitro and in vivo models to characterize possible mechanisms of these platelet-targeted autoimmune responses leading to thrombocytopenia. We show that IV but not intramuscular injection of ChAdOx1 nCov-19 triggers platelet-adenovirus aggregate formation and platelet activation in mice. After IV injection, these aggregates are phagocytosed by macrophages in the spleen, and platelet remnants are found in the marginal zone and follicles. This is followed by a pronounced B-cell response with the emergence of circulating antibodies binding to platelets. Our work contributes to the understanding of platelet-associated complications after ChAdOx1 nCov-19 administration and highlights accidental IV injection as a potential mechanism of platelet-targeted autoimmunity. Hence, preventing IV injection when administering adenovirus-based vaccines could be a potential measure against platelet-associated pathologies after vaccination.
Article activity feed
-
-
SciScore for 10.1101/2021.06.29.450356: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IRB: The study was approved by local ethics committee of Munich.
Consent: Human platelet isolation: Human blood for in-vitro assays was obtained after informed consent from healthy male and female donors aged 20-35.
Euthanasia Agents: Injection was performed either via tail vein or in the medial aspect of the thigh of isoflurane anesthetized mice.Sex as a biological variable Human platelet isolation: Human blood for in-vitro assays was obtained after informed consent from healthy male and female donors aged 20-35. Randomization Intravital imaging cell tracking was done manually on randomly selected cells which could be traced for at least 5 frames. Blinding not detected. Power Analysis not detected. Ta…
SciScore for 10.1101/2021.06.29.450356: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IRB: The study was approved by local ethics committee of Munich.
Consent: Human platelet isolation: Human blood for in-vitro assays was obtained after informed consent from healthy male and female donors aged 20-35.
Euthanasia Agents: Injection was performed either via tail vein or in the medial aspect of the thigh of isoflurane anesthetized mice.Sex as a biological variable Human platelet isolation: Human blood for in-vitro assays was obtained after informed consent from healthy male and female donors aged 20-35. Randomization Intravital imaging cell tracking was done manually on randomly selected cells which could be traced for at least 5 frames. Blinding not detected. Power Analysis not detected. Table 2: Resources
Antibodies Sentences Resources Sections were stained with antibodies against F4/80 (Biolegend, Cat. No. 123110), CD45R/B220 (Biolegend, Cat. No. 103212), ki67 (Abcam, Cat. No. ab15580) F4/80suggested: (BioLegend Cat# 123110, RRID:AB_893486)CD45R/B220suggested: (BioLegend Cat# 103212, RRID:AB_312997)ki67suggested: (Abcam Cat# ab15580, RRID:AB_443209)Secondary antibodies and nucleic acid stain included FITC goat anti mouse (Thermofisher, Cat. No. A11029) and Cy3 goat anti rabbit (Jackson Immunoresearch, Cat. No. 111-165-003) and Hoechst33342. anti mousesuggested: (Thermo Fisher Scientific Cat# A-11029, RRID:AB_2534088)anti rabbitsuggested: NoneHoechst33342suggested: NoneExperimental Models: Organisms/Strains Sentences Resources Mouse Strains: C57BL/6 mice were purchased from The Jackson Laboratory. C57BL/6suggested: NoneAID-/- sIgM-/- mice were bred and maintained at our animal facility. AID-/- sIgM-/-suggested: NoneSoftware and Algorithms Sentences Resources Flowjo (BD) was used for flowcytometric analysis, gating strategies are shown in Supplemental Figure 2. Flowjosuggested: (FlowJo, RRID:SCR_008520)Statistics were computed with Imaris, Meandering Index was defined as Net displacement/Total track length. Imarissuggested: (Imaris, RRID:SCR_007370)For area quantification, thresholds were taken with ImageJ and overlap quantified by percent area overlapping between thresholds. ImageJsuggested: (ImageJ, RRID:SCR_003070)For all statistical tests Prism (GraphPad) was used. GraphPadsuggested: (GraphPad Prism, RRID:SCR_002798)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 did not find any issues relating to the usage of bar graphs.
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.
-
