Conventional and regulatory CD4+ T cells that share identical TCRs are derived from common clones

Results from studies comparing the diversity and specificity of the TCR repertoires expressed by conventional (Tconv) and regulatory (Treg) CD4+ T cell have varied depending on the experimental system employed. We developed a new model in which T cells express a single fixed TCRα chain, randomly rearranged endogenous TCRβ chains, and a Foxp3-GFP reporter. We purified CD4+Foxp3- and CD4+Foxp3+ cells, then performed biased controlled multiplex PCR and high throughput sequencing of endogenous TCRβ chains. We identified >7,000 different TCRβ sequences in the periphery of 5 individual mice. On average, ~12% of TCR sequences were expressed by both conventional and regulatory populations within individual mice. The CD4+ T cells that expressed shared TCR sequences were present at higher frequencies compared to T cells expressing non-shared TCRs. Furthermore, nearly all (>90%) of the TCR sequences that were shared within mice were identical at the DNA sequence level, indicating that conventional and regulatory T cells that express shared TCRs are derived from common clones. Analysis of TCR repertoire overlap in the thymus reveals that a large proportion of Tconv and Treg sharing observed in the periphery is due to clonal expansion in the thymus. Together these data show that there are a limited number of TCR sequences shared between Tconv and Tregs. Also, Tconv and Tregs sharing identical TCRs are found at relatively high frequencies and are derived from common progenitors, of which a large portion are generated in the thymus

Asteroid Distributions in the Ecliptic

We present analysis of the asteroid surface density distribution of main belt asteroids (mean perihelion $\Delta \simeq 2.404$ AU) in five ecliptic latitude fields, -17 \gtsimeq \beta(\degr) \ltsimeq +15, derived from deep \textit{Large Binocular Telescope} (LBT) $V-$band (85% completeness limit $V = 21.3$ mag) and \textit{Spitzer Space Telescope} IRAC 8.0 \micron (80% completeness limit $\sim 103 \mu$Jy) fields enabling us to probe the 0.5--1.0 km diameter asteroid population. We discovered 58 new asteroids in the optical survey as well as 41 new bodies in the \textit{Spitzer} fields. The derived power law slopes of the number of asteroids per square degree are similar within each $\sim 5$\degr{} ecliptic latitude bin with a mean value of $-0.111 \pm 0.077$. For the 23 known asteroids detected in all four IRAC channels mean albedos range from $0.24 \pm 0.07$ to $0.10 \pm 0.05$. No low albedo asteroids ($p_{V}$ \ltsimeq 0.1) were detected in the \textit{Spitzer} FLS fields, whereas in the SWIRE fields they are frequent. The SWIRE data clearly samples asteroids in the middle and outer belts providing the first estimates of these km-sized asteroids' albedos. Our observed asteroid number densities at optical wavelengths are generally consistent with those derived from the Standard Asteroid Model within the ecliptic plane. However, we find an over density at \beta \gtsimeq 5\degr{} in our optical fields, while the infrared number densities are under dense by factors of 2 to 3 at all ecliptic latitudes.Comment: 35 pages including 5 figures, accepted to The Astronomical Journa

Quantifying Ca 2+ Current and Permeability in ATP-gated P2X7 Receptors

International audienceATP-gated P2X7 receptors are prominently expressed in inflammatory cells and play a key role in the immune response. A major consequence of receptor activation is the regulated influx of Ca(2+) through the self-contained cation non-selective channel. Although the physiological importance of the resulting rise in intracellular Ca(2+) is universally acknowledged, the biophysics of the Ca(2+) flux responsible for the effects are poorly understood, largely because traditional methods of measuring Ca(2+) permeability are difficult to apply to P2X7 receptors. Here we use an alternative approach, called dye-overload patch-clamp photometry, to quantify the agonist-gated Ca(2+) flux of recombinant P2X7 receptors of dog, guinea pig, human, monkey, mouse, rat, and zebrafish. We find that the magnitude of the Ca(2+) component of the ATP-gated current depends on the species of origin, the splice variant, and the concentration of the purinergic agonist. We also measured a significant contribution of Ca(2+) to the agonist-gated current of the native P2X7Rs of mouse and human immune cells. Our results provide cross-species quantitative measures of the Ca(2+) current of the P2X7 receptor for the first time, and suggest that the cytoplasmic N terminus plays a meaningful role in regulating the flow of Ca(2+) through the channel

Osteoclast Activated FoxP3+ CD8+ T-Cells Suppress Bone Resorption in vitro

BACKGROUND: Osteoclasts are the body's sole bone resorbing cells. Cytokines produced by pro-inflammatory effector T-cells (T(EFF)) increase bone resorption by osteoclasts. Prolonged exposure to the T(EFF) produced cytokines leads to bone erosion diseases such as osteoporosis and rheumatoid arthritis. The crosstalk between T-cells and osteoclasts has been termed osteoimmunology. We have previously shown that under non-inflammatory conditions, murine osteoclasts can recruit naïve CD8 T-cells and activate these T-cells to induce CD25 and FoxP3 (Tc(REG)). The activation of CD8 T-cells by osteoclasts also induced the cytokines IL-2, IL-6, IL-10 and IFN-γ. Individually, these cytokines can activate or suppress osteoclast resorption. PRINCIPAL FINDINGS: To determine the net effect of Tc(REG) on osteoclast activity we used a number of in vitro assays. We found that Tc(REG) can potently and directly suppress bone resorption by osteoclasts. Tc(REG) could suppress osteoclast differentiation and resorption by mature osteoclasts, but did not affect their survival. Additionally, we showed that Tc(REG) suppress cytoskeletal reorganization in mature osteoclasts. Whereas induction of Tc(REG) by osteoclasts is antigen-dependent, suppression of osteoclasts by Tc(REG) does not require antigen or re-stimulation. We demonstrated that antibody blockade of IL-6, IL-10 or IFN-γ relieved suppression. The suppression did not require direct contact between the Tc(REG) and osteoclasts. SIGNIFICANCE: We have determined that osteoclast-induced Tc(REG) can suppress osteoclast activity, forming a negative feedback system. As the CD8 T-cells are activated in the absence of inflammatory signals, these observations suggest that this regulatory loop may play a role in regulating skeletal homeostasis. Our results provide the first documentation of suppression of osteoclast activity by CD8 regulatory T-cells and thus, extend the purview of osteoimmunology

CD4+ T-cell development in a mouse expressing a transgenic TCR derived from a Treg

CD4(+)Foxp3(+) regulatory T cells (Tregs) maintain peripheral tolerance and influence immune responses to foreign antigens. The thymus is an important source of Tregs, but controversy exists as to whether T cells are selected into the Treg lineage based on signals received through TCRs specific for self-peptides. To examine the specificity of TCRs expressed by Tregs and its effect on CD4(+) T cell development, we generated Treg-TCR transgenic mice. Deletion of >90% of CD4(+) T cells in RAG sufficient mice, and nearly 100% deletion in RAG(−/−) mice expressing this TCR indicate that the TCR is specific for an unknown, naturally expressed peptide in the thymus. Deletion occurs late in development, suggesting this peptide is presented by APCs in the thymic medulla. These studies are the first to describe the effects of expressing a Treg-TCR on CD4(+) T cell development. The implications of our data for models of Treg selection are discussed

Regulation of Gastric Carcinogenesis by Inflammatory CytokinesSummary

Chronic inflammation caused by infection with Helicobacter pylori and autoimmune gastritis increases an individualâs risk of developing gastric cancer. More than 90% of gastric cancers are adenocarcinomas, which originate from epithelial cells in the chronically inflamed gastric mucosa. However, only a small subset of chronic gastritis patients develops gastric cancer, implying a role for genetic and environmental factors in cancer development. A number of DNA polymorphisms that increase gastric cancer risk have mapped to genes encoding cytokines. Many different cytokines secreted by immune cells and epithelial cells during chronic gastritis have been identified, but a better understanding of how cytokines regulate the severity of gastritis, epithelial cell changes, and neoplastic transformation is needed. This review summarizes studies in both human and mouse models, describing a number of different findings that implicate various cytokines in regulating the development of gastric cancer. Keywords: Gastric Cancer, Inflammation, Cytokine

Cutting Edge: The Relative Distribution of T Cells Responding to Chemically Dominant or Minor Epitopes of Lysozyme Is Not Affected by CD40-CD40 Ligand and B7-CD28-CTLA-4 Costimulatory Pathways

Abstract We examined the frequencies and specificities of the CD4+ T cell responses to the protein hen egg white lysozyme in mice deficient in the CD40-CD40 ligand or B7-CD28 costimulatory pathways. The frequency of T cells was decreased by between 3- and 4-fold in CD40−/− mice, and 12-fold in B7-1/B7-2−/− mice, but surprisingly, the relative distribution of T cells responding to peptides that were presented at levels that differed by &amp;gt;250-fold was similar. We also examined the CD4 response after blocking the regulatory molecule CTLA-4 during immunization. We observed no difference in either the frequency or specificity of the CD4+ T cell response if CTLA-4 was blocking during priming. Thus, the T cell response was generated toward the constellation of chemically dominant and subdominant epitopes as a whole, and did not discriminate among them based on their relative abundance.</jats:p