Polycystic Kidney Disease:The Cyst-ematic Destruction of Renal Function

Polycystic kidney disease is the most common genetic, life-threatening disease, affecting more than 12.5 million people worldwide. Fluid-filled renal cysts that eventually destroy renal tissue and renal function altogether are characteristic of polycystic kidney disease. The autosomal dominant form of the disease which is also the most common form, ADPKD, is linked to mutations in the genes PKD1 and PKD2. The complete normal function of PKD1 and PKD2 is unknown, but most research suggests that they play some role in cell signaling and controlling the cell cycle. The diseased phenotype is thought to be caused by mutations in these genes that cause misregulation of the cell cycle leading to proliferation. The recessive form of polycystic kidney disease, ARPKD, is triggered by a mutation in the gene PKHD1 and is manifested more severely than the dominant form. ARPKD has not been as widely studied as ADPKD because it affects fewer people than the dominant form of the disease. Currently, there is no treatment for any of the forms of PKD, though some studies have shown some promise in effectively attenuating renal cystic growth

IL-10 Dysregulation Underlies Chemokine Insufficiency, Delayed Macrophage Response, and Impaired Healing in Diabetic Wounds.

Persistent inflammation is a major contributor to healing impairment in diabetic chronic wounds. Paradoxically, diabetic wound environment during the acute phase of healing is completely different because it exhibits a reduced macrophage response owing to inadequate expression of CCL2 proinflammatory cytokine. What causes a reduction in CCL2 expression in diabetic wounds early after injury remains unknown. In this study, we report that in contrast to prolonged exposure to high glucose, which makes monocytes proinflammatory, short-term exposure to high glucose causes a rapid monocyte reprogramming, manifested by increased expression and secretion of IL-10, which in an autocrine/paracrine fashion reduces glucose uptake and transforms monocytes into an anti-inflammatory phenotype by dampening signaling through toll-like receptors. We show that IL-10 expression is significantly increased in diabetic wounds during the acute phase of healing, causing significant reductions in toll-like receptor signaling and proinflammatory cytokine production, delaying macrophage and leukocyte responses, and underlying healing impairment in diabetic wounds. Importantly, blocking IL-10 signaling during the acute phase of healing improves toll-like receptor signaling, increases proinflammatory cytokine production, enhances macrophage and leukocyte responses, and stimulates healing in diabetic wounds. We posit that anti-IL-10 strategies have therapeutic potential if added topically after surgical debridement, which resets chronic wounds into acute fresh wounds

CD32 is enriched on CD4dimCD8bright T cells.

CD4dimCD8bright T cells, a genuine population of CD8+ T cells, are highly activated and cytolytic. Recently, the low affinity IgG Fc fragment receptor CD32a was described as marker of HIV latency while others reported that CD32a is associated with T cell activation. Given that we have previously established that CD4dimCD8bright T cells are highly activated, mediate anti-HIV responses, and are infected by HIV, we assessed here CD32 expression on CD4dimCD8bright T cells in context of HIV. CD32 frequency on peripheral CD4dimCD8bright and CD4+ T cells was determined by flow cytometry among HIV negative and HIV positive patients. We report that among HIV- individuals, mean CD32 percent expression was 60% on CD4dimCD8bright T cells and 17% on CD4+ T cells (p<0.01). Among HIV+ patients, mean CD32 percent expression was 54% on CD4dimCD8bright T cells and 12% on CD4+ T cells (p<0.001). CD32 expression on CD4dimCD8bright T cells did not correlate with CD4 count and viral load and was not different by HIV serostatus. CD32 was also higher on other double positive T cell populations in both HIV negative and HIV positive donors in comparison to their single positive T cell counterpart. Together, these studies indicate that CD32 is enriched on double positive T cells regardless of HIV serostatus. The functional role of CD32 on these double positive T cells remains to be elucidated

HIV_EC and HIV_NEG have comparable Bregs frequency.

<p>PBMC from HIV_EC (n = 15), HIV_ART (n = 20), HIV_NEG, (n = 20) and HIV_VIR, (n = 17) were cultured for 48H and during the final 5H supplemented with PMA (25 ng/ml), Ionomycin (1 ug/ml), Brefeldin A (1∶100) and by flow cytometry, the (<b>a,b</b>) frequency of CD19⁺CD24^hiCD38^hi Bregs and (<b>c</b>) IL-10-positive Bregs (intracellular cytokine staining) determined. p values for differences as calculated by Mann Whitney test two-tailed t test (Graphpad Prism software) are indicated; * = p<0.05, ** = p<0.01, lines indicate mean with SEM.</p

SAHA-treated Breg-depleted PBMC from HIV_EC and HIV_ART exhibit upregulated expression of antigen-presenting molecules and proliferation of CD4+ T cells.

<p>After 4 days in culture, (<b>a</b>) the expression of MHC-II and MHC-I/II on B cells and dendritic cells (LIN⁻CD11c⁺HLA-DR⁺) respectively was determined by flow cytometry in (<b>b</b>) SAHA-treated total or Breg-depleted PBMC from HIV_EC, (n = 4, upper panel) and HIV_ART, (n = 4, lower panel). The gating strategy and representative histogram overlays are depicted in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0092934#pone-0092934-g003" target="_blank">Figure 3a</a>. (<b>c</b>) VPD450-proliferation dye labeled total or Breg-depleted PBMC were stimulated with SAHA (500 nM, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0092934#pone-0092934-g001" target="_blank">Figure 1b</a> right panel, n = 5) and after 4 days in culture proliferation of CD4⁺ T cells was determined by flow cytometry. p values for differences in CD4⁺ T cell proliferation as calculated by paired two-tailed t test (Graphpad Prism software) are indicated.</p

Association between elevated frequency of CTL–competent T cells, clearance of infected CD4⁺ T cells and reduced viral DNA.

<p>(<b>a</b>) 500 nm SAHA-treated total or Breg-depleted PBMC from HIV_EC (n = 4) and HIV_ART (n = 5) were cultured for 4 days and the frequency of infected CD4⁺ T cells was determined by binding to KC57-antibody. (<b>b</b>) In HIV_ART subjects (n = 5), relative levels of HIV DNA between SAHA-treated total or Breg-depleted PBMC were determined by qPCR with LTR hybridizing primers after 4 days in culture. p values as calculated by paired two-tailed t test (Graphpad Prism software) are indicated.</p

SAHA treated Breg-depleted PBMC from HIV_EC and HIV_ART exhibit higher frequencies of anti-HIV CTL-competent CD8⁺ T cells.

<p>(<b>a</b>) 500 nM SAHA-treated total or Breg-depleted PBMC from HIV_EC (n = 4) and HIV_ART (n = 6) were cultured for 4 days and the frequency CD107a⁺CD8⁺ T cells determined by flow cytometry. (<b>b</b>) After 4 days in culture, by flow-cytometry using an HLA-A*0201 MHC-I HIV Dextramer® (Immudex), the frequency of HIV_gagSL9⁺ CD8⁺ T cells was determined; left panel depicts representative dot-plots demonstrating specific binding and right panel shows the summary of results (A2^pos = HLA-A*2 positive, A2^neg = HLA-A*2 negative). p values for differences as calculated by paired one-tailed t test (Graphpad Prism software) are indicated.</p

An Efficient Humanized Mouse Model for Oral Anti-Retroviral Administration

HIV anti-retrovirals (ARVs) have vastly improved the life expectancy of people living with HIV (PLWH). However, toxic effects attributed to long-term ARV use also contribute to HIV-related co-morbidities such as heart disease, bone loss and HIV-associated neurocognitive disorders (HAND). Unfortunately, mouse models used to study the effects of ARVs on viral suppression, toxicity and HIV latency/tissue reservoirs have not been widely established. Here, we demonstrate an effective mouse model utilizing immune-compromised mice, reconstituted with infected human peripheral blood mononuclear cell (PBMCs). ARVs areincorporated into mouse chow and administered daily with combination ARV regimens includingAtripla (efavirenz, tenofovir disoproxil fumarate, and emtricitabine) and Triumeq (abacavir, dolutegravir and lamivudine). This model measures HIV-infected human cell trafficking, and ARV penetration throughout most relevant HIV organs and plasma, with a large amount of trafficking to the secondary lymphoid organs. Furthermore, the HIV viral load within each organ and the plasma was reduced in ARV treated vs. untreated control. Overall, we have demonstrated a mouse model that is relatively easy and affordable to establish and utilize to study ARVs’ effect on various tissues, including the co-morbid conditions associated with PLWH, such as HAND, and other toxic effects