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C Vink

Publications and source records attributed to C Vink.

At least 19 recordsLinked to original sources
PubMed2006

A viral conspiracy: hijacking the chemokine system through virally encoded pirated chemokine receptors.

Several herpesviruses and poxviruses contain genes encoding for G protein-coupled receptor (GPCR) proteins that are expressed on the surface of infected host cells and/or the viral envelope. Most of these membrane-associated proteins display highest homology to the subfamily of chemokine receptors known to play a key role in the immune system. Virally encoded chemokine receptors have been modified through evolutionary selection both in chemokine binding profile and signaling capacity, ultimately resulting in immune evasion and cellular reprogramming in favor of viral survival and replication. Insight in the role of virally encoded GPCRs during the viral lifecycle may reveal their potential as future drug targets.

Animals↗
PubMed2006

Molecular fingerprinting of the intestinal microbiota of infants in whom atopic eczema was or was not developing.

BACKGROUND: The rise in atopic diseases has been linked to disturbances in the intestinal microbiota composition. OBJECTIVE: The purpose of this study was to investigate the intestinal microbiota composition in infants in whom atopic (IgE-associated) eczema was or was not developing, using a molecular fingerprinting technique. METHODS: Within a prospective birth cohort study, fecal samples have been collected at the infant's age of 1 month. Within the context of this cohort, we conducted a nested case-control study comparing fecal samples of 26 infants who became sensitized and developed eczema within the first year of life with 52 non-sensitized non-eczematous infants. The composition of the fecal samples was examined using PCR combined with denaturing gradient gel electrophoresis. Using real-time PCR, total bacterial counts and bifidobacterial counts were enumerated. RESULTS: Neither total bacterial profiles nor the type and proportion of bifidobacteria in the feces were associated with the development of atopic eczema. The similarity of bacterial profiles was low; mean similarity was approximately 33% in both infants with or without atopic eczema. The prevalence of one specific band in total bacterial profiles was significantly higher in infants with atopic eczema compared with controls (96% vs. 71%, P = 0.01). Identification of this band revealed that it represented Escherichia coli. CONCLUSION: Although no association was found between the development of IgE-associated eczema and the dominant gut microbiota as a whole or with the bifdobacterial microbiota, the association with E. coli indicates that differences in gut microbiota do precede the development of atopy.

Animals↗
PubMed2005

Identification and characterization of two antisense transcripts from the major immediate early region of rat cytomegalovirus.

We have identified and characterized two antisense transcripts from the rat cytomegalovirus (RCMV) major immediate early (MIE) region. These transcripts, designated IE-AS1 and IE-AS2, are complementary to part of the sense IE1 transcript. The IE-AS transcripts were first detected in peripheral blood leukocytes (PBL) of RCMV-infected rats at 7 days post-infection (pi) in the absence of IE1 transcription. Nevertheless, both the IE1 and IE-AS transcripts were found at the same time in the salivary glands of RCMV-infected rats at 7 and 120 days pi as well as in RCMV-infected rat embryo fibroblasts (REFs) at 48 h pi.

Amino Acid Sequence↗
PubMed2003

Virally encoded G protein-coupled receptors: targets for potentially innovative anti-viral drug development.

Various herpes- and poxviruses contain DNA sequences encoding proteins with homology to cellular chemokine receptors, which belong to the family of G protein-coupled receptors (GPCRs). Since GPCRs play a crucial role in cellular communication and chemokine receptors play a prominent role in the immune system, the virally encoded GPCRs may be crucial determinants of viral action. The Kaposi's sarcoma-associated herpesvirus (KSHV, or human herpesvirus 8), implicated in the pathogenesis of Kaposi's sarcoma (KS), a highly vascularized tumor, encodes a GPCR, referred to as ORF74. This virally encoded receptor was found to induce tumorigenesis and transgenic expression of ORF74 induces an angioproliferative disease resembling KS. Cytomegalovirus (CMV), suggested to play a role in atherosclerosis, encodes four GPCRs, among which US28. This virally encoded GPCR is able to induce migration of smooth muscle cells, a feature essential for the development of atherosclerosis. Remarkably, the KSHV and some CMV-encoded GPCRs display constitutive activity, while their cellular homologs do not. It remains to be determined whether this phenomenon contributes to the pathogenesis of viral action. Also, the family of poxviruses encodes GPCRs of which the function is not clear yet. In this review we will give an overview of the different virally encoded GPCRs, and discuss their putative role in viral action and potential as drug target.

Antiviral Agents↗
PubMed2002

A rat cytomegalovirus strain with a disruption of the r144 MHC class I-like gene is attenuated in the acute phase of infection in neonatal rats.

We previously generated an RCMV strain in which the r144 gene, encoding a major histocompatibility complex class I homolog, had been deleted (RCMVdelta r144). To investigate the role of r144 during acute infection of neonatal rats, we infected three days-old neonatal rats with either RCMVdelta r144 or wild type (wt) RCMV and the presence of infectious virus as well as viral DNA in various organs was determined at either 3, 5 or 21 days p.i.. In addition, we assessed both type and number of inflammatory cells in these organs. Interestingly, a significantly lower concentration of infectious virus as well as viral DNA was found in spleens of RCMVdelta r144-infected rats than in those of wt RCMV-infected animals at 3 days p.i.. At the same time point, a significantly lower amount of infiltrating NK cells and monocytes/macrophages was seen in the spleens of RCMVdelta r144-infected rats than in spleens of rats infected with wt RCMV. At 21 days p.i., RCMVdelta r144-infected rats were found to have lower virus titers in the salivary glands than wt RCMV-infected animals. Significant differences between RCMVdelta r144- and wt RCMV-infected rats were detected neither at other time points nor at other sites. We conclude that after infection of neonatal rats, the replication of RCMVdelta r144 is severely restricted compared to wt RCMV.

Acute Disease↗
PubMed2001

Rat cytomegalovirus open reading frame R44 is an early-late gene that encodes a nuclear protein.

Rat cytomegalovirus (RCMV) open reading frame R44 is the homolog of human cytomegalovirus gene UL44, which encodes the DNA polymerase accessory protein. Here, we show that R44 is transcribed as a 3.6-kb mRNA within the early and late phases of infection in vitro. In order to find potential monoclonal antibodies (MoAbs) directed against the R44-encoded protein (pR44), a panel of anti-RCMV MoAbs was screened for binding to pR44 recombinant proteins. Thus, an anti-pR44 MoAb, termed RCMV8, was identified. By using this MoAb, pR44 could be detected as early as 8 h after RCMV infection in vitro. The pR44 protein was determined to have a molecular mass of approximately 55 kDa and was found to be localized to the nucleus of RCMV-infected cells.

Animals↗
PubMed2001

The role of cytomegalovirus-encoded homologs of G protein-coupled receptors and chemokines in manipulation of and evasion from the immune system.

Cytomegaloviruses (CMVs) have the ability to persist lifelong within the infected host. This ability implies that these viruses are highly adapted to their hosts. Most importantly, they will have to employ strategies to remain hidden from the host's immune system. Virus genes predicted to be involved in these strategies include genes encoding homologs of cellular immune effector or regulatory proteins, such as chemokine (CK) receptor-like G protein-coupled receptors (GPCRs), CKs and MHC class I molecules. These genes may have been pirated by the virus during the long co-evolution of pathogen and host. In light of the crucial roles that GPCRs, CKs and MHC class I molecules play in the normal physiology of the host, it is to be expected that the CMV homologs of these proteins may have a profound impact on this physiology and, at the same time, serve vital functions in maintenance as well as replication of the virus within the infected host. As a consequence, these viral homologs can be envisaged as attractive targets for novel anti-viral strategies. The aim of this report is to present an overview of the current state of knowledge on the (putative) functions of the CMV homologs of GPCRs and CKs.

Amino Acid Sequence↗
PubMed2001

Different profiles of cytomegalovirus RNA transcripts and anti-cytomegalovirus IgM antibodies in renal transplant recipients.

BACKGROUND: A difference in anti-cytomegalovirus IgM antibody profile has been found between sera from acutely cytomegalovirus (CMV)-infected patients and sera from CMV-infected patients with subclinical infection. OBJECTIVES: The aim of this study is to investigate whether such different IgM antibody responses are correlated with differences in the expression of CMV immediate early and late mRNAs. STUDY DESIGN: We have investigated the anti-CMV IgM response in 46 renal transplant recipients by employing two commercially available IgM kits (AxSYM and IMX) as well as two novel enzyme-linked immunosorbent assays (ELISAs), which were developed using recombinant ppUL32 (pp150) and pUL80a (p38), respectively. The results were compared with four direct CMV diagnostic tests: pp65 antigenemia, viral culture and nucleic acid sequence-based amplification (NASBA), detecting either CMV immediate early 1 (IE1) mRNA (IE1-NASBA), or CMV pp67 (late) mRNA (pp67-NASBA). RESULTS: Analysis of all CMV-infected recipients (n=28) showed that in 16 recipients (group I) more than one direct test became positive after transplantation, while in the other 12 recipients (group II), IE1-NASBA was the only direct test to become positive. In group I, 100, 81, 100 and 50% of the recipients were IgM-positive with AxSYM, IMX, p38 and pp150, respectively. In group II, 100, 83, 17 and 83% of the recipients were IgM-positive with AxSYM, IMX, p38 and pp150, respectively. CONCLUSIONS: Our data indicate that the IgM-response against p38 and pp150 differs significantly (P<0.01) between group I recipients with productive CMV infection, and group II recipients with a non-productive CMV infection which may be of diagnostic and prognostic relevance.

Antibodies, Viral↗
PubMed2000

Rat cytomegalovirus R89 is a highly conserved gene which expresses a spliced transcript.

In all sequenced herpesvirus genomes, a homolog of the herpes simplex virus type 1 UL15 gene has been identified. This gene encodes a protein that is involved in viral genome maturation. Although transcription of the alphaherpesvirus UL15 gene has been analyzed in detail, not much is known about the expression of its betaherpesvirus homologs. We therefore set out to characterize transcription of the rat cytomegalovirus counterpart of UL15, R89. Here we report that R89 consists of two exons separated by a 4.7-kb intron. The spliced R89 transcript, which is expressed at late times postinfection (p.i.), has the capacity to encode a protein of 670 amino acids with a calculated molecular mass of 77.1 kDa. The predicted amino acid sequence of this protein is highly similar to that of the proteins predicted to be encoded by the human cytomegalovirus UL89 and murine cytomegalovirus M89 genes (64.3 and 84.5% overall identity, respectively). The region between R89 exon 1 and exon 2 was found to contain five additional genes, r90, R91, R92, R93 and R94, the latter two of which are conserved among all herpesviruses. We show that these genes are transcribed in a highly complex fashion, resulting in numerous mono- and polycistronic mRNAs.

Amino Acid Sequence↗
PubMed2000

Diagnostic implications of human cytomegalovirus immediate early-1 and pp67 mRNA detection in whole-blood samples from liver transplant patients using nucleic acid sequence-based amplification.

Nucleic acid sequence-based amplification (NASBA) was used for detection of the human cytomegalovirus (CMV) immediate early-1 (IE) and the late pp67 mRNA in 353 blood samples collected from 34 liver transplant patients. The diagnostic value of these assays was compared to that of the pp65 antigenemia assay. Overall, 95 and 42% of the antigenemia-positive samples were IE NASBA and pp67 NASBA positive, respectively. Although the results from pp67 NASBA and the antigenemia assay appeared to correspond poorly, a clear correlation was seen between pp67 NASBA-negative results and low numbers of pp65 antigen-positive cells. Twenty patients (59%) were treated with ganciclovir after the diagnosis of symptomatic CMV infection. Before initiation of the antiviral therapy, the antigenemia assay detected the onset of symptomatic infection in all patients, whereas 95 and 60% of these patients were IE NASBA and pp67 NASBA positive, respectively. Although the sensitivity of IE NASBA was very high, the positive predictive value (PPV) of this assay for the onset of a symptomatic infection was only 63%. The PPV of the antigenemia assay as well as pp67 NASBA was considerably higher (80 and 86%, respectively). Thus, the detection of IE mRNA using NASBA appears to be particularly useful as a marker for early initiation of antiviral therapy in patients at high risk for the development of a symptomatic infection. Also, IE NASBA was found to be more sensitive than the antigenemia assay for monitoring CMV infection during antiviral therapy. On the contrary, pp67 NASBA did not appear to have additional diagnostic value compared to the antigenemia assay.

Adult↗
PubMed2000

Complete DNA sequence of the rat cytomegalovirus genome.

We have determined the complete genome sequence of the Maastricht strain of rat cytomegalovirus (RCMV). The RCMV genome has a length of 229,896 bp and is arranged as a single unique sequence flanked by 504-bp terminal direct repeats. RCMV was found to have counterparts of all but one of the open reading frames (ORFs) that are conserved between murine CMV (MCMV) and human CMV (HCMV). Like HCMV, RCMV lacks homologs of the genes belonging to the MCMV m02 glycoprotein gene family. However, RCMV contains 15 ORFs with homology to members of the MCMV m145 glycoprotein gene family. Four ORFs are predicted to encode homologs of host proteins; R33 and R78 both putatively encode G protein-coupled receptors, whereas r144 and r131 encode homologs of major histocompatibility class I heavy chains and CC chemokines, respectively. An intriguing feature of the RCMV genome is the presence of an ORF, r127, with similarity to the rep gene of parvoviruses as well as ORF U94 of human herpesvirus 6A (HHV-6A) and HHV-6B. Counterparts of these ORFs have not been found in the other sequenced herpesviruses.

Amino Acid Sequence↗
PubMed2000

The r144 major histocompatibility complex class I-like gene of rat cytomegalovirus is dispensable for both acute and long-term infection in the immunocompromised host.

The rat cytomegalovirus (RCMV) r144 gene encodes a polypeptide homologous to major histocompatibility complex class I heavy chains. To study the role of r144 in virus replication, an RCMV r144 null mutant strain (RCMVDeltar144) was generated. This strain replicated with efficiency similar to that of wild-type (WT) RCMV in vitro. Additionally, WT RCMV and RCMVDeltar144 were found not to differ in their replication characteristics in vivo. First, the survival rate was similar among groups of immunosuppressed rats infected with either RCMVDeltar144 or WT RCMV. Second, the dissemination of virus did not differ in either RCMVDeltar144- or WT RCMV-infected, immunosuppressed rats, either in the acute phase of infection or approximately 1 year after infection. These data indicate that the RCMV r144 gene is essential neither for virus replication in the acute phase of infection nor for long-term infection in immunocompromised rats. Interestingly, in a local infection model in which footpads of immunosuppressed rats were inoculated with virus, a significantly higher number of infiltrating macrophage cells as well as of CD8(+) T cells was observed in WT RCMV-infected paws than in RCMVDeltar144-infected paws. This suggests that r144 might function in the interaction with these leukocytes in vivo.

Amino Acid Sequence↗
PubMed1999

Deletion of the R78 G protein-coupled receptor gene from rat cytomegalovirus results in an attenuated, syncytium-inducing mutant strain.

The rat cytomegalovirus (RCMV) R78 gene belongs to an uncharacterized class of viral G protein-coupled receptor (GCR) genes. The predicted amino acid sequence of the R78 open reading frame (ORF) shows 25 and 20% similarity with the gene products of murine cytomegalovirus M78 and human cytomegalovirus UL78, respectively. The R78 gene is transcribed throughout the early and late phases of infection in rat embryo fibroblasts (REF) in vitro. Transcription of R78 was found to result in three different mRNAs: (i) a 1.8-kb mRNA containing the R78 sequence, (ii) a 3.7-kb mRNA containing both R77 and R78 sequences, and (iii) a 5.7-kb mRNA containing at least ORF R77 and ORF R78 sequences. To investigate the function of the R78 gene, we generated two different recombinant virus strains: an RCMV R78 null mutant (RCMVDeltaR78a) and an RCMV mutant encoding a GCR from which the putative intracellular C terminus has been deleted (RCMVDeltaR78c). These recombinant viruses replicated with a 10- to 100-fold-lower efficiency than wild-type (wt) virus in vitro. Interestingly, unlike wt virus-infected REF, REF infected with the recombinants develop a syncytium-like appearance. A striking difference between wt and recombinant viruses was also seen in vivo: a considerably higher survival was seen among recombinant virus-infected rats than among RCMV-infected rats. We conclude that the RCMV R78 gene encodes a novel GCR-like polypeptide that plays an important role in both RCMV replication in vitro and the pathogenesis of viral infection in vivo.

Animals↗
PubMed1999

Molecular mimicry by cytomegaloviruses. Function of cytomegalovirus-encoded homologues of G protein-coupled receptors, MHC class I heavy chains and chemokines.

Cytomegaloviruses (CMVs) are well known for their high prevalence rate within host populations as well as their ability to induce lifelong infections. To maintain a persistent and stable relationship with their host, CMVs have evolved various molecular mechanisms to both control host cell metabolism and evade immune surveillance. Among the viral gene products that are likely to be involved in these processes are homologues of cellular G protein-coupled receptors, MHC class I molecules and chemokines. The viral genes encoding these homologues have probably been pirated by the viruses during a long pathogen/host coevolution. In this report, we will discuss the possible functions of these homologues in the pathogenesis of CMV infections.

Animals↗
PubMed1998

The Maastricht strain and England strain of rat cytomegalovirus represent different betaherpesvirus species rather than strains.

The major immediate early (MIE) locus of the Maastricht strain of rat cytomegalovirus (RCMV) was found to comprise five exons of which the first is noncoding. The first three exons are spliced to either exon 4, generating IE1, or exon 5, generating IE2. An additional splicing event unique to RCMV (Maastricht) was identified in exon 5, resulting in a 466-bp deletion. IE1 transcripts were detected exclusively during the IE phase of infection in vitro, whereas IE2 transcripts were detected during both the IE and late phase of infection. The similarities between amino acid sequences derived from the MIE gene of RCMV (Maastricht) and murine cytomegalovirus are low (22 and 37% for IE1 and IE2, respectively). Surprisingly, the similarities between the MIE proteins of RCMV (Maastricht) and the England strain of RCMV are also low (23 and 32% for IE1 and IE2, respectively). This suggests that these RCMV strains represent different betaherpesvirus species rather than strains. This is underscored by the difference between both viruses in genome size as well as growth characteristics. The existence of two different RCMV-like species might have important implications for the use of these viruses as models for human cytomegalovirus.

Animals↗
PubMed1998

The R33 G protein-coupled receptor gene of rat cytomegalovirus plays an essential role in the pathogenesis of viral infection.

We have identified a rat cytomegalovirus (RCMV) gene that encodes a G-protein-coupled receptor (GCR) homolog. This gene (R33) belongs to a family that includes the human cytomegalovirus UL33 gene. R33 was found to be transcribed during the late phase of RCMV infection in rat embryo fibroblasts. Unlike the mRNAs from all the other members of the UL33 family that have been studied to date, the R33 mRNA is not spliced. To study the function of the R33 gene, we constructed an RCMV strain in which the R33 open reading frame is disrupted. The mutant strain (RCMV deltaR33) did not show differences in replication from wild-type RCMV upon infection of several rat cell types in vitro. However, marked differences were seen between the mutant and wild-type strain in the pathogenesis of infection in immunocompromised rats. First, the mutant strain induced a significantly lower mortality than the wild-type virus did. Second, in contrast to wild-type RCMV, the mutant strain did not efficiently replicate in the salivary gland epithelial cells of immunocompromised rats. Although viral DNA was detected in salivary glands of RCMV deltaR33-infected rats up to 14 days postinfection, it could not be detected at later time points. This indicates that although the strain with R33 deleted is probably transported to the salivary glands in a similar fashion to that for wild-type virus, the mutant virus is not able to either enter or replicate in salivary gland epithelial cells. We conclude that the RCMV R33 gene plays a vital role in the pathogenesis of infection.

Amino Acid Sequence↗