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Biomedical subjects

J B Hudson

Publications and source records attributed to J B Hudson.

At least 19 recordsLinked to original sources
PubMed1992

Biosynthesis of human papillomavirus from a continuous cell line upon epithelial differentiation.

The study of the human pathogen papillomaviruses (HPVs) has been hampered by the inability to propagate the virus in tissue culture. The addition of 12-O-tetradecanoyl phorbol-13-acetate to the media of organotypic (raft) cultures increased expression of physiological markers of keratinocyte differentiation and concomitantly induced production of virions. Capsid production was detected in differentiated suprabasal cells. Virions approximately 54 nanometers in size were observed by electron microscopy in raft tissue cross sections in the suprabasal layers. Virions purified through isopycnic gradients were found to contain type 31b DNA and exhibited an icosahedral shape similar to that of papillomaviruses found in clinical samples.

Capsid↗
PubMed1992

Structure-activity studies of photoactivated antiviral and cytotoxic tricyclic thiophenes.

The photoactivated antiviral and cytotoxic activities of the naturally occurring thiophene, alpha-terthienyl (1), and 15 synthetic analogues were evaluated against murine cytomegalovirus and Sindbis virus, and murine mastocytoma cells. After irradiation with near UV light, alpha-terthienyl and most of its analogues had significant toxicity, with minimum inhibitory concentrations in the range of 0.02-40 microM. In the absence of near UV irradiation, only one analogue had antiviral activity and five were cytotoxic. The most active analogues were those containing carboxylic acid, hydroxyl, or cyano substituents. Quantitative structure-activity relationship analysis of thiophene phototoxicity suggested that the rate of singlet oxygen production is the primary determinant of antiviral and cytotoxic activities. For phototoxicity against murine cytomegalovirus, a significant role for hydrophobicity was also demonstrated. Tricyclic thiophenes show significant potential for photochemotherapy of viral infections and cancer, and further evaluation in animal models is recommended.

3T3 Cells↗
PubMed1992

Differentiation-induced and constitutive transcription of human papillomavirus type 31b in cell lines containing viral episomes.

The expression of viral genes during the productive life cycle of human papillomaviruses (HPV) is tightly coupled to the differentiation program of epithelial cells. We have examined transcription of HPV as a function of differentiation in an in vitro organotypic raft culture system which allows for epithelial stratification at the air-liquid interface. When CIN612 cells, which contain episomal copies of HPV type 31b (HPV31b), were allowed to stratify in raft cultures, they differentiated in a manner which was histologically similar to that seen in a cervical intraepithelial neoplasia I biopsy lesion. In monolayer cultures of CIN612 cells, two major polycistronic HPV31b transcripts of 1.7 kb which encode (i) E6, E7, E1-E4, and E5 and (ii) E6*, E7, E1-E4, and E5 were identified. These RNAs initiated at a promoter, P97, in the upstream regulatory region of the virus. Following differentiation in raft cultures, the relative abundance of RNAs initiated at P97 was unchanged. In contrast, the expression of a 1.3-kb RNA encoding an E1-E4 fusion protein and E5 was found to increase substantially following differentiation. This transcript was initiated at a novel promoter within the E7 gene (P742). These studies have therefore identified a constitutive viral promoter which is active throughout stratified epithelium as well as a novel promoter which is induced upon epithelial cell differentiation.

Amino Acid Sequence↗
PubMed1991

Therapeutic potential of plant photosensitizers.

Many bioactive phytochemicals have been shown in recent years to be photosensitizers, i.e. their toxic activities against viruses, micro-organisms, insects or cells are dependent on or are augmented by light of certain wavelengths. These activities are often selective, and this has led to the concept of therapeutic prospects in the control of infectious diseases, pests and cancer. Reaction mechanisms commonly involve singlet oxygen and radicals, which are thought to cause photodamage to membranes or macromolecules. The main classes of plant photosensitizers reviewed here are polyyines (acetylenes, thiophenes and related compounds); furanyl compounds; beta-carbolines and other alkaloids; and complex quinones. We propose that within each group of phytochemicals there are several representatives that merit further study for therapeutic abilities in appropriate animal models.

Animals↗
PubMed1991

Antiviral activities of hypericin.

Hypericin, a photodynamic plant quinone, readily inactivated murine cytomegalovirus (MCMV), Sindbis virus, and human immunodeficiency virus type 1 (HIV-1), especially on exposure to fluorescent light. Sindbis virus was significantly more sensitive than MCMV. The inactivated MCMV, when used to infect cells, was incapable of synthesizing early or late viral antigens. In addition to this direct virucidal effect, when hypericin was added to cells infected with viable MCMV, inhibition was also observed, particularly when the compound was added in the first two hours of infection. Again the antiviral effect was augmented by visible light. At effective antiviral concentrations, there were no discernible adverse effects on cultured cells. Thus hypericin appears to have two modes of antiviral activity: one directed at the virions, possibly on membrane components (although other virion targets cannot be ruled out), and the other directed at virus-infected cells. Both activities are substantially enhanced by light. Other recent studies on the antiviral activities of hypericin have not considered the role of light, and it is conceivable that apparent discrepancies between their results may have reflected different conditions of light exposure.

Animals↗
PubMed1991

Screening of medicinal plants from Yunnan Province in southwest China for antiviral activity.

In an ethnopharmacological screening of medicinal plants used in Yunnan province of China, ethanol extracts from 31 plant species were assayed for inhibition of murine cytomegalovirus and Sindbis virus infections. Parallel assays were carried out with and without exposure to UVA radiation to test for photo-mediation of activity. Antiviral activity was observed with 16 of the plant extracts. Eight plant extracts have been selected for further studies, with the objective of characterizing the antiviral constituents.

Antiviral Agents↗
PubMed1991

Ultraviolet-dependent biological activities of selected polyines of the asteraceae.

A series of eight polyines, isolated from several plants and containing various furanyl, thiophene, and spiro groups, were evaluated for antibiotic, antiviral, and cytotoxic activities in the presence and absence of long wavelength ultraviolet light, UVA. One compound, a furanopolyine, had impressive activity against a range of microorganisms, and membrane-containing viruses, as well as cytotoxic activity. All of these properties were UVA-dependent. Three other similar compounds had different patterns of UV-dependent activity against the various organisms, but some of these could also be of use in therapy of infectious diseases. In contrast, the dehydromatricaria esters and the two spiro compounds had relatively little biological activity.

Alkenes↗
PubMed1991

Antiviral activity of the photoactive plant pigment hypericin.

The polycyclic compound hypericin, a known photodynamic agent, was investigated for antiviral activity in the presence and absence of light. The three viruses tested: murine cytomegalovirus; Sindbis virus; and human immunodeficiency virus type 1, were all susceptible to hypericin; but these antiviral activities were considerably enhanced, in a dose-dependent manner, by exposure to light.

Animals↗
PubMed1991

Amplification of human papillomavirus genomes in vitro is dependent on epithelial differentiation.

Human papillomaviruses (HPVs) infect squamous epithelium and establish their genomes as episomes in proliferating basal cells. As infected cells differentiate, the viral DNA is amplified to high copy number and infectious virus is produced. Viral production has not yet been observed in vitro due to the inability of standard culture methods to duplicate most stages of epithelial differentiation. In this study, we have examined a cell line derived from a low-grade cervical lesion and found that it contained episomal copies of an HPV-31 subtype, HPV-31b, at approximately 50 copies per cell. When allowed to stratify at the air-liquid interface of in vitro raft cultures, this cell line differentiates in a manner which histologically resembles a low-grade cervical lesion in vivo. We have observed the amplification of HPV-31b genomes in distinct foci in the upper portion of the in vitro-stratified epithelium similar to that found in productive HPV infections in vivo. Although transcripts from the late region of HPV-31b were also detected specifically in stratified raft cultures, no capsid protein was found. These studies duplicate one important aspect of a productive HPV infection in vitro, the differentiation-dependent amplification of papillomavirus genomes.

Blotting, Northern↗
PubMed1990

Immortalization and altered differentiation of human keratinocytes in vitro by the E6 and E7 open reading frames of human papillomavirus type 18.

The E6-E7 region of human papillomavirus types 16 and 18 is selectively retained and expressed in cervical carcinoma cells. In cultured human keratinocytes, expression of the E6 and E7 open reading frames of human papillomavirus type 18, under the control of its homologous promoter, resulted in high-frequency immortalization. Furthermore, by using a system that allows for stratification of keratinocytes in vitro (raft system), we observed that the morphological differentiation of these E6-E7 immortalized cells was altered such that parabasal cells extended throughout most of the epithelium, with abnormal nuclei present in the upper regions. Examination of E6-E7-expressing cell lines in the raft system at a later passage revealed that complete loss of morphological differentiation had occurred. E7 alone was a much less effective immortalizing agent than E6 and E7 together and acted only minimally to alter morphological differentiation in vitro. No such activities were found for E6 alone. High-frequency transformation of human epithelial cells thus appears to require expression of both E6 and E7 gene products.

Cell Differentiation↗
PubMed1990

In vitro differentiation of epithelial cells from cervical neoplasias resembles in vivo lesions.

Cell lines derived from cervical neoplasias, as well as cells from normal cervix and neonatal foreskin were examined in an in vitro culture system (raft system) that allows for stratification and differentiation of epithelial cells at an air-liquid interface. Epithelial cells from human foreskin and ectocervix were observed to differentiate in a manner histologically similar to normal epithelium in vivo as indicated by a single layer of basal cells with a defined mid and upper zone. In contrast, cells expanded from cervical squamous carcinoma explants showed total absence of normal differentiation in the raft system with numerous cells in the upper portion of the stratum exhibiting mitotic figures. Cell cultures derived from low grade cervical intraepithelial neoplasia and condyloma acuminata exhibited partial differentiation in addition to perinuclear clearing, binucleate cells and individual cell keratinization. These studies show that in the raft system, cultured cells derived from tissue biopsies can duplicate many of the histological features observed in cervical neoplasias. In addition, epithelial cells derived from normal fetal ectocervix and electroporated with cloned human papillomavirus (HPV) type 16 DNA exhibited abnormal differentiation patterns similar to those of cervical intraepithelial neoplasia in vivo. This model system will thus be useful for examining the effects of HPV infection on epithelial maturation and for investigating the role of other factors in the progression of cervical malignancies.

Cell Differentiation↗
PubMed1989

Properties of the immunosuppressive factor induced by murine cytomegalovirus.

Murine cytomegalovirus infection of spleen cultures induced the production of a small (less than 10,000 molecular weight) immunosuppressive factor (VISF), which suppressed concanavalin-A mitogenesis in fresh mouse spleen cells, and in fresh human peripheral blood leukocytes. The factor did not affect the growth of two murine T-cell lines or of mouse fibroblasts. A similar factor was also found in the serum of infected mice, at the time of maximum immune suppression. The properties of VISF indicate that the mechanism of MCMV immune suppression is different from that caused by several other viruses which are important in human and veterinary medicine.

Animals↗
PubMed1989

Plant photosensitizers with antiviral properties.

Many antiviral compounds obtained from plants are photosensitizers, i.e., their biological properties are dependent upon or augmented by light of specific wavelengths, commonly long wave ultraviolet, UVA. Three groups of chemically distinct plant photosensitizers have been investigated in some detail in regard to antiviral properties. These are (a) thiophenes and polyacetylenes; (b) furyl compounds; (c) certain alkaloids. Some of the thiophenes and their acetylenic derivatives possess extremely potent phototoxic activities toward membrane-containing viruses. These activities are markedly affected by the chemical structures of these compounds. Inactivated virus retains its integrity, however, and penetrates cells, but does not replicate. Their mechanism of action is believed to occur via singlet-oxygen damage to the membranes, although other targets cannot be ruled out. In contrast, the antiviral activities of plant furyl compounds (such as psoralens and furanochromones) appear to depend on UVA-mediated covalent adduct formation with the viral genomes. Some of the photoactive beta-carboline alkaloids also have impressive antiviral activities, especially against viruses with single-stranded genomes. These and other types of alkaloids appear to work by mechanisms that do not require covalent bonding to nucleic acids, and may also involve other target molecules as well. Some of these compounds have potent antiviral activities at concentrations well below cytotoxic levels, and accordingly should be tested in animal models.

Alkaloids↗
PubMed1989

The antiviral action of lignans.

A total of 18 purified lignans was evaluated for antiviral activity against murine cytomegalovirus (CMV) and Sindbis virus, by means of different treatment regimens. Podophyllotoxin and alpha-peltatin were the most potent compounds, and they apparently inhibited murine CMV at an essential early step in the replication cycle after the adsorption of virus to the cells. On the other hand, justicidin B and the diphyllin derivatives were much more effective against Sindbis virus, and 12 of the lignans had no demonstrable effect at all, despite their known activities in other bioassays.

Antiviral Agents↗
PubMed1988

The DNA-binding properties of immediate-early and early proteins of murine cytomegalovirus.

The DNA-binding properties of the immediate early (IE) and early proteins induced by the murine cytomegalovirus (Smith strain) in permissively infected 3T3-L 1 cells and nonpermissively infected J774A.1 cells were characterized. Eight of 10 IE proteins and 4 of 7 early proteins had affinity for DNA (native or denatured). A 96K IE protein was observed to have affinity for denatured DNA-cellulose when synthesized in J774A.1 cells but not when synthesized 3T3-L 1 cells.

Antigens, Viral↗
PubMed1988

Analysis in vitro of two biologically distinct strains of murine cytomegalovirus.

The two biologically distinct strains of murine cytomegalovirus, "Smith" and "K181", showed slightly different restriction-endonuclease profiles; but we could not detect strain differences in the numbers or sizes of the major immediate-early, early or late viral proteins. It is possible however that some of the minor late proteins may differ.

Antigens, Viral↗
PubMed1988

Further characterization of the murine cytomegalovirus induced early proteins in permissive and nonpermissive cells.

Some of the properties of the immediate-early (IE) and early proteins induced by the murine cytomegalovirus (Smith strain) were examined in permissively infected 3 T 3-L 1 cells, and in non-permissively infected J 774A.1 (mouse macrophage) and human fibroblast cells, in order to determine differences that could account for the restriction in virus replication in the latter two cell lines. The different virus induced proteins had distinctive partitioning characteristics between nuclear and cytoplasmic fractions. The 96 K major IE protein had an exclusively nuclear association, as did the most abundant early proteins of 39 K and 36 K. The other viral proteins however were evenly distributed between nucleus and cytoplasm. In general these patterns were also seen in the infected non-permissive cells. Several proteins showed more than one charge isomer on two-dimension gels, and in addition five IE proteins and two early proteins were phosphorylated. Only two differences between the permissive and nonpermissive infections were observed; the IE proteins of 100 K and 89 K when synthesized in the human cells had a stronger affinity for the nuclear fraction; also a phosphorylated form of the 30 K IE protein was not detected in MCMV infected J 774 A.1 cells.

Antigens, Viral↗