Demographic characteristics.

<p>Demographic characteristics.</p

Homologue sequences of EBNA6_740 to human proteins.

<p>Homologue sequences of EBNA6_740 to human proteins.</p

Reactivity against EBNA-6, LPO and TPO in CFS patients.

<p>A) Median intensity of IgG response against EBNA-6 peptide 740 in seroarray cohort (CFS, n = 93 and healthy, n = 50) and a subsequent validation cohort (CFS, n = 227 and healthy, n = 47) was analysed by multiwell assay. B) Optical density (OD) and Interquartile Range (IQR) of IgG response against EBNA6_740, a negative control peptide, LPO, and TPO in healthy controls (n = 115) and CFS patients (n = 162) by peptide ELISA. C) Correlation of ELISA OD values of EBNA6_740 and LPO for all patients and controls. D) IgG responses against EBNA-6 protein in in healthy controls (n = 40) and CFS patients (n = 40) detected by Odyssey Infrared Imaging System as arbitrary fluorescence units (AFU), and correlation of EBNA-6 protein and EBNA-6 peptide as well as LPO peptide IgG in CFS patients. Statistical analysis by two-tailed Mann-Whitney-U test with * p<0.05, ** p<0.01, *** p <0.001, **** p<0.0001, r-spearman coefficient.</p

Primer and probe for EBV load and type PCR.

<p>Primer and probe for EBV load and type PCR.</p

DataSheet_3_Primary ChAdOx1 vaccination does not reactivate pre-existing, cross-reactive immunity.pdf

Currently available COVID-19 vaccines include inactivated virus, live attenuated virus, mRNA-based, viral vectored and adjuvanted protein-subunit-based vaccines. All of them contain the spike glycoprotein as the main immunogen and result in reduced disease severity upon SARS-CoV-2 infection. While we and others have shown that mRNA-based vaccination reactivates pre-existing, cross-reactive immunity, the effect of vector vaccines in this regard is unknown. Here, we studied cellular and humoral responses in heterologous adenovirus-vector-based ChAdOx1 nCOV-19 (AZ; Vaxzeria, AstraZeneca) and mRNA-based BNT162b2 (BNT; Comirnaty, BioNTech/Pfizer) vaccination and compared it to a homologous BNT vaccination regimen. AZ primary vaccination did not lead to measurable reactivation of cross-reactive cellular and humoral immunity compared to BNT primary vaccination. Moreover, humoral immunity induced by primary vaccination with AZ displayed differences in linear spike peptide epitope coverage and a lack of anti-S2 IgG antibodies. Contrary to primary AZ vaccination, secondary vaccination with BNT reactivated pre-existing, cross-reactive immunity, comparable to homologous primary and secondary mRNA vaccination. While induced anti-S1 IgG antibody titers were higher after heterologous vaccination, induced CD4+ T cell responses were highest in homologous vaccinated. However, the overall TCR repertoire breadth was comparable between heterologous AZ-BNT-vaccinated and homologous BNT-BNT-vaccinated individuals, matching TCR repertoire breadths after SARS-CoV-2 infection, too. The reasons why AZ and BNT primary vaccination elicits different immune response patterns to essentially the same antigen, and the associated benefits and risks, need further investigation to inform vaccine and vaccination schedule development.</p

DataSheet_4_Primary ChAdOx1 vaccination does not reactivate pre-existing, cross-reactive immunity.pdf

Currently available COVID-19 vaccines include inactivated virus, live attenuated virus, mRNA-based, viral vectored and adjuvanted protein-subunit-based vaccines. All of them contain the spike glycoprotein as the main immunogen and result in reduced disease severity upon SARS-CoV-2 infection. While we and others have shown that mRNA-based vaccination reactivates pre-existing, cross-reactive immunity, the effect of vector vaccines in this regard is unknown. Here, we studied cellular and humoral responses in heterologous adenovirus-vector-based ChAdOx1 nCOV-19 (AZ; Vaxzeria, AstraZeneca) and mRNA-based BNT162b2 (BNT; Comirnaty, BioNTech/Pfizer) vaccination and compared it to a homologous BNT vaccination regimen. AZ primary vaccination did not lead to measurable reactivation of cross-reactive cellular and humoral immunity compared to BNT primary vaccination. Moreover, humoral immunity induced by primary vaccination with AZ displayed differences in linear spike peptide epitope coverage and a lack of anti-S2 IgG antibodies. Contrary to primary AZ vaccination, secondary vaccination with BNT reactivated pre-existing, cross-reactive immunity, comparable to homologous primary and secondary mRNA vaccination. While induced anti-S1 IgG antibody titers were higher after heterologous vaccination, induced CD4+ T cell responses were highest in homologous vaccinated. However, the overall TCR repertoire breadth was comparable between heterologous AZ-BNT-vaccinated and homologous BNT-BNT-vaccinated individuals, matching TCR repertoire breadths after SARS-CoV-2 infection, too. The reasons why AZ and BNT primary vaccination elicits different immune response patterns to essentially the same antigen, and the associated benefits and risks, need further investigation to inform vaccine and vaccination schedule development.</p

DataSheet_1_Primary ChAdOx1 vaccination does not reactivate pre-existing, cross-reactive immunity.pdf

Currently available COVID-19 vaccines include inactivated virus, live attenuated virus, mRNA-based, viral vectored and adjuvanted protein-subunit-based vaccines. All of them contain the spike glycoprotein as the main immunogen and result in reduced disease severity upon SARS-CoV-2 infection. While we and others have shown that mRNA-based vaccination reactivates pre-existing, cross-reactive immunity, the effect of vector vaccines in this regard is unknown. Here, we studied cellular and humoral responses in heterologous adenovirus-vector-based ChAdOx1 nCOV-19 (AZ; Vaxzeria, AstraZeneca) and mRNA-based BNT162b2 (BNT; Comirnaty, BioNTech/Pfizer) vaccination and compared it to a homologous BNT vaccination regimen. AZ primary vaccination did not lead to measurable reactivation of cross-reactive cellular and humoral immunity compared to BNT primary vaccination. Moreover, humoral immunity induced by primary vaccination with AZ displayed differences in linear spike peptide epitope coverage and a lack of anti-S2 IgG antibodies. Contrary to primary AZ vaccination, secondary vaccination with BNT reactivated pre-existing, cross-reactive immunity, comparable to homologous primary and secondary mRNA vaccination. While induced anti-S1 IgG antibody titers were higher after heterologous vaccination, induced CD4+ T cell responses were highest in homologous vaccinated. However, the overall TCR repertoire breadth was comparable between heterologous AZ-BNT-vaccinated and homologous BNT-BNT-vaccinated individuals, matching TCR repertoire breadths after SARS-CoV-2 infection, too. The reasons why AZ and BNT primary vaccination elicits different immune response patterns to essentially the same antigen, and the associated benefits and risks, need further investigation to inform vaccine and vaccination schedule development.</p

Total IgG responses against EBV, EBNA-1 and enterovirus peptides in the different cohorts.

<p>A) The signal intensity of IgG responses against all EBV peptides (upper row), EBNA-1 peptides (middle row) and enterovirus (lower row) in CFS, healthy controls and MS patients. B) Comparison of patients with SLE and Hodgkin’s lymphoma in disease remission with cancer-related fatigue (HD_NF) or without fatigue (HD_FAT) analysed in another seroarray screening experiment. Statistical analysis by Wilcoxon rank sum test with * p<0.05, *** p <0.001.</p

Seroarray analysis of IgG antibody reactivity against 15-mer EBV peptides which cover 14 EBV proteins with sequences of EBV type I (B-95.8) in 50 healthy donors.

<p>IgG responses of each sample against each peptide are depicted. Responses are defined as a signal intensity above 5,000. In the left graph the % of samples with a response against each single peptide of the indicated proteins are shown. The right graph shows the mean intensity of these IgG responses (defined as a signal intensity above 5,000) against the single peptides. EA- early antigen; PM- polymerase; VCA- viral capsid antigen; gp- glycoprotein.</p

DataSheet_2_Primary ChAdOx1 vaccination does not reactivate pre-existing, cross-reactive immunity.pdf

Currently available COVID-19 vaccines include inactivated virus, live attenuated virus, mRNA-based, viral vectored and adjuvanted protein-subunit-based vaccines. All of them contain the spike glycoprotein as the main immunogen and result in reduced disease severity upon SARS-CoV-2 infection. While we and others have shown that mRNA-based vaccination reactivates pre-existing, cross-reactive immunity, the effect of vector vaccines in this regard is unknown. Here, we studied cellular and humoral responses in heterologous adenovirus-vector-based ChAdOx1 nCOV-19 (AZ; Vaxzeria, AstraZeneca) and mRNA-based BNT162b2 (BNT; Comirnaty, BioNTech/Pfizer) vaccination and compared it to a homologous BNT vaccination regimen. AZ primary vaccination did not lead to measurable reactivation of cross-reactive cellular and humoral immunity compared to BNT primary vaccination. Moreover, humoral immunity induced by primary vaccination with AZ displayed differences in linear spike peptide epitope coverage and a lack of anti-S2 IgG antibodies. Contrary to primary AZ vaccination, secondary vaccination with BNT reactivated pre-existing, cross-reactive immunity, comparable to homologous primary and secondary mRNA vaccination. While induced anti-S1 IgG antibody titers were higher after heterologous vaccination, induced CD4+ T cell responses were highest in homologous vaccinated. However, the overall TCR repertoire breadth was comparable between heterologous AZ-BNT-vaccinated and homologous BNT-BNT-vaccinated individuals, matching TCR repertoire breadths after SARS-CoV-2 infection, too. The reasons why AZ and BNT primary vaccination elicits different immune response patterns to essentially the same antigen, and the associated benefits and risks, need further investigation to inform vaccine and vaccination schedule development.</p