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Human polyomavirus JC and BK persistent infection.

Primary contact with the human polyomaviruses (HPV) is followed by lifelong persistence of viral DNA in its host. The most prominent organs affected are the kidney, the Central Nervous System (CNS)and the hematopoietic system. Under impairment of immune competence limited activation of virus infection can be followed by prolonged virus multiplication, severe destruction of tissue and disease. The mechanisms responsible for activation episodes of the asymptomatic persistent infection are not understood and questions on cellular localization, routes of dissemination of HPV infection and its activation are controversially discussed. The type of interaction of HPVs with target organs and patients groups is highly differentiated. Organ-specific activation above basic level argues for strong dependence on the respective immune states of risk group patients. However, since immune impairment generally plays an important role in the activation of polyomavirus infection, amplification of virus deoxyribonucleic acid (DNA) and activation of virus replication is also a normal event that is probably subject to immunomodulation in the healthy individual. It also becomes clear that BKV and JCV infection is differentially regulated by mechanisms depending on the balance of immune control as well as on organ-specific signalling.

BK Virus↗

Persistent infections by Leishmania (Viannia) braziliensis.

Here we review the phenomenon of persistency in Leishmania (Viannia) braziliensis infections. In other Leishmania species where appropriate animal models exist, considerable advances in the understanding of basic immunologic mechanisms of persistency have been made; for a review see Aebisher (1994). On the contrary, the evidences of persistence in infections with L. braziliensis rest on studies of human clinical cases many of which we summarized and discussed in this work.

Animals↗

General primer-mediated polymerase chain reaction for detection of enteroviruses: application for diagnostic routine and persistent infections.

The aim of this study was to determine the applicability of the polymerase chain reaction (PCR) for routine diagnostic use and for the detection of persistent enteroviral infections. To this end, general primers were selected in the highly conserved part of the 5'-noncoding region of the enteroviral genome. They were tested on 66 different enterovirus serotypes. A specific fragment was amplified from 60 of 66 serotypes. An amplification product was not observed from coxsackievirus types A11, A17, and A24 and echovirus types 16, 22, and 23. Enteroviral RNA was detected by the PCR in routinely collected throat swabs and stool specimens that were found to be positive for enterovirus by isolation in tissue culture. Enteroviral RNA was detected in one of five myocardial biopsy specimens from patients with dilated cardiomyopathy, implicating virus persistence. No amplification product was obtained from eight control samples. Our results demonstrate the significance of the PCR for the detection of enteroviral RNA and, in particular, for the demonstration of persistent enteroviral infections.

Base Sequence↗

Unique amino acid substitutions in the capsid proteins of foot-and-mouth disease virus from a persistent infection in cell culture.

Maintenance of a persistent foot-and-mouth disease virus (FMDV) infection in BHK-21 cells involves a coevolution of cells and virus (J. C. de la Torre, E. Martínez-Salas, J. Díez, A. Villaverde, F. Gebauer, E. Rocha, M. Dávila, and E. Domingo, J. Virol. 62:2050-2058, 1988). The resident FMDV undergoes a number of phenotypic changes, including a gradual decrease in virion stability. Here we report the nucleotide sequence of the P1 genomic segment of the virus rescued after 100 passages of the carrier cells (R100). Only 5 of 15 mutations in P1 of R100 were silent. Nine amino acid substitutions were fixed on the viral capsid during persistence, and three of the variant amino acids are not represented in the corresponding position of any picornavirus sequenced to date. Cysteine at position 7 of VP3, that provides disulfide bridges at the FMDV fivefold axis, was substituted by valine, as determined by RNA, cDNA, and protein sequencing. The modified virus shows high buoyant density in cesium chloride and depicts the same sensitivity to photoinactivation by intercalating dyes as the parental FMDV C-S8c1. Amino acid substitutions fixed in VP1 resulted in altered antigenicity, as revealed by reactivity with monoclonal antibodies. In addition to defining at the molecular level the alterations the FMDV capsid underwent during persistence, the results show that positions which are highly invariant in an RNA genome may change when viral replication occurs in a modified environment.

Amino Acid Sequence↗

Characterization of human cells persistently infected with cytomegalovirus and exposed to a chemical carcinogen.

A non-productive, persistent human cytomegalovirus (CMV) infection was developed in human embryo lung (LU) cells by initially using supraoptimal temperatures to restrict the replication of a recent CMB isolate (76-24). Although CMV replication was observed in some cells within a few weeks of shifting the cultures to 37 degrees C and subculturing, CMV cytopathic effects and infectious CMV were not detected with further subculturing during the next 5 weeks. CMV-specific nuclear antigens were, however, observed in most cells (designated LU-76-24) for 38 additional culture passages made during the following 12 months. Although the LU-76-24 cells were morphologically distinct from LU cell cultures carried in parallel as controls, the LU-76-24 were contact-inhibited in spite of a somewhat disoriented growth pattern. Treatment of LU-76-24 cells with 0.05 microgram/ml of the chemical carcinogen 4-nitroquinoline 1 oxide (NQO) induced a lytic CMV infection from which a clone of cells (designated LUC-NQO) was isolated. No effect was seen when LU cells were treated with the same dose of NQO. LUC-NQO cells were morphologically altered from LU-76-24 cells and had lost contact inhibition. Even though CMV-specific nuclear antigens were observed in LUC-NQO cells, infectious CMV was not detected in lysates of these cells, nor was CMV rescued by co-cultivation of these cells with susceptible cells or by induction with 5-iodo-2'-deoxyuridine. LUC-NQO, but not LU or LU-76-24 cells, plated with good efficiency in semi-solid medium. The results indicate that long-term, non-productive, persistent CMV infections can be established in vitro and suggest that such cells may be more susceptible than non-infected cells to the action of chemical carcinogens.

4-Nitroquinoline-1-oxide↗

Pituitary dwarfism in mice persistently infected with lymphocytic choriomeningitis virus.

Most strains of mice injected intracerebrally with lymphocytic choriomeningitis virus grow to adulthood maintaining a persistent virus infection associated with chronic virus-induced immune complex disease. However, mice on a k background are highly susceptible to neonatal infection and develop the clinical syndrome of pituitary dwarfism and hypoglycemia. Examination of pituitary tissue fails to reveal morphologic alteration by light and electron microscopy. Within the pituitary, viral antigens are exclusively distributed within the cells of the adenohypophysis. Using ultrastructural colloidal gold-labeling techniques, we demonstrate the presence of mature virus particles budding from the surface of growth hormone containing cells from the pituitary. This study indicates that persistent lymphocytic choriomeningitis virus infection of the growth hormone cells in susceptible mice is associated with pituitary dwarfism without producing visible structural damage.

Animals↗

Lack of neutralizing antibodies to caprine arthritis-encephalitis lentivirus in persistently infected goats can be overcome by immunization with inactivated Mycobacterium tuberculosis.

The pathogenesis of the persistent progressive diseases of sheep (visna-maedi) and goats (arthritis-encephalitis) is dependent on continuous replication of the causative lentiviruses. One subgroup of these viruses, Icelandic visna virus, accomplishes this form of replication by undergoing antigenic mutation. Mutant viruses arising late in the infection escape neutralization by antibodies directed to the parental virus. In contrast, we show here that viruses obtained from persistently infected sheep and goats with natural disease in this country do not induce virus-neutralizing antibodies, although antibodies to virus core proteins were produced. The lack of neutralizing antibodies was not overcome by hyperimmunization of animals with concentrated preparations of live or inactivated virus. Thus, failure to produce these specific antibodies was not due to lack of sufficient antigen or interference with the immune response because of the ability of these viruses to infect macrophages. The hyporesponsive state, however, was overcome by immunization of animals with virus and large amounts of inactivated Mycobacterium tuberculosis. Induction of agglutinating and neutralizing antibodies by this method was probably due to a unique form of antigen processing by macrophages activated by M. tuberculosis. Neutralizing antibodies were produced for the first time against the caprine arthritis-encephalitis virus by this method. These antibodies have similar biological properties to those induced by Icelandic visna virus. They belong to the immunoglobulin G1 subclass, they are effective against a narrow range of caprine arthritis-encephalitis viruses, and they identify (for the first time) antigenic variants among these caprine agents.

Animal Diseases↗

Genomic polymorphism of human papillomavirus type 52 predisposes toward persistent infection in sexually active women.

We investigated the role of human papillomavirus (HPV) type 52 polymorphism in the persistence of HPV infection, which is a predictor for cervical lesions. Cervical samples obtained at 6-month intervals were tested for HPV-52 in 1055 women; 41, 12, and 58 women had persistent, transient, and unclassified HPV-52 infections, respectively. HPV-52 isolates were analyzed by polymerase chain-reaction sequencing of the long control region (LCR), E6, and E7 genes. Although age (odds ratio [OR], 0.90 [95% confidence interval [CI], 0.81-0.99]), nonprototypic LCR (OR, 9.26 [95% CI, 2.1-41.7]), and E6 variant (OR, 7.04 [95% CI, 1.4-37]) were associated, in univariate analysis, with the persistence of HPV-52 infection, a nonprototypic LCR variant was the only independent predictor of it (OR, 14.1 [95% CI, 1.1-200]). In the latter variants, the loss of a binding site for a repressor of HPV expression was associated with the persistence of HPV infection (OR, 7.25 [95% CI, 1.67-31.25]).

Adult↗

Cloning of the rhesus lymphocryptovirus viral capsid antigen and Epstein-Barr virus-encoded small RNA homologues and use in diagnosis of acute and persistent infections.

Epstein-Barr virus (EBV) is the most common cause of infectious mononucleosis and is associated with the development of several human malignancies. A closely related herpesvirus in the same lymphocryptovirus (LCV) genera as EBV naturally infects rhesus monkeys and provides an important animal model for studying EBV pathogenesis. We cloned the small viral capsid antigen (sVCA) homologue from the rhesus LCV and developed a peptide enzyme-linked immunosorbent assay (ELISA) to determine whether epitopes in the rhesus LCV sVCA are a reliable indicator of rhesus LCV infection. In order to define a "gold standard" for rhesus LCV infection, we also cloned the EBV-encoded small RNA 1 (EBER1) and EBER2 homologues from rhesus LCV and developed a reverse transcription (RT)-PCR assay to detect persistent LCV infection in rhesus monkey peripheral blood lymphocytes. Animals from a conventional and a hand-reared colony were studied to compare the prevalence of rhesus LCV infection in the two groups. There was a 100% correlation between the peptide ELISA and EBER RT-PCR results for rhesus LCV infection. In addition, specificity for LCV infection and exclusion of potential cross-reactivity to the rhesus rhadinovirus sVCA homologue could be demonstrated using sera from experimentally infected animals. These studies establish two novel assays for reliable diagnosis of acute and persistent rhesus LCV infections. The rhesus LCV sVCA peptide ELISA provides a sensitive and reliable assay for routine screening, and these studies of the hand-reared colony confirm the feasibility of raising rhesus LCV-naive animals.

Amino Acid Sequence↗

Woodchuck gamma interferon upregulates major histocompatibility complex class I transcription but is unable to deplete woodchuck hepatitis virus replication intermediates and RNAs in persistently infected woodchuck primary hepatocytes.

Gamma interferon (IFN-gamma) is an important mediator with multiple functions in the host defense against viral infection. IFN-gamma, in concert with tumor necrosis factor alpha (TNF-alpha), leads to a remarkable reduction of intrahepatic replication intermediates and specific mRNAs of hepatitis B virus (HBV) by a noncytolytic mechanism in the transgenic mouse model. Thus, it is rational to evaluate the potential value of IFN-gamma for the treatment of chronic HBV infection. In the present study, we expressed recombinant woodchuck IFN-gamma (wIFN-gamma) in Escherichia coli and mammalian cells. wIFN-gamma protected woodchuck cells against infection of murine encephalomyocarditis virus in a species-specific manner. It upregulated the mRNA level of the woodchuck major histocompatibility complex class I (MHC-I) heavy chain in permanent woodchuck WH12/6 cells and regulated differentially the gene expression. However, the level of the replication intermediates and specific RNAs of woodchuck hepatitis virus (WHV) in persistently WHV-infected primary woodchuck hepatocytes did not change despite a treatment with 1,000 U of wIFN-gamma per ml or with a combination of wIFN-gamma and woodchuck TNF-alpha. Rather, hepatocytes derived from chronic carriers had an elevated level of the MHC-I heavy-chain mRNAs, most probably due to the exposure to inflammatory cytokines in vivo. Treatment with high doses of wIFN-gamma led to an abnormal cell morphology and loss of hepatocytes. Thus, wIFN-gamma regulates the gene expression in woodchuck hepatocytes but could not deplete WHV replication intermediates and mRNAs in persistently infected hepatocytes. The cellular response to wIFN-gamma may be changed in hepatocytes from chronically WHV-infected woodchucks. It should be clarified in the future whether the continuous exposure of hepatocytes to inflammatory cytokines or the presence of viral proteins leads to changes of the cellular response to wIFN-gamma.

Animals↗

Possible role of infectious simian foamy virus DNA in persistent infection.

Simian foamy virus type 7 (SFV-7) can persist in peripheral blood leukocytes of SFV-7-infected rabbits. Rabbit peripheral blood leukoyctes cultured in vitro did not support the growth of SFV-7, but low titer of virus could be detected after 4 to 5 days incubation. The DNA derived from SFV-7 infected primary rabbit kidney cells is infectious. The presence of SFV-7 DNA provirus in infected rabbit kidney cells during replication may contribute one of the mechanisms of persistance.

Animals↗

Infection of human fibroblast-like synovial cells with Chlamydia trachomatis results in persistent infection and interleukin-6 production.

Recent studies have shown that the urogenital pathogen Chlamydia trachomatis to be a major bacterium triggering reactive arthritis (ReA), and is able to induce interleukin-6 (IL-6) production in human fibroblast-like synovial cells (FSC) in vitro. In the present study, we examined the correlation between IL-6 production and multiplication of chlamydia in FSC. All FSC from five patients secreted highly increased quantities of IL-6 in a dose-dependent and time-dependent fashion. Heat and UV inactivated chlamydia failed to enhance production of IL-6. When azithromycin was added to infected cultures of FSC at 0 or 48 h after infection, the level of IL-6 production was very low. Transmission electron microscopy of such infected cultures revealed many abnormal forms of chlamydia within the inclusions in FSC. From one step-growth curve experiments, it was suggested that C. trachomatis hardly multiplied in FSC. In contrast, in C. trachomatis infected HeLa 229 cells, chlamydia multiplied as usual, but little IL-6 production were found. These observations indicated that live chlamydia and the persistence of chlamydia may be essential for stimulating the synthesis of IL-6 in FSC.

Adult↗

Arachidonic acid immunoregulation in lambs persistently infected with border disease virus.

To evaluate arachidonic acid-related immunoregulatory mechanisms during long-term persistent pestivirus infection, we measured plasma contents of leukotriene C4 (LTC4), prostaglandin D2 (PGD2) and their plasma fatty acid (FA) precursor, arachidonic acid (AA), in six lambs with congenital border disease (BD). Significantly elevated average plasma LTC4 during the first half year of life was associated with increased PDG2 when compared to uninfected control lambs. Significantly elevated total plasma esterified AA stores suggest an effective BDV-mediated prostenoid immunostimulation. However, at 1 yr old, prostenoid secretion had fallen to normal (LTC4) or below normal (PGD2) levels. In contrast, there remained significantly elevated plasma esterified AA, present as available substrate for formation of these anti-viral immunoregulatory agents. These results suggested that preventing mobilization of AA from lipid stores for effective immune responses may be a viral strategy of BD virus that is associated with long term border disease effects.

Age Factors↗

Slow persistent infection caused by visna virus: role of host restriction.

Proviral DNA has been demonstrated by in situ hybridization in foci of cells of a lamb infected with the RNA slow virus visna. A few of these cells also contain the major virion structural antigen p30. This restriction in virus gene expression in the infected animal provides a mechanism for persistence of virus in this chronic infection.

Animals↗

Localization of virus and antibody response in mice infected persistently with MHV-JHM.

Suckling mice infected intranasally with MHV-JHM and nursed by immunized dams develop a late onset demyelinating encephalomyelitis. Analysis by in situ hybridization revealed that MHV-JHM entered the central nervous system (CNS) via the olfactory and trigeminal nerves and spread over the next two weeks to the spinal cord, prior to amplification at this site. Serial measurements of neutralizing antibody titers showed that the late onset disease developed in some mice at levels of antibody which protected mice from the fatal, acute encephalitis, supporting the notion that cell-mediated and not humoral immunity is important in protecting mice from MHV-JHM persistence.

Animals↗

Viral and immunologic examination of human immunodeficiency virus type 1-infected, persistently seronegative persons.

Persons who were human immunodeficiency virus type 1 (HIV-1)-infected but who remained persistently seronegative (HIPS) on HIV-1 antibody tests were examined through AIDS case surveillance. Six such individuals (HIPS-1 to -4, -7, and -9) were examined to determine whether their persistent seronegativity was attributable to immune dysfunction or infection with atypical HIV. Of the 6, 4 had antibody titers to at least 1 other common pathogen. In vitro stimulation of peripheral blood mononuclear cells from HIPS-4 and HIPS-7 with pokeweed mitogen or phosphorothioate oligodeoxynucleotide (direct B cell mitogen) did not produce HIV-1-specific antibody. Reconstitution experiments with recombinant interleukin (rIL)-4 and rIL-12 also had no impact on antibody production. Virus isolates from HIPS-4 and -9 were R5X4-tropic, whereas HIPS-7 was CCR5-tropic only. Sequence analysis of long terminal repeat, p24, and env gp41 did not reveal any specific mutation, and phylogenetic analysis confirmed that all 6 virus specimens were HIV-1 subtype B. These data suggest that the lack of a detectable antibody response in these patients may be the result of immune dysfunction.

Amino Acid Sequence↗

Ducks: a new experimental host system for studying persistent infection with avian leukaemia retroviruses.

Long-term persistence of the avian leukosis virus (ALV), the transformation-defective mutant of Prague strain Rous sarcoma virus subgroup C (td PR-C) was established in heterologous duck hosts after infection in mid-embryogenesis. Transient viraemia was observed for about 4 weeks after hatching and was lost in most of the infected ducks by about 6 months. Loss of viraemia was accompanied by the increasing synthesis of virus-neutralizing antibodies. In spite of strong virus-neutralizing antibodies, virus was detected by the cocultivation assay in duck tissues throughout the observation period up to 5 years. In the viraemic phase of infection, we found integrated proviruses in various tissues, preferentially in stomach muscle tissue and in the thymus. The long-term persistence of virus was frequently accompanied by liver necrosis and neoplastic diseases. Injection of td PR-C virus into early embryos resulted in more pronounced infection accompanied by an increased copy number of viral DNA per cell, high mortality and remarkable atrophy of thymus tissue in infected ducklings.

Animals↗

Persistent infection with mouse hepatitis virus of low virulence in nude mice.

A persisting type of infection with wasting syndrome was established in congenitally athymic nude mice after intraperitoneal inoculation with a mouse hepatitis virus which was not fully pathogenic for heterozygous haired littermates. From the liver, spleen, lymph nodes, and brain of most infected nude mice, the virus was detected at high titers during aperiod from 6 to 35 days postinfection, occurrence of degenerative and necrotic lesions being correlated with virus titers in these organs. The titer of serum neutralizing antibody remained undetectable or very low in most diseases nude mice, whereas some animals resisting the infection could produce antibody at a later stage. In heterozygous haired mice, some lesions were detectable at a very early stage of infection in the spleen and liver, but they seemed to disappear with a marked elevation of the neutralizing antibody titer. Nude mice were able to resist the virus infection when they had previously received transfer of thymocytes from weanling heterozygous littermates.

Animals↗