Varicella-zoster Antigen Research Articles

The duration of in vitro stimulation with recall antigens determines the subset distribution of interferon-γ-producing lymphoid cells: A kinetic analysis using the Interferon-γ Secretion Assay™

Analyses of cellular immune responses during natural infections and following vaccination with established or candidate vaccines are becoming increasingly important and so are the research tools used to achieve this goal. During a recent evaluation of the analytical performance characteristics of one of these techniques, the interferon-γ secretion assay, we noticed that following overnight incubation of PBMC with recall antigens (varicella-zoster antigen, Candida albicans antigen or hepatitis B surface antigen) NK cells are frequently the most predominant interferon-γ-producing cell population. In this study, we monitored the subset distribution of interferon-γ-producing cells following more extended in vitro culture periods and found that, irrespective of the antigen applied, the contribution of NK cells decreased whereas the importance of T cells and NKT cells rose. Analysis of the subset distribution showed that HBsAg stimulated CD4 cells predominantly whereas Candida antigen and varicella-zoster antigen were better inducers of CD8 responses. No correlation was found between the kinetics of total number of interferon-γ-producing cells and the changes of concentrations of interferon-γ in the culture supernatants. Interferon-γ levels in culture supernatants correlated strongly with the kinetics of T H lymphocytes (CD3 +, CD4 +), CTL (CD3 +, CD8 +), and NKT cells (CD3 +, CD56 +). These observations lead us to conclude that methods that enumerate cytokine-secreting cells without determining their phenotype should be interpreted with great care and that an ‘elispot’ should not be directly considered as the footprint of a T lymphocyte.

Avidity distribution of antibodies against peripheral nerve myelin in patients with polyneuropathy associated with IgM monoclonal gammopathy and in healthy controls

To elucidate the role and nature of antibodies against peripheral nerve myelin (PNM) we studied their avidity distribution. Twelve patients with demyelinating polyneuropathy associated with IgM monoclonal gammopathy were compared with 12 healthy blood donors previously found to have anti-PNM antibodies of IgM isotype. For comparison, the avidity distribution of IgM antibodies against the varicella zoster antigen in 10 patients with herpes zoster infection was also studied. Microtitre plates containing antibody bound to antigen were exposed to increasing concentrations of sodium thiocyanate (NaSCN) followed by an ELISA assay. NaSCN changes the ion strength and the pH, and thereby the critical conditions for antibody-antigen binding. Resistance to NaSCN was used as a measure of antibody avidity. Anti-PNM antibodies from patients with monoclonal gammopathy were of predominantly low avidity whereas antibodies from blood donors were of predominantly high avidity. Avidity index, representing the molar concentration of NaSCN required to reduce the initial absorbance values by 50%, was on average 11.7 times higher in blood donors (range 0.24-2.65, mean = 0.82) than in patients with monoclonal gammopathy (range 0.04-0.10, mean = 0.07) ( p = 0.002). On the other hand, patients with monoclonal gammopathy had on average a 100-fold higher relative concentration of antibodies against PNM compared to blood donors (range 4.1–392.6 AU, mean 85.0 AU, and range 0.2-1.7 AU, mean 0.85 AU, respectively) ( p = 0.002). Antibodies against the varicella zoster antigen from patients with herpes zoster showed a high avidity index (range 0.25-2.6, mean = 1.24). Using Western blot, several 14–30 kDa proteins in PNM were found to be the target antigen for IgM anti-PNM antibodies in both patients with monoclonal gammopathy and polyneuropathy, and in blood donors. Additional bands in the 43–67 kDa and > 94 kDa regions were commoner in blood donors than in patients with monoclonal gammopathy and polyneuropathy. These differences in staining pattern may reflect differences in avidity distribution or a qualitative difference at antigen binding sites of the anti-PNM antibodies. In conclusion, we found a strikingly low avidity of anti-PNM antibodies from patients with monoclonal gammopathy and polyneuropathy, and their pathogenetic role could apparently be questioned. However, low avidity implies reduced systemic elimination, with an increased risk of pathological depositions. This together with complement activation is an attractive pathogenetic model.

Effect of Hypnotic Suggestion on the Delayed-Type Hypersensitivity Response

To determine whether individuals selected for good general health, high hypnotizability, and the ability to alter skin temperature under hypnotic suggestion can influence the delayed-type hypersensitivity (DTH) response to varicella-zoster (VZ) antigen under hypnotic suggestion. A blinded clinical trial using a repeated measures design with subjects serving as their own controls. Subjects were randomly assigned to undergo a predetermined sequence of four different experimental conditions, occurring at weekly intervals, with each condition including VZ skin testing: (1) hypnosis with suggestions to enhance the DTH response to VZ antigen; (2) hypnosis with suggestions to suppress the DTH response; (3) hypnosis with suggestions for relaxation only; and (4) skin testing without hypnosis. A National Institutes of Health-supported clinical research center in a teaching hospital. A stratified sample of 24 ambulatory, healthy, highly hypnotizable, volunteer college students selected for their above-average ability to alter skin temperature after hypnotic suggestions and their positive baseline responses to VZ antigen. There were 11 males and 13 females with a mean +/- SD age of 22 +/- 6 years. The mean +/- SD hypnotizability score (Harvard Group Scale of Hypnotic Susceptibility) was 11 +/- 1. Intradermal skin testing with VZ antigen (Mantoux method) and hypnotic suggestion. Areas of induration of the DTH response measured at 24 and 48 hours after injection of antigen. The area of the DTH response was not affected by the experimental interventions. The area of erythema was likewise unaffected. Our subjects were unable to alter their DTH responses using hypnotic suggestion.

Atypical varicella/zoster infection in AIDS.

A case of atypical varicella zoster in a 33-year-old AIDS patient is reported. The patient had had two attacks of herpes zoster within a year and was given high-dose acyclovir several times. Thereafter he developed small keratotic pellucid papules on fingers, wrists and face, which were found to contain varicella-zoster antigen by the ELISA test. Skin biopsy showed acanthosis and lack of vesication, as is usually seen in herpes infections. The atypical varicella-like lesions persisted despite repeated doses of acyclovir but cleared temporarily when the patient was given foscarnet. We believe that the prolonged therapy may have allowed selection of acyclovir-resistant varicella-zoster strains, resulting in the atypical clinical course.

Individual NK Cell Clones Lyse Both Tumor Cell Targets and Herpes Simplex Virus-Infected Fibroblasts in the Absence of Interferon

The target specificity of natural killer (NK) cells for either tumor cells or virus-infected cells has been investigated. Lymphocyte clones with the surface phenotype of NK cells (CD3-, CD16+) were obtained by limiting dilution of peripheral blood mononuclear cells stimulated with PHA, Herpes simplex virus type 1 (HSV-1), or Varicella-Zoster antigens. Clones were maintained in media with recombinant interleukin 2 (IL-2). Both NK-sensitive (K562 cells) and NK-resistant (Raji cells) targets were lysed by three cloned lines of NK cells. The ability to lyse NK-resistant target cells was largely lost when the cloned lymphocytes were cultured overnight in the absence of IL-2. Effector cells from all three clones were also capable of specifically lysing HSV-1 infected human fibroblasts in comparison with uninfected fibroblasts. We also showed that lysis of HSV-1 infected targets by NK cloned cells was independent of interferons in the culture system.

HLA-DR3 and -DR4 control T-lymphocyte responses to mumps and Coxsackie B4 virus: studies on patients with Type 1 (insulin-dependent) diabetes and healthy subjects

To study the relationships between the responses to viral antigens and the HLA-DR3 and -DR4 associations in Type 1 (insulin-dependent) diabetes mellitus, the frequency of T-lymphocyte proliferating in response to mumps, Coxsackie B4 and varicella-zoster antigens was determined. A decreased frequency was found in T lymphocytes able to respond to mumps or Coxsackie B4 when presented together with DR3, as compared with the frequency of T lymphocytes able to respond to these viruses together with other DR determinants. This was not found for varicella-zoster or purified protein derivative of tuberculin. In contrast, an increased frequency was found in T lymphocytes responding to mumps or Coxsackie B4 together with DR4, compared with other DR determinants. The results were similar in Type 1 diabetic and healthy individuals. The results suggest that elements on the DR3 and DR4 molecules may control T-lymphocyte responses to mumps and Coxsackie B4 viruses.

Varicella-Zoster Virus Transformation of Hamster Embryo Cells

Varicella-zoster virus infection of primary hamster embryo cells resulted in oncogenic transformation. These transformed cells exhibited virus-specific antigens by immunofluorescence and developed surface Fc receptors. They induced aggressive fibrosarcomas when injected into inbred hamsters. The tumour-bearing hamsters develop antibodies to varicella-zoster antigens.

Selective decline in cellular immune response to varicella‐zoster in the elderly

The incidence of herpes zoster rises markedly in the aged. We evaluated the hypothesis that cellular immunity to varicella-zoster (VZ) viral antigens may be impaired in aged subjects. We found that the lymphocyte proliferation to VZ antigen was less in older asymptomatic individuals than in normal young controls. In contrast, responses to other antigens did not differ. Antibody titers to VZ were similar in both young and old subjects. Impairment of cellular immunity to VZ, on a population basis, may contribute to the increased risk of herpes zoster in the elderly.

Cellular and humoral immunity in the pathogenesis of recurrent herpes viral infections in patients with lymphoma.

86 patients with lymphoma were evaluated prospectively for clinical and laboratory evidence of recurrent varicella-zoster, herpes simplex, and cytomegalovirus infections during the first 16 mo of treatment. Cellular immunity to the viral antigens was measured by in vitro lymphocyte transformation and interferon production. Antibody titers and nonspecific measures of cellular immunity, including T-cell quantitation and transformation to phytohemagglutinin, were also assessed. The patients treated with radiation and chemotherapy had the highest incidence of reactivation of each of the viruses (15-19%). Greater susceptibility to herpes viral reactivation in these patients correlated with suppression of cell-mediated immunity to the specific virus. In individual patients, suppression of cellular immunity to the specific herpes viral antigen preceded each episode of reactivation, but recurrent infection did not occur in all patients with diminished specific lymphocyte transformation. Absence of the response appears to be a necessary but not a sufficient condition for the recrudescence of latent infection. Better preservation of cellular immunity to herpes simplex antigen during treatment was associated with infrequent reactivation of herpes simplex. In 25 patients with acute herpes zoster, uncomplicated recovery from the infection was accompanied by the development of lymphocyte transformation and interferon production to varicella-zoster antigen. Quantitation of T-cell numbers and phytohemagglutinin transformation did not correlate with the presence of viral cellular immunity in treated patients. Responses returned while T-cell numbers were low, and the recovery of phytohemagglutinin transformation often preceded recovery of the responses to viral antigens. Although some patients had deficiencies in viral cellular immunity at diagnosis, the duration of the suppression of specific antiviral responses resulting from treatment appears to be the most important factor predisposing to the recurrence of herpes infections in lymphoma patients.

Circulating immune complexes and complement-fixing antibodies in patients with varicella-zoster infection. Relationship to début of the disease.

Serum samples from 32 varicella-zoster (VZ)-infected patients were examined for circulating immune complexes (IC) within 1 1/2 week after début of the skin rash as well as during the following 5 weeks. Three test systems were used: a complement consumption (CC) assay, a C1q solid phase (CU1-SP) assay and a polyclonal rheumatoid factor-binding (pRF-I) assay. The CC and pRF-I assays were the most effective in detecting large IgG aggregates of IC, while aggregates of intermediate and small size were more easily demonstrated in the C1q-SP assay. IC were demonstrated in at least 2 assays simultaneously in 16 patients. Four of these patients were IC-positive in both observation periods, while 11 converted to IC-positive in the second observation interval. In 3 patients circulating IC were demonstrable as late as 8--10 weeks after the appearance of the skin rash. Maximal levels of complement-fixing antibodies to VZ antigens were observed 2 to 5 weeks after the appearance of the rash.

Transfer factor and cellular reactivity to varicella-zoster antigen in childhood leukemia

The present studies were designed to evaluate possible efficacy of specific human transfer factor (TF) in preventing or attenuating varicella-zoster (VZ) infection in children with acute lymphocytic leukemia (ALL). TF was prepared following leukapheresis of adult donors who were convalescing from chicken pox. Recipients were VZ seronegative children with ALL, 12 in remission and 3 in relapse; a single injection of TF was given equivalent to 10 8 lymphocytes per 7 kg body wt. The following VZ-specific parameters were measured: lymphocyte blastogenesis, cytotoxicity and leukocyte inhibitory factor (LIF) production, and indirect fluorescent and CF antibody titers. No patients in relapse converted immune response while 10 12 in remission developed positive reactivity in at least one assay of cell-mediated immunity (CMI); 3 12 were positive in all three parameters of CMI and 8 12 in two assays. Cytotoxicity was the most consistently positive test following TF administration. No patients developed VZ antibody. Seventeen months follow-up has demonstrated persistent cellular immunity.

Nervous system complications of herpes zoster: immunofluorescent demonstration of varicella-zoster antigen in CSF cells

In nine out of 11 patients with herpes zoster and neurological symptoms varicella-zoster antigen was demonstrated in CSF cells by indirect immunofluorescent staining. Cerebrospinal fluid cytology was in most cases highly suggestive of viral infection. This study demonstrates the value of both immunofluorescent staining on CSF cells and CSF cytology, as corroboration of an otherwise exclusively serologically based virological diagnosis.

Cell-mediated immunity to varicella-zoster virus infection in subjects with lymphoma or leukemia

Normal subjects and patients with lymphoma or leukemia were tested for the levels of lymphocytes, E-rosette--forming T-cells, serum and vesicle fluid interferon, and specific in vitro proliferative response to varicella-zoster antigen after clinical varicella or herpes zoster illness. The effect of polyinosinic acid/polycytidilic acid on the immune response was also evaluated. The development of VZ specific cell-mediated response in normal subjects was characterized by intense proliferative activity eight to ten days after the onset of illness, with significant decline 70 to 80 days later. The responses in subjects with lymphoma or leukemia were much lower. Few subjects with chickenpox or zoster with lymphoma or leukemia died during the infection. Death was associated with significant depletion of E-rosette--forming T-cells, and grossly deficient specific cellular response to VZ antigen. Treatment with Poly IC frequently induced elevations in serum as well as vesicle fluid interferon levels, and increased the proliferative activity of lymphocytes against VZ antigen.

Varicella and Acute Cerebellar Ataxia

In two cases of varicella-associated cerebellar ataxia, varicella-zoster antigens in CSF cells were shown by an indirect immunofluorescent technique. Direct viral invasion in CNS disease complicating varicella plays an important part in pathogenesis and rules out a single immune-mediated mechanism.

Disseminated Varicella Infection: Treatment with Transfer Factor in a Patient with Hodgkin's Disease

Treatment with transfer factor in a patient with disseminated varicella infection complicating stage IV Hodgkin's disease is described. The patient, a 24-year-old woman, showed transient clinical improvement and restoration of immune response to varicella-zoster antigen after receiving transfer factor. Though she later died from septicaemia, further trials of treatment of disseminated viral infection in patients with Hodgkin's disease with transfer factor are indicated.

Herpes zoster and impaired cell-associated immunity to the varicella-zoster virus in patients with Hodgkin's disease

Thirty-two patients with Hodgkin's disease and 12 normal donors were studied for their in vitro lymphocyte responsiveness to a membrane-associated varicella-zoster (VZ) antigen. When compared to the normal donors, patients with Hodgkin's disease in whom radiotherapy was recently completed and those with active, recurrent disease had markedly impaired cell-associated immunity to VZ antigen. In addition, there was a suggestion that patients in long-term remission who had received primary combined modality therapy (radiotherapy plus chemotherapy) had an impaired response when compared to normal persons or to patients who had received single modality therapy. Newly diagnosed, untreated patients with Hodgkin's disease did not differ significantly from normal persons as a group but two of six were unresponsive to the VZ antigen whereas all normal subjects were responsive. Most patients in remission for at least one year following therapy had normal in vitro responsiveness. In two patients herpes zoster developed after the demonstration of absent in vitro lymphocyte reactivity to the VZ antigen.

The demonstration of cell-associated immunity to viruses. In vitro lymphocyte responsiveness to varicella-zoster antigen

The demonstration of in vitro lymphocyte responsiveness to common pediatric viruses has previously been fraught with many technical and conceptual problems. Based upon our prior experience in demonstrating cell-associated immunity to mumps, rubella and measles viruses we illustrate our methodology and conceptual framework by documenting in vitro lymphocyte responsiveness to the Varicella-zoster virus, another ubiquitous virus of childhood. We discuss our approach to the problems of reactivity, specificity and reliability in the use of membrane-associated viral antigens.

Zoster, reinfection or activation of latent virus?: Observations on the antibody response

An epidemiologic investigation of 108 patients with zoster revealed that (1) only four had a possible recent exposure to varicella or zoster, (2) there was no increase in zoster cases corresponding to the seasonal peak of varicella, and (3) the fewest cases of zoster were contributed by subjects with the greatest likelihood of being exposed to varicella. These observations indicate that reinfection did not play a significant role in the causation of zoster. Patients with localized zoster had an anamnestic type of antibody response, confirming the putative thesis that these subjects had been exposed previously to varicella-zoster antigen. The character of the response suggested that passive immunization of these patients would not be indicated. Two of the five patients with generalized zoster, in contrast, had a delay in the appearance of antibody. The failure to observe a delay in all five patients, however, suggested that this may be but a single manifestation of a more profound immunologic defect in these subjects. An epidemiologic investigation of 108 patients with zoster revealed that (1) only four had a possible recent exposure to varicella or zoster, (2) there was no increase in zoster cases corresponding to the seasonal peak of varicella, and (3) the fewest cases of zoster were contributed by subjects with the greatest likelihood of being exposed to varicella. These observations indicate that reinfection did not play a significant role in the causation of zoster. Patients with localized zoster had an anamnestic type of antibody response, confirming the putative thesis that these subjects had been exposed previously to varicella-zoster antigen. The character of the response suggested that passive immunization of these patients would not be indicated. Two of the five patients with generalized zoster, in contrast, had a delay in the appearance of antibody. The failure to observe a delay in all five patients, however, suggested that this may be but a single manifestation of a more profound immunologic defect in these subjects.

The etiologic agents of varicella and herpes zoster; serologic studies with the viruses as propagated in vitro.

The preparation of antigenic materials capable of specific fixation of complement in the presence of convalescent phase sera from patients with varicella and herpes zoster is described. Satisfactory antigens were obtained by the repetitive harvest and subsequent concentration of the pooled nutrient fluids from bottle cultures of human embryonic skin-muscle tissue or of monkey kidney tissue infected with varicella or herpes zoster viruses. Specific fixation of complement was also demonstrated with antigens prepared from extracts of the infected cell sheets harvested from bottle cultures. In individuals with varicella, complement-fixing antibody usually appeared in the serum 4 or 5 days after the development of the exanthem and further significant increases in titer were characteristically observed during the 2nd week of illness. The complement-fixing antibody response in herpes zoster tended to follow the same pattern as in varicella, with the exception that in sera from some individuals relatively high titers were present in the acute phase specimen. Complement-fixing antigens prepared from varicella strains or from a zoster strain reacted to essentially the same degree with convalescent sera from the homologous and the heterologous clinical entities. The varicella-zoster antigens did not fix complement in the presence of paired sera obtained from a limited number of individuals with primary infections due to herpes simplex virus or from individuals with generalized vaccinia infection. Specific inhibition in vitro of the focal cytopathic process produced by the varicella-zoster viruses was demonstrated. This was accomplished by the incorporation of the sera under test as constituents of nutrient media of the cultures, either prior to or at the time of their inoculation with virus. The neutralization of focal cytopathogenicity thus obtained was relative in degree and never absolute; it was therefore assayed by repetitive counts of the number of focal lesions per culture in the various test groups. Inhibition of varicella-zoster viral cytopathogenicity occurred in the presence of convalescent serum from either clinical entity. The results of the immunologic studies with the viruses of herpes zoster and varicella as propagated in vitro are considered as providing further evidence in support of the concept of the close relationship and probable identity of the two agents.