IL22 Regulates Human Urothelial Cell Sensory and Innate Functions through Modulation of the Acetylcholine Response, Immunoregulatory Cytokines and Antimicrobial Peptides: Assessment of an In Vitro Model

Abstract

Human urinary disorders are generally studied in rodent models due to limitations of functional in vitro culture models of primary human urothelial cells (HUCs). Current HUC culture models are often derived from immortalized cancer cell lines, which likely have functional characteristics differ from healthy human urothelium. Here, we described a simple explant culture technique to generate HUCs and assessed their in vitro functions. Using transmission electron microscopy, we assessed morphology and heterogeneity of the generated HUCs and characterized their intercellular membrane structural proteins relative to ex vivo urothelium tissue. We demonstrated that our cultured HUCs are free of fibroblasts. They are also heterogeneous, containing cells characteristic of both immature basal cells and mature superficial urothelial cells. The cultured HUCs expressed muscarinic receptors (MR1 and MR2), carnitine acetyltransferase (CarAT), immunoregulatory cytokines IL7, IL15, and IL23, as well as the chemokine CCL20. HUCs also expressed epithelial cell-specific molecules essential for forming intercellular structures that maintain the functional capacity to form the physiological barrier of the human bladder urothelium. A subset of HUCs, identified by the high expression of CD44, expressed the Toll-like receptor 4 (TLR4) along with its co-receptor CD14. We demonstrated that HUCs express, at the mRNA level, both forms of the IL22 receptor, the membrane-associated (IL22RA1) and the secreted soluble (IL22RA2) forms; in turn, IL22 inhibited expression of MR1 and induced expression of CarAT and two antimicrobial peptides (S100A9 and lipocalin-2). While the cellular sources of IL22 have yet to be identified, the HUC cytokine and chemokine profiles support the concept that IL22-producing cells are present in the human bladder mucosa tissue and that IL22 plays a regulatory role in HUC functions. Thus, the described explant technique is clearly capable of generating functional HUCs suitable for the study of human urinary tract disorders, including interactions between urothelium and IL22-producing cells.

Female urinary microbiota

“…Understanding the roles these peptides play in the lower urinary tract requires significantly more study; however, they are likely to play a key role in microbial community regulation, resilience following dysbiotic/infection episodes and response to treatment for lower urinary tract disorders. Similarly, the recent detection of IL-22 receptors in the urothelium allows consideration of interactions between the microbial community, known urothelial cholinergic receptors, the non-barrier role of the urothelium, and AMPs [23]. Interpretation of these interactions may provide new insights into important immune functions of the bladder.…”

The urinary microbiota: a paradigm shift for bladder disorders?

“…They also determined that AMP activity was significantly more robust in individuals whose day-of-surgery standard urine cultures were positive for typical uropathogens or in individuals whose standard urine cultures became positive following surgery [16]. In an attempt to understand mechanism, another team reported that the cytokine interleukin-22 regulates human urothelial cell sensory and innate functions by modulating AMP synthesis [25]. Given the new knowledge that the FUM exists and the detection of AMPs within the adult urinary tract [26], it will be essential to clarify the physiologic role of urinary AMPs in health and disease.…”

Correlation of Colony Lactobacillus spp. with The Incident of Overactive Bladder with OBSS Score at Pauh Primary Health Center Padang

“…and Streptococcus is a genus that most widely reported as urinal microbiota and exist in the whole study that published this moment as an acid lactate bacteria genus that related with some body tissue especially urinary tract where they play a role as aprotector against pathogens. 12 The correlation between the amount of Lactobacillus spp colony with overactive bladder events with statisticstests done in this study obtained the result was not significant. This events could due by other factor that cause the occurrence of OAB as myocyte damage in the destrusor muscle, damage in inhibitor path in brain and marrow spine or damage insensitivity terminal afferents in biological urinary, UTI, estrogen deficiency and systemic abnormalities.…”