IL-7 improves neural cell-survival.

<p>(<b>A</b>) Bromodeoxyuridine (BrdU) treatment experimental procedure. (<b>B</b>) BrdU positive cells in the ARC of PBS- (□) or IL-7- (▪) treated mice. Animals were sacrificed one day after BrdU treatment (post-natal day 8 (P8)) for neural cell proliferation analysis (n = 5 per group), or 29 days after BrdU treatment (P36) for neural cell survival analysis (n = 5 per group). Data were represented as mean ± SEM of BrdU-positive cells within the ARC per animal and per slice. *<i>p</i><0.05.</p

IL-7 modulates hypothalamic neuropeptide mRNA gene expression.

<p>PBS ( = 5, □) or IL-7 (n = 5, ▪) was i.p. injected in 2-month-old C57BL/6 mice. The expression of hypothalamic neuropeptides was assessed by real-time PCR, in fed (<b>A</b>) or in re-fed (<b>B</b>) condition. The effects of IL-7 treatment were evaluated by calculating the relative expression levels as follows: 2^ΔCt (ΔCt = mean Ct genes of interest - mean Ct GAPDH), using the raw cycle-threshold (Ct) values. *<i>p</i><0.05, *** <i>p</i><0.001. POMC: pro-opiomelanocortin, CART: cocaine-amphetamine related peptide, NP-Y: neuropeptide-Y, AgRP: Agouti-related peptide.</p

IL-7 activates hypothalamic cells and IL-7 receptor is expressed on hypothalamic cells.

<p>(<b>A</b>) Representative microphotographs of Fos expression induced by peripheral IL-7 injection in C57BL/6 mice. Fos-immunoreactive cells in arcuate nucleus (ARC) in PBS-treated and IL-7-treated mice. Arrows indicate Fos-imunoreactive cells. 3^rd V: third ventricule; VMH: ventromedial hypothalamic nucleus. Scale bars represent 100 µm. (<b>B</b>) mRNA expression of IL-7Rα and γ_c chains in the whole hypothalamus of 2-month-old C57BL/6 mice (n = 3). Lanes (1, 2, 3) represent one animal. The - lane represents the negative control <i>i</i>.<i>e</i>. without retro-transcription. (<b>C</b>) Representative microphotographs of frontal hypothalamic frozen sections from 2-month-old mice showing fluorescent immunostaining of IL-7Rα and γ_c chains in the arcuate nucleus. Frozen section from IL-7Rα KO were used as control for the IL-7Rα specific staining. ARC: arcuate nucleus, ME: median eminence, 3^rd V: third ventricule; VMH: ventromedial hypothalamic nucleus. (<b>D</b>) Mean number of phosphorylated (p)-STAT5 and p-STAT3 immunoreactive (IR) cells in the hypothalamic arcuate nucleus of C57BL/6 mice injected with recombinant IL-7 (▪; n = 5) or with PBS (□; n = 5). Results are expressed as mean of immunoreactive cells ±SEM per animal and per slice. *<i>p</i><0.05. (<b>E</b>) Representative microphotographs of p-STAT5 and p-STAT3-IR cells in the ARC in PBS-treated and IL-7-treated mice. Scale bar represent 100 µm. ARC: arcuate hypothalamic nucleus; ME: median eminence; 3rd: third ventricle.</p

Role of c-Jun-target genes in infected macrophage adhesion to fibronectin.

<p><b>A</b>, Infected macrophage adhesion is downregulated in the vaccine line (V-Delta 169) compared to virulent (V) Jed 4. RT-qPCR determined levels of genes (<i>cd69</i>, <i>icam-1</i>, <i>cd49</i> and <i>itgb5</i>) potentially involved in mediating adhesion. <b>B</b>, Virulent Jed 4 binds at high density to fibronectin compared to the V-Delta 169 vaccine line; 70% of fibronectin binding capacity is lost upon ablation of c-Jun.</p

Primer sequences used for real-time PCR.

<p>Primer sequences used for real-time PCR.</p

Dissemination of Jed 4 macrophages in Rag2/γC mice.

<p><b>A (Virulent line) & B (Engineered attenuated line)</b>, The Ts4 micro-satellite locus was amplified from DNA extracted from the lungs (tracks 1–7), kidneys (tracks 8–11) and spleens (tracks 12–18) of 18 mice injected with virulent Jed 4 macrophages. Apart from a Ts4 specific band in the positive control no parasite DNA was amplified from extracts of lungs (tracks 1–8), kidneys (tracks 9–11) and spleens (tracks 12–18) of 18 mice injected with V-Delta 169 macrophages. (+) = Positive control DNA, (−) = negative control water, MW = molecular-weight size marker (200 bp ladder).</p

Suppression of c-Jun leads to down-regulation of AP-1 activity.

<p><b>A</b>, Flag-tagged dominant-negative mutant of c-Jun, lacking the first 169 amino acid transactivation domain (Flag-Δ169) was introduced into <i>T. annulata</i>-infected macrophages. <b>B</b>, Suppression of c-Jun reduces AP-1 activity in <i>T. annulata</i>-infected macrophages, as measured by 3XTRE-driven luciferase activity (TRE-luc). The activity of AP-1 is 20 times lower in the vaccine line (V-Delta 169) compared to the control line (V). <b>C</b>, Western blot analysis of the expression of two different AP-1 family members in vaccine and control lines showing that expression of c-Jun is decreased and ATF2 increased in vaccine line. In spite of an increase in ATF2 protein the level of phospho-ATF2 (p-ATF2) is markedly reduced. Protein amounts were compared to actin and decreases in p-ATF2 and c-Jun quantified using Image J (C1 and C2).</p

(A) mice were treated with DSS in the drinking water for 6 d, followed by water for 10 d

This protocol was repeated for a total of three cycles. After the last cycle, cells isolated from the colon were restimulated in vitro with PMA/ionomycin for 5 h and subjected to intracellular staining for GFP, IL-17, and Foxp3. Histograms (from left to right) report percent GFPTCR-β cell subsets in total T cells (see ), total numbers of RORγt Tαβ cells present in the organ, and the ratio of IL-17–producing to Foxp3 cells within RORγt Tαβ cells (see ). Right panels show immunofluorescence histology of a colon from a healthy or a treated mouse. Bar, 100 μm. (B) mice were infected intranasally with 100 PFUs of influenza A virus for 7 d. Cells were then isolated from the lung and processed as in A. Right panels show immunofluorescence histology of a lung from healthy or an infected mouse. Bar, 50 μm. (C) Cells were isolated from the mesenteric LNs of a 4-mo-old × mouse and processed as in A. Right panels show immunofluorescence histology of a mesenteric LN from a normal or a tumor-bearing mouse. Bar, 100 μm. Data shown are representative of at least three independent experiments. Three to four mice were analyzed per group. *, P < 0.05 as compared with control (mock-treated or WT mice).<p><b>Copyright information:</b></p><p>Taken from "In vivo equilibrium of proinflammatory IL-17 and regulatory IL-10 Foxp3 RORγt T cells"</p><p></p><p>The Journal of Experimental Medicine 2008;205(6):1381-1393.</p><p>Published online 9 Jun 2008</p><p>PMCID:PMC2413035.</p><p></p

MACS-sorted naive (CD62L) CD4 T cells from the spleens of mice were stimulated in duplicates with anti-CD3 and anti-CD28 in the presence of blocking anti–IFN-γ and anti–IL-4 antibodies and the indicated cytokines or RA

After different periods of time, cells were restimulated with PMA/ionomycin for 5 h and analyzed by flow cytometry for the expression of GFP, Foxp3, IL-17, and IL-10. All plots are gated on TCR-β cells, except plots for IL-10 that are gated on GFPTCR-β cells. Numbers indicate percent cells in quadrants. Data are representative of three independent experiments.<p><b>Copyright information:</b></p><p>Taken from "In vivo equilibrium of proinflammatory IL-17 and regulatory IL-10 Foxp3 RORγt T cells"</p><p></p><p>The Journal of Experimental Medicine 2008;205(6):1381-1393.</p><p>Published online 9 Jun 2008</p><p>PMCID:PMC2413035.</p><p></p

A Novel Mouse Model for Stable Engraftment of a Human Immune System and Human Hepatocytes

<div><p>Hepatic infections by hepatitis B virus (HBV), hepatitis C virus (HCV) and <i>Plasmodium</i> parasites leading to acute or chronic diseases constitute a global health challenge. The species tropism of these hepatotropic pathogens is restricted to chimpanzees and humans, thus model systems to study their pathological mechanisms are severely limited. Although these pathogens infect hepatocytes, disease pathology is intimately related to the degree and quality of the immune response. As a first step to decipher the immune response to infected hepatocytes, we developed an animal model harboring both a human immune system (HIS) and human hepatocytes (HUHEP) in BALB/c Rag2^-/- IL-2Rγc^-/- NOD.<i>sirpa</i> uPA^tg/tg mice. The extent and kinetics of human hepatocyte engraftment were similar between HUHEP and HIS-HUHEP mice. Transplanted human hepatocytes were polarized and mature <i>in vivo</i>, resulting in 20–50% liver chimerism in these models. Human myeloid and lymphoid cell lineages developed at similar frequencies in HIS and HIS-HUHEP mice, and splenic and hepatic compartments were humanized with mature B cells, NK cells and naïve T cells, as well as monocytes and dendritic cells. Taken together, these results demonstrate that HIS-HUHEP mice can be stably (> 5 months) and robustly engrafted with a humanized immune system and chimeric human liver. This novel HIS-HUHEP model provides a platform to investigate human immune responses against hepatotropic pathogens and to test novel drug strategies or vaccine candidates.</p></div