Abstract A newly recognized link between the complement system and adaptive immunity is that decay accelerating factor (DAF), a cell surface C3/C5 convertase regulator, exerts control over T responses. Extending these results, we show cultures of Marilyn TCR-transgenic cells stimulated with DAF-deficient (Daf1−/−) APCs produce significantly more IL-12, C5a, IFN-γ compared containing wild-type APCs. DAF-regulated IL-12 production subsequent differentiation into IFN-γ-producing effectors was...

Abstract IFN-γ- and IL-17-producing T cells autoreactive across myelin components are central to the pathogenesis of multiple sclerosis. Using direct in vivo, adoptive transfer, vitro systems, we show this study that generation these effectors oligodendrocyte glycoprotein35–55-induced experimental autoimmune encephalomyelitis depends on interactions locally produced C3a/C5a with APC cell C3aR/C5aR. In absence surface C3/C5 convertase inhibitor decay-accelerating factor (DAF), but not...

Abstract Induction of proinflammatory T cell immunity is augmented by innate dendritic (DC) maturation commonly initiated TLR signaling. We demonstrate that ligation TLR3, TLR4, and TLR9 induces murine DC production complement components local the anaphylatoxin C5a. In vitro, ex vivo, in vivo analyses show TLR-induced maturation, as assessed surface phenotype, expression profiling gene array, functional ability to stimulate responses, requires autocrine C3a receptor C5a (C3ar1/C5ar1) Studies...

The transcription factor Kruppel-like 4 (KLF4) regulates the expression of immunosuppressive and anti-thrombotic proteins. Despite its importance in maintaining homeostasis, signals that control mechanism transactivation remain unclarified. CD55 [aka decay accelerating (DAF)], now known to be a regulator T B cell responses, biases between pro- anti-inflammatory processes by controlling autocrine C3a C5a receptor (C3ar1/C5ar1) signaling cells. similarity CD55’s KLF4’s regulatory effects...

SUMMARY Human retinal pigment epithelial (RPE) cells form part of the blood–retina barrier where they potentially can regulate leucocyte function. RPE are known to secrete several cytokines in response stimulation by other cytokines. Anaphylatoxin C5a, a potent inflammatory mediator produced during complement activation, binds G-protein coupled C5a receptors (C5aR) on monocytes/macrophages and releases various from cells. We previously reported that human cell line ARPE-19 possesses C5aR...

The involvement of complement in B2 cell responses has been regarded as occurring strictly via components plasma. In this study, we show that Ab production and class switch recombination (CSR) depend on autocrine C3a C5a receptor (C3ar1/C5ar1) signaling cells. CD40 upregulation, IL-6 production, growth response to BAFF or APRIL, AID/Bcl-6 expression, well follicular CD4+ CD21 all depended signal transduction. OVA immunization C3ar1-/-C5ar1-/- mice elicited IgM but no other isotypes, whereas...

ABSTRACT Purified vascular endothelial cell (EC) growth factor receptor-2 (VEGFR2) auto-phosphorylates upon VEGF-A occupation in vitro, arguing that VEGR2 confers its mitotic and viability signaling of itself. Herein, we show that, ECs, VEGFR2 function requires concurrent C3a/C5a receptor (C3ar1/C5ar1) IL-6 (IL-6R)–gp130 co-signaling. C3ar1/C5ar1 or IL-6R blockade totally abolished auto-phosphorylation, downstream Src, ERK, AKT, mTOR STAT3 activation, EC cycle entry. augmented production...

Escherichia coli promoters for transcription of ribosomal and tRNAs are greatly activated by an A+T-rich "UP" element upstream the −35 region. These same have also been found to otherwise deviate in several respects from consensus promoter sequence. Here we present results a kinetic characterization interaction RNA polymerase with UP element-containing which virtue or near-consensus sequence features should be among most optimal that can encountered polymerase. We show such promoters, (1)...

Strand separation in promoter DNA induced by Escherichia coli RNA polymerase is likely initiated at a conserved A residue position −11 of the nontemplate strand. Here we describe use fluorescence techniques to study interaction with base. Forked templates were employed, containing fluorescent base, 2-aminopurine (2AP), substituted single-stranded tail comprising nucleotides on strand which base pairing disrupted an polymerase-promoter complex. We demonstrate that presence 2AP instead has no...

To investigate whether the presence of decay-accelerating factor (or CD55), an intrinsic complement regulator, protects against development vascular disease, given that activation can affect leukocytes and platelets.Leukocyte-platelet complexes are critical for initiation progression atherosclerosis restenosis; however, mechanism by which these processes promote injury is incompletely defined. We performed femoral artery wire in Daf1(-/-) mice their wild-type controls. Leukocyte...

ABSTRACT The p R and RM promoters of bacteriophage lambda direct transcription in divergent directions from start sites separated by 83 phosphodiester bonds. We had previously shown that the presence an RNA polymerase at interfered with open complex formation this effect was alleviated deletion 10 bp between two promoters. Here we present a detailed characterization dependence interference on interpromoter distance. It found reduced is unique to 10-bp deletion. relief demonstrated be due...

How receptor tyrosine kinase (RTK) growth signaling is controlled physiologically incompletely understood. We have previously provided evidence that the survival and mitotic activities of vascular endothelial cell factor receptor-2 (VEGFR2) are dependent on C3a/C5a (C3ar1/C5ar1) IL-6 (IL-6R)-gp130 joint in a physically interactive platform. Herein, we document platelet derived epidermal receptors (PDGFR EGFR) regulated by same interconnection clarify mechanism underlying dependence. show...

ABSTRACT One prerequisite for generating recombinant antibodies therapeutic and non-therapeutic purposes from hybridoma clones is the reliable, efficient, cost-effective sequencing of clone that produces desired antibody. However, many fusion partners produce aberrant endogenous mRNA transcripts most which resemble kappa chains. These chain mRNAs can interfere with or even prevent determination functional murine antibody cDNA sequences during PCR amplification step. In this paper, we report...

Abstract Background Tumor necrosis factor receptor 2 (TNFR2) is considered an appealing target due to its low-level expression on immune cells, but it could be upregulated regulatory T cells (Tregs) in the tumor microenvironment which plays key roles Treg proliferation and function. It has been demonstrated that Tregs with high TNFR2 were most suppressive subsets among all populations tumor. While early studies showed co-stimulates naïve revealed later also limits CD8+ T-cell-mediated viral...