Biomedical subjects

V Zachar

Publications and source records attributed to V Zachar.

60 records · Page 4Linked to original sources

PubMedDate not supplied

Physiological oxygen tension is relevant to MHC-1 expression, spontaneous transformation, and interferon response of in vitro aging murine fibroblasts.

Working from the hypothesis that modest deviations from physiological oxygen tension will influence cell characteristics important for infections/immunity and tumor development, cells were studied at three oxygen tensions during in vitro aging. Primary mouse embryo fibroblasts were established and subsequently passaged at 3, 6, and 18 kPa oxygen tension (6 representing normal tissue tension and 18 being the conventionally tension at in vitro cultures). The growth rate was slightly higher at 6 than 3 and 18 kPa. All cultures eventually stopped growing and subsequently transformed to nonmalignant cells with unlimited growth capacity. Cells kept at 3 kPa reached the highest number of cell doublings before crisis. Stimulation with PolyI:C resulted in detectable interferon response only at the high oxygen tension, and after transformation none of the cultures responded with interferon production. Expression of the major histocompatibility complex H-2K was elevated above and below physiological oxygen tension, indicating regulatory processes optimizing MHC expression at about physiological oxygen tension.

Animals↗

PubMedDate not supplied

Differential HIV replication and HIV-induced interferon production in mononuclear phagocytes: relationship to cell maturation.

We have investigated the replication of human immunodeficiency virus (HIV) and HIV-induced interferon (IFN) production in human mononuclear phagocytes at 2 different stages of in vitro maturation. Blood monocytes and monocyte-derived macrophages from 6 healthy, HIV-seronegative donors were challenged with HIV1IIIB and HIV2ROD. Freshly separated monocytes produced IFN when inoculated with both HIV types. In these cultures, an inverse correlation was observed between the amount of IFN production and the rate of HIV replication. In contrast to the monocytes, 5-day-old monocyte-derived macrophages did not produce IFN when challenged with HIV, but a significant replication of HIV1IIIB and HIV2ROD was found in all cultures.

Acquired Immunodeficiency Syndrome↗

PubMedDate not supplied

Clonal analysis suggests provirus expression in a subpopulation of human malignant trophoblast cells harbouring the human T cell lymphotropic virus type-I genome.

Previous studies have indicated that the villous trophoblast may be involved in intrauterine HTLV-I infection. Although the data furnished by our group (Liu et al., 1995) have demonstrated that the human trophoblast-derived malignant cell lines JAR and JEG-3 are susceptible to HTLV-I, the infection, even after thorough analysis, appeared to be limited to expression at the transcriptional level. In the present report, we sought to explore virus expression at the single cell level using eight clonally selected cell lines which were derived by limiting dilution from the previously infected parental cultures. Of the three cell lines JAR-H2, JAR-H3, and JEG-H3, all of which harboured full-length provirus, only in two (JAR-H2 and JEG-H3) were the virus-specific tax/rex and env transcripts demonstrated using RT-PCR. When compared with MT-2 cells, the detected steady-state levels of HTLV-I mRNA appeared to be lower by three orders of magnitude. Viral Tax protein displaying a typical intranuclear localization was found in 1-2% of JAR-H2 and JEG-H3 cells. Moreover, an altered phenotype characterized by multinucleated syncytia was observed in these cell cultures with the same frequency as Tax transactivator, implying a fusogenic activity of env protein. Infectious virus, however, could not be rescued from JAR-H2 or JEG-H3 clones by coculture with cord blood mononuclear cells. Our data suggest that trophoblast represents a susceptible, albeit a slightly permissive, host system for HTLV-I.

Base Sequence↗

PubMedDate not supplied

Differential interferon production in human first and third trimester trophoblast cultures stimulated with viruses.

Stimulation of human placental first and third trimester trophoblast and syncytiotrophoblast cultures with viruses [Newcastle Disease Virus (NDV) and Sendai virus] led to a high interferon (IFN) production. The magnitude of the production was dependent on the gestational age of the trophoblast, type of inducer and the stage of differentiation of the trophoblast. The data obtained indicated that the first trimester trophoblast cultures produced five to sixfold more IFN than the third trimester trophoblast on per cell basis whereas syncytiotrophoblast at term produced twice as much IFN than the mononuclear term trophoblast when stimulated with the viruses. NDV and Sendai virus produced different levels and composition of IFN-alpha and -beta in both first and third trimester trophoblast and syncytiotrophoblast cultures. Purification of the virus-induced trophoblast interferons (tro-IFNs) by tandem high-performance affinity chromatography resulted in specific activities between 0.7 and 2.7 x 10(8) IU/mg of protein when assayed on human amniotic WISH cells. The tro-IFN-alpha protected both human and bovine MDBK cells from virus infection whereas the tro-IFN-beta protected only the human cell lines tested. The possible roles of the tro-IFNs are discussed in light of the observed differences in trophoblast IFN response.

Cells, Cultured↗

PubMedDate not supplied

In vitro infection of human placental trophoblast by wild-type vaccinia virus and recombinant virus expressing HIV envelope glycoprotein.

Short-time (< or = 7 days) cultures of trophoblast mononuclear cells isolated from term placentae were challenged with vaccinia virus. Cytopathic effects were induced in crude placental cell preparations as well as in cultures established after negative immunosorting of major histocompatibility complex class I epitope-expressing cells, i.e. cultures exclusively derived from villous cytotrophoblast according to our present state of knowledge. The trophoblast in vitro supported a full replicative cycle of both wild-type viruses and a recombinant clone serving as a vector for the human immunodeficiency virus type 1 envelope gene. Results may shed light on mechanisms involved in the rarely observed foetal damage caused by smallpox vaccination during pregnancy.

Animals↗

PubMedDate not supplied

Biological characterization of three novel variants of IFN-alpha 13 produced by human placental trophoblast.

Interferon (IFN)-alpha from the human placenta was cloned and expressed with the aim to study the antiviral, antiproliferative, and immunostimulatory activities. In the present study, we describe three previously unknown sequence variants of IFN-alpha 13 originating from the villous trophoblast. The first variant differed from IFN-alpha 13 by a Cys99Arg substitution and a 10-amino acid C-terminal deletion, which led to a severe reduction of the antiviral and antiproliferative potential. The second variant with a Glu32Tyr substitution also displayed diminished antiviral and antiproliferative properties, but to a lesser extent than the first clone. For the third variant, a Ser25Pro substitution in the N-terminal part of the protein and two substitutions in the C-terminal part of the protein, Arg126Gly and Ala140Gly, resulted in diminished antiviral but not antiproliferative properties. Regardless of the altered antiviral and antiproliferative properties, all sequence variants demonstrated natural killer (NK) cell stimulatory potentials paralleling that of prototype IFN-alpha 13. Further studies are needed to gain a better understanding of the functional significance of different IFN-alpha subtypes at the maternal-fetal interface, in particular in light of the controversial role the NK cells play in the positive outcome of pregnancy.

Animals↗