A variety of human skin equivalents (HSEs) has been designed for clinical use or for exploratory skin research. In vitro HSE models have been used to target relationships between the skin and nervous or immune systems but have not yet considered the neuro-immuno-cutaneous (NIC) system. In this study, HSEs are described, with and without neural and immune components, to discern these types of effects. These systems are composed of only primary human cells and contain an epidermis, dermis, hypodermis (with immune cells), and human induced neural stem cells for the neuronal component. RNA sequencing is utilized to confirm differences between sample groups and to identify unique or important genes with respect to sample type. Only samples with both neural and immune components result in the upregulation of genes in all the key biological pathways explored. The analysis of protein secretion confirms that this group has measurable functions related to all key cell types. Overall, this novel skin tissue system confirms that designing HSEs that include the NIC system results in a tissue model that reflects key functions. These systems could be used to identify selected targets of interest in skin research related to healthy or diseased states.