PT  - JOURNAL ARTICLE
AU  - Seth Currlin
AU  - Harry S. Nick
AU  - Marda Jorgensen
AU  - Jerelyn A. Nick
AU  - Maigan A. Brusko
AU  - Hunter Hakimian
AU  - Jesus Penaloza-Aponte
AU  - Natalie Rodriguez
AU  - Miguel Medina-Serpas
AU  - Mingder Yang
AU  - Irina Kusmartseva
AU  - Todd M. Brusko
AU  - Kevin Otto
AU  - Amanda L. Posgai
AU  - Clive H. Wasserfall
AU  - Mark A. Atkinson
TI  - 3D-mapping of human lymph node and spleen reveals integrated neuronal, vascular, and ductal cell networks
AID  - 10.1101/2021.10.20.465151
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.10.20.465151
4099  - http://biorxiv.org/content/early/2021/10/21/2021.10.20.465151.short
4100  - http://biorxiv.org/content/early/2021/10/21/2021.10.20.465151.full
AB  - As secondary lymphoid organs, the spleen and lymph node represent important hubs for both innate and adaptive immunity1,2. Neuroanatomical and tracing data, largely derived from rodents, suggest that lymph nodes contain sensory and sympathetic innervation3,4, whereas the spleen contains postganglionic sympathetic innervation5, with conflicting views regarding the existence of cholinergic or vagal innervation6. Herein, we map the neuronal, vascular, and sinus cell networks from human spleen and lymph node using highly multiplexed CODEX (CO-Detection by indEXing) and 3D light sheet microscopy of cleared tissues. These data demonstrate striking delineation of two distinct layers within the lymph node subcapsular sinus—the ceiling defined by Podoplanin expression and floor by LYVE1, which overlays the lymph node follicles. Within the lymph node interior, we observed a mesh-like vessel network innervated with GAP43 and β3-tubulin. Dense perivascular innervation occurred in both tissues, including a subset of axonal processes expressing choline acetyl transferase (ChAT). Four neuronal markers (ChAT, PGP9.5, tyrosine hydroxylase, and β3-tubulin) localized to the arterial tunica externa suggest expression in the nervi vasorum while GAP43 was expressed within the internal elastic membrane of arteries. These data represent highly novel 3D visualization of perivascular and periductal autonomic innervation within these two key human organs.Competing Interest StatementThe authors have declared no competing interest.