Immune context unmasks regulatory effects of Neanderthal and Denisovan introgression

Neanderthal and Denisovan introgression have left a pervasive footprint in the human genome, yet its regulatory consequences remain poorly understood. Here we use a massively parallel reporter assay to characterize the cis-regulatory activity of 4,161 high-frequency introgressed variants across respiratory (A549), hepatic (HepG2), and hematopoietic (K562) cells exposed to immune and infectious stimuli. We find that ~18% of variants show differential activity between archaic and modern alleles, including 94 whose effects are revealed or modulated by stimulation, often in a cell type-specific manner. We identify loci, including STAT2, IL23A, and RNF41, where clusters of introgressed alleles exert coordinated regulatory effects consistent with adaptive programs. Finally, we dissect the mechanisms underlying the association between Neanderthal introgression and COVID-19 severity and show that the risk allele rs17713054-A, which displays the strongest effect in our assay, increases activity of a TNF--responsive enhancer in lung epithelial cells, directly upregulating SLC6A20.Together, these findings reveal widespread context-dependent regulatory effects of archaic introgression, with broad evolutionary and biomedical implications.