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. 2021 Sep 9;100(5):1135–1136. doi: 10.1016/j.kint.2021.09.001

Figure 1.

Figure 1

Humoral and cellular response following the third vaccination in kidney transplant recipients in whom the primary vaccination failed. Renal transplant recipients with failed seroconversion after BNT162b2 (Pfizer–BioNTech) prime-boost vaccination were subjected to the third vaccination by mRNA-1273 (Moderna). Humoral and cellular immune responses before (red) and 2 weeks after (blue) the third vaccination are presented. Enzyme-linked immunosorbent assay (ELISA) was performed for the assessment of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S)-protein–binding antibodies, and neutralizing antibody capacity was assessed by a pseudovirus system bearing the SARS-CoV-2 S-protein. S-protein–reactive T cells were analyzed by flow cytometry following an overnight stimulation of peripheral blood mononuclear cells with overlapping peptide pools (OPPs) spanning the S-protein of SARS-CoV-2. Activation markers CD154 and CD137 were used for the assessment and quantification of S-protein–reactive T cells within CD3+ T cells. Expression levels of cytokines, interferon gamma (IFNγ), tumor necrosis factor alpha (TNFα), and interleukin 2 (IL-2), as well as the effector molecule granzyme B (GrB), were analyzed among activated CD4+ T cells by intracellular staining and flow cytometry after stimulation with OPPs spanning the whole S-protein of SARS-CoV-2. Differences between the subcohorts were analyzed using the paired, 2-sided t test. The significance threshold was set at 0.05. Box plots depict the median and first and third quartiles of a variable; the maximum length of the whiskers corresponds to 1.5× the interquartile range. (a) Titers of anti–SARS-CoV-2 S-protein–binding antibodies assessed by ELISA. (b) Neutralizing antibody titers for the SARS-CoV-2 S-protein. (c) Frequencies of S-reactive CD4+ T cells, as defined by CD154+CD137+ expression. (d) Frequencies of S-reactive CD8+ T cells, as defined by CD137 expression and cytokine production. (e) Frequencies of S-reactive follicular CD4+ T-helper cells, as defined by the expression of CXC chemokine receptor 5 (CXCR5). (f–i) Frequencies of S-reactive CD4+ T cells producing (f) GrB, (g) IFNγ, (h) IL2, and (i) TNFα. AU, arbitrary unit.