Reactivity of varicella-zoster virus subunit antigens in enzyme-linked immunosorbent assay to sera from varicella,zoster, and herpes simplex virus infections

Serological responses to varicella-zoster virus (VZV) subunit antigens, such as capsid, envelope, and soluble (S) antigens, in patients with VZV and herpes simplex virus (HSV) infections were studied by comparing with responses to virion (V) antigens using an enzyme-linked immunosorbent assay (ELISA). S antigen, prepared by concentrating supernatant of VZV or HSV type 1 (HSV-1)-infected cell culture fluid, reacted strongly to sera from patients with secondary infection but reacted poorly to those from patients with a primary infection of VZV or HSV. Antibody titers to VZV-S antigen persisted for a long period in patients with VZV infections. Patients infected with VZV showed antibody increase to HSV-1, when tested by complement fixation or complement-enhanced neutralization test, in cases with a history of prior HSV infection. However, such a cross-reaction was only observed to a minor extent in ELISA test using S antigen. S antigen reaction was stronger in secondary infections in tests with various subunit antigens. Almost no cross-reactivity was observed in an immunoblotting test with S antigen. Differentiation between infections with either varicella or zoster or HSV can be made by comparison of antibody responses to V and S antigens.     

Stimulation of human lymphocytes by Herpes simplex virus antigens.

Lymphocytes from individuals with laboratory evidence of prior infection with herpes simplex virus (HSV) type 1 or type 2 demonstrated transformation (av antigens. Higher stimulation indexes were obtained when lymphocytes were incubated with the homologous as compared with the heterologous antigen. Higher mean lymphocyte stimulation indexes were also demonstrated in seropositive as compared with seronegative individuals. Lymphocytes from children with HSV-1 stomatitis usually became responsive to HSV-1 antigen within 2 to 6 weeks after the onset of illness. Lymphocytes from infants with neonatal HSV-2 infection were stimulated by HSV-2 antigen.     

Pathogenesis of Herpes simplex virus infections in guinea pigs.

The pathogenesis of herpes simplex virus types 1 and 2 has been studied in guinea pigs after inoculation by various routes (subcutaneous and intradermal infection in footpads and vaginal infection). Clinical observations as well as virus isolation studies are reported. Herpes simplex virus type 2 infection by all three routes of inoculation led to acute primary and recurrent lesions. Virus persisted in the nervous system, particularly in sensory ganglia, and locally at the site of inoculation. Herpes simplex virus type 1 infection induced no or very mild primary symptoms. Recurrent lesions were only observed after intradermal inoculation. Invasion of the nervous system and consequent establishment of latent ganglionic infection was less efficient than after herpes simplex virus type 2 infection. Peripheral persistence was, however, equally common.     

Herpes simplex virus, cytomegalovirus and Epstein-Barr virus antibody titres in sera from schizophrenic patients

Serum antibody titres to herpes-simplex (HSV-1, 2), cytomegalovirus (CMV), and Epstein-Barr virus capsid antigen (EBV-VCA) were determined in 38 unrelated chronic schizophrenic patients, 11 nuclear families with at least 2 schizophrenic members, and 2 control groups. The distributions of antibody titres to herpes simplex and cytomegalovirus were similar among all groups. Patients had higher anti-EBV-VCA titres than non-hospitalized controls; however, hospital staff members in contact with the patients also had significantly higher antibody titres to EBV-VCA. Antibodies to EBV early antigen (EBV-EA) were also determined for 27 unrelated patients and 24 mental hospital employees. The schizophrenic patients had significantly higher antibody titres to EBV-EA than the hospital worker control group. These data do not support the hypothesis that herpes viruses are associated with the aetiology of schizophrenia. Although elevated anti-EBV early antigen titres may suggest persistent active EBV infection, it is unlikely to be related to the aetiology of the disorder, since discordance for EBV seropositivity was present among sibling pairs concordant for schizophrenia.     

Herpes simplex and varicella-zoster virus infections during pregnancy: current concepts of prevention,diagnosis and therapy. Part 1: Herpes simplex virus infections

Primary herpes simplex virus (HSV) infection may lead to severe illness in pregnancy and may be associated with transplacental virus transmission and fetal infection. The consequences may be abortion, stillbirth and congenital malformations. In neonates, the clinical findings after intrauterine HSV infection are characterized by skin lesions, diseases of the eye and neurologic damage. Herpes genitalis of pregnant women at the time of labor may result in life-threatening neonatal herpes. Currently, neither active nor passive immunization is available to prevent HSV infections during pregnancy and in the newborn infant. Therefore, antiviral treatment using aciclovir and/or valaciclovir must be considered in all primary episodes of genital herpes as well as in neonates who show signs of either infection. Clinical herpes lesions of the genitalia and/or positive test for virus detection at the time of delivery are an indication for cesarean section. However, this surgical intervention may be reduced by suppressive treatment of recurrent genital herpes with aciclovir or valaciclovir.     

Complement-fixing antibody in human sera reactive with viral and soluble antigens of cytomegalovirus.

Antibody titers to cytomegalovirus were determined in 204 single-serum specimens and in 138 serum pairs, utilizing as antigen extracts of infected cells prepared by either freezing and thawing cells (FT antigen) or extracting them with a glycine buffer (GE antigen). With the serum pairs, 40% of the early sera and 44% of the late sera showed a fourfold or higher antibody titer with the GE antigen compared with those with the FT antigen, whereas 39% of the single sera had a fourfold or higher titer with the GE antigen. Sucrose density gradient centrifugation of the two antigen preparations indicated that the GE antigen contained two peaks of complement-fixing activity, one at a density of 1.18 to 1.22 g/cm3 and one at 1.06 to 1.10 g/cm3. The FT antigen had only one discernible peak of complement-fixing activity at 1.04 to 1.10 g/cm3. On electron microscopy, the more dense peak contained mainly nucleocapsids with some dense bodies and occasional enveloped virions, and the lighter peak was composed of amorphous material. Antibody titers obtained with the crude GE antigen and the more dense peak (designated viral antigen) were in agreement, whereas titers with the FT antigen agreed with those obtained with the lighter peak (soluble antigen). The antibody detected with both antigens was mainly immunoglobulin G.     

Immune response to herpes simplex virus infections: virus-specific antibodies in sera from patients with recurrent facial infections.

Radioimmunoprecipitation assays were used to identify antibodies against a number of herpes simplex virus type 1-specific antigens in serum samples from individuals with recurrent facial herpes virus infections and from seropositive individuals without recurrent infections. Individuals with recurrent infections contributed three sequential serum samples each: immediately after the appearance of lesions, 3 weeks later, and 3 months later. Antibodies against at least 18 viral polypeptides were present in all positive sera: these included antibodies against the major nucleocapsid polypeptide (approximate molecular weight, 150,000) and against two glycopolypeptides with molecular weights of 115,000 to 130,000. No significant differences were observed between the serum samples in regard to their virus-specific antibody composition. The high-molecular-weight glycopolypeptides were partially purified and used in quantitative titration experiments. All sera tested were equally reactive with this material. It was concluded that under the experimental conditions an individual's susceptibility to recurrent herpetic infections could not be correlated with quantitative or qualitative changes in the levels of virus-specific antibodies.     

Herpes simplex virus type-selective enzyme-linked immunosorbent assay with Helix pomatia lectin-purified antigens

Helix pomatia lectin-purified antigens with specific reactivity to herpes simplex virus type 1 (HSV-1) and HSV-2 antibodies in human sera were used in an enzyme-linked immunosorbent assay. The type specificity of the antigens was assessed by double immunodiffusion precipitation in gel against rabbit HSV-1 and HSV-2 hyperimmune sera, and by enzyme-linked immunosorbent assay with human reference sera containing antibodies to either type of HSV. Fifty-two sera from patients with documented infection with either HSV-1 or HSV-2 were assayed for HSV type-specific immunoglobulin G antibodies. The reactivity of the sera against lectin-purified antigens correlated completely with the results of virus typing. We conclude that HSV type-specific immunoglobulin G antibodies can easily be measured by enzyme-linked immunosorbent assay with the use of Helix pomatia lectin-purified HSV-1 and HSV-2 antigens.     

Herpes simplex virus infections and current methods for laboratory detection

Herpes simplex virus (HSV) is endemic in all societies throughout the world and produces year-round infections in all age groups. While orofacial and genital skin infections predominate, a significant minority of individuals develop more serious herpetic disease in the eye, meninges, and brain tissue. Perinatal infection of the newborn has a high likelihood of dissemination to multiple visceral organs, and immunocompromised patients can develop aggressive necrotizing skin lesions, as well as disseminated disease. HSV is not fastidious and is easily propagated in vitro; virus culture methods are straightforward and currently are offered in many community hospital microbiology laboratories. However, amplified nucleic acid probe assays have now been developed that are significantly more sensitive than culture, have fewer specimen collection and transport constraints, and can generate reliable results the same day of testing. Recent innovations in HSV antibody assays that use serotype 1- and serotype 2- specific reagents can now provide accurate serological separation of HSV-1 and HSV-2 infections by ELISA and Western blot (immunoblot) methods. Community hospitals should anticipate more frequent requests for these more sensitive and specific diagnostic tests. When reliable commercial assays become available, new serological and molecular tests may be within the scope of the community hospital laboratory.     

Herpes simplex virus-1 and varicella virus infections in familial dysautonomia patients

Familial dystautonomia (FD) patients are deficient in type C fibers, suggesting that there may be a different pattern of infection and clinical presentation when infected by Herpes simplex virus type 1 (HSV-1) or Varicella-Zoster virus (VZV). These viruses infect and are reactivated in the periphery of the body through type C sensory nerve fibers. HSV-1 infects epithelial cells, penetrates into type C fibers, and migrates to the ganglia to generate latent infection. In reactivation, the viral DNA migrates through type C fibers, infecting the epidermis at the entry site. VZV infects through the respiratory tract, causing systemic viral infection and latency in the ganglia, from which it is reactivated and reaches the skin. The study was carried by clinical questionnaire and by HSV and VZV IgG antibodies on fifty-one FD patients and eighty matched controls. The questionnaire revealed that no FD patient had a history of clinical HSV-1 infection, compared to 15% in the control group (P < 0.05), while 50% FD patients had been infected by varicella, compared to 66% in the VZV control group. However in FD, VZV clinical manifestations were mild in comparison to controls. There was no difference in infection rates for some other viral diseases. HSV-1 antibodies were detected in 24% of the FD patients, compared to 38% in the control group (P < 0.1). VZV antibodies were similar in FD and controls (66%, 63%). We concluded that the rate of HSV infection in FD is low and clinical reactivation is rare. The rate of varicella infection appears to be the same for patients and controls, but in FD the clinical presentation is mild. We suggest that these differences are due to the lack of type C fibers in FD patients. J. Med. Virol. 54:158–161, 1998. © 1998 Wiley-Liss, Inc.     

Herpes simplex virus DNA isolation from infected cells with a novel procedure.

Herpes Simplex Virus type 1-infected cells were extracted in the presence of 0.25% Triton X-100-0.2 M NaCl. Viral DNA associated with proteins was found in the supernatant after low-speed centrifugation. Only viral DNA was recovered by this procedure, as shown by CsCl density analysis after deproteinization. Full-length viral DNA molecules were observed in the electron microscope.     

Virus-specific antibodies in sera from patients with genital herpes simplex virus infection.

Virus-specific antibodies against a number of herpes simplex virus type 2 antigens were determined by radioimmunoprecipitation assays in sequential serum samples obtained from 12 patients with initial genital herpes simplex virus infection. The progressive appearance of antibodies to virus-specific antigens was observed; antibodies against a 130,000-molecular-weight glycoprotein complex appeared first, followed by antibodies against the major nucleocapsid polypeptide and then antibodies against a number of other viral antigens, including a polypeptide with a molecular weight of 62,000. Patients who developed a wide variety of antibodies to viral polypeptides shortly after resolution of their initial episode seemed to experience more severe initial infections and more recurrences than did those who reacted poorly with these virus-specific antigens. This was most apparent with respect to antibodies to virus-specific polypeptides with molecular weights between 30,000 and 43,000. Antibody specificity did not change during the course of follow-up regardless of whether serum samples were taken shortly before, during, or after recurrent episodes. Glycoprotein-specific antibodies were quantitated with the purified 130,000-molecular-weight glycoprotein material. No significant fluctuations in these antibody titers were observed before or after recurrences of the disease.     

Herpes simplex virus infections in guinea pigs deficient in the fourth component of complement.

Separate groups of normal and C4-deficient guinea pigs were inoculated with herpes simplex virus by intradermal (i.d.) and intraperitoneal (i.p.) routes. Virus infection, confirmed by clinical, virological, and serological criteria, did not last longer and was not more severe in C4-deficient guinea pigs than in normal guinea pigs. Serum C component levels were measured before, during, and after herpes simplex virus infection. In normal gruinea pigs there was no evidence for C4 utilization after either i.d. or i.p. inoculation. In both normal and C4-deficient guinea pigs, C1 and C3-9 levels remained unchanged in spite of i.d. or i.p. infection. These data suggested that C4 and the classical C pathway were not important for virus clearance.     

Antibody-dependent enhancement of dengue virus infection in vitro by undiluted sera from monkeys infected with heterotypic dengue virus

Two monkeys (Macaca fascicularis) each were infected with dengue virus type 1 (DENV-1) and type 2 (DENV-2). High levels of neutralizing antibody to homotypic serotype were detected from day 10 to week 58 after infection. Levels of cross-reactive neutralizing antibody to other serotypes were at lower levels or undetectable. Serum samples collected from day 10 to week 58 enhanced infection by homotypic and heterotypic serotypes of DENV when diluted, demonstrating antibody-dependent enhancement (ADE). The ADE activities to heterotypic and homotypic dengue virus infections peaked at dilutions of 1:10–1:100 and 1:100–1:1,000, respectively. Serum samples collected enhanced heterotypic dengue virus infection without any dilution. The results indicate that sera from infected monkeys have an ability to enhance heterotypic dengue virus infection in vitro without dilution, although some of these sera also possess neutralizing activity.     

Herpes simplex and varicella-zoster virus infections during pregnancy: current concepts of prevention,diagnosis and therapy. Part 2: Varicella-zoster virus infections

Varicella during pregnancy can be associated with severe illnesses for both the mother and her neonate. Varicella pneumonia must be regarded as a medical emergency, since pregnant women are at risk of life-threatening ventilatory compromise and death. After maternal chickenpox in the first and second trimesters, congenital varicella syndrome may occur in nearly 2% of the cases. The characteristic symptoms consist of skin lesions in dermatomal distribution, neurological defects, eye diseases and skeletal anomalies. If the mother develops varicella rashes between day 4 (5) antepartum and day 2 postpartum, generalized neonatal varicella leading to death in about 20% of the cases has to be expected. Normal zoster has not been shown to be associated with maternal pneumonia, birth defects or problems in the perinatal period. On the basis of the clinical consequences of varicella-zoster virus infections during pregnancy, the present paper summarizes the currently available concepts of prevention, diagnosis and therapy.     

Herpes simplex virus infection of human T-cell subpopulations.

The ability of herpes simplex virus type 1 to productively infect human T-cell subpopulations was examined. Unstimulated helper/inducer (T4+) and cytotoxic/suppressor (T8+) lymphocytes limited herpes simplex virus replication as effectively as unseparated peripheral blood T cells (T3+). Phytohemagglutinin stimulation before infection resulted in equivalently productive herpes simplex virus infections in the three cell fractions.     

Protective effect of an oral infection with Herpes simplex virus type 1 against subsequent genital infection with Herpes simplex virus type 2

The problem of whether oral Herpes simplex virus type 1 (HSV-1) infection provides protection against subsequent genital infection by Herpes simplex virus type 2 (HSV-2) was investigated. Mice were used as models. Following conditions in man, both the oral and genital infections applied were noninjurious. Mice infected orally with HSV-1 were weakly protected against virus take following vaginal challenge with HSV-2. Genital takes were found in 67% of the immunized mice, as compared with 83% of the controls (protection rate 20%,P=0.002). The course of genital infection in the immunized mice, however, was relatively mild: Lethality decreased from 97% in the controls to 35% in the immunized mice (protection rate 63%,P<0.001). Furthermore, local and neurologic symptoms occurred less frequently. Attempts to isolate the virus from homogenized brain and spinal cord of immunized mice that died after genital challenge with HSV-2 failed in most cases. Also virus could not be recovered from the liver of infected mice, irrespective of the experimental group.