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Donata Vercelli

Publications and source records attributed to Donata Vercelli.

27 records · Page 2Linked to original sources
PubMed2002

Polymorphisms in toll-like receptor 4 are not associated with asthma or atopy-related phenotypes.

Toll-like receptor 4 (TLR4) is the principal receptor for bacterial endotoxin recognition, and functional variants in the gene confer endotoxin-hyporesponsiveness in humans. Furthermore, there is evidence that endotoxin exposure during early life is protective against the development of atopy and asthma, although this relationship remains poorly understood. It is therefore possible that genetic variation in the TLR4 locus contributes to asthma susceptibility. In this study we characterize the genetic diversity in the TLR4 locus and test for association between the common genetic variants and asthma-related phenotypes. In a cohort of 90 ethnically diverse subjects, we resequenced the TLR4 locus and identified a total of 29 single nucleotide polymorphisms. We assessed five common polymorphisms for evidence of association with asthma in two large family-based cohorts: a heterogeneous North American cohort (589 families), and a more homogenous population from northeastern Quebec, Canada (167 families). Using the transmission-disequilibrium test, we found no evidence of association for any of the polymorphisms tested, including two functional variants. Furthermore, we found no evidence for association between the TLR4 variants and four quantitative intermediate asthma- and atopy-related phenotypes. Based on these results, we found no evidence that genetic variation in TLR4 contributes to asthma susceptibility.

Asthma↗
PubMed2002

Inhibition of CXCR4-tropic HIV-1 infection by lipopolysaccharide: evidence of different mechanisms in macrophages and T lymphocytes.

Bacterial LPS protects primary human macrophages from infection by CCR5-tropic HIV-1 isolates through the release of the CC chemokines RANTES and macrophage inflammatory protein-1 alpha and -1 beta. Here, we show that LPS also suppresses infection of macrophages by CXCR4-tropic HIV-1 isolates. A marked down-regulation of both CD4 and CXCR4 expression was associated with this effect. Furthermore, a soluble factor(s) released by macrophages upon LPS treatment inhibited infection with CXCR4-tropic HIV-1 isolate viruses in both macrophages and T lymphocytes. Infection of both cell types appeared to be blocked at the level of viral entry and was independent of stromal cell-derived factor-1, the only known natural ligand of CXCR4. Moreover, the suppressive effect of LPS was unrelated to the release of IFN-alpha and -beta, macrophage-derived chemokine, leukemia inhibitory factor, or TNF-alpha. These results suggest the existence of potent HIV-1 inhibitory factor(s), uncharacterized to date, released by activated cells of the mononuclear phagocytic system.

Anti-HIV Agents↗
PubMed2002

c-Rel is a selective activator of a novel IL-4/CD40 responsive element in the human Ig gamma4 germline promoter.

Induction of isotype switching to a specific C(H) gene correlates with the transcriptional activation of the same gene in germline (GL) configuration. Expression of correctly spliced GL transcripts is necessary to target a switch region for recombination. In human B cells, the IgE and IgG4 isotypes are both induced by IL-4 through sequential switching, but are functionally antagonistic because IgG4 appears to have IgE-blocking activity. In order to understand the molecular mechanisms that regulate IgG4 production, we undertook a systematic analysis of the gamma4 GL promoter. A HindIII/NaeI region (-421/+474) highly conserved in the human gamma locus mediated the synergistic activation of a reporter gene in response to IL-4 and CD40 cross-linking. STAT6 binding to the proximal gamma4 GL promoter was essential for both IL-4-induced activation and CD40-dependent enhancement of transcription. Of note, a 45bp region (-76/-32) centered around the STAT6 binding motif drove robust synergistic activation of a heterologous fos promoter upon stimulation with IL-4 and CD40 cross-linking. This finding suggested that the (-76/-32) region may contain a novel IL-4/CD40 responsive element (RE). Electrophoretic mobility shift assays (EMSA) analysis using BL-2 nuclear extracts and in vitro translated NF-kappaB/Rel family proteins revealed the presence of a motif that overlaps the 5' end of the STAT6 element and binds selectively c-Rel. A mutation that abrogated c-Rel, but not STAT6, binding strongly impaired the CD40-induced enhancement of IL-4-dependent gamma4 GL transcription in reporter assays. These results indicate that c-Rel is selectively involved in the CD40-dependent activation of the IL-4/CD40 RE in the proximal gamma4 GL promoter.

Base Sequence↗
PubMed2002

Myb proteins repress human Ig epsilon germline transcription by inhibiting STAT6-dependent promoter activation.

Cytokine-dependent induction of correctly spliced germline (GL) transcripts is required to target the appropriate switch region for class switch recombination. GL transcription is linked to the cell cycle and the number of cell divisions through mechanisms that have not been defined. The human proximal epsilon GL promoter contains an IL-4 responsive element (IL-4RE) that binds STAT6 and is sufficient to confer IL-4 inducibility to a heterologous promoter in transient transfection studies. We show herein that the IL-4RE contains a novel Myb binding motif that overlaps the 3' end of the STAT6 palindrome. EMSA analysis showed binding to the IL-4RE of endogenous Myb proteins expressed in BL-2 B cells and Jurkat T cells. However, double occupancy of a probe spanning both STAT6 and Myb binding motifs could not be detected. Thus, binding of either factor may prevent protein/DNA interactions at the other site, raising the possibility that Myb binding may interfere with STAT6-dependent activation of the IL-4RE. Indeed, cotransfection of A-Myb or c-Myb expression vectors in HEK293 and BL-2 cells suppressed STAT6-dependent transcription from a reporter construct containing four copies of the IL-4RE cloned upstream of a minimal thymidine kinase promoter. Most importantly, overexpression of A-Myb was sufficient to suppress IL-4-induced endogenous epsilon GL transcription in BL-2 cells. Our results indicate that Myb proteins, which are known to act as cell cycle sensors, may play an important mechanistic role in the in vivo regulation of epsilon GL transcription in human B cells.

B-Lymphocytes↗
PubMed2002

Single nucleotide polymorphisms in innate immunity genes: abundant variation and potential role in complex human disease.

Under selective pressure from infectious microorganisms, multicellular organisms have evolved immunological defense mechanisms, broadly categorized as innate or adaptive. Recent insights into the complex mechanisms of human innate immunity suggest that genetic variability in genes encoding its components may play a role in the development of asthma and related diseases. As part of a systematic assessment of genetic variability in innate immunity genes, we have thus far have examined 16 genes by resequencing 93 unrelated subjects from three ethnic samples (European American, African American and Hispanic American) and a sample of European American asthmatics. Approaches to discovering and understanding variation and the subsequent implementation of disease association studies are described and illustrated. Although highly conserved across a wide range of species, the innate immune genes we have sequenced demonstrate substantial interindividual variability predominantly in the form of single nucleotide polymorphisms (SNPs). Genetic variation in these genes may play a role in determining susceptibility to a range of common, chronic human diseases which have an inflammatory component. Differences in population history have produced distinctive patterns of SNP allele frequencies, linkage disequilibrium and haplotypes when ethnic groups are compared. These and other factors must be taken into account in the design and analysis of disease association studies.

Animals↗
PubMed2002

The functional genomics of CD14 and its role in IgE responses: an integrated view.

Several studies in recent years have suggested that there is a strong genetic component in the pathogenesis of IgE-mediated diseases. Epidemiologic studies have identified a number of genes that carry single base changes (single nucleotide polymorphisms) associated with parameters of allergy. What remain to be established are the mechanisms whereby genetic variation results in dysregulation of IgE-mediated responses. This is the task of functional genomics. In this article, some of the most powerful approaches that have been devised to provide a mechanistic explanation for the effects of genetic variation on the regulation of gene expression and function are discussed. Recent data on the impact of genetic variation on the regulation of CD14 are explored in the context of the potential role played by this gene in the pathogenesis of allergy. Also discussed is the notion that taken individually, each instance of variation might result in small effects. It is the combination of variations in the same gene and/or in genes arrayed along one functional pathway that might eventually lead to dysregulation strong enough to cause disease. In this scenario, the environment is likely to play an essential role in determining the functional outcome of genetic variation.

Gene Expression Regulation↗
PubMed2002

Novel insights into class switch recombination.

The discovery of activation-induced cytidine deaminase, a putative RNA editing enzyme essential for both class switch recombination and somatic hypermutation, marked the field of isotype switching studies in the year 2000. More recent work from the same group now highlights some essential mechanistic aspects of the switch recombination process. In particular, much has been learnt about the relationship between transcription and recombination and the transcriptional competence of intronic promoters on looped-out circular DNA uncoupled from the proximal and distal enhancers. These findings have far-reaching implications, particularly for studies of class switch recombination in tissues. Although these important advances do not directly relate to interleukin-4-dependent immunoglobulin E switching, the conceptual and experimental tools developed through these studies are certain to foster progress in the immunoglobulin E field.

Animals↗
PubMed2002

Genetics of IL-13 and functional relevance of IL-13 variants.

Studies in both humans and mice clearly show that interleukin (IL)-13 is a central regulator of allergic inflammation. Numerous single nucleotide polymorphisms have been recently identified in the IL-13 gene and have been found to be associated with allergic and/or asthmatic phenotypes in different populations throughout the world. Furthermore, functional genomics studies are highlighting mechanistic pathways that may link genetic variation in IL-13 and disease. IL-13 is therefore an ideal model to study the impact of genetic variation on the regulation of gene expression and function in complex diseases.

Animals↗