The endocrine system dynamically controls tissue differentiation and homeostasis, but has not been studied using dynamic tissue culture paradigms. Here we show that a microfluidic system supports murine ovarian follicles to produce the human 28-day menstrual cycle hormone profile, which controls human female reproductive tract and peripheral tissue dynamics in single, dual and multiple unit microfluidic platforms (Solo-MFP, Duet-MFP and Quintet-MPF, respectively). These systems simulate the in vivo female reproductive tract and the endocrine loops between organ modules for the ovary, fallopian tube, uterus, cervix and liver, with a sustained circulating flow between all tissues. The reproductive tract tissues and peripheral organs integrated into a microfluidic platform, termed EVATAR, represents a powerful new in vitro tool that allows organ–organ integration of hormonal signalling as a phenocopy of menstrual cycle and pregnancy-like endocrine loops and has great potential to be used in drug discovery and toxicology studies.

Highlights d Ovarian tumors separate into two groups: high T and low T cell infiltration groups d Granulysin-expressing CD4 + T cells are present in the high T infiltration cell group d MKI67-expressing plasmablasts are identified in high T cell infiltration tumors d Correlation of CD8 and Tox in this study with immunoreactive subtype in TCGA data

Clinical efficacy in the treatment of rheumatoid arthritis with anti-CD20 (Rituximab)-mediated B-cell depletion has garnered interest in the mechanisms by which B cells contribute to autoimmunity. We have reported that B-cell depletion in a murine model of proteoglycan-induced arthritis (PGIA) leads to an increase in regulatory T (Treg) cells that correlates with decreased autoreactivity. Here, we demonstrate that the increase in Treg cells after B-cell depletion is due to an increase in the differentiation of naïve CD4+ T cells into Treg cells. Since the development of PGIA is dependent on IFN-γ and B cells are reported to produce IFN-γ, we hypothesized that B-cell-specific IFN-γ plays a role in the development of PGIA. Accordingly, mice with B-cell-specific IFN-γ-deficiency were as resistant to the induction of PGIA as mice that were completely IFN-γ-deficient. Importantly, despite a normal frequency of IFN-γ-producing CD4+ T cells, B-cell-specific IFN-γ-deficient mice exhibited a higher percentage of Treg cells compared with that in wild type (WT) mice. These data indicate that B-cell IFN-γ production inhibits Treg-cell differentiation and exacerbates arthritis. Thus, we have established that IFN-γ, specifically derived from B cells, uniquely contributes to the pathogenesis of autoimmunity through prevention of immunoregulatory mechanisms.

Three-dimensional (3D) in vitro models have been established to study the physiology and pathophysiology of the endometrium. With emerging evidence that the native extracellular matrix (ECM) provides appropriate cues and growth factors essential for tissue homeostasis, we describe, a novel 3D endometrium in vitro model developed from decellularized human endometrial tissue repopulated with primary endometrial cells. Analysis of the decellularized endometrium using mass spectrometry revealed an enrichment of cell adhesion molecules, cytoskeletal proteins, and ECM proteins such as collagen IV and laminin. Primary endometrial cells within the recellularized scaffolds proliferated and remained viable for an extended period of time in vitro. In order to evaluate the hormonal response of cells within the scaffolds, the recellularized scaffolds were treated with a modified 28-day hormone regimen to mimic the human menstrual cycle. At the end of 28 days, the cells within the endometrial scaffold expressed both estrogen and progesterone receptors. In addition, decidualization markers, IGFBP-1 and prolactin, were secreted upon addition of dibutyryl cyclic AMP indicative of a decidualization response. This 3D model of the endometrium provides a new experimental tool to study endometrial biology and drug testing. Summary SentencePrimary endometrial cells within a recellularized scaffold respond to a 28-day menstrual cycle.

T helper (Th) cytokines IFN-γ and IL-17 are linked to the development of autoimmune disease. In models of rheumatoid arthritis (RA) i.e. proteoglycan (PG) -induced arthritis (PGIA), IFN-γ is required whereas in collagen-induced arthritis, IL-17 is necessary for development of arthritis. Here we show that the route of immunization determines the requirement for either IFN-γ or IL-17 in arthritis. Intraperitoneal (i.p.) immunization with PG induces a CD4+ T cell IFN-γ response with little IL-17 in the spleen and peripheral lymph nodes. However, s.c. immunization induces both an IFN-γ and an IL-17 CD4+ T cell response in spleen and LNs. The failure to induce a CD4+ T cell IL-17 response after i.p. immunization is associated with T cell priming as naïve T cells activated in vitro were fully capable of producing IL-17. Moreover, PGIA is converted from an IFN-γ to an IL-17-mediated disease by altering the route of immunization from i.p. to s.c. The histological appearance of joint inflammation (cellular inflammation and bone erosion) are similar in the i.p. versus s.c. immunized mice despite the presence of CD4+ T cells producing IL-17 in joint tissues only after s.c. immunization. These data indicate a critical role for the site of initial T cell priming and the Th cytokines required for susceptibility to arthritis. Our findings suggest that T cell activation at different anatomical sites in RA patients may skew the T cells towards production of either IFN-γ or IL-17.