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Supplementary Figure 3: Gating strategy for a representative sample

Supplementary Figure 2: Virus neutralization test with 50% neutralization titers (NT50) defined as the sample dilution (reciprocal titer) conveying 50% neutralization in SARS-CoV-2 (strains 2019-nCoV-Italy-INMI1 and VLD20211207) infected wells.

Supplementary Tables S1-S2

Purpose:Patients with cancer display reduced humoral responses after double-dose COVID-19 vaccination, whereas their cellular response is more comparable with that in healthy individuals. Recent studies demonstrated that a third vaccination dose boosts these immune responses, both in healthy people and patients with cancer. Because of the availability of many different COVID-19 vaccines, many people have been boosted with a different vaccine from the one used for double-dose vaccination. Data on such alternative vaccination schedules are scarce. This prospective study compares a third dose of BNT162b2 after double-dose BNT162b2 (homologous) versus ChAdOx1 (heterologous) vaccination in patients with cancer.Experimental Design:A total of 442 subjects (315 patients and 127 healthy) received a third dose of BNT162b2 (230 homologous vs. 212 heterologous). Vaccine-induced adverse events (AE) were captured up to 7 days after vaccination. Humoral immunity was assessed by SARS-CoV-2 anti-S1 IgG antibody levels and SARS-CoV-2 50% neutralization titers (NT50) against Wuhan and BA.1 Omicron strains. Cellular immunity was examined by analyzing CD4+ and CD8+ T-cell responses against SARS-CoV-2–specific S1 and S2 peptides.Results:Local AEs were more common after heterologous boosting. SARS-CoV-2 anti-S1 IgG antibody levels did not differ significantly between homologous and heterologous boosted subjects [GMT 1,755.90 BAU/mL (95% CI, 1,276.95–2,414.48) vs. 1,495.82 BAU/mL (95% CI, 1,131.48–1,977.46)]. However, homologous-boosted subjects show significantly higher NT50 values against BA.1 Omicron. Subjects receiving heterologous boosting demonstrated increased spike-specific CD8+ T cells, including higher IFNγ and TNFα levels.Conclusions:In patients with cancer who received double-dose ChAdOx1, a third heterologous dose of BNT162b2 was able to close the gap in antibody response.

Supplementary Figure 6: Correlation between CD4+ and CD8+ T cell responses and SARS-CoV-2 binding antibodies post homologous or heterologous booster vaccination in cancer patients.

Supplementary Figure 4: Spike-specific CD4+ T cell responses post homologous or heterologous booster vaccination in subcohorts.

Supplementary Figure 5: Spike-specific CD8+ T cell responses post homologous or heterologous booster vaccination in subcohorts.