Simplex Antigen Research Articles

Herpes simplex antigen in immune complexes of patients with erythema multiforme: presence following recurrent herpes simplex infection

Herpes simplex antigen in immune complexes of patients with erythema multiforme: presence following recurrent herpes simplex infection

In vitro stimulation of rabbit T lymphocytes by cells expressing herpes simplex antigens.

Lymphocyte stimulation responses to herpes antigens were studied using virus-infected X-irradiated cells. Rabbits were immunized with herpes simplex virus type 1 (strain HFEM) grown in RK 13 cells. For in vitro stimulation assay BHK21 cells were X-irradiated (15 000 rad) and infected with a high m.o.i. of a temperature-sensitive (ts) mutant (N102) of HFEM strain at the non-permissive temperature (38.5 degrees C) of virus. Virus antigens were expressed on the infected cells and there was no leakage of infectious virus into the medium at 38.5 degrees C. T lymphocytes from rabbits immunized with herpes simplex virus were specifically activated by herpesvirus-infected X-irradiated cells; lymph node cells from rabbits immunized with RK13 cells and from non-immune rabbits showed no proliferative response.

Abolished production of interferon by leucocytes of patients with the acquired cytogenetic abnormalities 5q -- or -- 5 in secondary and de-novo acute non-lymphocytic leukaemia.

Interferon production by leucocytes on stimulation with inactivated antigens from herpes simplex virus, varicella zoster virus and cytomegalovirus was determined in 19 patients with preleukaemia or acute non-lymphocytic leukaemia, in 15 following treatment for other tumours. Production of interferon was abolished in 10 patients and preserved in nine cases after stimulation with herpes simplex antigen. Cytogenetic studies of bone marrow demonstrated an abnormal karyotype in 13/15 patients with secondary preleukaemia or leukaemia. Characteristics were (a) hypodiploid cell lines demonstrated in eight cases, (b) a B-chromosome defect found in five cases and verified as 5q -- in four, and (c) defects in C-group chromosomes in 10, verified as monosomy 7 in nine. All four patients with de-novo preleukaemia or leukaemia had abnormal cytogenetic findings, two B-chromosome abnormalities verified as monosomy 5. A relationship between abnormalities in chromosome no. 5 and abolished production of interferon was demonstrated, as only one of seven patients with 5q --, --5 or --B produced interferon by the leucocytes, compared with eight of a total of 12 patients with other cytogenetic defects or a normal karyotype (P=0.04). The results are remarkable, as chromosome no. 5 has recently been discovered to contain a gene for interferon production. Furthermore abolished production of interferon by leucocytes seemed correlated to previous irradiation and to disease-related fever.

In vitro studies on the human cellular immune response to commercially available herpes simplex antigens.

Patients with recurrent herpes labialis and genitalis as well as normal controls were tested for the in vitro responsiveness of their lymphocytes to mitogens, to several herpes antigens and to carrier medium alone (containing chorioallantois membrane antigens). No statistically significant differences were found in the results with the direct and indirect migration inhibitory test (MIF) and in studies of the blastogenic response of lymphocytes (LTT) among the three groups. These findings held up when correlations were drawn to severity or course of the disease and suggest that immunological modulation can play at most a minor role in inducing recurrent herpes simplex infections.

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.

Familial herpes simplex infection associated with activation of the complement system

A patient with severe recurrent herpes infection was evaluated for immunologic analysis including a profile of complement components. The peripheral blood lymphocytes from the patient responded by proliferation to herpes simplex antigen but failed to produce leukocyte migration inhibition factor. Herpes simplex antibody titers increased during active infection. Total hemolytic complement (TCH50), the third (C3), fifth (C5), sixth (C6) and seventh (C7) components of complement, and factor B were dramatically reduced; the first (C1), second (C2) and fourth (C4) components of complement were within normal limits. In family members with a history of recurrent herpes simplex, one or more of the later complement components (C5, C6 or C7) was reduced. This study demonstrates the activation of complement in these serums via the alternative pathway.

Rapid diagnosis of viral neuroinfections by immunofluorescent and immunoperoxidase technics.

The results of immunofluorescent (IF) and immunoperoxidase (IP) technics applied for the detection of antigen in cerebrospinal fluid (CSF) cells in patients with mumps, herpes zoster and herpes simplex meningitis and meningoencephalitis are presented. Thirty patients were under study. The detection of mumps and herpes zoster viral antigen in CSF cells was possible in 100% of cases investigated. Herpes simplex virus antigen was detected in four of seven cases with symptoms of severe meningoencephalitis. Complement fixation (CF) antibodies to herpes simplex virus (type I) and positive seroconversion were detected in the four latter patients. The diagnostic value of the methods used for the detection of mumps, herpes simplex and herpes zoster viral antigens in CSF cells of patients is discussed.

An in vitro study of depressed cell-mediated immunity and of T and B lymphocytes in atopic dermatitis

Lymphocyte transformation tests, E binding(T) rosette assay and immunofluorescent preparations of B cells were studied in patients with atopic dermatitis, and also in healthy controls. Most of the patients were found to have high levels of IgE in serum. The patients were studied both during severe bouts of dermatitis and also when the dermatitis was almost healed. Lymphocyte transformation tests showed that patients were hyporeactive to PPD and herpes simplex antigen in vitro, both during periods of severe dermatitis and also when the dermatitis was in remission. The response to PHA in the patients was normal in vitro. No factors which could reduce cell-mediated immunity in vitro were found in patients' sera. A decreased number of T lymphocytes and a slight increase in the number of immunoglobulin-bearing lymphocytes (B cells) were demonstrated in the patients, both during remission and during recurrence of severe dermatitis. In 3 of 8 patients, increased numbers of IgE-bearing lymphocytes were found to be present when the patients had severe dermatitis. A possible correlation between high serum levels of IgE and depressed cell-mediated immunity in patients with atopic dermatitis is discussed.

Lymphocyte transformation and interferon production in human mononuclear cell microcultures for assay of cellular immunity to herpes simplex virus

Interferon production and transformation in response to herpes simplex virus antigen were studied in microcultures of human mononuclear cells. Mononuclear cells consisting of monocytes and both T and B lymphocytes were purified by Ficoll-Hypaque gradients. Lymphocytes, predominantly T with 5% B, were obtained by passage of buffy-coat cells through nylon fiber columns. For some experiments, autochthonous macrophages and column-purified lymphocytes were stimulated with herpesvirus antigen. The effect of specific antibody and cell concentration on reactivity is described. Crude and purified antigens were compared as cell culture stimulants. Significant differences in transformation and interferon were observed between donors with a history of herpes labialis and donors with no detectable antibody, both in cultures prepared by Ficoll-Hypaque gradients and by column purification of lymphocytes. Cultures from seronegative donors prepared by Ficoll-Hypaque gradients produced interferon but did not transform when stimulated by herpes simplex antigen. "Immune" interferon production, that is, type II as opposed to type I, occurred only with autochthonous macrophage and column-purified lymphocyte cultures. Interferon produced by Ficoll-Hypaque-purified mononuclear cultures was type I, and its production was unrelated to immune status. Similarly, column-purified lymphocytes responded to herpes simplex virus antigen with type I interferon if obtained from a seropositive donor.

Cellular immunity and circulating antibody to herpes simplex virus in subjects with recurrent herpes simplex lesions and controls as measured by the mixed leukocyte migration inhibition test and complement fixation

Cellular immune responses and circulating antibody levels to herpes simplex virus were examined in patients with recurrent herpes simplex (HSV) infections and controls. Mixed leukocyte migration inhibition by herpes simplex antigen was less in affected patients than in controls but serum antibody levels were higher. There was no significant difference in leukocyte migration between patients with active or recent lesions and other herpes subjects, and the response in the mixed leukocyte migration test to phytohaemagglutinin (PHA) was similar in patients and controls. The data presented suggest that a localized defect in cell mediated immunity to herpes simplex virus may exist and be responsible for recurrent herpes simplex infections.

Use of Immunoperoxidase on Brain Tissue for the Rapid Diagnosis of Herpes Encephalitis

An indirect immunoperoxidase method to detect herpes simplex viral antigen in brain cell suspensions from patients suspected to have herpes encephalitis is described. The method is rapid, reliable and specific, and successfully identified the herpes virus infected cells in four of five culture-proven cases. There was one false-negative reaction, but no false-positive. The immunoperoxidase technic offers a number of advantages over immunofluorescence for routine diagnosis.

An in vivo and in vitro study of cell-mediated immunity in atopic dermatitis

Cell-mediated immunity was studied in patients with atopic dermatitis. Tuberculin skin tests were performed on 70 patients and on 18 controls. The patients with "severe dermatitis" and those with high IgE levels were hyporeactive. Lymphocyte transformation tests were carried out in 33 patients and 21 controls. This showed that the patients were hyporeactive to PPD and herpes simplex antigen and this was most prominent in the patients having high IgE values. However, there was no clear, statistically significant difference between patients and controls with regard to their in vitro response to PHA.

Immune Specific Production of Interferon by Human T Cells in Combined Macrophage-Lymphocyte Cultures in Response to Herpes Simplex Antigen

Human peripheral blood lymphocytes, highly enriched for T cells, were obtained by passing gravity-sedimented leukocytes through nylon wool columns. The eluted cells were cultured with autologous macrophages and the mixture was studied for its capacity to produce interferon in vitro in response to stimulation with herpes simplex virus antigen. The interferon produced by the combined macrophage-lymphocyte cultures was shown to depend upon the presence of T cells; elimination of these cells by treatment with an anti-T cell serum plus complement greatly diminished the amount of interferon produced. The memory for the immune-specific release of interferon also appeared to be carried by the T lymphocytes rather than the glass-adherent macrophages. Furthermore, the results suggest that under our conditions of culture immune-specific interferon originates from T cells.

Cell-mediated immunity to herpes simplex virus in man.

The immune response to herpes simplex virus was assessed by complement fixation tests, in vitro stimulation of lymphocytes, and intradermal skin tests on 63 healthy volunteers and 26 patients receiving antilymphocyte globulin (ALG) and/or adrenocorticotropic hormone. Lymphocytes from subjects with frequent recurrent herpes infections all showed significant stimulation by herpes simplex antigen, but only four of 37 without a history of previous herpes were positive. Skin tests with herpes simplex antigen demonstrated intact delayed hypersensitivity in the 10 patients tested. Lymphocyte responses to phytohemagglutinin (PHA), purified protein derivative (PPD), streptokinase, or vaccinia were similar in subjects with or without previous herpes. We were, therefore, unable to demonstrate any defect in these parameters of the immune response in patients with recurrent herpes simplex infections. Therapy with ALG totally abrogated both in vitro and in vivo evidence of cell-mediated immunity and was associated with herpes labialis in seven of the 21 courses. These infections were more severe than usual, and it may be that the cell-mediated response is more important in the localization of recurrent herpetic infections than in their prevention.

Precipitating antibodies to herpes simplex virus in human sera: prevalence of antibody to common antigen (band II).

The sera of 140 female subjects were investigated for precipitating antibody to type 1 and type 2 herpes simplex antigen. 73 of these sera formed a common precipitin line against type 1 and type 2 vir

Cell-mediated immunity to herpes simplex virus in man.

The immune response to herpes simplex virus was assessed by complement fixation tests, in vitro stimulation of lymphocytes, and intradermal skin tests on 63 healthy volunteers and 26 patients receiving antilymphocyte globulin (ALG) and/or adrenocorticotropic hormone. Lymphocytes from subjects with frequent recurrent herpes infections all showed significant stimulation by herpes simplex antigen, but only four of 37 without a history of previous herpes were positive. Skin tests with herpes simplex antigen demonstrated intact delayed hypersensitivity in the 10 patients tested. Lymphocyte responses to phytohemagglutinin (PHA), purified protein derivative (PPD), streptokinase, or vaccinia were similar in subjects with or without previous herpes. We were, therefore, unable to demonstrate any defect in these parameters of the immune response in patients with recurrent herpes simplex infections. Therapy with ALG totally abrogated both in vitro and in vivo evidence of cell-mediated immunity and was associated with herpes labialis in seven of the 21 courses. These infections were more severe than usual, and it may be that the cell-mediated response is more important in the localization of recurrent herpetic infections than in their prevention.

Patterns of cytomegaloviral complement-fixing antibody activity: a longitudinal study of blood donors.

Fifty donors undergoing plasmapheresis were the subjects of an 18-month longitudinal study of patterns of the activity of complement-fixing (CF) antibody to cytomegalovirus (CMV). Sera, collected monthly whenever possible, were tested with each of three preparations of antigens obtained from the Davis, AD169, and Esp strains of virus. Of 510 sera examined, 321 were reactive with one or more of the antigens; 96% of the reactive sera fixed complement with Esp, 94% with Davis, and 83% with AD169. Patterns of reactivity were determined for 20 of the 39 reactive donors. Multiple fluctuations in titer were observed in 19 of the 20 donors, and 11 individuals vacillated at least once between significant and undetectable (>1:8 to <1:4) levels. A degree of CMV-strain specificity was observed in some antibody patterns. Donors with labile CF reactivity to CMV did not exhibit concurrent fluctuations in titer on examination with herpes simplex antigen. The antibody patterns suggest that the CMV-host relationship is more dynamic than heretofore appreciated. While not necessarily applicable to the normal adult population, the findings emphasize the necessity for critical interpretation of the clinical significance of titers of CF to CMV.

Comparison of herpes simplex antigens prepared from various tissues.

Herpes simplex virus was propagated in various tissues, and the antigens prepared from each of them were compared for complement-fixing components. The amount of viral antigen component exceeded that of soluble antigen in the antigens prepared from tissue cultures but the reverse was true with chorioallantoic membrane antigen. The viral antigen component was more sensitive than the soluble antigen for the determination of herpes simplex antibodies.

Immunopathology of oral leukoplakia.

The lymphocyte transformation test was performed with autologous saline homogenates of leukoplakia. A negative correlation was established (P<0.05) between the (14)C thymidine uptake of lymphocytes in vitro and the non-pyroninophilic mononuclear cell infiltration in biopsies. A depression of lymphocyte transformation was revealed in patients with carcinoma or carcinoma in situ and some with epithelial atypia, as compared with those showing only hyperkeratosis and to a less extent those with acanthosis. A corresponding depression was not found when lymphocytes were stimulated with phytohaemagglutinin, Candida albicans or Herpes simplex antigens. The Pyroninophilic cell count was raised in biopsies with carcinoma or carcinoma in situ and in some with epithelial atypia or acanthosis. These result suggest that in leukoplakia the carcinomatous transformation may be associated with some immunological changes.

Varicelliform eruptions in herpes zoster--some clinical and serological observations.

AbstractIn a hospital series of 100 consecutive cases of herpes zoster, studied prospectively, a secondary appearance of chickenpox-like eruptions was noted in half of the 12 patients with a diagnosed malignant disease but also in as many as 33% of the other patients. In the latter cases the eruptions were usually much fewer. A serological study of 11 patients showed a rising CF titer against varicella antigen in 5 out of 7 patients from whom the first serum specimen was obtained within 1 week of illness. One of the patients had a response also against herpes simplex antigen. No CF antibodies against varicella antigen were detected in 2 patients; one of them was 92 years old and the other suffered from lymphatic leukemia treated with corticosteroids.