[The immune suppression syndrome in homosexual men. An epidemiological study from the Cancer Research Institute in Arhus].
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Adult↗
Biomedical subjects
B F Vestergaard
Publications and source records attributed to B F Vestergaard.
Detection and characterization of EBV antigens by micro-ELISA and chromatofocusing.
A micro-ELISA technique was developed for the detection of Epstein-Barr virus (EBV)-determined antigens. The enzyme-linked immunosorbent assay (ELISA) was applied with peroxidase-protein A to detect the antigens adsorbed to micro-ELISA plates. Human and rabbit antisera containing antibodies to known EBV components were used as reagents. The early antigen (EA) complex, associated with the viral cycle, was readily detected in extracts of n-butyrate- or n-butyrate + TPA-induced cells. The nuclear antigen, EBNA, could be unequivocally detected only after the partial purification of the antigen by DNA cellulose chromatography. EA (and VCA) could be separated by chromatofocusing of induced cell extracts into several fractions detected by the micro-ELISA technique. This indicates that the purification of individual antigens of the EA complex can be monitored by ELISA.
Animals↗
Herpesvirus antibodies and vascular complications in essential hypertension.
Antibodies against herpes simplex virus (HSV) and cytomegalovirus (CMV) were examined in sera from 132 patients with essential hypertension and 54 normotensive healthy subjects of the same age and sex. Prevalences of HSV and CMV antibodies (titre greater than or equal to 4) were equal in patients and controls. A HSV antibody tire greater than or equal to 64 was found in 39.5% (17/53) of patients with WHO stage III hypertension, in 26.2% (22/85) of patients with stage I-II hypertension, and in only 9.4% (5/54) of normotensive subjects (p less than 0.0005). The HSV antibodies were mainly of type 1. No association between CMV antibodies and vascular complications could be demonstrated.
Adult↗
A prospective study of the relationship between genital herpes and carcinoma of the uterine cervix. I. Seroepidemiology of herpes simplex virus type 1 and type 2 in women with a previous history of clinically diagnosed genital herpes.
The development of a simple, sensitive and reliable enzyme-linked immunosorbent assay (ELISA) for the exact titration of herpes simplex virus type 1 -- and type 2 -- specific antibodies in human serum has made it possible to perform large serological HSV type-specific diagnosis of clinical cases of post and present genital HSV infections. The distribution of HSV type-specific IgG antibody in 473 sera from women with a first episode of symptomatic genital herpes (group A), 602 sera from women with a previous history of symptomatic genital herpes (group B) and 945 sera from non-selected adult Danes (controls, group C) gave the following results: 34 percent of the women in group B had antibodies only to HSV type compared to 6 and 4 percent in group A and C respectively. 64 percent of women in group A were sero-negative, while only 9 and 18 percent sero-negative were found in group B and C respectively. A majority of the sero-negative in group A seroconverted. One third developed antibodies to HSV type 1 and two thirds to HSV type 2.
Antibodies, Viral↗
A geographical study of antibodies to membrane antigens of HSV-2-infected cells and HSV-2-specific antibodies in patients with cervical cancer.
Sera was obtained from patients with squamous carcinoma of the cervix from Great Britain (29), Sri Lanka (32), Malawi (27), and Sudan (27), and from controls from all countries except Sudan. Controls were matched for ethnic origin, age and social class. Sera were tested by indirect immunofluorescence (IF) for IgG and IgA antibodies to membrane antigens (MA) and IgA antibodies to VCA of HSV-2 infected cells. Compared with controls, IgA antibodies to MA (IgA anti-MA) were detected more frequently and at higher titres in all groups of patients. However, there was no significant difference in prevalence of these antibodies at titres greater than or equal to 1:4 between Malawian patients and controls, although a significantly higher proportion of patients had IgA anti-MA titres of greater than or equal to 1:16. In contrast, IgA anti-VCA did not distinguish patients from controls. More than 90% of both patients and controls from all countries had IgG antibodies to MA (IgA anti-MA). Malawian patients had a significantly higher geometric mean titre (GMT) of IgG anti-MA than controls and both patients and controls had significantly higher GMTs than their counterparts from other countries. The variation between herpes IgG anti-MA titres in subjects from different countries did not reflect differences in serum immunoglobulin levels and similar variations in titre were not seen in rubella and measles HAI titres. Among the patients there was a geographical variation in the prevalence of HSV-2 specific antibodies detected by ELISA, which varied from 52% among British and Sudanese patients to 73% among Malawian patients. Even when adjustment was made for possible false negative results, there were between 10 and 31% patients without HSV-2-specific antibodies, although only 2 of 103 (1.9%) patients had neither HSV-1 nor HSV-2 antibodies. The association of HSV-2 with cervical carcinoma appeared to vary with age.
Adult↗
Optimized enzyme-linked immunosorbent assay for detection of human and bovine rotavirus in stools: Comparison with electron-microscopy, immunoelectro-osmophoresis, and fluorescent antibody techniques.
Detection of human and bovine rotavirus in stools is described using a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) with poly-styrene microtest plates as solid phase, immunoglobulin fraction of rabbit antiserum to rotavirus (human) as catching antibody, and the same reagent labelled with horseradish peroxidase as conjugate. The ELISA has been optimized with regard to simplicity, rapidity, sensitivity, and specificity. In a comparative study, stool specimens from 81 infants and children and 92 neonatal calves with diarrhoea were tested for rotavirus by ELISA, electron microscopy (EM), immunoelectro-osmophoresis (IEOP), and fluorescent antibody technique (FA). The relative sensitivity of the different assays for human and bovine rotavirus was: EM 68%, 76%; IEOP 80%, 76%; FA not determined, 85%; and ELISA 86%, 98%, respectively. Less than 1 ng of purified human rotavirus could be detected in ELISA, whereas 100 ng was the minimal amount detected by IEOP. It is concluded that the developed ELISA is a simple, rapid, reliable, and sensitive method for the diagnosis of human and bovine rotavirus infections.
Animals↗
Production of monospecific antibodies to varicella zoster virus antigen in rabbits tolerant to human IgG and immunized with antigen immunoprecipitated with human IgG from zoster reconvalescent sera.
A method for production of antisera specific to a single antigen of a human infectious agent is described with varicella zoster virus (VZV) as a model. Microisolation of immunochemically pure antigen contained in defined immunoprecipitates was achieved by crossed immunoelectrophoresis of a crude antigenic preparation (detergent solubilized, VZV infected cells) into agarose gel containing zoster reconvalescent immunoglobulin. Immunization of rabbits made tolerant to human IgG with immunoprecipitates resulted in the production of antibodies to the antigen part of the immunoprecipitate. The method represents a short cut to the production of heterologous diagnostic antisera to defined antigens of human infectious agents.
Animals↗
In vitro susceptibilities of normal human skin fibroblasts to oncoviruses, and the decreased susceptibility to HSV of fibroblasts from untreated Hodgkin's patients.
Fibroblast cultures established from the skin of 56 healthy controls and 15 untreated Stages I and II Hodgkin's patients (HD) were studied in their 3rd, 4th and 5th in vitro passage with respect to transformation with Simian sarcoma virus (SSV) and SV40 and with respect to replication of herpes simplex virus (HSV) Types 1 and 2, pox virus and interferon release. Susceptibility to the 5 viruses varied independently, except for an inverse correlation between susceptibility to SSV and HSV. HD cultures showed a depressed replication of both types of HSV. There was a borderline (P = 0.02) correlation between magnitude of HSV replication and presence of HL-A type B-w44, but this does not explain the HD control difference. Furthermore, the level of serum antibodies to HSV common antigen was not related to magnitude of in vitro replication. The results thus speak against generally enhanced cellular susceptibility to HSV as a reason for the high titres of serum antibodies to HSV in HD patients.
Adolescent↗
Type-specific herpes-simplex-virus antibodies in patients with recurrent duodenal ulcer.
The prevalences of herpes-simplex-virus (HSV) type-1-specific antibodies in 52 patients with recurrent duodenal ulcer, 60 controls, and 32 patients with recurrent cold sores were 94%, 80%, and 100% respectively. Seropositive ulcer patients had significantly higher antibody levels than seropositive controls. The cold-sore group had higher antibody levels than the controls, but this difference did not reach statistical significance. Since all three groups had similar HSV type-2-specific antibody levels, the results demonstrate an unusual activity of HSV type 1 in duodenal-ulcer patients, compatible with the hypothesis that herpesvirus is an aetiological agent in some cases of this disorder.
Adolescent↗
The influence of the major histocompatibility complex (H-2) on experimental diabetes in mice.
Mice with different histocompatibility loci on an identical background genome (congenic resistant lines of mice) were used to study the possible influence of the histocompatibility complex on experimental diabetes. The major histocompatibility complex (H-2) was not found to influence the diabetogenic effect of encephalomyocarditis (EMC) virus. In contrast the glucose intolerance following heterologous and homologous immunization with pancreatic antigens appeared H-2 influenced. Antibodies against cell surface components on viable B-cells were present in serum from mice with glucose intolerance induced by homologous immunization. The results suggest that the susceptibility to experimental autoimmune diabetes in mice is influenced by the H-2 complex.
Animals↗
ELISA for herpes simplex virus (HSV) type-specific antibodies in human sera using HSV type 1 and type 2 polyspecific antigens blocked with type-heterologous rabbit antibodies.
Crude antigenic preparations made from rabbit cornea cells infected with either HSV type 1 or type 2 could be used in ELISA for titration of HSV type-specific antibodies in human sera. After immunochemical binding of the crude HSV antigens in microtitre wells by use of type-homologous rabbit antibodies, type-common antigenic sties were blocked with type-heterologous rabbit antibodies. Titration of human sera in this system showed that high concentrations of type heterologous rabbit antibodies were capable of completely blocking type-common antigenic sites, while leaving type-specific antigenic sites unblocked and capable of reacting with human antibodies. Thus HSV type-specific antibodies in human sera could be measured in ELISA without the use of purified typespecific antigens.
Animals↗
Quantitative immunoelectrophoretic analysis of human antibodies against herpes simplex virus antigens.
By use of crossed immunoelectrophoresis with intermediate gel, antidbody titers against six individual herpes simplex virus (HSV) glycoproteins and two nonglycosylated proteins were determined in 100 human sera. High antibody titers were found against two different HSV type-common glycoproteins designated Ag8 and Ag11 (containing glycosylated polypeptides D and B, respectively). The anti-Ag8 and -Ag11 titers correlated with HSV neutralizing antibody titers. Most of the serological cross-reactivity between HSV type 1 and type 2 was probably caused by antibodies to Ag8 and Ag11. Human antibodies against one HSV type 1-specific glycoprotein (Ag6, containing glycosylated polypeptide C) and two HSV type 2 glycoproteins (Ag4 and Ag9) were also demonstrated, and the titers correlated better with neutralizing antibody titers of the homologous than of the heterologous virus type. The data presented can be directly applied to the further development of diagnostic reagents.
Antibodies, Viral↗
Isolation of the major herpes simplex virus type 1 (HSV-1)-specific glycoprotein by hydroxylapatite chromatography and its use in enzyme-linked immunosorbent assay for titration of human HSV-1-specific antibodies.
A 131,000 molecular weight herpes simplex virus type 1 (HSV-1) glycoprotein designated antigen number 6 (Ag-6) was previously shown to possess almost exclusively HSV-1-specific antigenic sites. Fused rocket and crossed immunoelectrophoresis of fractions obtained from hydroxylapatite chromatography of crude HSV-1 antigen (Triton X-100-solubilized, infected tissue culture cells) showed that a subfraction of Ag-6 could be separated from the other HSV antigens. Enzyme-linked immunosorbent assay with the isolated Ag-6 showed that sera from rabbits infected with HSV-1 and HSV-1 human antisera contained antibodies to Ag-6, whereas sera from HSV-2-infected rabbits and sera from patients with primary HSV-2 infections did not react with Ag-6. Enzyme-linked immunosorbent assay of 852 human sera for antibodies to HSV type-common glycoproteins, Ag-6, and HSV 2-specific antigens showed that 139 sera which reacted negatively with HSV type-common glycoproteins also did not react with Ag-6 with HSV-2 specific antigens. The 713 sera reacting positively to HSV type-common antigens either reacted with Ag-6 (328 sera) or with HSV-2-specific antigens (31 sera) or both (354 sera). This means that Ag-6 might be useful in large-scale human serology for the detection of past infection with HSV-1, irrespective of whether or not past infection with HSV-2 has occurred.
Animals↗
Immunoelectrophoretic identification and purification of herpes simplex virus antigens released from infected cells in tissue culture.
Proteins released from herpes simplex virus type 1 (HSV-1)-and type 2 (HSV-2)-infected HEp-2 cells have been characterized by the crossed immunoelectrophoretic technique. Both HSV type-common and type-specific antigens were found in the tissue culture medium 24 h after infection. Antigen Ag-6, an HSV-1-specific antigen, was found in high concentration in the medium as compared to other HSV antigens released from HSV-1-infected cells. The HSV-2-specific antigens, Ag-4 and Ag-1, were released in molecular modifications with altered electrophoretic mobility as compared to their cellular counterparts. Purification of HSV antigens was performed by ion-exchange chromatography, and an HSV type-common antigen, Ag-11, and an HSV-2-specific antigen, Ag-4A, were isolated.
Antigens, Viral↗
Immunoelectron microscopic localization of herpes simplex virus antigens in infected cells using the unlabeled antibody-enzyme method.
Subcellular localization of viral antigens was demonstrated during viral morphogenesis using herpes simplex virus type 1 (HSV-1) infected monolayers of rabbit cornea cells. The localization was done by immunoelectron microscopy employing the peroxidase-antiperoxidase (PAP) immunocytochemical technique and the postembedding staining method. The localization of viral antigens was followed at time intervals during infection from 2 to 19 hr. After exposure of sections to either polyspecific antibodies against total HSV-1 antigens or monospecific antibodies against HSV-1 antigen No. 8, specific immunological reaction products were identified both in the cytoplasm and nucleus after 2 hr. The distribution and quantity of reaction products varied in the infected cells during the viral morphogenesis. The present results on the subcellular distribution of the HSV-1 antigens are related to current biochemical findings.
Animals↗
Analysis of herpes simplex virus type 1 antigens exposed on the surface of infected tissue culture cells.
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Antigens, Neoplasm↗
Production of rabbit antibodies against the viral capsid antigen (VCA) of the Epstein-Barr virus (EBV).
Rabbits were immunized with nuclear or cytoplasmic extracts of the Epstein-Barr virus (EBV)-producing marmoset cell line B95-8. Following extensive absorption with human EBV-negative cells (HeP-2, Ramos and BJAB), sera were obtained that no longer reacted with cellular or serum proteins of human origin, but gave a single precipitin band with extracts of the human EBV-producing line, P3HR-1. Immunofluorescence tests performed with appropriate parallel human serum controls showed that the rabbit serum contained no activity against EBNA + EA - VCA - Raji cells, or against P3HR-1 virus superinfected, cytosine arabinoside-treated Raji cells that contained EBNA and EA, but not VCA. The sera gave a brilliant indirect immunofluorescence reaction with the virus-producing (EBNA+ EA+ VCA+) P3HR-1 lines. Two-color fluorescence tests, performed with a direct TRITC-labelled VCA conjugate and indirect FITC-staining with the rabbit serum, showed that the same cells reacted in both red and green fluorescence, confirming that the rabbit serum was directed specifically against some antigen formed in the virus-producer cells. Since the synthesis of the relevant antigen was prevented by cytosine arabinoside it cannot be EA and must be a late antigen. The morphology and localization of the antigen support the conclusion that the antigen is VCA or some part of the VCA complex.
Animals↗
Crossed immunoelectrophoresis of a herpes simplex virus type 1-specific antigen: immunological and biochemical characterization.
Crossed immunoelectrophoresis of herpes simplex virus (HSV) antigens obtained from infected cells solubilized with the nonionic detergent Triton X-100 has shown that antigen number 6 (Ag-6) is found in preparations from HSV type 1-infected cells but not in HSV type 2 preparations. A monospecific antiserum was produced against Ag-6 by intracutaneous inoculation of the corresponding immunoprecipitate into rabbits. This antiserum neutralized HSV type 1 but not HSV type 2. The polypeptide composition of Ag-6 (molecular weight, 131,000 daltons) was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the immunoprecipitate, using radiolabeled antigen. The multitude of information obtained demonstrates that crossed immunoelectrophoresis can be used preparatively, since specific immunoprecipitates represent a short cut for the attainment of immunoreactive viral protein entities useful for further immunological and biochemical analyses.
Antigens, Viral↗