Unexpected Expression and Function of FcεRI in Immortalized Breast Cancer Cells: A Cautionary Null Study.

The high-affinity IgE receptor, FcεRI, is typically associated with type 2 effectors such as mast cells (MC). The relatively unique expression profile of FcεRI and accumulating evidence from pre-clinical and clinical settings, such as MC interactions with tumors, have led us to study MCs as a potential therapeutic target in breast cancer. Our work identified MCs interacting with tumor cells at primary sites using the 4T1 (BALB/c) adenocarcinoma model in vivo. However, this analysis was complicated by a surprising finding that the tumor cells intrinsically and strongly expressed FcεRI. We further studied the expression and function of FcεRI in breast cancer cells in vitro. The 4T1 cells expressed FcεRI to a level similar to mouse bone marrow-derived MC (BMMC). Additionally, two established breast cancer cultures derived from human T-47D cells, one estrogen-dependent (E3) and the other estrogen-withdrawn (EWD8), also expressed FcεRI with EWD8 cells showing the greatest abundance. Functional analyses indicated that IgE-mediated antigen stimulation did not elicit classic Ca2+ flux in breast cancer cells as seen in the respective species' MCs; however, FcεRI crosslinking could stimulate IL-6 production from the T-47D derivatives. Preliminary analysis of primary breast cancer biopsy datasets using R2: Genomics Analysis and Visualization Platform was discordant with our in vivo model and in vitro observations. Indeed, FcεRI mRNA abundance declined in metastatic breast cancers compared to non-cancerous breast tissue. Altogether, we report a previously unidentified and immunologically substantive difference between breast cancer models and human primary tumors. Investigators pursuing FcεRI-relevant therapeutics in this context should be aware of this translational barrier.

Berberine Delays Onset of Collagen-Induced Arthritis through T Cell Suppression.

There is evidence that berberine (BBR), a clinically relevant plant compound, ameliorates clinically apparent collagen-induced arthritis (CIA) in vivo. However, to date, there are no studies involving the use of BBR which explore its prophylactic potential in this model of rheumatoid arthritis (RA). The aim of this study was to determine if prophylactic BBR use during the preclinical phase of collagen-induced arthritis would delay arthritic symptom onset, and to characterize the cellular mechanism underlying such an effect. DBA/1J mice were injected with an emulsion of bovine type II collagen (CII) and complete Freund's adjuvant (day 0) and a booster injection of CII in incomplete Freund's adjuvant (day 18) to induce arthritis. Mice were then given i.p. injections of 1 mg/kg/day of BBR or PBS (vehicle with 0.01% DMSO) from days 0 to 28, were left untreated (CIA control), or were in a non-arthritic control group (n = 15 per group). Incidence of arthritis in BBR-treated mice was 50%, compared to 90% in both the CIA and PBS controls. Populations of B and T cells from the spleens and draining lymph nodes of mice were examined on day 14 (n = 5 per group) and day 28 (n = 10 per group). BBR-treated mice had significantly reduced populations of CD4+Th and CD4+CXCR5+ Tfh cells, and an increased proportion of Foxp3+ Treg at days 14 and 28, as well as reduced expression of co-stimulatory molecules CD28 and CD154 at both endpoints. The effect seen on T cell populations and co-stimulatory molecule expression in BBR-treated mice was not mirrored in CD19+ B cells. Additionally, BBR-treated mice experienced reduced anti-CII IgG2a and anti-CII total IgG serum concentrations. These results indicate a potential role for BBR as a prophylactic supplement for RA, and that its effect may be mediated specifically through T cell suppression. However, the cellular effector involved raises concern for BBR prophylactic use in the context of vaccine efficacy and other primary adaptive immune responses.

Beyond IgE: Alternative Mast Cell Activation Across Different Disease States.

Mast cells are often regarded through the lens of IgE-dependent reactions as a cell specialized only for anti-parasitic and type I hypersensitive responses. However, recently many researchers have begun to appreciate the expansive repertoire of stimuli that mast cells can respond to. After the characterization of the interleukin (IL)-33/suppression of tumorigenicity 2 (ST2) axis of mast cell activation-a pathway that is independent of the adaptive immune system-researchers are revisiting other stimuli to induce mast cell activation and/or subsequent degranulation independent of IgE. This discovery also underscores that mast cells act as important mediators in maintaining body wide homeostasis, especially through barrier defense, and can thus be the source of disease as well. Particularly in the gut, inflammatory bowel diseases (Crohn's disease, ulcerative colitis, etc.) are characterized with enhanced mast cell activity in the context of autoimmune disease. Mast cells show phenotypic differences based on tissue residency, which could manifest as different receptor expression profiles, allowing for unique mast cell responses (both IgE and non-IgE mediated) across varying tissues as well. This variety in receptor expression suggests mast cells respond differently, such as in the gut where immunosuppressive IL-10 stimulates the development of food allergy or in the lungs where transforming growth factor-ß1 (TGF-ß1) can enhance mast cell IL-6 production. Such differences in receptor expression illustrate the truly diverse effector capabilities of mast cells, and careful consideration must be given toward the phenotype of mast cells observed in vitro. Given mast cells' ubiquitous tissue presence and their capability to respond to a broad spectrum of non-IgE stimuli, it is expected that mast cells may also contribute to the progression of autoimmune disorders and other disease states such as metastatic cancer through promoting chronic inflammation in the local tissue microenvironment and ultimately polarizing toward a unique Th17 immune response. Furthermore, these interconnected, atypical activation pathways may crosstalk with IgE-mediated signaling differently across disorders such as parasitism, food allergies, and autoimmune disorders of the gut. In this review, we summarize recent research into familiar and novel pathways of mast cells activation and draw connections to clinical human disease.

Soluble transforming growth factor beta-1 enhances murine mast cell release of Interleukin 6 in IgE-independent and Interleukin 13 in IgE-dependent settings in vitro.

INTRODUCTION: For immune cells transforming growth factor beta-1 (TGF-ß1) can enhance or repress effector functions. Here, we characterize the effects of TGF-ß1 on IgE-mediated and IL-33-mediated activation of primary murine mast cells derived from hematopoietic stem cells (bone marrow derived mast cells; BMMC). We also investigated potential interactions between TGF-ß1 and stem cell factor (SCF). We conclude TGF-ß1 plays a selectively stimulatory role for mast cell cultures in vitro. METHODS: BMMCs from C57BL/6 mice were differentiated with IL-3 and then treated with TGF-ß1. BMMCs were exposed to TGF-ß1, primed with IgE, activated with antigen, and then IL-6 and IL-13 cytokine release was quantified using ELISA. Additionally, the effects of TGF-ß1 on both IgE and IL-33-mediated short term activation were observed via flow cytometric analysis of both surface LAMP-1 expression and intracellular IL-6. Receptor colocalization was visualized using fluorescence confocal microscopy and individual receptor expression levels were also quantified. RESULTS: Resting IL-6 production increased with TGF-ß1 but significance was lost following BMMC activation via IgE receptor (FcεRI) crosslinking. This was similar to a comparison effect due to SCF treatment alone, which also enhanced resting levels of IL-6. TGF-ß1 treatment enhanced release of IL-13 only with FcεRI-IgE-mediated activation. TGF-ß1 suppressed mobilization of IL-6 with short-term BMMC activation when stimulated with IL-33. Lastly, colocalization patterns of the SCF receptor (CD117) and FcεRI with IgE crosslinking were unaffected by TGF-ß1 treatment, but individual expression levels for FcεRI, CD117, and TGFßRII were all reduced following either IgE activation or TGF-ß1 treatment; this reduction was partially recovered in BMMCs that were both activated by IgE and treated with TGF-ß1. DISCUSSION: These data reveal a novel positive effect of soluble TGF-ß1 on mast cell activation in vitro, suggesting mast cells may be activated through a non-canonical pathway by TGF-ß1. Understanding this interaction will provide insight into the potential role of mast cells in settings where TGF-ß1 is produced in an aberrant manner, such as in and around high grade tumors.

Stronger sexual selection in warmer waters: the case of a sex role reversed pipefish.

In order to answer broader questions about sexual selection, one needs to measure selection on a wide array of phenotypic traits, simultaneously through space and time. Nevertheless, studies that simultaneously address temporal and spatial variation in reproduction are scarce. Here, we aimed to investigate the reproductive dynamics of a cold-water pipefish simultaneously through time (encompassing variation within each breeding cycle and as individuals grow) and space (by contrasting populations experiencing distinct water temperature regimes) in order to test hypothesized differences in sexual selection. Even though the sampled populations inhabited locations with very different water temperature regimes, they exhibited considerable similarities in reproductive parameters. The most striking was the existence of a well-defined substructure in reproductive activity, where larger individuals reproduce for longer periods, which seemed dependent on a high temperature threshold for breeding rather than on the low temperatures that vary heavily according to latitude. Furthermore, the perceived disparities among populations, such as size at first reproduction, female reproductive investment, or degree of sexual size dimorphism, seemed dependent on the interplay between seawater temperature and the operational sex ratio (OSR). Contrary to our expectations of an enhanced opportunity for sexual selection in the north, we found the opposite: higher female reproductive investment coupled with increased sexual size dimorphism in warmer waters, implying that a prolonged breeding season does not necessarily translate into reduced sexual selection pressure. In fact, if the limited sex has the ability to reproduce either continuously or recurrently during the entire breeding season, an increased opportunity for sexual selection might arise from the need to compete for available partners under strongly biased OSRs across protracted breeding seasons. A more general discussion on the effects of climate change in the pressure of sexual selection is also presented.