Background: Biosynthesis and metabolism of nicotinic acid adenine dinucleotide phosphate (NAADP) are unclear. Results: Alkaline phosphatase (AP) was identified and characterized as enzyme involved in NAADP degradation. Conclusion: In cells not expressing CD38, AP is a valid candidate enzyme for degradation of NAADP to nicotinic acid adenine dinucleotide. Significance: This is the first evidence for identity of the enzyme degrading NAADP in cells not expressing CD38.

Objectives Potential differences in the breadth, distribution and magnitude of CD4 + T‐cell responses directed against the SARS‐CoV‐2 spike glycoprotein between vaccinees, COVID‐19 patients and subjects who experienced both ways of immunisation have not been comprehensively compared on a peptide level. Methods Following virus‐specific in vitro cultivation, we determined the T‐cell responses directed against 253 individual overlapping 15‐mer peptides covering the entire SARS‐CoV‐2 spike glycoprotein using IFN‐γ ELISpot and intracellular cytokine staining. In vitro HLA binding was determined for selected peptides. Results We mapped 955 single peptide‐specific CD4 + T‐cell responses in a cohort of COVID‐19 patients ( n = 8), uninfected vaccinees ( n = 16) and individuals who experienced both infection and vaccination ( n = 11). Patients and vaccinees (two‐time and three‐time vaccinees alike) had a comparable number of CD4 + T‐cell responses (median 26 vs. 29, P = 0.7289). Most of these specificities were conserved in B.1.1.529 and the BA.4 and BA.5 sublineages. The highest magnitude of these in vitro IFN‐γ CD4 + T‐cell responses was observed in COVID‐19 patients (median 0.35%), and three‐time vaccinees showed a higher magnitude than two‐time vaccinees (median 0.091% vs. 0.175%, P < 0.0001). Twelve peptide specificities were each detected in at least 40% of subjects. In vitro HLA binding showed promiscuous presentation by DRB1 molecules for several peptides. Conclusion Both SARS‐CoV‐2 infection and vaccination prime broadly directed T‐cell responses directed against the SARS‐CoV‐2 spike glycoprotein. This comprehensive high‐resolution analysis of spike peptide specificities will be a useful resource for further investigation of spike‐specific T‐cell responses.