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Biomedical subjects

B Norrild

Publications and source records attributed to B Norrild.

At least 55 records · Page 3Linked to original sources
PubMed1986

Analyses of transplanted murine tumors for HSV DNA sequences.

Meignier et al. (1986) report the results of exposure of C57BL/6NCr mice to vaginal plugs containing live or inactivated herpes simplex virus 1 or 2 (HSV-1 or HSV-2) or recombinant viruses 5 times a week for up to 114 weeks. Genital organs showing abnormalities were transplanted into nude mice. Of 33 transplants, 13 produced subcutaneous tumors in nude mice and 12 were subsequently transplanted into C57BL/6NCr mice. We report that the DNA extracted from coded tumor tissues of nude mice and from normal viscera of the same rodents did not hybridize with HSV-1 and HSV-2 DNA probes representing the viral genomic regions shown previously to be capable of morphologically transforming cells in culture. The sensitivity of the assays was such that we could detect 0.5 copies of the HSV sequences of complexity equal to or greater than 1 Kbp per cell DNA equivalent. To control for the sensitivity of the assays in the actual hybridizations, the tumor-cell DNA was also hybridized with a beta-globin mouse DNA probe. A striking feature of these control hybridizations was the detection of beta-globin polymorphism in some nude mouse tumors. The beta-globin polymorphism allowed us to conclude that the analyzed tissues contained significant amounts of the tumor cells occurring in the C57BL/6NCr mice.

Animals↗
PubMed1986

Immunological reactivity of human sera with individual herpes simplex proteins: a comparative study of sera from patients with preinvasive or invasive cervical cancer and from controls.

Forty-three human sera collected from patients with preinvasive or invasive cervical carcinoma were analyzed for their repertoire of herpes simplex virus (HSV) specific antibodies reactive with individual viral HSV-1 and HSV-2 proteins. The reactivity was compared to that of sera from 27 control persons. The patients and controls were clinically and histologically characterized in a previous study we carried out, where the analysis of the HLA-antigen pattern was compared among the groups [Vass-Sørensen, 1984]. Immunoprecipitation analysis showed that only a subset of the infected cell proteins was precipitable by the human sera. The major proteins identified in the polyacrylamide gels were the glycoproteins B and D, the ICP-5 and ICP-8. There was no difference between the results obtained with patients and control sera. Immunoblot analysis showed that a different subset of HSV proteins reacted with the human sera, but the variability among individuals was significant. Rank data showed that sera from both patients and controls reacted most frequently with proteins belonging to the "35-family" [Braun et al, 1984] and with the glycoproteins B and D of HSV-1 and HSV-2.

Antibodies, Viral↗
PubMed1986

Organization of cytoskeleton elements during herpes simplex virus type 1 infection of human fibroblasts: an immunofluorescence study.

Cultured human fibroblasts showed a typical fibrillar organization of microtubules in immunofluorescence, including the vimentin type of intermediate filament as well as actin-containing microfilaments. During infection with herpes simplex virus type 1 (HSV-1), the vimentin organization was maintained whereas actin, myosin and tubulin showed a progressive association with the viral glycoproteins within juxtanuclear structures. These structures could also be revealed with fluorochrome-coupled wheat germ agglutinin. Disruption of the microtubules by demecolcine treatment or their stabilization by taxol treatment did not prevent the aggregation of viral proteins in the cytoplasm. Taxol stabilization of the microtubules allowed the juxtanuclear accumulation of the glycoproteins in HSV-infected cells whereas treatment with demecolcine led to an accumulation of the glycoproteins either in small vesicles in the cytoplasm or in the focal adhesion areas of the cells. Production of infectious intracellular virus particles was reduced in cells treated with demecolcine or with taxol before and during infection. The results of this study indicate that the normal intracellular transport and distribution of the HSV glycoproteins and the formation of infectious virus are dependent on the presence of intact microtubules.

Alkaloids↗
PubMed1985

Crossed immunoelectrophoretic analysis of herpes simplex virus type 2 proteins. Characterization of antigen-5.

Herpes simplex virus type 2 proteins extracted from infected cells and analysed by crossed immunoelectrophoresis identified a nonglycosylated antigen named Ag-5. The antigen contained two proteins when extracted from the agarose gel and the molecular weights were 128K and 91K. Both proteins are located in the nucleus of the infected cells and the 128K is identical to ICP-8. The 91K protein is based on the reactivity with monoclonal antibodies most likely the alkaline exonuclease mapped by Preston and Cordingly (25). Our data show that although the proteins ICP-8 and 91K coprecipitate they differ in both peptide composition and in immunological specificity.

Animals↗
PubMed1985

Changes of blood group antigens in premalignant and malignant lesions of the human exocervix.

In this preliminary study the distribution of blood group A- and H-antigens and their structural precursor N-acetyllactosamine in normal, pre-malignant and malignant squamous epithelium were investigated using the immunofluorescence technique. In normal cervical epithelium the basal cells and few parabasal cells expressed N-acetyllactosamine. The parabasal and lower spinous cells expressed the H-antigen and the upper spinous layer expressed the A-antigen. All pre-malignant and malignant lesions showed irregular decrease in contents of A- and H-antigens. N-acetyllactosamine was either completely lost or found to accumulate in these lesions, and it is suggested that this antigen could be a valuable tool in the diagnosis of cervical cancer.

ABO Blood-Group System↗
PubMed1985

Keratin proteins in the epithelium of the uterine cervix.

Keratin proteins in human cervical squamous epithelium were investigated by the immunofluorescence technique, using the monoclonal keratin antibodies AE1 and AE2. Although the exocervic is covered by a non-keratinizing, histologically homogenous, squamous epithelium, different staining patterns were found within a single histological section. This indicates a regional variation in the maturation pattern of stratified exocervical epithelium. Detection of morphological versus biochemical keratinization in the diagnosis of pathological cervical conditions is discussed.

Antibodies, Monoclonal↗
PubMed1984

Identification and preliminary mapping with monoclonal antibodies of a herpes simplex virus 2 glycoprotein lacking a known type 1 counterpart.

The properties of herpes simplex virus 2 (HSV-2)-specific proteins reactive with monoclonal antibody H966 derived from mice immunized with HSV-2 strain G are reported. The reactive proteins contained in infected cell lysates subjected to electrophoresis in denaturing gels and transferred to nitrocellulose sheets form a relatively sharp band characteristic of Mr 124,000 proteins and a diffuse, more slowly migrating band. Antigens reactive with H966 were detected on the surface of viable, unfixed cells. The electrophoretic mobility of the H966-reactive proteins made in the presence of tunicamycin was more rapid than that of the proteins made in the absence of the drug. Direct evidence that the HSV-2-specific antigen was a glycoprotein emerged from purification of [14C]glucosamine-labeled proteins with similar electrophoretic mobilities by immunoabsorption to H966 bound to Sepharose beads. Analyses of the reactivity of HSV-1 X HSV-2 recombinants indicated the gene specifying the glycoprotein maps in the S component of the DNA. The glycoprotein detected by H966 has no known counterpart in HSV-1 and corresponds to the glycoprotein previously designated as gC of HSV-2 and reported to map to the right of gC specified by HSV-1. Inasmuch as an HSV-2 gene colinear with HSV-1 gC has been reported to specify a glycoprotein currently designated as gC of HSV-2 by Para et al. [J. Virol. 45, 1223-1227 (1983)], the glycoprotein identified by H966 should be designated as gG.

Animals↗
PubMed1984

Prevalence of antibodies to herpes simplex virus and frequency of HLA antigens in patients with preinvasive and invasive cervical cancer.

Previous herpes simplex virus type 2 (HSV-2) infections are known to predispose women for the development of cervical cancer, but causal relationship between the virus and the cancer has never been proven. Forty-six patients with cervical carcinoma (13 with preinvasive lesions and 33 with various stages of invasive disease) were selected for the present study. Among the patients 96% were seropositive for the presence of antibodies to herpes simplex virus compared to 87% of the 30 controls. Antibodies specific for HSV-2 were found in the sera from 24% of the patients and 17% of the controls by the use of an immunoblotting test. Patients and controls were typed for HLA-A,B,C and D/DR antigens, but no significant associations were found.

Adult↗
PubMed1984

Biosynthesis of the D2 cell adhesion molecule: pulse-chase studies in cultured fetal rat neuronal cells.

D2 is a membrane glycoprotein that is believed to function as a cell adhesion molecule (CAM) in neural cells. We have examined its biosynthesis in cultured fetal rat brain neurones. We found D2-CAM to be synthesized initially as two polypeptides: Mr 186,000 (A) and Mr 136,000 (B). With increasing chase times the Mr of both molecules increased to 187,000-201,000 (A) and 137,000-158,000 (B). These were similar to the sizes of D2-CAM labeled with [14C]glucosamine, [3H]fucose and [14C]mannosamine, indicating that the higher Mr species are glycoproteins. In the presence of tunicamycin, which specifically blocks the synthesis of high mannose cores, Mr were reduced to 175,000 (A) and 124,000 (B). Newly synthesized A and B are susceptible to degradation by endo-beta-N-acetyl-glucosaminidase H, which specifically degrades high mannose cores, but they are resistant to such degradation after 150 min of posttranslational processing. Hence, we deduce that A and B are initially synthesized with four to five high mannose cores which are later converted into N-linked complex oligosaccharides attached to asparagine residues. However, no shift of [35S]methionine radioactivity between A and B was detected with different pulse or chase times, showing that these molecules are not interconverted. Thus, our data indicate that the neuronal D2-CAM glycoproteins are derived from two mRNAs.

Animals↗
PubMed1983

Requirements for transport of HSV-1 glycoproteins to the cell surface membrane of human fibroblasts and Vero cells.

The intracellular transport of the HSV-1 glycoproteins gA/gB, gC and gD has been followed by the indirect immunofluorescence technique (IIF). Infected tissue culture cells were stained with monoclonal antibodies made to the individual glycoproteins and with fluorochrome-coupled wheat germ agglutinin reacting specifically with Golgi apparatus of the cells. Staining of either infected, human fibroblasts or of VERO cells at 9 hours p.i. with antibodies to gA/gB showed a prominent ring-like nuclear fluorescence and distinct staining of the Golgi apparatus in the cells. Antibodies to gC and gD stained mainly the Golgi apparatus and areas close to or at the surface of the cells. By immunocytolysis of HSV-1-infected VERO cells the viral glycoproteins were demonstrable at the surface of cells but growth of infected cells in the presence of either TM or monensin inhibited the expression of most of the viral glycoproteins at the cell surface. Blocking of the glycosylation of the viral glycoproteins with tunicamycin (TM) was followed by accumulation of the core of the glycoproteins gA/gB and gD in granular structures close to the nucleus as seen by immunofluorescence microscopy. Antibodies to gC did also stain granules close to the nucleus but in addition the periphery of the cells were stained. Inhibition of intracellular transport from the Golgi apparatus by the carboxylic ionophore monensin was followed by accumulation of all the HSV-1 glycoproteins in vesicles derived from the Golgi apparatus in both human fibroblasts and VERO cells. Our data thus support the hypothesis that the HSV-1 glycoproteins are processed in the Golgi apparatus before the transport to and incorporation into the cell surface membrane of infected cells and into virion envelopes.

Animals↗
PubMed1983

Populations of herpes simplex virus glycoprotein gC with and without affinity for the N-acetyl-galactosamine specific lectin of Helix pomatia.

Two fractions of herpes simplex virus glycoprotein gC were isolated and characterized by means of immunosorbent-purification with monoclonal antibodies against gC and Helix pomatia lectin (HPA) affinity chromatography. About 25 per cent of the glycoprotein gC population demonstrated affinity for the lectin, compatible with presence of N-acetylgalactosamine as terminal sugar of the oligosaccharide. The HPA-binding populations of gC appeared as two electrophoretic bands with lower molecular weights than the non-binding gC. The gC subfraction without affinity for the HPA was subjected to treatments aiming to desialylize the carbohydrate moiety. Only 5 per cent of the initially non-reactive fraction of gC became reactive to HPA after the treatments, suggesting that masking of penultimate N-acetylgalactosamine by sialic acid was not a main reason for lack of HPA affinity. Results of treatment with alkaline Na BH4 demonstrated presence of oligosaccharide-peptide linkages sensitive to beta-elimination suggesting O-glycosidic type of linkage. The subfraction of gC demonstrating affinity for HPA as well as gC devoid of HPA binding capacity both revealed affinity for Con A. Therefore N-glycosidically as well as O-glycosidically linked oligosaccharides seemed to be present on the one and same glycoprotein. On the basis of the results presented we assume that the glycosylation of HSV glycoprotein gC may lead to, at least, two populations of the glycoprotein gC, one with terminal N-acetylgalactosamine residues of oligosaccharides O-glycosidically linked to the polypeptide and the other without affinity for HPA. However, both populations of gC contain similar proportions of oligosaccharides of the high mannose or complex types with N-glycosidic carbohydrate-peptide linkages as indicated by their affinity for Con A.

Acetylgalactosamine↗
PubMed1983

Accumulation of herpes simplex virus type 1 glycoprotein D in adhesion areas of infected cells.

By indirect immunofluorescence microscopy (IIF) we have localized glycoprotein D (gD) of herpes simplex virus-infected cells to the vinculin-containing junctional areas and to the focal adhesion sites on the ventral surface of the cells. Double-staining IIF showed that gD and vinculin co-distributed in the infected cells and treatment of the infected cells 4 h post-infection with rabbit antibodies to gD prevented attachment of the cells to the growth substratum. Our results therefore support the hypothesis that gD plays a central role in the social behaviour of infected cells.

Biological Transport↗
PubMed1983

Quantitation and estimation of cooperation between target-binding sites in natural and antibody-dependent cell-mediated cytotoxicity by use of a mathematical transformation. Influence of interferon.

We have studied the use of a mathematical transformation, the Hill transformation, in natural and antibody-dependent cell-mediated cytotoxicity as a function of effector cell concentration. It can be concluded that when adherent cells are removed from the effector cell suspension, cooperation between binding sites for effector cells on the target cell changes from negative to zero. After stimulation of nonadherent cells with alpha-interferon, the binding sites still remain noninteracting. This is the case both in natural and in antibody-dependent cell-mediated cytotoxicity experiments. The Hill transformation seems to be an easy and easily reproducible way of quantitating cytotoxicity.

Antibody-Dependent Cell Cytotoxicity↗
PubMed1983

Colonization of murine ganglia by a superinfecting strain of herpes simplex virus.

We report on the colonization of murine trigeminal ganglia after sequential infection of mice by herpes simplex viruses (HSVs). In preliminary studies, we have established that whereas the HSV-1(F) strain efficiently colonizes ganglia when inoculated by either the ear or eye routes, the HSV-1 X HSV-2 recombinant C7D colonizes ganglia when inoculated by the eye route only. The experimental design consisted of inoculating the right eye with C7D on day 1 and with HSV-1(F) in both left and right eyes on day 26. Both right and left trigeminal ganglia were removed and analyzed independently for latent virus on day 52. Our studies indicate that HSV-1(F) viruses were recovered from all left trigeminal ganglia but from only a small number of right trigeminal ganglia. Some right trigeminal ganglia yielded no viruses, whereas others yielded both C7D and HSV-1(F) viruses identified on the basis of plaque morphology and restriction enzyme cleavage patterns of viral DNA. The results indicate that more than one virus may colonize the same ganglion and that trigeminal ganglia may be protected from colonization by a superinfecting virus by determinants acting at a local level in the absence of demonstrable virus.

Animals↗
PubMed1983

Application of denatured, electrophoretically separated, and immobilized lysates of herpes simplex virus-infected cells for detection of monoclonal antibodies and for studies of the properties of viral proteins.

We report the use of herpes simplex virus type 1 (HSV-1)- and HSV-2-infected cell polypeptides (ICPs) separated by electrophoresis in polyacrylamide gels and transferred to nitrocellulose to (i) detect monoclonal antibodies to viral polypeptides and to (ii) study the properties of the proteins with the monoclonal antibodies. Our results were as follows. (i) When the antigens were electrophoretically separated in denaturing gels and then immobilized on nitrocellulose strips, we detected a greater diversity of monoclonal antibodies to viral proteins than when we used the technique of immune precipitation of soluble, nondenatured viral antigens. The primary advantage of the technique is in the detection of nonprecipitating antibody and of antibody to poorly soluble antigens not available for reaction in preparations cleared by high-speed centrifugation before immune reaction. (ii) Studies of the viral polypeptides reactive with three monoclonal antibodies indicated that the technique can be used to investigate several properties of the antigens. Specifically, monoclonal antibody to ICP 4 confirmed the accumulation of viral protein in the nucleus and the mapping of the gene in the S component. The results showed, however, that HSV-1 and HSV-2 ICP 4 do have common antigenic determinants. The reaction of a nonprecipitating monoclonal antibody with electrophoretically separated, immobilized polypeptides contained in cytoplasmic and nuclear fractions, those chemically deglycosylated, or those specified by specific HSV-1 x HSV-2 intertypic recombinants identified the antigens reactive with the second monoclonal antibody as various forms of glycoprotein gC. Of particular interest was a set of four antigens, 39,000 to 46,500 in apparent molecular weight, reactive with each of several monoclonal antibodies. These studies showed that two polypeptides partition in the cytoplasm and two in the nucleus and that all comap with the previously mapped ICPs 35 and 37 in the region of the genome defined by the viral thymidine kinase gene on the left and the glycoprotein gA/B gene on the right. Unlike ICP 4 and gC, the four polypeptides are linked by intermolecular bisulfide bonds, inasmuch as the polypeptides were not at the expected locations upon denaturation and electrophoresis in the absence of reducing agents.

Animals↗
PubMed1982

Effect of tunicamycin on the synthesis of herpes simplex virus type 1 glycoproteins and their expression on the cell surface.

Herpes simplex virus specifies five glycoproteins which have been found on the surface of both the intact, infected cells and the virion envelope. In the presence of the drug tunicamycin, glycosylation of the herpes simplex virus type 1 glycoproteins is inhibited. We present in this report evidence that the immunologically specificity of the glycoproteins designated gA, gB, and gD resides mainly in the underglycosylated "core" proteins, as demonstrated by the immunoblotting technique. We showed also that tunicamycin prevented exposure of the viral glycoproteins on the cell surface, as the individual glycoproteins lost their ability to participate as targets for the specific antibodies applied in the antibody-dependent, cell-mediated cytotoxicity test. Immunocytolysis was reduced between 73 and 97%, depending on the specificity of the antibodies used. The intracellular processing of the herpes simplex virus type 1-specific glycoprotein designated gC differed from the processing of gA, gB, and GD, as evidenced by the identification of an underglycosylated but immunochemically modified form of gC on the surface of infected cells grown in the presence of tunicamycin.

Animals↗