interleukin-8 and Cytomegalovirus-Infections

Human cytomegalovirus (HCMV) infection is frequently associated with AIDS patients and immunocompromised recipients of organ transplants. The progression of HCMV infection is related to a complex interrelation of virus replication with the host immune system, including soluble and cellular factors. A chemokine, interleukin-8 (IL-8), is essentially involved in neutrophil-mediated tissue injury. Moreover, several chemokine receptors are co-receptor for HIV entry. Hence, we investigated the effects of IL-8 on HCMV replication in human embryonic fibroblasts, MRC-5 cells. IL-8 augmented both infectious virus production and replication of HCMV, with concomitant increases in the levels of both the HCMV pp71 genome and the synthesis of the HCMV late antigen. The enhancing effect of IL-8 was observed at concentration from 0.1 ng to 10 ng of IL-8/ml, showing a dose-response relationship similar to that observed in the neutrophil chemotactic activity of IL-8. IL-8 did not enhance the growth of MRC-5 cells, indicating that IL-8 enhanced HCMV replication and virus production without affecting the proliferation of host cells. We also found that HCMV selectively induced transcripts of CXCR-1 in fibroblasts by RT-PCR, but significant numbers of binding sites could not be detected on HCMV infected cells by using 125I-labeled IL-8. Thus, IL-8 may enhance HCMV replication in fibroblasts through interaction with small number of CXCR-1. Furthermore, HCMV infection induced IL-8 gene transcription in a human monocytic cell line, THP-1, leading to IL-8 secretion. It is unlikely that HCMV infection enhanced IL-8 production indirectly by inducing the production of some soluble factors, because virus-free filtrated HCMV or UV-irradiated HCMV infected supernatants failed to induce IL-8 production. The functional analysis of the IL-8 gene revealed that both AP-1 and NF-kB factor-binding element were involved in conferring the responsiveness to HCMV. Moreover, electrophoretic mobility shift assay demonstrated that the formation of AP-1 and NF-kB complex was observed upon HCMV infection. These results suggest that IL-8 produced upon HCMV infection, may aggravate HCMV infection by enhancing its replication. Thus, IL-8 and CXCR-1 might be a novel target for intervention therapy for opportunistic HCMV infection.

Topics: Antigens, CD; Cells, Cultured; Cytomegalovirus; Cytomegalovirus Infections; Fibroblasts; Humans; Interleukin-8; Receptors, Interleukin; Receptors, Interleukin-8A; Transcription, Genetic; Virus Replication

Evidence is accumulating that interleukin-8 (IL-8), discovered as a potent neutrophil chemotactic factor, plays a crucial role in establishing acute neutrophil-mediated inflammation by exerting various activities against non-leukocytic as well as leukocytic cells. Various types of cells can rapidly produce a large amount of IL-8 upon stimulation with inflammatory stimuli, such as lipopolysaccharide, IL-1, and tumor necrosis factor (TNF). However, IL-8 production is tightly regulated at several levels, particularly at the transcriptional levels to prevent aberrant production. Our previous experiments demonstrated that IL-8 gene transcription requires the cooperative activation of a transcription factor, NF-kappa B, with an additional transcription factor, AP-1 or NF-IL6, depending on types of cells and stimuli. In addition, we recently observed that infection with Helicobacter pylori or cytomegalovirus activated NF-kappa B, therapy inducing IL-8 protein secretion as well as IL-8 gene transcription. Moreover, IL-8, in turn, enhanced cytomegalovirus replication and infectious virion production. Collectively, these results suggest the potential involvement of IL-8 in the pathology of bacterial or viral infection.

Topics: Cytomegalovirus; Cytomegalovirus Infections; Gene Expression Regulation; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; NF-kappa B; Transcription Factors; Transcription, Genetic; Virus Replication

To observe the clinical efficacy of treating infant cytomegalovirus (CMV) hepatitis with ganciclovir and impact on cytokines.. 76 patients with CMV hepatitis infant were randomly divided into treatment group and control group. The patients of two groups were treated using conventional therapy, on the basis of conventional therapy the treatment group was treated using induction and maintenance treatment of ganciclovir. Before and after treatment the growth and decline in jaundice, liver function, side effects and IL-8 and IFN-γ levels of the patients were detected, clinical efficacy was evaluated.. The clinical total effective rate of the treatment group was 91.4%, it was significantly higher than that of the control group (71.4%), compared with the control group the difference was significant (P < 0.05). After treatment, the growth and decline of jaundice (TBIL and DBIL), liver enzymes (ALT and AST) and cytokines (IL-8 and IFN-γ) levels increased, compared with the pre-treatment and the control group the difference was significant (P < 0.05). The treatment group was no adverse reaction.. The induction and maintenance treatment of ganciclovir in treatment of infant CMV hepatitis can make the body to restore balance of specific cellular immunity, and can significantly improve the symptoms of jaundice, liver function and clinical efficay and it is worthy to be popularized.

Topics: Antiviral Agents; Cytokines; Cytomegalovirus Infections; Female; Ganciclovir; Hepatitis, Viral, Human; Humans; Infant; Interferon-gamma; Interleukin-8; Male

Human cytomegalovirus (CMV) is associated with aspergillosis, but the simultaneous presence of CMV viral interleukin-10 (cmvIL-10) and aspergillosis has never been investigated. CmvIL-10 is produced by CMV-infected cells and acts as an immune modulator during CMV infection. The aim of this study was to evaluate cmvIL-10 levels in peripheral blood and its influence on the clinical outcomes of Aspergillus infection.. Patients who visited or were admitted to the hospital with suspected Aspergillus infection, including invasive aspergillosis (IA) and chronic pulmonary aspergillosis (CPA), were prospectively enrolled. The cmvIL-10, human IL-10 (hIL-10), IL-1B, IL-6, IL-8, IFN-γ, and TNF-α levels in peripheral blood were measured.. Patients with Aspergillus infection had a higher level of cmvIL-10 than the control group (158 ± 305 vs 27.9 ± 30.4 pg/ml, p < .05). The level of cmvIL-10 was not correlated with CMV viremia or end-organ disease. The cmvIL-10 but not hIL-10 level was positively correlated with the IFN-γ level (p < .05) and marginally negatively correlated with IL-1B and IL-8 levels (p < .1). In patients with CPA, a high level of cmvIL-10 (≥100 pg/ml) was a poor prognostic factor for long-term survival (p < .05). In contrast, CMV viremia or end-organ disease was associated with poor survival in patients with IA (p = .05).. Aspergillus infection was associated with CMV coinfection with cmvIL-10 in blood. A cmvIL-10 concentration ≥100 pg/ml was a predictor for unfavourable outcome in CPA patients.

Topics: Aspergillosis; Cytomegalovirus; Cytomegalovirus Infections; Humans; Interleukin-10; Interleukin-8; Viral Proteins; Viremia

Human cytomegalovirus (HCMV) infects the chorioamnion, but whether these infections cause fetal membrane dysfunction remains poorly understood. We sought to assess whether guinea pig cytomegalovirus (GPCMV) infects amnion-derived cells in vitro, compare the inflammatory response of amnion cells to GPCMV and HCMV, and determine if GPCMV infects the amnion in vivo. We found that GPCMV replicates in primary guinea pig amnion derived cells and HPV16 E6/E7-transduced amniotic epithelial cells (AEC[E6/E7]s). HCMV and GPCMV infection of amnion cells increased the transcription of the chemokines CCL5/Ccl5, CXCL8/Cxcl8, and CXCL10/Cxcl10. Myd88-knockdown decreased Ccl5 and Cxc8 transcription in GPCMV-infected AEC[E6/E7]s. GPCMV was detected in the guinea pig amnion after primary maternal infection, revealing that guinea pigs are an appropriate model to study fetal membrane physiology after cytomegalovirus infection. As inflammation is known to cause fetal membrane weakening, the amnion's response to cytomegalovirus infection may cause preterm birth and other adverse pregnancy outcomes.

Topics: Amnion; Animals; Chemokine CCL5; Chemokine CXCL10; Chemokines; Cytomegalovirus; Cytomegalovirus Infections; Female; Guinea Pigs; Humans; Interleukin-8; Pregnancy; Pregnancy Complications

Porcine cytomegalovirus (PCMV) causes lifelong latent infections in swine. The pathogen is occasionally associated with inclusion body rhinitis and pneumonia in piglets, reproductive disorders in pregnant sows and respiratory disease complex in older pigs. Immunosuppressive potential of PCMV infection is discussed. Macrophages were recognised as one of target cell types where propagation of virus occurs. The aim of present study was to set up model PCMV infection of monocyte derived macrophages (MDMs) in vitro for PCMV immunobiology research. Obtained results showed that PCMV is able to infect and propagate in MDMs. Possible immunosuppressive effect of PCMV on infected macrophages was evaluated by measurement of immune relevant gene expression in MDMs. Infection decreased expression of IL-8 and TNF-α (pro-inflammatory cytokines) and increased expression of IL-10 (anti-inflammatory cytokine) on mRNA transcription level. Obtained data support hypothesis that higher sensitivity of animals to coinfection with other swine pathogens and its more severe clinical manifestations could potentially be the consequence of PCMV infection.

Topics: Animals; Cytokines; Cytomegalovirus; Cytomegalovirus Infections; Gene Expression; Immunity, Innate; Interleukin-10; Interleukin-8; Macrophages; Swine; Swine Diseases; Tumor Necrosis Factor-alpha

This study aims to perform comprehensive longitudinal immune factor analysis of aqueous humor in relation to the aqueous CMV viral load and systemic CD4 counts during treatment of patients with co-infection of HIV and CMVR.. Aqueous humor samples were collected from 17 HIV-positive patients with CMVR scheduled to undergo weekly intravitreal ganciclovir therapy as part of the prospective CMV Retinitis Intravitreal Ganciclovir Singapore Study (CRIGSS) over the course of 1year. Full data across all the 4 time points was obtained and analyzed for CMV DNA viral load, 41 cytokine and chemokine factors using real-time PCR with the FlexMAP 3D (Luminex®) platform and assessed using the Milliplex Human Cytokine® kit.. The following immune factors (Spearman correlation coefficient r value in parenthesis, p<0.05) showed strong correlation with CMV DNA load in the aqueous - MCP-1 (0.80, IFN-g (0.83), IP-10 (0.82), IL-8 (0.81), fractalkine (0.73), RANTES (0.68) - while the following showed moderate correlation - PDGF-AA (0.58), Flt-3L (0.59) and G-CSF (0.53). Only PDGF-AA revealed a statistically significant negative correlation with serum CD4 levels (r=-0.74).. Immune factors that correlate with intraocular CMV DNA load are identified. They are indicative of a Th1 and monocyte-macrophage mediated response, and exhibit a decreasing trend longitudinally through the course of treatment. These factors may be an important new consideration in individualizing the treatment of patients with CMVR.

Topics: Adult; Antiviral Agents; Aqueous Humor; CD4-Positive T-Lymphocytes; Coinfection; Cytomegalovirus; Cytomegalovirus Infections; Cytomegalovirus Retinitis; Female; Ganciclovir; Granulocyte Colony-Stimulating Factor; HIV; HIV Infections; Humans; Immunologic Factors; Interleukin-8; Longitudinal Studies; Male; Middle Aged; Prospective Studies; Singapore

Human cytomegalovirus (HCMV), a β-herpesvirus, has evolved many strategies to subvert both innate and adaptive host immunity in order to ensure its survival and propagation within the host. Induction of IL-8 is particularly important during HCMV infection as neutrophils, primarily attracted by IL-8, play a key role in virus dissemination. Moreover, IL-8 has a positive effect in the replication of HCMV. This work has identified an HCMV gene (UL76), with the relevant property of inducing IL-8 expression at both transcriptional and protein levels. Up-regulation of IL-8 by UL76 results from activation of the NF-kB pathway as inhibition of both IKK-β activity or degradation of Ikβα abolishes the IL-8 induction and, concomitantly, expression of UL76 is associated with the translocation of p65 to the nucleus where it binds to the IL-8 promoter. Furthermore, the UL76-mediated induction of IL-8 requires ATM and is correlated with the phosphorylation of NEMO on serine 85, indicating that UL76 activates NF-kB pathway by the DNA Damage response, similar to the impact of genotoxic drugs. More importantly, a UL76 deletion mutant virus was significantly less efficient in stimulating IL-8 production than the wild type virus. In addition, there was a significant reduction of IL-8 secretion when ATM -/- cells were infected with wild type HCMV, thus, indicating that ATM is also involved in the induction of IL-8 by HCMV. In conclusion, we demonstrate that expression of UL76 gene induces IL-8 expression as a result of the DNA damage response and that both UL76 and ATM have a role in the mechanism of IL-8 induction during HCMV infection. Hence, this work characterizes a new role of the activation of DNA Damage response in the context of host-pathogen interactions.

Topics: Ataxia Telangiectasia Mutated Proteins; Cell Line; Cell Nucleus; Cytomegalovirus; Cytomegalovirus Infections; DNA Damage; Genes, Reporter; Host-Pathogen Interactions; Humans; I-kappa B Kinase; Interleukin-8; Mutation; Protein Transport; Proteolysis; Recombinant Proteins; RNA Interference; Signal Transduction; Trans-Activators; Transcription Factor RelA; Up-Regulation; Virus Replication

Cervical shedding of HIV-1 DNA may influence HIV-1 sexual transmission. HIV-1 DNA was detected in 250 (80%) of 316 and 207 (79%) of 259 cervical cytobrush specimens from 56 US and 80 Kenyan women, respectively. Plasma HIV-1 RNA concentration was associated with increased HIV-1 DNA shedding among US and Kenyan women. Kenyan women had higher cervicovaginal concentrations of proinflammatory interleukins (IL)-1β, IL-6, IL-8, and anti-inflammatory secretory leukocyte protease inhibitor compared with US women (all P < 0.01). HIV-1 DNA shedding was associated with increased concentrations of IL-1β and IL-6 and lower secretory leukocyte protease inhibitor among US women but not Kenyan women.

Topics: Adult; Anti-Retroviral Agents; CD4 Lymphocyte Count; Cervix Uteri; Cytomegalovirus Infections; DNA, Viral; Female; Herpes Genitalis; HIV Infections; HIV-1; Humans; Interleukin-1; Interleukin-1beta; Interleukin-8; Kenya; Middle Aged; Prospective Studies; Reproductive Tract Infections; RNA, Viral; United States; Uterine Cervicitis; Vagina; Vaginitis; Viral Load

In vitro studies suggest that human cytomegalovirus (CMV) modulates the functions of dendritic cells (DCs). However, there are limited data on DC homeostasis in CMV-infected patients.. The aim of this study was to characterize circulating DCs and plasma cytokine levels in immunocompetent patients with primary, symptomatic CMV infections.. The study population consisted of 14 patients suffering of CMV mononucleosis and 14 healthy volunteers (11 CMV-seropositive and 3 CMV-seronegative subjects) included as controls. Peripheral blood mononuclear cells were isolated and used to characterize DCs and to quantify CMV in the blood. Plasma levels of pro-inflammatory and anti-inflammatory cytokines were also measured.. We observed that patients who were developing CMV mononucleosis presented lower myeloid and plasmacytoid DC counts in peripheral blood compared with healthy controls. We also noted elevated levels of inflammatory mediators, of which tumor necrosis factor-α (TNF-α)-which activates DCs and endothelial cells-was the highest. Notably, the decrease in blood DCs correlated with high TNF-α and IL-8 levels by a hyperbolic function.. Our results suggest that increased levels of inflammatory factors facilitate alterations in DC homeostasis during primary CMV infection, which may contribute to viral-induced modulation of host immunity.

Topics: Adult; Aged; Antigen-Presenting Cells; Blood; Cytokines; Cytomegalovirus; Cytomegalovirus Infections; Dendritic Cells; Female; Humans; Infectious Mononucleosis; Inflammation; Interleukin-8; Leukocytes, Mononuclear; Male; Middle Aged; Tumor Necrosis Factor-alpha; Up-Regulation

The immune system undergoes profound age-related changes, including a gradual increase in the production and circulation of proinflammatory cytokines. Despite the known capacity of fibroblasts to produce cytokines, little is known so far about the inflammatory response of fibroblasts to cellular stress such as viral and/or bacterial infection in the context of aging. Therefore the aim of this study was to analyze the levels of IL6 and IL8 secretion in supernatants of human skin fibroblasts from young and elderly persons. Cytokine and chemokine secretion was analyzed before and after in vitro infection of the cells with Cytomegalovirus (CMV) and/or stimulation with Lipopolysaccharide (LPS). The exposure of fibroblasts to these agents caused inflammatory changes, reflected by the secretion of both the cytokine IL6 and the chemokine IL8 by fibroblasts from young as well as elderly persons. The cytokine/chemokine production induced by either agent alone could be further increased by co-stimulation of the cells with both stimuli. The level of protein secretion was dependent on the chronological age of the fibroblasts. Stimulated human skin fibroblasts from elderly donors produced higher amounts of IL6 as well as IL8 than fibroblasts from young donors. These differences were more pronounced for IL6 than for IL8. The inflammatory response of fibroblasts to stimulation differed among donors and did not correspond to the responsiveness of whole blood derived from the same person. In summary lifelong CMV-infection may act as an in vivo trigger for inflammatory changes by increasing the inflammatory response to bacterial products such as LPS. It may thus contribute to age-related inflammatory processes, referred to as 'inflamm-aging'.

Topics: Adult; Aged, 80 and over; Aging; Blotting, Western; Cells, Cultured; Cytomegalovirus Infections; Female; Fibroblasts; Humans; Inflammation; Interleukin-6; Interleukin-8; Lipopolysaccharides; Male; Young Adult

Pulmonary cytomegalovirus (CMV) infection causes fatal CMV pneumonia (CMVp) in immunocompromised patients; however, the mechanisms underlying CMV-infection-induced pulmonary lesion development remain largely unknown. We examined the relationship between CMVp patterns and intrapulmonary viral tropism, including expression of inflammatory cytokines and related molecules. Double immunohistochemistry of CMV antigen and cellular markers showed that epithelial tropism was associated with a diffuse alveolar damage (DAD) pattern (CMVp-DAD) while stromal tropism was associated with a predominantly interstitial inflammation/fibrosis (IIF) (CMVp-IIF) or a combination of DAD and IIF (CMVp-complex). Transforming growth factor (TGF)-β1 expression was relevant to CMV-induced tissue injury, and its expression was higher in CMVp-complex and CMVp-IIF than in CMVp-DAD. Expression of integrin β6 (ITGB6), an adhesion molecule and important activator of TGF-β1 in interstitial pneumonia, was lost in CMV-infected pneumocytes, especially CMVp-DAD, whereas CMV-negative pneumocytes in CMVp-complex and CMVp-IIF showed overexpression. Diffuse interleukin (IL)-8 up-regulation and strong expression were present in both CMV-infected pneumocytes and stromal cells only in CMVp-IIF cases with marked interstitial neutrophilic infiltration. On the basis of viral tropism and the expression of TGF-β1, ITGB6, and IL-8, we conclude that CMV-infected pulmonary cells play an important role in the development of diverse CMVp patterns.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Viral; Cytokines; Cytomegalovirus; Cytomegalovirus Infections; Fatal Outcome; Female; Humans; Integrin beta Chains; Interleukin-8; Male; Middle Aged; Pneumonia, Viral; Transforming Growth Factor beta1; Viral Tropism

Human cytomegalovirus (HCMV) is linked to the acceleration of vascular diseases such as atherosclerosis and transplant vasculopathy. One of the hallmarks of these diseases is angiogenesis (AG) and neovessel formation. Endothelial cells (ECs) are an integral part of AG and are sites of HCMV persistence. AG requires multiple synchronous processes that include EC proliferation, migration, and vessel stabilization. Virus-free supernatant (secretome) from HCMV-infected ECs induces AG. To identify factor(s) involved in this process, we performed a human cytokine array. Several cytokines were significantly induced in the HCMV secretomes including interleukin-6 (IL-6), granulocyte macrophage colony-stimulating factor, and IL-8/CXCL8. Using in vitro AG assays, neutralization of IL-6 significantly reduced neovessel formation. Addition of the HCMV secretome to preformed vessels extended neovessel survival, but this effect was blocked by neutralization of IL-6. In these cells, IL-6 prevented apoptosis by blocking caspase-3 and -7 activation through the induction of survivin. Neutralization of IL-6 receptor on ECs abolished the ability of HCMV secretome to increase survivin expression and activated effector caspases. Moreover, survivin shRNA expression induced rapid regression of tubule capillary networks in ECs stimulated with HCMV secretome and activated effector caspases. These observations may explain how CMV accelerates vascular disease despite limited infection in tissues.

Topics: Angiogenesis Inducing Agents; Apoptosis; Blotting, Western; Caspase 3; Cell Adhesion; Cell Movement; Cell Proliferation; Cells, Cultured; Cytomegalovirus; Cytomegalovirus Infections; Endothelium, Vascular; Humans; Inhibitor of Apoptosis Proteins; Interleukin-6; Interleukin-8; Macrophage Colony-Stimulating Factor; Microtubule-Associated Proteins; Receptors, Interleukin-6; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Small Interfering; Survivin; Umbilical Veins

DC-SIGN is a member of the C-type lectin family. Mainly expressed by myeloid DCs, it is involved in the capture and internalization of pathogens, including human CMV. Several transcripts have been identified, some of which code for putative soluble proteins. However, little is known about the regulation and the functional properties of such putative sDC-SIGN variants. To better understand how sDC-SIGN could be involved in CMV infection, we set out to characterize biochemical and functional properties of rDC-SIGN as well as naturally occurring sDC-SIGN. We first developed a specific, quantitative ELISA and then used it to detect the presence sDC-SIGN in in vitro-generated DC culture supernatants as cell-free secreted tetramers. Next, in correlation with their inflammatory status, we demonstrated the presence of sDC-SIGN in several human body fluids, including serum, joint fluids, and BALs. CMV infection of human tissues was also shown to promote sDC-SIGN release. Based on the analysis of the cytokine/chemokine content of sDC-SIGN culture supernatants, we identified IFN-γ and CXCL8/IL-8 as inducers of sDC-SIGN production by MoDC. Finally, we demonstrated that sDC-SIGN was able to interact with CMV gB under native conditions, leading to a significant increase in MoDC CMV infection. Overall, our results confirm that sDC-SIGN, like its well-known, counterpart mDC-SIGN, may play a pivotal role in CMV-mediated pathogenesis.

Topics: Body Fluids; Cell Adhesion Molecules; Cloning, Molecular; Cytomegalovirus; Cytomegalovirus Infections; Dendritic Cells; Enzyme-Linked Immunosorbent Assay; Exosomes; Female; HEK293 Cells; Humans; Inflammation; Interferon-gamma; Interleukin-8; Lectins, C-Type; Matrix Metalloproteinases; Mucous Membrane; Myeloid Cells; Protein Isoforms; Protein Multimerization; Protein Processing, Post-Translational; Receptors, Cell Surface; Reproducibility of Results; Signal Transduction; Solubility; Titrimetry; Up-Regulation

Human cytomegalovirus (HCMV), the β-herpesvirus prototype, has evolved a wide spectrum of mechanisms to counteract host immunity. Among them, HCMV uses cellular captured genes encoding molecules capable of interfering with the original host function or of fulfilling new immunomodulatory tasks. Here, we report on UL7, a novel HCMV heavily glycosylated transmembrane protein, containing an Ig-like domain that exhibits remarkable amino acid similarity to CD229, a cell-surface molecule of the signalling lymphocyte-activation molecule (SLAM) family involved in leukocyte activation. The UL7 Ig-like domain, which is well-preserved in all HCMV strains, structurally resembles the SLAM-family N-terminal Ig-variable domain responsible for the homophilic and heterophilic interactions that trigger signalling. UL7 is transcribed with early-late kinetics during the lytic infectious cycle. Using a mAb generated against the viral protein, we show that it is constitutively shed, through its mucine-like stalk, from the cell-surface. Production of soluble UL7 is enhanced by PMA and reduced by a broad-spectrum metalloproteinase inhibitor. Although UL7 does not hold the ability to interact with CD229 or other SLAM-family members, it shares with them the capacity to mediate adhesion to leukocytes, specifically to monocyte-derived DCs. Furthermore, we demonstrate that UL7 expression attenuates the production of proinflammatory cytokines TNF, IL-8 and IL-6 in DCs and myeloid cell lines. Thus, the ability of UL7 to interfere with cellular proinflammatory responses may contribute to viral persistence. These results enhance our understanding of those HCMV-encoded molecules involved in sustaining the balance between HCMV and the host immune system.

Topics: Animals; Antibodies, Monoclonal; Antigens, CD; Base Sequence; Cell Line; Cytokines; Cytomegalovirus; Cytomegalovirus Infections; Dendritic Cells; Humans; Immune Evasion; Immunologic Factors; Interleukin-6; Interleukin-8; Leukocytes; Lymphocyte Activation; Metalloproteases; Mice; Molecular Sequence Data; Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; Signaling Lymphocytic Activation Molecule Family; Tumor Necrosis Factors; Viral Envelope Proteins; Viral Matrix Proteins

Appendicitis is a very common surgical diagnosis with unclear pathology. Human cytomegalovirus (HCMV) can modulate our immune system and has been associated with inflammatory bowel disease (IBD) and various other inflammatory diseases.. We investigated the association between HCMV and acute appendicitis in 14 immunocompetent patients. Tissue sections from 10 AIDS patients with verified HCMV infection were used as positive controls, and uninflamed intestinal tissue sections from 12 patients were used as negative controls.. Cells double positive for HCMV early antigens and IL-6/IL-8 were observed in the appendices of 64.3% of appendicitis patients (9 of 14) by immunohistochemical analysis. HCMV late antigen was found in the appendices of 42.9% of the acute appendicitis patients (6 of 14). Latent HCMV appendix infection, as verified by in situ hybridization, as well as HCMV IgG, was observed in 78.6% of patients (11 of 14). The study samples from all 6 healthy appendices were negative for HCMV early and late antigens, although 50% (3 of 6) were HCMV IgG and HCMV DNA positive.. We have shown that HCMV infection of the appendix is associated with acute appendicitis (P = 0.002) and possibly with the severity of the disease. Our study identified HCMV as a pathogen to be sought for in the appendicitis patient group, possibly allowing further medical treatment of these patients.

Topics: Adult; Aged; Antigens, Viral; Appendicitis; Case-Control Studies; Cytomegalovirus Infections; Disease Progression; Female; Humans; Immunocompetence; Interleukin-6; Interleukin-8; Male; Middle Aged; Prevalence; Sweden

Cytomegalovirus (CMV) is the most common cause of congenital infection worldwide and occurs as a result of transplacental transmission of the virus. The human neonate is highly susceptible to infection due to a combination of immaturity of the immune system and antigenic inexperience. This study uses the in vivo model of congenital CMV to examine both the humoral and cell-mediated immune responses in vertically infected neonates and their mothers. Ten pairs of matched neonates and their mothers were evaluated for specific IgM responses to three immunodominant CMV antigens: pp38 (pUL80a), pp52 (pUL44) and pp150 (pUL32). In contrast to conventional enzyme immunoassay (EIA) testing for CMV-specific IgM, which found five of the mothers and four of the neonates to be positive, Western immunoblotting showed all 10 adults and nine newborns to be positive. Eight mothers and nine newborns had serological evidence of primary infection. All neonates showed a response to pp38, an assembly protein, nine responded to the pp52 immediate early antigen but only four had reactivity to the pp150 tegument associated protein. Of the mothers, eight had pp38 reactivity, 10 showed a response to the pp52 antigen and seven to the pp150 antigen. T cell-mediated immunity was assessed by measuring cytokines using a multiplex microarray assay. Levels of interferon (IFN)-gamma were high in both groups [mean +/- standard error of the mean (s.e.m.): neonates = 657 +/- 238 pg/ml, mothers = 1072 +/- 677 pg/ml, pNS]; however, neonates had significantly higher levels of interleukin (IL)-8 (316 +/- 136 pg/ml versus 48 +/- 28 pg/ml, P < 0.005). Similar levels of IL-2, IL-7, IL-10 and IL-12 were measured in both groups, but levels of IL-1alpha, IL-1beta, IL-4, IL-6 and tumour necrosis factor (TNF)-alpha were either absent or low. In response to CMV, neonates and adults mount a predominant T helper 1 (Th1) response, as evidenced by the presence of IL-2, IL-8, IL-12 and IFN-gamma with concomitant lack of IL-4. These findings suggest that the neonate, when presented with infection in utero, is capable of mounting an individual response; however, the lower IFN-gamma and higher IL-8 levels suggest reduced immune responsiveness when compared to their adult counterparts.

Topics: Adolescent; Adult; Antibodies, Viral; Antibody Specificity; Antigens, Viral; Blotting, Western; Cytomegalovirus; Cytomegalovirus Infections; Female; Humans; Immunity, Cellular; Immunoglobulin M; Infant, Newborn; Infectious Disease Transmission, Vertical; Interferon-gamma; Interleukin-8; Pregnancy; Pregnancy Complications, Infectious; Th1 Cells

Human cytomegalovirus (HCMV) and human immunodeficiency virus type-1 (HIV-1) infect the female genital tract. A human cervical explant model was developed to study single and dual infection by these viruses in the genital compartment. An HCMV strain expressing green fluorescent protein, and two clinical HCMV strains produced peak viral DNA copies at 14 to 21 days post-infection. Peak levels of HIV-1(Ba-L) p24 antigen occurred at 7 days post-infection. HIV-1(Ba-L) appeared to enhance HCMV in co-infected tissues. Singly and dually infected explants produced increased levels of cytokines IL-6, IL-8, and GRO-alpha in culture supernatants. Immunohistochemical and flow cytometric analysis showed HCMV infection of leukocytes with the phenotype CD45+/CD1a+/CD14+/HLA-DR+ but not stromal or endothelial cells. Cells expressing both GFP and HIV-1 p24 antigen were detected in co-infected tissues. The cervical explants provide an ex vivo human model for examining mechanisms of virus-virus interaction and pathogenesis in clinically relevant tissue.

Topics: Cervix Uteri; Chemokine CXCL1; Culture Media; Cytomegalovirus; Cytomegalovirus Infections; Endothelial Cells; Female; Genes, Reporter; Green Fluorescent Proteins; HIV Core Protein p24; HIV Infections; HIV-1; Humans; Interleukin-6; Interleukin-8; Leukocytes; Organ Culture Techniques; Stromal Cells

The aim of this study was to examine whether chronic infections and genetic factors of the host play roles in the pathophysiology of acute noncardioembolic ischemic stroke. Blood samples from 59 subjects with ischemic stroke and 52 control patients were investigated by nested PCR for the presence of C. pneumoniae DNA, HCMV DNA and enterovirus RNA, by ELISA for the levels of antibodies to C. pneumoniae, HCMV, HSV, HHV-6, EBV and the inflammatory chemokine IL-8, and by PCR for promoter polymorphism of the IL-8 and CD14 host genes. Associations of stroke with the HCMV IgG and HSV-1 IgA antibody levels were observed. No association of stroke was detected with the presence of C. pneumoniae, HCMV or enterovirus nucleic acids in the peripheral blood, C. pneumoniae IgM, IgG and IgA, the HSV IgG, the EBV IgG, or HHV-6 IgG antibody levels, the pathogen burden, the IL-8 or CD14 promoter polymorphisms, or with the serum levels of IL-8 in the overall study population. These results are consistent with the hypothesis that certain pathogens are involved in the development of ischemic stroke.

Topics: Adult; Aged; Antibodies, Viral; Chlamydophila Infections; Chlamydophila pneumoniae; Chronic Disease; Cytomegalovirus; Cytomegalovirus Infections; DNA, Bacterial; DNA, Viral; Enterovirus; Enterovirus Infections; Enzyme-Linked Immunosorbent Assay; Genetic Predisposition to Disease; Herpesviridae; Herpesviridae Infections; Herpesvirus 1, Human; Humans; Interleukin-8; Ischemia; Lipopolysaccharide Receptors; Middle Aged; Polymerase Chain Reaction; Polymorphism, Genetic; Promoter Regions, Genetic; Risk Factors; RNA, Viral; Stroke

We sought to determine whether cytomegalovirus infection of extravillous trophoblast cells induces inflammatory changes that lead to cell death.. Extravillous trophoblast (HTR-8/SVneo) cells were infected with cytomegalovirus. Cell death assays were performed 8 to 48 hours after infection. The expression and secretion of cytokines (interleukin-6 and -8) preceding cell death was measured by polymerase chain reaction and enzyme-linked immunosorbent assay.. Cell viability (lactate dehydrogenase assay) was reduced approximately 20%, and rates of apoptosis (measured by TdT-mediated dUTP-X nick end labeling [TUNEL] assay) were increased approximately 40% at 16 to 48 hours after infection. Significantly elevated levels of caspase-3 mRNA levels were observed before increased cell death. Interleukin-6 and -8 mRNA expression and protein secretion were up-regulated 8 to 16 hours after cytomegalovirus infection.. Placental exposure to cytomegalovirus induces an inflammatory response that precedes invasive trophoblast cell death. Cytomegalovirus may prevent normal placental invasion, which results in adverse reproductive outcomes that are associated with placental dysfunction.

Topics: Apoptosis; Caspase 3; Caspases; Cell Death; Cell Survival; Cytomegalovirus Infections; Female; Humans; In Situ Nick-End Labeling; In Vitro Techniques; Interleukin-6; Interleukin-8; L-Lactate Dehydrogenase; Placenta; Trophoblasts

Human cytomegalovirus virus (CMV) is a major cause of morbidity and mortality in immunocompromised individuals. Recently, a novel group of cytokines [interleukin (IL)-28A/B and IL-29, also termed interferon (IFN)-lambdas] has been described. Here, we demonstrate that intestinal epithelial cell (IEC) lines as well as murine and human colonic tissue express the IFN-lambda receptor subunits IL-28R and IL-10R2. IL-28A and IL-29 binding to their receptor complex activates ERK-1/2 and stress-activated protein kinase/c-Jun NH2-terminal kinase MAPKs and Akt, resulting in increased IL-8 protein expression. IFN-lambdas also induce phosphorylation of signal transducer and activator of transcription 1 and significantly increase mRNA expression of suppressor of cytokine signaling 3 and the antiviral proteins myxovirus resistance A and 2',5'-oligoadenylate synthetase. These signals result in an up to 83% reduction of cells positive for human CMV immediate-early protein after human CMV infection. In mice, IL-28A mRNA expression is upregulated after infection with murine CMV in vivo. Both IL-28A and IL-29 significantly decrease cell proliferation but have no effect on Fas-induced apoptosis. In conclusion, IECs express functional receptors for IFN-lambdas, which mediate antiviral and antiproliferative signals in IECs, suggesting a potential for therapeutic use in certain viral infections and as (antiproliferative) anticancer therapy.

Topics: 2',5'-Oligoadenylate Synthetase; Animals; Apoptosis; Cell Line, Tumor; Cell Proliferation; Colon; Cytokines; Cytomegalovirus Infections; Epithelial Cells; Gene Expression Regulation; GTP-Binding Proteins; Humans; Interferons; Interleukin-8; Interleukins; Intestinal Mucosa; Mice; Mice, Inbred C57BL; Mitogen-Activated Protein Kinases; Myxovirus Resistance Proteins; Receptors, Cytokine; Signal Transduction

Recently, we reported that thrombin specifically stimulates protease-activated receptor-1 (PAR-1) signaling in RPE entailing inhibition of Sp1 dependent HCMV replication. We now studied whether thrombin modulates the expression of the proinflammatory cytokine/chemokines IL-6 and IL-8 in mock- and cytomegalovirus-infected human retinal pigment epithelial cells (RPE). Our data show that thrombin/PAR-1 stimulates IL-6 and IL-8 gene transcription and protein secretion in both mock- and HCMV-infected RPE. Thrombin/PAR-1-mediated signaling stimulated PKC and NF-kappaB-dependent IL-6 and IL-8 gene expression via phosphoinositide 3-kinase and further downstream via p42/44 and p38 MAPKs. Thus, thrombin/PAR-1-mediated IL-6/IL-8 gene expression is uncoupled from Sp1 inhibition and may support proinflammatory pathomechanisms probably involved in hemorrhage/HCMV retinitis progression.

Topics: Cytomegalovirus; Cytomegalovirus Infections; Humans; Interleukin-6; Interleukin-8; Pigment Epithelium of Eye; Receptor, PAR-1; Signal Transduction; Thrombin

The probable risk factors leading to aortic valve calcification are not clearly defined. The cross-sectional study of 85 patients with vascular and valvular calcification was performed. Correlations between the immune tests and aortic stenosis severity were investigated. The predictors of aortic valve calcification were probably C-reactive protein and interleukin-6. The predictors of aortic stenosis progression were interleukin-8, antibodies of Chlamydia pneumoniae and cytomegalovirus, and dysregulation of complement's components. Implication of immune reactivity could influence aortic valve calcification.

Topics: Aged; Aged, 80 and over; Antibodies, Bacterial; Antibodies, Viral; Aortic Valve Stenosis; C-Reactive Protein; Calcinosis; Chlamydia Infections; Chlamydophila pneumoniae; Cross-Sectional Studies; Cytomegalovirus; Cytomegalovirus Infections; Disease Progression; Humans; Interleukin-6; Interleukin-8; Male; Risk Factors; Russia

Monocytes play a prominent role in inflammation, coagulation and atherosclerosis by their ability to produce tissue factor (TF) and cytokines. The aim of the present study was to establish whether virus-infected monocytes initiate coagulation. In addition, the production of cytokines by monocytes may accelerate the chronic process of atherosclerosis and may contribute to coronary syndromes by eliciting plaque instability.. Monocytes were isolated by Vacutainer(R), BD Biosciences, Alphen aan den Rijn, Netherlands and subsequent magnetic cell sorting (MACS(R), Milteny Biotec, Bergish Gladbach, Germany). Coagulation times in normal pooled plasma and Factor VII-deficient plasma were measured after infection with cytomegalovirus (CMV), Chlamydia pneumoniae (Cp) and influenza A\\H1N1. Anti-TF antibodies were added to neutralize TF expressed on monocytes. Interleukins (IL) 6, 8 and 10 were measured in the supernatants.. Chlamydia pneumoniae- and CMV-infected monocytes decreased the clotting time by 60%, and influenza-infected monocytes by 19%, as compared to uninfected monocytes. Procoagulant activity was absent when Factor VII-deficient plasma or anti-TF antibodies were used. Monocytes produced both IL-6 and IL-8 after infection with CMV (317 pg mL-1 and 250 pg mL-1) or Cp (733 pg mL-1 and 268 pg mL-1). Similar results were obtained for influenza virus-infected monocytes, but the levels of both cytokines were 3-5-fold higher (1797 pg mL-1 and 725 pg mL-1). Interleukin-10 was not produced by infected monocytes.. The procoagulant activity of virus-infected monocytes is TF-dependent. Although influenza infection did not generate a significant reduction in clotting time, the pronounced expression of IL-6 and IL-8 may induce local and/or systemic inflammatory reactions, which may be associated with plaque rupture and atherosclerosis. The lack of production of the anti-inflammatory cytokine IL-10 may even accelerate these processes.

Topics: Antibodies; Chlamydophila Infections; Coronary Artery Disease; Cytomegalovirus Infections; Humans; Influenza A virus; Influenza, Human; Interleukin-10; Interleukin-6; Interleukin-8; Monocytes; Thromboplastin; Virus Diseases; Whole Blood Coagulation Time

Human cytomegalovirus (CMV) infection has been linked to chronic heart disease. The mechanism of CMV dissemination to the heart remains unknown. CMV antigens and nucleic acid sequences have been detected in endothelial cells (ECs) in vivo, and ECs are fully permissive hosts to CMV replication in vitro. This report examines the characteristics of CMV replication in primary cultures of human heart microvascular ECs (HHMECs).. Capillary ECs were isolated from heart tissue biopsies of six patients at the time of heart surgery. HHMECs were infected with CMV and viral antigens were detected by immunofluorescence assay using monoclonal antibodies as specific reagents. Cytokine and chemokine release in the supernatant of sham- and CMV-infected cells was quantitated by ELISA. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to analyse expression of mRNA for adhesion molecules.. CMV was found to productively infect HHMECs without cytolytic effects. Infected cultures released high levels of pro-inflammatory chemokines and enhanced their adhesion molecule expression.. Our data provide new insights into the mechanism of CMV dissemination to the heart, signalling the need for further investigation of the pathogenetic role of this virus in cardiac disorders.

Topics: Adult; Aged; Antigens, Viral; Cells, Cultured; Chemokine CCL2; Cytomegalovirus; Cytomegalovirus Infections; E-Selectin; Endothelium, Vascular; Humans; Intercellular Adhesion Molecule-1; Interleukin-6; Interleukin-8; Microcirculation; Microscopy, Fluorescence; Microscopy, Phase-Contrast; Middle Aged; Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; Vascular Cell Adhesion Molecule-1; Virus Replication

The possible contribution of cytomegalovirus (CMV) to pathogenetic events associated with atherosclerotic lesion establishment and progression is still controversial. We evaluated the possibility that active ongoing CMV infection could be correlated to evolution of unstable atheromatous lesion, by analyzing patients suffering from unstable angina (n=61), acute myocardial infarction (n=43), stable angina (n=26) and peripheral arteriopathy (n=22) as compared to healthy subjects (n=30). Particularly, we assessed: past exposure to CMV by evaluating anti-CMV IgG antibodies; ongoing CMV infection by evaluating anti-CMV IgM antibodies and circulating interleukin (IL)-8 in serum; and CMV DNAemia in peripheral blood mononuclear cells (PBMC). Mean IgG values were significantly increased in patients from all groups, as compared to healthy subjects. CMV-specific IgM, as well as CMV DNAemia, were undetectable in both controls and patients. Circulating IL-8, significantly elevated in a group of individuals experiencing active CMV infection, was not significantly higher in cardiovascular disease patients, as compared to control subjects. These findings confirm previous evidence from the increased exposure to CMV infection in patients with atheromatous lesions. However, they provide further evidence against a direct implication of active systemic CMV infection in the pathogenesis of cardiovascular diseases, particularly those involving plaque instability.

Topics: Aged; Antibodies, Viral; Arteriosclerosis; Cytomegalovirus; Cytomegalovirus Infections; DNA, Viral; Female; Gene Dosage; Humans; Immunoglobulin G; Immunoglobulin M; Interleukin-8; Male; Middle Aged; Reference Values

Glial cells function as sensors for infection within the brain and produce cytokines to limit viral replication and spread. We examined both cytokine (TNF-alpha, IL-1beta, and IL-6) and chemokine (MCP-1, MIP-1alpha, RANTES, and IL-8) production by primary human glial cells in response to cytomegalovirus (CMV). Although CMV-infected astrocytes did not produce antiviral cytokines, they generated significant quantities of the chemokines MCP-1 and IL-8 in response to viral infection. On the other hand, supernatants from CMV-stimulated purified microglial cell cultures showed a marked increase in the production of TNF-alpha and IL-6, as well as chemokines. Supernatants from CMV-infected astrocyte cultures induced the migration of microglia towards chemotactic signals generated from infected astrocytes. Antibodies to MCP-1, but not to MIP-1alpha, RANTES, or IL-8, inhibited this migratory activity. These findings suggest that infected astrocytes may use MCP-1 to recruit antiviral cytokine-producing microglial cells to foci of infection. To test this hypothesis, cocultures of astrocytes and microglial cells were infected with CMV. Viral gene expression in these cocultures was 60% lower than in CMV infected purified astrocyte cultures lacking microglia. These results support the hypothesis that microglia play an important antiviral role in defense of the brain against CMV. The host defense function of microglial cells may be directed in part by chemokines, such as MCP-1, produced by infected astrocytes.

Topics: Astrocytes; Brain; Cells, Cultured; Chemokine CCL3; Chemokine CCL4; Chemokine CCL5; Chemotaxis; Coculture Techniques; Cytomegalovirus; Cytomegalovirus Infections; Encephalitis, Viral; Fetus; Gene Expression Regulation, Viral; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Macrophage Inflammatory Proteins; Microglia; RNA, Messenger; Tumor Necrosis Factor-alpha; Virus Replication

Human cytomegalovirus (HCMV) infection is associated with excessive proinflammatory immune responses such as cytokine/chemokine production or upregulation of adhesion molecules on the host cells. It is assumed that these features of HCMV-related immunopathology can not be treated effectively with currently available anti HCMV drugs. In the present study the efficacy of ganciclovir (GCV), foscarnet (PFA), cidofovir (HPMPC), and ISIS 2922, an antisense oligonucleotide complementary to HCMV immediate-early (IE) mRNA, was investigated on HCMV-induced secretion and functional activity of the C-X-C chemokine IL-8 and the expression of the intercellular adhesion molecule-1 (ICAM-1). As compared with mock-infected cells IL-8 production was increased up to 9-fold and ICAM-1 expression up to 4-fold in HCMV-infected fibroblasts. Treatment of infected cells with GCV (40 microM), PFA (200 microM) or HPMPC (2 microM) suppressed completely virus replication as demonstrated by quantification of late (L) antigen expression and infectious virus production. These drugs, however, failed to inhibit IE antigen expression and did not prevent HCMV-induced upregulation of IL-8 and ICAM-1. In contrast, ISIS 2922 (1 microM) suppressed both IE and L antigen expression by 99% and inhibited infectious virus production by 10(4)-fold. Moreover, ISIS 2922 significantly suppressed HCMV-induced upregulation of both IL-8 and ICAM-1 expression on the transcriptional and on the protein level. Our results indicate that ISIS 2922 but not inhibitors of HCMV DNA prevents HCMV-induced upregulation of IL-8 and ICAM-1, both hallmarks of inflammatory processes. Thus, inhibition of HCMV IE expression with ISIS 2922 may be an important strategy for the treatment of HCMV-related immunopathogenesis.

Topics: Antiviral Agents; Cells, Cultured; Chemotaxis, Leukocyte; Cidofovir; Cytomegalovirus Infections; Cytosine; Fibroblasts; Foscarnet; Ganciclovir; Humans; Immunohistochemistry; Intercellular Adhesion Molecule-1; Interleukin-8; Neutrophils; Oligonucleotides, Antisense; Organophosphonates; Organophosphorus Compounds; Reverse Transcriptase Polymerase Chain Reaction; Thionucleotides; Up-Regulation

Cytomegalovirus (CMV) infection is associated with leucocyte infiltration in various organs, which supports a role for chemokines and adhesion molecules in the pathogenesis of CMV infection. In a prospectively conducted study of renal transplant recipients, 10 patients with CMV disease, five patients with asymptomatic CMV infection and 10 patients who did not have any CMV infection were included. During CMV infection, and in particular during CMV disease, plasma levels of the chemokines IL-8, macrophage inflammatory protein-1alpha (MIP-1alpha) and monocyte chemotactic protein-1 (MCP-1) and the soluble adhesion molecules vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1) and L-selectin increased and were positively correlated with the degree of CMV pp65 antigenaemia. Furthermore, a decrease in plasma levels of these chemokines and adhesion molecules was observed following ganciclovir therapy in the patients with CMV disease. This could suggest a role for these molecules in the pathogenesis of CMV infection.

Topics: Adult; Aged; Chemokine CCL2; Chemokine CCL3; Chemokine CCL4; Cytomegalovirus Infections; Female; Humans; Intercellular Adhesion Molecule-1; Interleukin-8; Kidney Transplantation; L-Selectin; Macrophage Inflammatory Proteins; Male; Middle Aged; Phosphoproteins; Prospective Studies; Solubility; Vascular Cell Adhesion Molecule-1; Viral Matrix Proteins

To assess the relationship between serum cytokines and cytomegalovirus (CMV) reactivation, 75 allogeneic bone marrow transplant patients underwent weekly measurements of interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-alpha, CMV blood cultures, and antigenemia tests. Of the patients, 44 (58.7%) developed CMV infection, and 19 (25.3%) developed clinical CMV disease. The mean maximum levels of all three cytokines were significantly increased in patients with CMV infection compared with levels in those without. Maximum levels of IL-6 were significantly higher in patients with active CMV disease than in those who did not develop CMV disease (281.2+/-85.5 vs. 95.7+/-15.0 pg/mL; P=.034). Levels of IL-8 and TNF-alpha were also elevated in patients who developed active disease. In a multivariate logistic regression model, IL-6 levels were independently associated with CMV disease (odds ratio=1.70 per 100-pg/mL increase in IL-6; P=.009). Cytokines may play an important role in the pathogenesis of CMV after bone marrow transplantation and may be a useful predictor for CMV.

Topics: Adult; Bone Marrow Transplantation; Cytokines; Cytomegalovirus Infections; Humans; Interleukin-6; Interleukin-8; Middle Aged; Multivariate Analysis; Tumor Necrosis Factor-alpha; Virus Activation

Human cytomegalovirus (HCMV) is one of the most frequent opportunistic agents causing severe illness in chronic human T cell leukemia-lymphoma virus type I (HTLV-I) infection. Our previous studies have shown that coinfection of macrophages with HCMV and HTLV-I significantly enhances HCMV replication, resulting in release of infectious HCMV from dually infected cells. We found that double infection of macrophages with HCMV and HTLV-I induced a rapid production of substantial amounts of interleukin-8 (IL-8) and transforming growth factor-beta1 (TGF-beta1). Results of transfection studies demonstrated that the tax gene product of HTLV-I was able to induce secretion of IL-8 and TGF-beta1. In addition to its cytokine-inducing effect, the Tax protein could interact with HCMV synergistically to result in production of much higher levels of IL-8 and TGF-beta1 than expected on the basis of their separate activities. Treatment of dually infected macrophage cultures with neutralizing antibodies to IL-8 and TGF-beta1 led to a nearly 1000-fold decrease in release of infectious HCMV from coinfected cells. Similar results were obtained when anti-IL-8 and anti-TGF-beta1 treatments were combined in macrophage cultures transfected with the tax gene before HCMV infection. Our results suggest that the stimulatory effect of HTLV-I Tax protein on HCMV replication in coinfected macrophages is largely mediated by high levels of IL-8 and TGF-beta1 production.

Topics: Antibodies, Monoclonal; Cell Line; Cytomegalovirus Infections; Gene Products, tax; HTLV-I Infections; Humans; Interleukin-8; Macrophages; Transforming Growth Factor beta; Virus Replication

Cytomegalovirus (CMV) disease is a major problem in renal transplant recipients, but few predictive markers of the disease are known. Several immunologic parameters of potential relevance for the defense against CMV were measured after renal transplantation in 25 patients before any manifestations of CMV infection occurred. In 10 patients who later developed CMV disease, plasma levels of interleukin-8 were significantly higher, whereas the levels of macrophage inflammatory protein-1alpha (MIP-1alpha) were significantly lower than in 15 patients who did not develop CMV disease. Also, lower numbers of CD4+ and CD8+ lymphocytes were observed in patients who later had CMV disease. These findings were independent of previous rejection therapy and were particularly pronounced in patients with primary CMV infection. Interleukin-8 and MIP-1alpha may be predictive markers of CMV disease and could be of potential use in selecting patients for prophylactic treatment.

Topics: Adult; Aged; Causality; Cell Adhesion Molecules; Chemokine CCL3; Chemokine CCL4; Cytomegalovirus Infections; Female; Humans; Interleukin-10; Interleukin-8; Kidney Transplantation; Macrophage Inflammatory Proteins; Male; Middle Aged; Monocytes; Receptors, Tumor Necrosis Factor; T-Lymphocyte Subsets; Tumor Necrosis Factor-alpha

Human cytomegalovirus (CMV) infection results in pneumonitis in bone-marrow and lung-transplant recipients. The source of CMV infection contributing to the onset of pneumonitis is unclear, but may involve infection of the lung endothelium in the presence of infiltrating mononuclear cells. Viral infection stimulates the host cell to express chemokines as signals to recruit specific immune cells to the site of injury. CMV encodes a chemokine receptor that may function to reduce host cell expression of chemokines. In the study reported here we found that extracellular concentrations of the chemokine regulated on activation, normal T cell expressed and secreted (RANTES) are depleted during productive infection of primary endothelial cells with CMV strain 4010, an endothelial-adapted strain of CMV. Utilizing adenovirus-transformed human kidney epithelial cells (type 293 cells) that stably express the CMV-encoded chemokine receptor US28, we found that depletion of extracellular RANTES during infection is attributable to US28, which binds and internalizes extracellular RANTES.

Topics: Cell Division; Cell Line; Chemokine CCL2; Chemokine CCL4; Chemokine CCL5; Cytomegalovirus Infections; Endothelium, Vascular; Humans; Interleukin-8; Macrophage Inflammatory Proteins; Receptors, CCR2; Receptors, Chemokine; Time Factors

Although syncytiotrophoblast (ST) cells can be infected by human cytomegalovirus (HCMV), in vitro studies have indicated that ST cells do not support the complete viral reproductive cycle, or HCMV replication may occur in less than 3% of ST cells. The present study tested the possibility that placental macrophages might enhance activation of HCMV carried in ST cells and, further, that infected ST cells would be capable of transmitting virus to neighboring macrophages. For this purpose, we studied HCMV replication in ST cells grown alone or cocultured with uninfected placental macrophages. Our results demonstrated that HCMV gene expression in ST cells was markedly upregulated by coculture with macrophages, resulting in release of substantial amounts of infectious virus from HCMV-infected ST cells. After having become permissive for viral replication, ST cells delivered HCMV to the cocultured macrophages, as evidenced by detection of virus-specific antigens in these cells. The stimulatory effect of coculture on HCMV gene expression in ST cells was mediated by marked interleukin-8 (IL-8) and transforming growth factor-beta1 (TGF-beta1) release from macrophages, an effect caused by contact between the different placental cells. Our findings indicate an interactive role for the ST layer and placental macrophages in the dissemination of HCMV among placental tissue. Eventually, these interactions may contribute to the transmission of HCMV from mother to the fetus.

Topics: Antigens, Viral; Coculture Techniques; Cytokines; Cytomegalovirus Infections; Humans; Interleukin-8; Macrophages; Phosphoproteins; Placenta; Transforming Growth Factor beta; Trophoblasts; Viral Matrix Proteins; Virus Replication

The observations that several types of viruses induced interleukin (IL)-8 production prompted us to investigate the interrelationship between IL-8 and cytomegalovirus (CMV) infection. CMV infection caused IL-8 production in a human monocytic cell line, THP-1, in dose- and time-dependent manners. Moreover, CMV induced IL-8 gene expression by concurrently activating transcription factors, NF-kappaB and AP-1. Furthermore, CMV infection of human fibroblast cell lines increased gene expression of a specific receptor for IL-8, CXCR1. IL-8 in turn enhanced CMV replication in a human embryonic fibroblast, MRC-5, in dose- and time-dependent manners. Augmented replication eventually culminated in the increased production of infectious CMV virions. Moreover, IL-8 can attenuate the antiviral activity of interferon (IFN), particularly that of alpha-type against picornaviruses such as encephalomyocarditis virus and poliovirus. The inhibitory effects were associated with reduced 2',5'-A oligoadenylate synthetase activity. These results would imply that CMV can induce IL-8, which can augment CMV replication directly and indirectly by counteracting antiviral activity of IFN.

Topics: Antiviral Agents; Cell Line; Cytomegalovirus; Cytomegalovirus Infections; Humans; Interferon-alpha; Interleukin-8; Monocytes; Virus Replication

The human cytomegaloviruses (HCMVs) appear to have the potential to disrupt production of hematopoietic cytokines. We examined the production of granulocyte/macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-8 by cultured and CMV-infected human umbilical vein endothelial cells (HUVECs) and compared this production with that of uninfected cells. Endothelial cells are, among other things, an integral component of human bone marrow stroma, and are responsible for production of factors that modulate the proliferation and differentiation of human hematopoietic progenitors. HCMV infection increased the production of GM-CSF in IL-1-primed HUVECs without altering GM-CSF levels in infected but unprimed HUVECs. However, this same virus was capable of causing increased production of the inhibitory cytokine IL-8. Both the viral pellet and the cleared viral supernatant appeared to contribute equally to the increased IL-8 and GM-CSF production, because each of these preparations alone was capable of exerting only half the effect seen with whole virus preparations. That both live virus and soluble protein factors within the viral stock contributed to the enhancement in GM-CSF and IL-8 production was further confirmed by inactivation with either ultraviolet or heat treatment of the viral stocks. Although the identity of the factor within the HCMV stock that contributes to this effect remains unknown, studies conducted in the presence of neutralizing antibodies or polymyxin B ruled out a role for tumor necrosis factor-alpha, IL-6, or endotoxin, all known inducers of GM-CSF. These studies indicate that HCMVs can exert both direct and indirect effects on the production of the hematopoietic factor GM-CSF and the inflammatory/inhibitory cytokine IL-8.

Topics: Cells, Cultured; Cytomegalovirus; Cytomegalovirus Infections; Endothelium, Vascular; Granulocyte-Macrophage Colony-Stimulating Factor; Hot Temperature; Humans; Interferon-gamma; Interleukin-1; Interleukin-8; Ultraviolet Rays; Umbilical Veins

Virus-induced alterations in the cellular expression of chemokines may be important in directing the migration of specific leucocyte subsets to sites of infection, thereby playing a pivotal role in viral pathogenesis. We show here that cytomegalovirus (CMV) infection of human fibroblasts resulted in significantly increased expression of the C-X-C or alpha-chemokine interleukin-8 (IL-8), at both the mRNA and protein levels. Increased IL-8 production was seen following infection with the high passage laboratory CMV strains AD169, Towne, or Davis, as well as the low passage clinical CMV isolates Toledo or C1F. The increase in IL-8 production had functional consequences, as demonstrated by the ability of supernatants from CMV-infected fibroblasts to significantly enhance neutrophil transendothelial migration. The latter was independent of alterations in adhesion molecule expression on the endothelial cells, and was abrogated by neutralizing antibodies specific for IL-8. Direct infection of endothelium with the endothelial cell-tropic CMV strain C1FE, also resulted in enhanced neutrophil transendothelial migration. Neutrophils play an important role in the dissemination of CMV throughout the body, and thus CMV-induced neutrophil recruitment would be expected to enhance CMV dissemination. Increased production of chemokines in response to CMV infection could also disrupt the fine balance between a beneficial and a destructive immune response, thereby potentially contributing to pathology.

Topics: Blotting, Northern; Cell Adhesion Molecules; Cell Culture Techniques; Chemotaxis, Leukocyte; Cytomegalovirus Infections; Endothelium, Vascular; Fibroblasts; Humans; Interleukin-8; Neutrophils; RNA, Messenger; Up-Regulation

The study's objective was to investigate serum levels of interleukin-8 (IL-8) after liver transplantation and to correlate these findings with tumor necrosis factor alpha serum levels and various clinical parameters. This was a prospective observation study conducted at the University Hospital of Innsbruck with 19 patients studied after orthotopic liver transplantation. Serum levels of IL-8 were analyzed by a solid-phase double ligand ELISA method. Serum TNF-alpha concentrations were measured by means of a commercially available radio immunoassay (IRE-Medgenix, Fleurus, Belgium). Three patients with an uneventful recovery after transplantation showed IL-8 levels below the detection limit. IL-8 serum levels markedly increased in patients with acute graft rejection, bacterial infection, and CMV disease. Increments of serum IL-8 preceded clinical complications in all patients. Highest levels were observed in bacterial infection, lowest in acute rejection. A statistically significant positive correlation was demonstrated between IL-8 and TNF-alpha serum levels in the context of bacterial infection and CMV disease. Elevated IL-8 serum levels represent a feature of alloimmune and infectious complications following liver transplantation. IL-8 can thus be considered a further indicator molecule in the heterogenous group of acute-phase reactants that accompany various inflammatory responses and do not permit the underlying clinical complication to be specified.

Topics: Adult; Bacterial Infections; Cytomegalovirus Infections; Female; Humans; Interleukin-8; Liver Transplantation; Male; Middle Aged; Tumor Necrosis Factor-alpha