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N J Schmidt

Publications and source records attributed to N J Schmidt.

At least 19 recordsLinked to original sources
PubMed1990

Epitopes functional in neutralization of varicella-zoster virus.

By competition neutralization assay using monoclonal antibodies (MAbs) to varicella-zoster virus (VZV) glycoproteins (gps), we attempted to determine the topographical relationship of epitopes which are functional in VZV neutralization. MAbs against gpI interfered moderately to strongly with neutralization of MAbs against gpIII, and one antigenic domain with two distinct epitopes was identified on gpIII. Competition neutralization assays performed with MAbs to gpI revealed at least three distinct antigenic domains: the first contained two complement-dependent neutralizing epitopes; the second contained five complement-dependent neutralizing, overlapping epitopes and one nonneutralizing, nonoverlapping epitope; and the third contained one complement-enhanced neutralizing epitope. Competition neutralization assays performed with MAbs to gpIV showed one antigenic domain with two distinct epitopes which competed with nonneutralizing gpI MAbs. gpII did not interfere with neutralization of gpI, gpIII, or gpIV. Our data suggest that neutralizing and nonneutralizing MAbs can interfere with the action of viral neutralization either by inhibition or by enhancement. This report describes the epitope mapping of VZV gps by a functional biological assay.

Antibodies, Monoclonal↗
PubMed1986

Sensitivity of different assay systems for immunoglobulin M responses to varicella-zoster virus in reactivated infections (zoster).

An immunoglobulin M response to varicella-zoster virus was detected in 70% of zoster patients by solid-phase radioimmunoassay, in 52% by indirect immunofluorescence, in 48% by neutralization on sucrose density gradient fractions, and in 27% by an antibody class capture enzyme immunoassay. The patients showed marked variations in their varicella-zoster virus immunoglobulin M responses detectable in the different assays.

Antibodies, Viral↗
PubMed1985

Racial differences in incidence of juvenile-onset type 1 diabetes: epidemiologic studies in southern California.

We have studied the epidemiologic characteristics of insulin-dependent (Type 1) diabetic patients aged 0-19 in a city (San Diego, southern California, USA) characterized by an impressive racial diversity and especially mild and constant climatic conditions. Ascertainment was through retrospective review of medical records in 19 hospitals. For the 3 years 1978-1981 the mean annual incidence of diabetes was 7.3 cases/100,000, with no statistical difference between the sexes. The observed incidence rates in the various ethnic groups was significantly different from expected (p less than 0.03), with an excess of cases among Caucasians and fewer than expected cases among Mexicans, Blacks and Orientals. There was no identifiable seasonal trend. Some of the clinical characteristics at diagnosis differed between the sexes: males were slightly older (9.3 +/- 5.2 years versus 8.8 +/- 3.9 for females), had a shorter duration of diabetes-related symptoms and a higher frequency of infections both at the time of diabetes diagnosis and in preceding months. Females tended to have a higher frequency of Type 1 diabetes in first-degree relatives. This study documents for the first time that, among multiple racial groups living in the same environment, Caucasians are at the highest risk of developing juvenile-onset Type 1 diabetes.

Asian People↗
PubMed1985

Diabetogenic potential of coxsackie B viruses in nature.

Thirty-seven clinical isolates of coxsackievirus (CV) serotypes B-1, B-3, B-4, and B-5 were inoculated into male SJL mice. Twelve strains resulted in minor abnormalities of glucose metabolism in one or more of six infected mice (Tables 1 and 2). Sequential infection of male SJL mice with CVB-3, CVB-4, and CVB-5 resulted in abnormal glucose metabolism in 25 percent of the mice (Fig. 1). The glucose index of the abnormal animals was similar to that produced by sequential infection with reovirus and cytomegalovirus but less than that seen with more severe beta cell tropic agents such as streptozotocin or encephalomyocarditis virus. Infection of autoimmune New Zealand (NZB X NZW) F1 male mice with CBV-3, CVB-4, and CVB-5 resulted in transient elevation of the blood glucose concentration associated with acute acinar pancreatitis (Fig. 2). In spite of recent evidence that infection with the coxsackie B viruses can result in human diabetes mellitus, the diabetogenic potential of CVB field strains appears to be limited. Diabetes mellitus may occur as a rare event, limited to genetically susceptible hosts. Autoimmune mechanisms or repeated infection with other CVB serotypes may convert minimal beta-cell destruction into clinically overt disease.

Animals↗
PubMed1985

Enzyme immunoassay versus plaque neutralization and other methods for determination of immune status to measles and varicella-zoster viruses and versus complement fixation for serodiagnosis of infections with those viruses.

Results by an enzyme immunoassay method (EIA) performed at one serum dilution and results by indirect immunofluorescence (IFA) and hemagglutination inhibition (HI) tests performed at step dilutions were correlated with results by a neutralization test (50% plaque neutralization [PN]) performed at step dilutions on single serum samples for serologic evaluation of immunity status to measles virus. PN results were taken as true indicators of immunity, and the other tests were evaluated on that basis. The predictive value of a positive result being positive also by PN was 95.3% for HI and 93.3% for EIA and IFA. The predictive value of a negative result being negative also by PN was 81.1% for HI, 100% for EIA, and 75.0% for IFA. A similar study on immunity status to varicella-zoster virus by EIA and by an anticomplement immunofluorescence test versus PN showed a 100% predictive value of a positive or negative result by EIA. By the anticomplement immunofluorescence test, the predictive value of a positive result was 97.7%, and that of a negative result was 88.5%. Studies on the comparative ability of EIA versus complement fixation (CF) to detect significant changes in antibody concentration between acute-phase and convalescent-phase serum samples indicative of a current infection were also done. Both tests were satisfactory for the serodiagnosis of measles or varicella-zoster virus infections. However, EIA was preferable to CF because it was less technically difficult, less labor intensive, and could be performed on sera that were anticomplementary in CF reactions.

Antigens, Viral↗
PubMed1985

Rapid detection of herpes simplex virus DNA in human brain tissue by in situ hybridization.

A commercial system for detection of herpes simplex virus (HSV) DNA by in situ hybridization gave positive results on 16 of 17 stored human brain specimens that were positive for HSV on initial testing by virus isolation and immunofluorescence staining, and the hybridization system gave negative results on 13 brain specimens that showed no evidence of HSV by isolation or immunofluorescence staining.

Animals↗
PubMed1984

Class-specific antibody responses to early and late antigens of varicella and herpes simplex viruses.

Antibody responses to early antigens of varicella-zoster virus (VZV), simian varicella virus, and herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) were studied in primary, secondary, and latent infections. IgG antibody responses to the early antigens occurred in primary and secondary VZV and HSV infections, and antibodies to early antigens were also demonstrable in healthy individuals with latent VZV and HSV infections, indicating that the presence of antibodies to early antigens cannot be taken as evidence of active infection with the viruses. Patients with current VZV or HSV infections showed heterotypic IgG antibody responses to early antigens of VZV and HSV to the same extent as to late antigens. In all groups of patients, IgG antibody titers to early antigens were similar to those against the corresponding late antigens, and no difference was seen in the reactivity of early antigens produced with four different blocking agents (cytosine arabinoside, bromodeoxyuridine, trisodium phosphonoformate, and cycloheximide). Antibodies of the IgM and IgA classes reacted with both early and late antigens of HSV, but only with late antigens of VZV and simian varicella virus, suggesting that these antibodies may be directed against late proteins that are expressed to a greater extent in HSV-infected cells treated with blocking agents than they are expressed in treated VZV-infected cells. Homologous IgM antibody responses occurred in both primary and secondary VZV infections, but only in primary HSV infections. Heterotypic IgM responses to HSV-2 antigen were noted in a few VZV patients who did not have demonstrable IgG antibody to HSV, suggesting that even in patients without prior experience with HSV, a VZV infection may stimulate the production of IgM antibodies that react with antigens that are shared by VZV and HSV-2. IgA antibodies to late antigens of VZV and HSV were demonstrable in latent, as well as active, infections with these viruses.

Animals↗
PubMed1984

Persistent infection of mouse fibroblasts with Coxsackievirus.

Infection of fibroblast cell lines initiated from BALB/c or NFR mice with coxsackievirus B3 (CBV-3) or B4 (CBV-4) resulted in infections which persisted for a limited number of subpassages of the infected cells in most cases, but for over a year in one case. In all instances primary acute infections were characterized by cytopathology and release of infectious virus progeny. Viral antigen could be detected during the acute phase of infection, but not in subcultured infected cells. Infectious center assays showed that every cell was infected during the acute phase of infection, but that from the first subcultivation on, the numbers of cells which were able to initiate infection were greatly reduced. The long term persistent CBV-3 infection was characterized by wide fluctuations in titers of virus released into the supernatant fluids. Interferon did not appear to play a role in maintenance of the persistent infection. Information derived from studies on mechanisms of CBV persistence in the in vitro model may help to elucidate the role of CBV in chronic human diseases such as myocarditis.

Animals↗
PubMed1984

Differing cardiotropic and myocarditic properties of group B type 4 coxsackievirus strains.

A murine model system for evaluation of myocarditic and cardiotropic properties of strains of group B, type 4 coxsackievirus (CBV-4) was developed in male BALB/c mice 4 weeks of age. Differing cardiotropic and myocarditic properties could be identified among field strains within the CBV-4 serotype. These properties were consistent for the virus strains, and were independent of the infecting virus dose. Virus replication in the heart appeared to be essential for development of myocarditis, but some infected hearts with high levels of infectious virus did not show myocarditis. Two of the myocarditic strains showed different histopathology in infected hearts; with one strain (Mil) the myocarditis was characterized by a marked inflammatory reaction with occasional accompanying myofiber necrosis. With the other strain (Bol), necrosis was the predominant finding, with a much lesser degree of inflammation. These findings suggest that there may be various pathogenic or immunopathogenic mechanisms by which CBV-4 can produce myocarditis.

Age Factors↗
PubMed1984

Coxsackievirus B3 persistence and myocarditis in NFR nu/nu and +/nu mice.

NFR nude (nu/nu) and euthymic (+/nu) littermates were infected with coxsackievirus B3 (CBV-3) and assayed for virus persistence in the heart, pancreas and spleen, for development of myocarditis and for antibody production. The virus grew to higher titer and persisted longer in hearts of nu/nu mice. In both types of mice there was comparable myocarditis with a mononuclear cell inflammatory response, and there was some evidence of chronic lesions for up to 21 days in +/nu and 28 days in nu/nu mice. Antibody of the IgM class was present at 7 days in both strains of mice. Thereafter, +/nu mice produced high titers of virus-specific IgG antibody, while nu/nu mice produced little or no viral IgG antibody. The persistence of virus for up to 28 days in NFR nude mice is longer than has been reported previously, and offers an opportunity for further study of the role of T-cells in virus persistence and myocarditis.

Animals↗
PubMed1984

Monoclonal antibodies for study of antigenic variation in coxsackievirus type B4: association of antigenic determinants with myocarditic properties of the virus.

Monoclonal antibodies were produced to a field strain ( Mil ) of group B coxsackievirus type 4 (CBV-4), and to the prototype JVB strain. Nine were neutralizing antibodies and four were non-neutralizing antibodies with virus-specific activity in indirect immunofluorescence (IF) staining. On the basis of reactivity with the panel of monoclonal antibodies, nine different strains of CBV-4 were found to fall into five distinct antigenic groups. Antigenic variants were produced by using the neutralizing monoclonal antibodies to select variants from the Mil virus stock which were no longer susceptible to the selecting antibody. A high frequency of antigenic variation was seen. By using the variants in cross-neutralization and IF tests with the monoclonal antibodies, it was possible to identify five tentative antigenic sites functional in neutralization; one site appeared to be complex and possibly to consist of overlapping epitopes. Reactivity of the monoclonal antibodies was similar, but not necessarily identical, by neutralization and by IF staining. The antigenic variants were found to differ from the parent Mil strain, and from one another, in their myocarditic and cardiotropic properties in a murine model. Two of the variants produced more extensive cardiac pathology, and two produced higher virus titres in the heart than was produced by the parent strain. One variant was notable for extensive production of necrotic lesions in the myocardium. Four of the variants showed less histopathology and three produced less virus in the heart than was produced by the parent strain.

Animals↗
PubMed1984

Coxsackievirus B3 persistence and myocarditis in N:NIH(S) II nu/nu and +/nu mice.

N:NIH(S) II nu/nu (athymic) and +/nu (euthymic) mice were inoculated with coxsackievirus B3 (CBV-3) and examined at various times after infection for virus titres in the heart, myocarditis and serum neutralizing antibodies. Virus was recovered from the hearts of nu/nu mice for up to 94 days post-inoculation, but was not recovered from the hearts of any +/nu mice beyond 14 days. Inflammatory cell infiltration and necrosis were present in the hearts of +/nu mice at all harvest times (7, 14, 21 and 28 days). Inflammation and necrosis did not become evident in nu/nu mice until 14 days post-inoculation, and was the present in mice from each harvest until the end of the experiment (94 days). In athymic mice, myocarditis showed a strong correlation with persistence of CBV-3 in the heart. In N:NIH(S) II mice, the presence (+/nu) or absence (nu/nu) of a thymus had a major influence on the clearance of virus from the heart and on the development of myocarditis.

Animals↗
PubMed1984

Antibody class capture assays for varicella-zoster virus.

Pooled monoclonal antibodies to varicella-zoster virus (VZV) were used as "detector" antibodies in a four-phase enzyme immunofluorescence assay for determination of immunoglobulin M (IgM), IgA, and IgG antibodies to VZV. Polyclonal antisera specific for heavy chains of human IgM, IgA, and IgG were employed as "capture" antibodies on the solid phase. The antibody class capture assay (ACCA) for VZV IgM antibody detected high titers of virus-specific IgM in all patients with varicella and in 5 of 10 zoster patients. VZV IgM antibody was not detected in patients with primary herpes simplex virus infections or in other individuals without active VZV infection, with one exception, a patient with encephalitis who had other serological findings compatible with a reactivated VZV infection. VZV-specific IgA and IgG antibody titers demonstrable by ACCA were compared with those measured by solid-phase indirect enzyme immunofluorescence assay (EIFA). VZV IgA antibody titers detected in patients with varicella and zoster were variable and could not be considered to be reliable markers of active VZV infection. IgA antibody titers detected by ACCA tended to be higher than those demonstrated by solid-phase indirect EIFA in varicella and zoster patients. VZV IgG antibody titers detected by ACCA in patients with varicella, and to a lesser extent in zoster patients, were as high as or higher than those demonstrated by solid-phase indirect EIFA. However, ACCA was totally insensitive in detecting VZV IgG antibody in individuals with past infections with VZV and would not be a suitable approach for determination of immunity status to VZV.

Antibodies, Monoclonal↗
PubMed1984

Varicella-zoster viral glycoproteins analyzed with monoclonal antibodies.

Monoclonal antibodies to varicella-zoster virus were used to study viral glycoproteins by immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Based on the viral glycoproteins immunoprecipitated, the five monoclonal antibodies fell into three groups. Two antibodies, 4B7 and 8G9 (group 1), immunoprecipitated a single glycoprotein of molecular weight (MW) 118,000 (118K glycoprotein) and had high neutralizing activity in the absence of complement. One antibody, 3C7 (group 2), which lacked neutralizing activity, immunoprecipitated two glycoproteins of MWs 120,000 and 118,000 and a glycoprotein giving a diffuse band in the region of 64,000 to 65,000. Pulse-chase experiments and experiments with monensin as an inhibitor of glycosylation suggested that the 120K polypeptide was derived by glycosylation of the 118K polypeptide and that a 43K antigen was processed into the 64 to 65K glycoprotein. Two antibodies, 3G8 and 4E6 (group 3), both had neutralizing activity only in the presence of complement, and both immunoprecipitated at least five polypeptides, with MWs ranging from 50,000 to 90,000. Antibody 3G8 was isotype immunoglobulin G2b (IgG2b), and its immunoprecipitating activity was stronger than that of 4E6, which was isotype IgG1. Pulse-chase experiments with antibody 3G8 showed that lower-MW glycopeptides chased into three polypeptides of MWs 90,000, 80,000, and 60,000 by 24 h. Immunoprecipitation experiments with antibody 3G8 on infected cells treated with glycosylation inhibitors 2-deoxyglucose, monensin, and tunicamycin, suggested that a prominent, early-appearing 70K polypeptide may have been processed into the glycoproteins of higher MWs and that the 60K polypeptide may have been derived by glycosylation of polypeptides of lower MWs.

Antibodies, Monoclonal↗
PubMed1983

Association of herpes simplex virus with platelets of experimentally infected mice.

In mice infected with herpes simplex virus (HSV) type 1 by the intracerebral route or with HSV type 2 by the vaginal route, virus was shown to be present in greater quantities in purified platelets than in leucocytes or other blood components. Results of the studies described suggest that platelets may play a major role in the hematogenous dissemination of HSV infection.

Animals↗
PubMed1983

Rapid viral diagnosis.

There has been much interest and activity in the development of techniques for rapid viral diagnosis which would allow successful intervention in the treatment of patients or their contacts or in the control of viral diseases in the community. The greatest emphasis has been on techniques that permit viral detection directly in the clinical specimen, since these avoid the need to cultivate the agent, are feasible for detection of viruses that cannot be cultivated, and can detect virus in specimens in which the agent is no longer infectious. Direct methods used for viral detection include electron microscopy and various immunoassays which are based on demonstrating reactivity of viral antigen in the specimen with known viral antisera. The use of immunoassays for more rapid identification of viruses isolated in laboratory host systems and for selective detection of viral antigen in inoculated cell cultures even before the agents produce an observable effect has been an important advance in viral diagnosis by the approach of isolation and identification. The reliability of all specific viral identification procedures depends on the use of high quality viral antisera. Some of the problems previously encountered in preparing satisfactory viral immune reagents are being overcome through the availability of highly specific monoclonal antibodies produced by cell hybridization techniques. Virus-specific IgM antibody assays for rapid diagnosis have been improved greatly through the use of a "capture" technique in which antibody to the human mu chain is used in the solid phase to separate IgM from other serum components which might compete with IgM antibody or give nonspecific reactivity, and also through the availability of highly specific monoclonal antibodies to the human mu chain. A variety of simple assays for determination of viral antibody status have been developed, and many are commercially available. The reliability of some of these antibody assays has been improved through the incorporation of more suitable controls and through better definition of interpretations which should be made from test results.

Antibodies, Monoclonal↗