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Biomedical subjects

J J Docherty

Publications and source records attributed to J J Docherty.

At least 37 records · Page 2Linked to original sources
PubMed1982

Inapparent genital herpes simplex virus infection in college women.

During a six-month period, 600 gynecological samples were collected from 585 women with typical herpes lesions, women with non-herpes symptoms (ie, vaginitis, moniliasis, trichomoniasis, etc), and normal women seen at the student health center gynecological clinic and processed for herpes simplex virus (HSV) isolation. From these specimens, 29 samples from 25 of the 585 women (4.3%) were positive for HSV. When these isolates were typed using plaque diameter in chick cells, heat stability of viral thymidine kinase (T.K.), and restriction endonuclease patterns it was found that 18 samples (15 patients or 60%) were HSV-2 and 11 samples (10 patients or 40%) were HSV-1. Inapparent HSV infections constituted 20.0% of the virologically confirmed samples (5 of 25 patients) and represented 0.9% of the total patients studied (5 of 585). The inapparent infections were about equally divided between the two HSV types (2 were HSV-2 and 3 were HSV-1), and 4 of 5 occurred in the presence of clinically diagnosed monilia.

Adolescent↗
PubMed1981

Antibody responses in humans to individual proteins of herpes simplex viruses.

Sera from 231 women were used to examine their frequency of precipitation of various herpes simplex virus type 1 and 2 (HSV-1 and HSV-2) proteins and to determine if there was a rank order of immune responsiveness of humans to these HSV antigens. Radiolabeled viral proteins were reacted with serum and immune complexes isolated with staphylococcal protein A. Individual antigens were resolved by polyacrylamide gel electrophoresis and visualized by fluorography. As a group, these sera precipitated 31 HSV-1 and 27 HSV-2 proteins. HSV-1 polypeptides with molecular weights of 133,000, 99,000, and 82,000, as well as HSV-2 polypeptides with molecular weights of 131,000 and 101,000, were precipitated by essentially all sera that contained antibodies to HSV-1 and HSV-2. When attempts were made to order the viral proteins by constructing precipitation profiles ranking the antigens in patterns according to their frequency of precipitation, it was observed that the antigens were generally not ordered. Demographic analysis of the sera suggested that the differences in the number of proteins precipitated were associated with differences in age, education, age at first marriage, and income, which collectively may reflect the frequency of exposure to the virus.

Adult↗
PubMed1981

Identification of a virus-specific polypeptide associated with a transforming fragment (BglII-N) of herpes simplex virus type 2 DNA.

The BglII N fragment of herpes simplex virus type 2 (HSV-2) DNA (approximately 0.58 to 0.63 map unit) was examined for encoded products. Using plasmid pGZ59, which consists of BglII-N cloned in pAT153, in conjunction with hybrid arrested translation, mRNA selection, and in vitro protein synthesis, we found that the major translated product of this region has an approximate molecular weight of 37,800. By further mapping, coding sequences for this polypeptide were located within the region of BglII-N representing approximately 0.58 to 0.61 genome map unit. To demonstrate immunological specificity, we used staphylococcal A protein immunoprecipitation with rabbit anti-HSV-1 or HSV-2 sera and antigens from HSV-1 or HSV-2 total mRNA translated in vitro and BglII-N-selected mRNA. The results show that the 37,800-dalton polypeptide has HSV-2 immunological specificity, as it is precipitated with anti-HSV-2 sera but not with anti-HSV-1 or control sera.

Antigens, Viral↗
PubMed1980

Reaction patterns of herpes simplex virus type 1 and type 2 proteins with sera of patients with uterine cervical carcinoma and matched controls.

Serum from 105 individuals with diagnosed uterine cervical cancer and 231 matched controls were examined for their ability to react with a large number of herpes simplex virus type 1 or type 2 (HSV-1, HSV-2) proteins. Radiolabeled HSV-1 or HSV-2 proteins were mixed with test serum and immune complexes were isolated with staphylococcal protein A. Viral proteins in the immune complexes were resolved by polyacrylamide gel electrophoresis and visualized by fluorography. When the frequency of precipitation for cancer and control serum was calculated for each HSV-1 and HSV-2 protein, the results demonstrated that four HSV-1 and 11 HSV-2 proteins were precipitated more frequently by cases than by controls (p less than or equal to 0.05). However, since these results could be influenced by the presence or absence of HSV-2 specific antibodies as well as social, economic, and sexual history, the data were grouped and analyzed according to these parameters. This enabled all significant differences between case and control sera in the precipitation of HSV-1 or HSV-2 proteins to be abolished except for two HSV-2 proteins with molecular weights of 38,000 and 118,000. These two proteins appear to be tumor associated and not merely covariables of past infection or risk factors alone.

Adult↗
PubMed1979

Detection of herpes simplex virus infection of female genitalia by the peroxidase-antiperoxidase method alone or in conjunction with the Papanicolaou stain.

The sensitive peroxidase-antiperoxidase (PXAPX) method individually and in conjunction with the Papanicolaou (PAP) stain was used to detect herpes simplex virus (HSV) in specimens from human female genitalia. Initial studies using a model system of HSV-1 or HSV-2-infected Vero cells established (i) acetone as the most effective fixative, (ii) optimal dilutions of preimmunization and anti-HSV serum for differentiation of infected from noninfected cells, (iii) optimal concentration of 3,3'-diaminobenzidine tetrahydrochloride and H2O2 for maximal staining of infected cells with minimal background reaction, and (iv) removal of endogenous peroxidase with absolute MeOH. These various parameters, once established, were utilized in the PXAPX or PXAPX-PAP on human specimens from the vulva or cervix. In these specimens, examined by standard light microscopy, PXAPX-positive cells were dark brown with a single nucleus or multiple nuclei. By coupling the PAP to the PXAPX, detailed nuclear observations of PXAPX-positive cells were possible and revealed nuclear changes consistent with HSV infection, including syncytium formation, chromatin condensation, and an occasional Cowdry type A inclusion. The PXAPX and PXAPX-PAP correlated (r = 0.742) over a period of 72 h with HSV isolation from these samples.

Adolescent↗
PubMed1978

Steroid hormone alteration of herpes simplex virus type 1 replication.

The effect of steroid hormones on herpes simplex virus type 1 replication was examined. Virus replication studies revealed that various concentrations of prednisolone, hydrocortisone, dexamethasone, or progesterone could decrease virus yields up to a maximum of 99%. Using isopycnic centrifugation in CsCl to separate viral from cell DNA, it was found that virus-specific DNA synthesis was decreased by 30 to 100% depending on the hormone and concentration used. Cell-specific DNA synthesis was also adversely affected, but this did not alter cell viability or plating efficiency.

Cell Line↗
PubMed1978

Electrokinetic alteration of the surface of herpes simplex virus infected cells.

Changes induced in the ionic composition of the surface of herpes simplex virus (HSV) infected cells were evaluated by whole cell electrophoresis and isoelectric focusing. The kinetics of the progressive decline in cell electrophoretic mobility (a measurement of negative surface charge density) after infection corresponded to the production of infection virus. The observed HSV-induced increase in cell isoelectric focusing pH demonstrated that the electrokinetic cell surface became ionically more positive. The nature of the ionic changes were investigated using a series of specific reagents in conjunction with cell electrophoresis. Infected cells had 2.4 times surface binding sites for the amino-specific reagent 4-acetamide-4'-isothiocyanostilbene-2,2'-disulphonic acid. A relationship between these virus-induced amino groups and HSV surface antigens was indicated by the similar kinetics of their appearance and electrophoretic neutralization by HSV antibodies.

Antigens, Viral↗
PubMed1977

Incidence and distribution of herpes simplex virus types 1 and 2 from genital lesions in college women.

During a 9-month period, 9,772 women were treated at the student health center's gynecology clinic. Herpes simplex virus was isolated from 30 of 57 patients clinically diagnosed as suffering from a herpetic or herpetic-like genial infection for a virological incidence rate of 0.31%. Using virus plaque diameter in chick embryo cells and heat stability of viral thymidine kinase, 37% of the isolates were classified as herpes simplex virus type 1 and 63% were classified as herpes simplex virus type 2.

Adolescent↗
PubMed1977

Detection of antibodies specific for herpes simplex virus in human sera by the enzyme-linked immunosorbent assay.

An enzyme-linked immunosorbent assay (ELISA) was used for the detection in human sera of antibody to herpes simplex virus antigens. Development and standardization of the assay suggested that antigenic purity, temperature of reactions, concentration of enzyme-conjugated antiglobulin, and concentrations of test sera are all critical parameters of a successful ELISA procedure. When ELISA titers of 30 human sera were compared to micro-complement fixation titers of the same sera, a significant degree of correlation was observed, but quantitatively ELISA was found to be up to 200 times more sensitive. The sensitivity of the assay and its adaptability to automation should provide an additional method for study or diagnosis of infection with herpes simplex virus.

Animals↗
PubMed1976

Comparative diagnostic aspects of herpes simplex virus tumor-associated antigens.

Sera from cancer patients and healthy individuals, obtained from two independent sources, were examined for their abilities to react with herpes simplex virus-associated tumor antigens, AG-4 and NVA-TAA (nonvirion antigen-tumor-associated antigen). Both antigens were prepared by infection of HEp-2 cells with herpes simplex virus type 2, and all antigen-antibody interactions were measured by the micro-complement fixation test. Of sera from 16 patients with cancer of the uterine cervix, 81% (P less than 0.01) reacted with NVA-TAA, whereas 78% (P less than 0.001) of 18 sera examined reacted with AG-4. These values differed significantly from those for normal sera, of which 14% reacted with NVA-TAA and 13% with AG-4. Of sera for 8 patients with squamous cell carcinoma of head and neck or vulva, 75% (P less than 0.02) reacted with NVA-TAA, whereas 63% (P less than 0.05) reacted with AG-4. As a group, other cancers (including adenocarcinoma of lung, breast, ovary, and cervix; liposarcoma; sarcoma; melanoma; and carcinoma of the endometrium) did not differ significantly from controls in reactive patterns with AG-4 or NVA-TAA. These studies partly supported the reported preferential reactivity of AG-4 and NVA-TAA with sera of patients with squamous cell carcinoma, especially of the uterine cervix.

Adult↗
PubMed1976

Quantitative cytophotometry evaluated as a method for analyses of herpes simplex viral deoxyribonucleic acid synthesis.

Feulgen deoxyribonucleic acid (F-DNA) microspectrophotometry was evaluated as a potential tool for quantification of herpes simplex virus type 1 (HSV-1) and 2(HSV-2) DNA synthesis in single cells. Since HSV DNA synthesis has been extensively studied using incorporation of radioactive precursors into viral and cellular DNA, microspectrophotometric measures were correlated with biochemical data obtained using tritium-labeled thymidine ([3H]TdR). It was established that: (i) viral-induced increaae in F-DNA can be cytophotometrically detected between 1 and 6 h postinjection (p.i.), which corresponds to the initial incorporation of [3H) TdR into viral DNA; (ii) peak F-DNA levels occurred 8 h p.i., which supported cytological observations of a more rapid development of an inclusion body in HSV-2-infected nuclei. Despite the fact that F-DNA cytophotometry is unable to distinguish between cell and viral DNA, the overall study supports the existence of a good correlation between data obtained using microspectrophotometric and conventional isotope methods. Furthermore, cytophotometry complements biochemical evaluations in that it permits analyses of DNA changes on a single-cell basis or the detection of infection in very small numbers of cells.

Cell Nucleus↗
PubMed1976

Cell electrophoresis research directed toward clinical cytodiagnosis.

The application of cell electrophoresis to cytodiagnosis requires that a scientifically established basis exists for identifying abnormal cells electrophoretically, that research to detect such differences in the cytodiagnostic setting is possible and that a rapid and simple method of cell electrophoresis is adaptable to the clinical setting. Data are presented indicating modifications of electrophoretic mobility due to herpes simplex virus type 1 infection and Rous sarcoma virus transformation of culture cells. A simple apparatus for electrophoretically separating cells on a density gradient and collecting them for subsequent analysis is described, and results of experiments with this apparatus are consistent with those obtained by microscopic electrophoresis. Laser-doppler spectroscopic electrophoresis is suggested as a rapid method adaptable to clinical application.

Cell Line↗
PubMed1976

Reaction of antigens isolated from herpes simplex virus-transformed cells with sera of squamous cell carcinoma patients.

Antigens isolated from herpes simplex virus type 1, herpes simplex virus type 2, or cytomegalovirus-transformed hamster cells were tested against 66 sera from non-cancer individuals or patients with different types of cancer. By use of the microcomplement fixation procedure to quantify all antigen-antibody interactions, it was observed that 94% (p less than 0.001) of all sera from patients with squamous cell carcinoma reacted with antigens from herpes simplex virus type 1-transformed cells, while 84% (p less than 0.001) of the same sera reacted with antigen preparations from herpes simplex virus type 2-transformed cells. When sera from patients with adenocarcinoma, sarcoma, liposarcoma, and melanoma were tested against these antigens, there was no significant difference in their reactivity from sera of noncancer patients. When sera from all individuals (normal and cancer) were tested against antigens from cytomegalovirus-transformed cells, no significant reaction pattern developed. These studies are the first to describe the isolation of a reactive tumor-associated protein from herpes simplex virus-transformed cells.

Adult↗
PubMed1975

Adenovirus-transformed cells restrict Herpes simplex virus replication.

A cell line that normally supports the replication of herpes simplex virus types 1 and 2 became resistant to these viruses after transformation by simian adenovirus 7. Kinetic studies of the mechanism of resistance demonstrated that both herpesviruses were able to attach to the transformed cells and express some early genomic functions, as demonstrated by the presence of low levels of viral thymidine kinase. However, isopycnic centrifugation studies of the abortive system failed to detect viral deoxyribonucleic acid synthesis, whereas indirect immunofluorescent studies of viral proteins revealed that less than 10 per cent of the cells contained these viral macromolecules at any given time. Collectively the data suggest that after transformation by simian adenovirus 7 these cells are altered so as to render them resistant or incapable of supporting the growth of herpes simplex virus types 1 and 2. The results further suggest that the block occurs after viral absorption and prior to viral deoxyribonucleic acid synthesis.

Adenoviridae↗
PubMed1975

Differential feulgen-deoxyribonucleic acid hydrolysis patterns of Herpes simplex virus type 1 and type 2 infected cells.

Infection of human embryonic lung cells with herpes simplex virus type 1 (HSV-1) and herpes simplex type 1 (HSV-2) resulted in: (a) qualitative (nuclear cytopathologic) alterations and quantitative (nuclear area) differences in infected compared to control nuclei; (b) increased Feulgen-deoxyribonucleic acid (F-DNA) amounts in infected cells, probably due to viral DNA; (c) higher F-DNA levels in HSV-2 infected cells; and (d) increased rates of F-DNA hydrolysis in viral-infected as compared to uninfected nuclei.

Cell Line↗