Decoding Immune Cell Doublets in Humans: From Artifacts to Insights

Although routinely detected in human blood and tissue samples, immune cell doublets are typically discarded during data acquisition or analysis, often dismissed as technical artifacts arising from ex vivo cell handling. However, emerging evidence suggests that these immune cell doublets reflect biologically meaningful interactions, offering critical insights into immune communication during health and disease. In this review, we evaluate the current evidence supporting the existence of biological immune cell doublets across various human tissues, highlighting their diverse phenotypes and functions that distinguish them from technical noise. We explore how existing single-cell datasets and analytical frameworks can be repurposed to study doublets, and we examine the primary challenge in the field, which is the lack of definitive markers to distinguish biological doublets from artifacts. Finally, we examine how novel technologies, namely high-throughput image-enabled sorting, are facilitating the next generation of research on this topic.