scholarly journals Sp1 mediates phorbol ester (PMA)-induced expression of membrane-bound guanylyl cyclase GC-A in human monocytic cells*

2018 ◽  
Vol 65 (3) ◽  
Author(s):  
Małgorzata Mitkiewicz ◽  
Bernadeta Bac ◽  
Marianna Kuropatwa ◽  
Ewa Kurowska ◽  
Janusz Matuszyk ◽  
...  
Keyword(s):  
Phorbol Ester ◽  
Dna Sequences ◽  
Mononuclear Cells ◽  
Cyclic Guanosine Monophosphate ◽  
Guanosine Monophosphate ◽  
Peripheral Blood Mononuclear ◽  
Induced Expression ◽  
Guanylyl Cyclases ◽  
Membrane Bound ◽  
Encoding Gene

Cyclic guanosine monophosphate (cGMP) is synthesized by two types of enzymes: particulate (membrane-bound) guanylyl cyclases (pGCs) and soluble (cytosolic) guanylyl cyclases (sGCs). sGCs are activated primarily by binding of nitric oxide to their prosthetic heme group while pGCs are activated by binding of peptide ligands to their extracellular domains. One of them, pGC type A (GC-A) is activated by atrial and brain natriuretic peptides (ANP and BNP, respectively). Human monocytes isolated from peripheral blood mononuclear cells have been shown to have sGC expression without concomitant expression of GC-A. However, GC-A activity appears in monocytes under certain conditions but a molecular mechanism of GC‑A expression is still poorly understood. In this report we show that phorbol ester (PMA) induced transcription of the gene encoding GC-A in human monocytic THP-1 cells. Moreover, PMA-treated THP-1 cells have been found to raise cGMP content following treatment with ANP. Studies using pharmacological inhibitors of protein kinases have suggested involvement of protein kinase C (PKC), mitogen extracellular kinases (MEK1/2), and extracellular signal-regulated kinases (ERK1/2) in PMA-induced expression of the GC-A encoding gene in THP-1 cells. Finally, we show that PMA stimulated binding of Sp1 transcription factor to GC-rich DNA sequences and mithramycin A (a selective Sp1 inhibitor) inhibited expression of the GC-A mRNA in PMA-treated THP-1 cells. Taken together, our findings suggest that the PMA/PKC/MEK/ERK signaling pathway induces Sp1-mediated transcription of the GC-A encoding gene in human monocytic THP-1 cells.

scholarly journals Kaposi's sarcoma (KS)-associated herpesvirus-like DNA sequences in peripheral blood mononuclear cells: association with KS and persistence in patients receiving anti-herpesvirus drugs

Blood ◽  
1996 ◽  
Vol 88 (1) ◽  
pp. 297-301 ◽  
Author(s):  
RW Humphrey ◽  
TR O'Brien ◽  
FM Newcomb ◽  
H Nishihara ◽  
KM Wyvill ◽  
...  
Keyword(s):  
Peripheral Blood Mononuclear Cells ◽  
Dna Sequences ◽  
Peripheral Blood ◽  
Kaposi's Sarcoma ◽  
Mononuclear Cells ◽  
Kaposi’S Sarcoma ◽  
Peripheral Blood Mononuclear ◽  
Therapeutic Implications ◽  
Blood Mononuclear Cells ◽  
Cd4 Lymphocyte

Abstract Herpesvirus-like DNA sequences (KSHV/HHV-8) have recently been described in AIDS-associated Kaposi's sarcoma (KS) lesions. Many questions remain regarding the role of this virus in KS and the therapeutic implications of this finding. In the current study, KSHV/HHV-8 DNA was detected in peripheral blood mononuclear cells (PBMCs) from human immunodeficiency virus (HIV)-infected patients with KS (34/98) more often than in HIV-infected individuals without KS (12/64, P = .03). The detection of KSHV/HHV-8 DNA did not correlate with the CD4 lymphocyte count. Five patients demonstrated KSHV/HHV-8 DNA in their PBMCs during administration of intravenous foscarnet and/or ganciclovir. The continued detection of KSHV/HHV-8 DNA in the PBMCs of patients receiving these anti-herpesvirus drugs has potential implications regarding the virus-cell relationship of KSHV/HHV-8, as well as for the value of these drugs in treating or preventing KS, but additional studies are needed.

scholarly journals Incomplete APOBEC3G/F Neutralization by HIV-1 Vif Mutants Facilitates the Genetic Evolution from CCR5 to CXCR4 Usage

2015 ◽  
Vol 59 (8) ◽  
pp. 4870-4881 ◽  
Author(s):  
Claudia Alteri ◽  
Matteo Surdo ◽  
Maria Concetta Bellocchi ◽  
Patrizia Saccomandi ◽  
Fabio Continenza ◽  
...  
Keyword(s):  
Amino Acid ◽  
Dna Sequences ◽  
Mononuclear Cells ◽  
Strongly Correlated ◽  
Peripheral Blood Mononuclear ◽  
Cxcr4 Usage ◽  
Risk Of Disease ◽  
Vif Protein ◽  
Defective Virus ◽  
Hiv 1

ABSTRACTIncomplete APOBEC3G/F neutralization by a defective HIV-1Vif protein can promote genetic diversification by inducing G-to-A mutations in the HIV-1 genome. The HIV-1 Env V3 loop, critical for coreceptor usage, contains several putative APOBEC3G/F target sites. Here, we determined if APOBEC3G/F, in the presence of Vif-defective HIV-1 virus, can induce G-to-A mutations at V3 positions critical to modulation of CXCR4 usage. Peripheral blood mononuclear cells (PBMC) and monocyte-derived macrophages (MDM) from 2 HIV-1-negative donors were infected with CCR5-using 81.A-VifWTvirus (i.e., with wild-type [WT] Vif protein), 81.A-VifE45G, or 81.A-VifK22E(known to incompletely/partially neutralize APOBEC3G/F). The rate of G-toA mutations was zero or extremely low in 81.A-VifWT- and 81.A-VifE45G-infected PBMC from both donors. Conversely, G-to-A enrichment was detected in 81.A-VifK22E-infected PBMC (prevalence ranging from 2.18% at 7 days postinfection [dpi] to 3.07% at 21 dpi in donor 1 and from 10.49% at 7 dpi to 8.69% at 21 dpi in donor 2). A similar scenario was found in MDM. G-to-A mutations occurred at 8 V3 positions, resulting in nonsynonymous amino acid substitutions. Of them, G24E and E25K strongly correlated with phenotypically/genotypically defined CXCR4-using viruses (P= 0.04 and 5.5e−7, respectively) and increased the CXCR4 N-terminal binding affinity for V3 (WT, −40.1 kcal/mol; G24E, −510 kcal/mol; E25K, −522 kcal/mol). The analysis of paired V3 and Vif DNA sequences from 84 HIV-1-infected patients showed that the presence of a Vif-defective virus correlated with CXCR4 usage in proviral DNA (P= 0.04). In conclusion, incomplete APOBEC3G/F neutralization by a single Vif amino acid substitution seeds a CXCR4-using proviral reservoir. This can have implications for the success of CCR5 antagonist-based therapy, as well as for the risk of disease progression.

Dipyridamole decreases inflammatory metalloproteinase-9 expression and release by human monocytes

2013 ◽  
Vol 109 (02) ◽  
pp. 280-289 ◽  
Author(s):  
Maria Annunziata Carluccio ◽  
Mariangela Pellegrino ◽  
Nadia Calabriso ◽  
Carlo Storelli ◽  
Giuseppe Martines ◽  
...  
Keyword(s):  
P38 Mapk ◽  
Mononuclear Cells ◽  
Nuclear Translocation ◽  
Human Peripheral Blood ◽  
Antiplatelet Agent ◽  
Guanosine Monophosphate ◽  
Human Monocytes ◽  
Peripheral Blood Mononuclear ◽  
Tnf Α ◽  
Anti Inflammatory

SummaryMatrix metalloproteinase (MMP)-9 plays an important role in stroke by accelerating matrix degradation, disrupting the blood-brain barrier and increasing infarct size. Dipyridamole is an antiplatelet agent with recognised benefits in ischaemic stroke prevention. In addition to its antiplatelet properties, recent studies have reported that dipyridamole also features anti-inflammatory and anti-oxidant properties. We therefore investigated whether dipyridamole can ameliorate the proinflammatory profile of human monocytes, a source of MMP-9 in stroke, in terms of regulation of MMP-9 activity and expression, and explored underlying mechanisms. Human peripheral blood mononuclear cells (PBMC) and U937 cells were treated with increasing concentrations of dipyridamole (up to 10 µg/ml) for 60 minutes before stimulation with tumour necrosis factor (TNF)-α or phorbol myristate acetate (PMA). Exposure of PBMC and U937 to dipyridamole reduced TNF-α- and PMA-induced MMP-9 activity and protein release as well as MMP-9 mRNA, without significantly affecting the release of TIMP-1. This inhibitory effect was independent of dipyridamole-induced cyclic adeno-sine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) increase. Correspondingly, dipyridamole also significantly inhibited TNF-α-induced nuclear factor (NF)-κB activation and nuclear translocation of the p65 NF-κB subunit through a mechanism involving the inhibition of IkBα degradation and p38 MAPK activation. In conclusion, dipyridamole, at therapeutically achievable concentrations, reduces the expression and release of MMP-9 through a mechanism involving p38 MAPK and NF-κB inhibition. These results indicate that dipyridamole exerts anti-inflammatory properties in human monocytes that may favourably contribute to its actions in the secondary prevention of stroke, independent of its antiplatelet properties.

scholarly journals Evidence for progenitors of chronic lymphocytic leukemia B cells that undergo intraclonal differentiation and diversification

Blood ◽  
1996 ◽  
Vol 87 (4) ◽  
pp. 1586-1594 ◽  
Author(s):  
M Dono ◽  
S Hashimoto ◽  
F Fais ◽  
V Trejo ◽  
SL Allen ◽  
...  
Keyword(s):  
B Cells ◽  
Chronic Lymphocytic Leukemia ◽  
B Cell ◽  
Dna Sequences ◽  
Mononuclear Cells ◽  
Lymphocytic Leukemia ◽  
Specific Primers ◽  
Region Gene ◽  
Peripheral Blood Mononuclear ◽  
Ancestral Gene

Peripheral blood mononuclear cells from five patients with IgG+ B-type chronic lymphocytic leukemia (B-CLL) were analyzed for the presence of clone-specific Ig H chain variable region gene mRNA transcripts linked to C mu and/or C alpha. This was assessed by (1) comparing the lengths of portions of the VHDJH of the IgG+ CLL clones with those of the mu and alpha isotype-expressing B cells, (2) performing clone-specific endonuclease digestion studies, and (3) determining the DNA sequences of the mu and alpha isotype-expressing cDNA. Thus, when B-cell mRNA from these five patients were reverse transcribed with C gamma-specific primers and then amplified by polymerase chain reaction, dominant cDNA were found with lengths corresponding to those of the IgG+ CLL B cell. In addition, in four cases, cDNA of lengths identical to those of the CLL B cell were detected when mRNA was reverse transcribed and amplified using c mu- and/or C alpha-specific primers, strongly suggesting clonal relatedness. These CLL-related mu- and alpha- expressing cDNA were present in greater amounts that unrelated (non- CLL) mu- and alpha-expressing cDNA from normal B cells that used genes of the same VH family. When the sequences of these CLL-related C mu- and C alpha-expressing cDNA were compared with those of the IgG+ CLL clones, it was clear that they were derived from the same ancestral gene as the IgG-expressing CLL B cell, thus documenting their common origin. Finally, nucleotide point mutations were observed in the mu- and alpha-expressing cDNA of certain patients, indicating divergence with the CLL. These data suggest that IgM+ B cells, which are precursors of the leukemic B cells, exist in increased numbers in the blood of most patients with IgG+ B-CELL and that these cells may differentiate, accumulate V genes mutations, and undergo isotype switching in vivo. In addition, the data are consistent with a sequential-hit model for the evolution of CLL.

scholarly journals Enhanced Adhesive Capacities of the Naturally Occurring Ile249–Met280Variant of the Chemokine Receptor CX3CR1

2004 ◽  
Vol 279 (19) ◽  
pp. 19649-19657 ◽  
Author(s):  
Mehdi Daoudi ◽  
Elise Lavergne ◽  
Alexandre Garin ◽  
Nadine Tarantino ◽  
Patrice Debré ◽  
...  
Keyword(s):  
G Protein ◽  
Calcium Release ◽  
Mononuclear Cells ◽  
Binding Capacity ◽  
Flow Chamber ◽  
Soluble Form ◽  
Peripheral Blood Mononuclear ◽  
Naturally Occurring ◽  
Aspiration Technique ◽  
Membrane Bound

It was recently shown that individuals carrying the naturally occurring mutant CX3CR1-Ile249–Met280(hereafter called CX3CR1-IM) have a lower risk of cardiovascular disease than individuals homozygous for the wild-type CX3CR1-Val249–Thr280(CX3CR1-VT). We report here that peripheral blood mononuclear cells (PBMC) from individuals with the CX3CR1-IM haplotype adhered more potently to membrane-bound CX3CL1 than did PBMC from homozygous CX3CR1-VT donors. Similar excess adhesion was observed with CX3CR1-IM-transfected human embryonic kidney (HEK) cell lines tested with two different methods: the parallel plate laminar flow chamber and the dual pipette aspiration technique. Suppression of the extra adhesion in the presence of pertussis toxin indicates that G-protein mediated the underlying transduction pathway, in contrast to the G-protein-independent adhesion previously described for CX3CR1-VT. Surprisingly, HEK and PBMC that expressed CX3CR1-IM and -VT were indistinguishable when tested with the soluble form of CX3CL1 for chemotaxis, calcium release, and binding capacity. In conclusion, only the membrane-anchored form of CX3CL1 functionally discriminated between these two allelic isoforms of CX3CR1. These results suggest that each form of this ligand may lead to a different signaling pathway. The extra adhesion of CX3CR1-IM may be related to immune defenses and to atherogenesis, both of which depend substantially on adhesive intercellular events.

scholarly journals Structure-Dependent Modulation of Alpha Interferon Production by Porcine Circovirus 2 Oligodeoxyribonucleotide and CpG DNAs in Porcine Peripheral Blood Mononuclear Cells

2007 ◽  
Vol 81 (10) ◽  
pp. 4919-4927 ◽  
Author(s):  
Frida Hasslung Wikström ◽  
Brian M. Meehan ◽  
Mikael Berg ◽  
Sirje Timmusk ◽  
Josefine Elving ◽  
...  
Keyword(s):  
Peripheral Blood Mononuclear Cells ◽  
Dna Sequences ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Alpha Interferon ◽  
Porcine Circovirus ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
Cpg Dinucleotide

ABSTRACT DNA sequences containing CpG motifs are recognized as immunomodulators in several species. Phosphodiester oligodeoxyribonucleotides (ODNs) representing sequences from the genome of porcine circovirus type 2 (PCV2) have been identified as potent inducers (ODN PCV2/5) or inhibitors (ODN PCV2/1) of alpha interferon (IFN-α) production by porcine peripheral blood mononuclear cells (poPBMCs) in vitro. In this study, the IFN-α-inducing or -inhibitory activities of specific phosphodiester ODNs were demonstrated to be dependent on their ability to form secondary structures. When a poly(G) sequence was added to a stimulatory self-complementary ODN, high levels of IFN-α were elicited, and the induction was not dependent on pretreatment with the transfecting agent Lipofectin. In addition, the IFN-α-inducing ODN required the presence of an intact CpG dinucleotide, whereas the inhibitory activity of ODN PCV2/1 was not affected by methylation or removal of the central CpG dinucleotide. Of particular significance, the IFN-α inhibition elicited by ODN PCV2/1 was only effective against induction stimulated by DNA control inducers and not RNA control inducers, indicating activity directed to TLR9 signaling. The PCV2 genome as a whole was demonstrated to induce IFN-α in cultures of poPBMCs, and the presence of immune modulatory sequences within the genome of PCV2 may, therefore, have implications with regard to the immune evasion mechanisms utilized by PCV2.

Low prevalence of Chlamydia psittaci infection in French patients with ocular adnexal lymphomas (OAL)

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 17536-17536
Author(s):  
D. Decaudin ◽  
A. Subtil ◽  
A. Ferreri ◽  
A. Vincent-Salomon ◽  
M. Ponzoni ◽  
...  
Keyword(s):  
B Cell ◽  
Dna Sequences ◽  
Mononuclear Cells ◽  
Lymphoid Hyperplasia ◽  
Lymphoproliferative Disease ◽  
Chlamydia Psittaci ◽  
Reactive Lymphoid Hyperplasia ◽  
Experimental Conditions ◽  
Peripheral Blood Mononuclear ◽  
Geographical Variations

17536 Background: A high prevalence of Chlamydia psittaci infection in both tumor tissues and peripheral blood mononuclear cells of Italian patients with OAL has been reported (Ferreri AJ, Guidoboni M, Ponzoni M, De Conciliis C, Dell’Oro S, Fleischhauer K, et al. Evidence for an association between Chlamydia psittaci and ocular adnexal lymphomas. J Natl Cancer Inst 2004;96(8):586–94.) This association has not been confirmed in some regions of the USA, and no data from other European countries are available. We therefore investigated the presence of DNA of C. psittaci, C. trachomatis, and C. pneumoniae DNA in French patients (pts) with OAL. Methods: Tumor samples from ophthalmologic biopsies of 16 OAL pts (10 conjunctiva, 6 orbit) were included in the study. Histologic type were MALT-type (n = 8), lymphoplasmocytic (n = 6), follicular (n = 1), and diffuse large B-cell (n = 1) lymphomas. Two other groups of lymphoproliferative disease were analyzed as controls. First, ten cases of nodal lymphomas (follicular and marginal zone B-cell lymphomas), and ten cases of reactive lymphoid hyperplasia were analyzed. A multiplex touchdown, enzyme time-release PCR designed to simultaneously detect C. psittaci, C. pneumoniae and C. trachomatis DNA sequences was performed. PCR analyses were performed in duplicate in an independent setting either inat the Curie Institut Curie in e, Paris, and in the National Cancer Institute, Aviano. Results: DNA of C. psittaci DNA was detected in the tumoral tissue of only one patient with follicular OAL. No DNA sequences of C. psittaci, C. pneumoniae and C. trachomatis DNA sequences were was detected in all any of the other OALs, or controls. Conclusions: The prevalence of C. psittaci infection in French pts with OAL was sensibly significantly lower than that reported in Italian series. Cross-controls between the two laboratories indicate that this finding is not due to different experimental conditions. Discrepancies may be explained by an heterogeneous epidemiological distribution of the bacterial infection. Large studies aimed to investigate geographical variations in the prevalence of this association are warranted. No significant financial relationships to disclose.

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