Human immunodeficiency virus infection of the liver.
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Acquired Immunodeficiency Syndrome↗
Biomedical subjects
A Kirn
Publications and source records attributed to A Kirn.
Interaction of human immunodeficiency virus with human macrovascular endothelial cells in vitro.
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Cells, Cultured↗
Altered pathogenicity in the liver induced by a mouse hepatitis virus type 3 thermosensitive mutant.
Intraperitoneal inoculation into sensitive BALB/c mice of D85, a thermosensitive (ts) mutant, provokes acute hepatitis followed by recovery of the mice. The ts mutant was able to replicate in the liver. However, the maximal viral titre was obtained 2 days later than was the case with the wild-type (wt) MHV 3 infection; the viral antigens remained localized within small foci and no invasion of the entire liver was observed. The hepatocytes infected with D85 showed strong steatosis similar to that induced by wt virus, but the other lesions induced by MHV 3 (closing of endothelial cell fenestrae and hepatocytolysis) were not seen. An important feature noticed with the D85 mutant concerned the establishment, in the surviving animals, of persistent infection: this phenomenon was demonstrated by the decrease of viral titre in the liver, viral RNA detection, and the fact that viral antigens gradually decreased until the 3rd month post-infection.
Animals↗
Assembly of nucleocapsidlike structures in animal cells infected with a vaccinia virus recombinant encoding the measles virus nucleoprotein.
A vaccinia virus recombinant containing the measles virus nucleoprotein gene was shown to induce the synthesis of a 60 kDa phosphorylated nucleoprotein similar to authentic measles virus nucleoprotein. Mammalian or avian cells infected with the recombinant virus displayed tubular structures reminiscent of viral nucleocapsids both in the cytoplasm and in the nucleus. Such structures could be labelled in situ by using an immunogold detection method specific for measles virus proteins. Electron microscopic examination of tubular structures purified from cells infected with the vaccinia virus recombinant indicated that they displayed most of the features of measles virus nucleocapsids, although their length was on the average shorter. These results demonstrate the spontaneous assembly of measles virus nucleocapsids in the absence of viral leader RNA and provide a means for a detailed molecular analysis of the requirements for nucleocapsid assembly. Furthermore, these findings raise the possibility of achieving complete assembly of measles virus particles, devoid of infectious RNA, by using a vaccinia virus vector.
Animals↗
Interaction of cultured human Kupffer cells with HIV-infected CEM cells: an electron microscopic study.
Evidence has accumulated indicating that macrophages could play a role in the physiopathology of AIDS. We recently demonstrated that cultured human liver macrophages, the so-called Kupffer cells (KC), are permissive for HIV. Their infection in vivo would lead these cells to constitute a target for the virus and a reservoir as well. Since they occupy a strategic position within the liver sinusoid, their opportunity to interact with blood-borne virus or already infected T lymphocytes may be very high. In the present study, we investigated the possibility for KC to be infected via HIV-infected CEM cells, a lymphoid cell line. Therefore, we cocultured both cell types for various times before fixing them for electron microscopy. Syncytia appeared within 20 h of infection as well as a large amount of virus particles. HIV in the way of budding was also easily observed. This has to be compared to the direct infection of KC with free virus which needs, at least, about 10 days to give the same results.
Cell Communication↗
Comparative ultrastructural study of rat livers preserved in Euro-Collins or University of Wisconsin solution.
University of Wisconsin solution greatly lengthens the time liver storage is possible compared with all previous solutions used. To test whether this improvement is related to better preservation of the endothelial cell, which is thought to be the most vulnerable cell type in cold storage, we compared time-related ultrastructural changes in rat livers stored in this solution or in Euro-Collins solution. Rat livers were harvested after combined arterial and portal perfusion with the cold-storage solution. They were then preserved for different lengths of time in the same solution at 4 degrees C before being perfusion-fixed and processed for light and electron microscopy. The first preservation damage was noted in endothelial cells; the time course of the lesions was similar in both solutions. After 2 hr of storage, enlarged and ruptured fenestrae with many gaps were observed. Swollen at 4 hr, the endothelial cells became stringlike at 10 hr, leading to stripped sinusoidal walls. Hepatocytes appeared better preserved in University of Wisconsin solution. The amount of glycogen, maintained near the control level at 24 hr in the latter, decreased dramatically between 0 and 4 hr in Euro-Collins solution, as ultrastructurally observed and biochemically confirmed. Furthermore, sinusoidal obstruction by blebs originating from the hepatocytes and quantified by image analysis on electron micrographs was markedly delayed. It was significantly less pronounced in University of Wisconsin solution at 24 hr than in Euro-Collins solution at 2 hr (p less than or equal to 0.05). Our findings confirm that endothelial cells are highly susceptible to preservation damage and show that University of Wisconsin solution does not improve preservation during storage.(ABSTRACT TRUNCATED AT 250 WORDS)
Adenosine↗
The nucleotide sequence of a delayed early gene (31K) of frog virus 3.
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Amino Acid Sequence↗
Inhibition in fat-storing cell multiplication by a factor produced by normal cultured murine hepatocytes.
In this study we demonstrate that hepatocytes isolated from normal mice may efficiently inhibit the multiplication of fat-storing cells (FSC) in culture, either in a coculture system where both cell types are separated by a filter of 0.45 microns pore size or via their conditioned medium. The inhibition may be completely reversed when the hepatocytes are removed and the culture medium is renewed. The inhibitory factor appears as early as 8 h in the medium with an almost maximum effect being reached after 24 h, as long as protein synthesis is allowed. It rapidly loses its efficiency through dilution. The inhibitory capacity of the conditioned medium is maintained after heating at 56 degrees C, dialysis of 100,000 x g centrifugation, but reduced after trypsin treatment. The infection of the hepatocytes by ectromelia virus causes an almost total suppression of the synthesis of the inhibitory factor. This latter result suggests that the multiplication of FSC, which may be inhibited by normal hepatocytes, would no longer be hindered in case of disregulation.
Animals↗
Permissivity of primary cultures of human Kupffer cells for HIV-1.
Kupffer cells (liver macrophages) represent the largest reservoir of fixed macrophages in the body. Accordingly, we have undertaken a study to evaluate their susceptibility to human immunodeficiency virus type 1 (HIV-1). Five-day-old primary cultures of Kupffer cells (KC) were infected with HIV-1, and as the infection progressed, syncytia appeared. Within the cells, viral proteins were detected by immunofluorescence using monoclonal antibodies directed against gp120 and p24. Electron microscopic examinations revealed the presence of typical Lentivirinae particles. The particles released from KC in the extracellular medium showed reverse transcriptase activity and p24 antigen; they could infect lymphocytic cells and were neutralized by a HIV+ patient's serum or an anti-gp120 monoclonal antibody. Our results thus demonstrate that the interaction of HIV-1 with KC in vitro leads to a productive infection. They suggest that the KC may be involved in the pathogenesis of HIV-1 infection and may (i) participate in the transmission of the infection to the peripheral blood cells, (ii) play a role in the depletion of uninfected CD4+ cells.
Acquired Immunodeficiency Syndrome↗
Isolation, culture and main characteristics of mouse fat-storing cells: interaction with viruses.
Fat-storing cells were isolated from 15-day-old mouse sinusoidal cell cultures (Kupffer or endothelial cells), where they had multiplied abundantly; they were then purified by a negative selection method based on the fact that they do not possess Fc receptors, as do both other types of cells. The fat-storing cells, which could be subcultured for at least 10 passages, have the main morphological characteristics already described in vivo, in particular, the rough endoplasmic reticulum and the lipid droplets, which become less and less apparent as the number of passages increases. Subcultured fat-storing cells, almost devoid of lipid droplets and vitamin A, were able to take up retinol, as the appearance of a typical autofluorescence indicated; the number of lipid droplets increased concomitantly. Furthermore, the cultured fat-storing cells were able to internalize one-micron-sized latex beads by phagocytosis. Infection of fat-storing cells with mouse hepatitis virus 3, ectromelia or Sindbis virus led to multiplication of the virus particles. There was a direct relation between the multiplication of mouse hepatitis virus 3 in cultured fat-storing cells and the susceptibility of the animals to the virus. In the case of Sindbis virus, interferon is produced, its production being independent of the presence of vitamin A.
Animals↗
Cell biology and kinetics of Kupffer cells in the liver.
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Animals↗
Antibodies directed against a synthetic peptide enable detection of a protein encoded by a vaccinia virus host range gene that is conserved within the Orthopoxvirus genus.
A vaccinia virus gene required for multiplication in some cell lines but not in others has been previously isolated and sequenced. A synthetic peptide predicted from the nucleotide sequence and corresponding to the carboxy-terminal 18 amino acids was used to raise antibodies in rabbits. The immune serum enabled detection of a 29-kilodalton (kDa) polypeptide by either immunoprecipitation or Western immunoblot assays. Synthesis of the 29-kDa polypeptide occurred immediately after infection and lasted for about 3 h. Shutoff of its synthesis was concomitant with the appearance of a delayed early polypeptide that may be antigenically related to the 29-kDa polypeptide. Analysis of cloned segments of the genomes of other orthopoxviruses by hybridization with the vaccinia virus host range gene demonstrates that it is well conserved within this genus.
Genes, Viral↗
[In vitro demonstration of a deficit in suppressor T-cell function in alcoholic cirrhosis. Role of hepatitis B virus infection].
The T lymphocyte suppressor cell activity has been evaluated in 33 alcoholic patients compared with 16 normal controls, using an in vitro test. Suppressor T cells were activated with concanavalin A, and suppressor effect was quantified by the inhibition of an autologous B cell culture response to Pokeweed Mitogen. When compared with controls, cirrhotic patients showed a significant defect of suppressor cell activity on B cell production of IgG (20 +/- 3 vs 46 +/- 5 p. 100, p less than 0.001) and IgM (26 +/- 4 vs 56 +/- 8 p. 100, p less than 0.05). In cirrhotic patients, defect of T cell suppressor function was independent of sex and severity of the cirrhosis (Child's staging). This defect was more marked in cirrhotics with serological markers of hepatitis B virus (HBV) infection (n = 11) than in cirrhotics without markers (n = 22) (9 +/- 5 vs 25 +/- 3 p. 100, p less than 0.05; 16 +/- 6 vs 30 +/- 5 p. 100, p less than 0.05 respectively for IgG and IgM production suppression). These results suggest that HBV and lymphocytes interact directly. This interaction could increase the T suppressor cell defect, and explain the promoting role of HBV infection in the constitution of the cirrhosis in alcoholics even when viral replication is not serologically apparent.
Female↗
Temperature-sensitive mutants of mouse hepatitis virus type 3 (MHV-3): isolation, biochemical and genetic characterization.
Mouse hepatitis virus 3 (MHV-3) is highly hepatotropic in sensitive mice. Temperature-sensitive mutants (ts mutants) induced by N-methyl-N'-nitrosoguanidine and 5-fluorouracil were isolated. Twelve mutants which were able to induce the formation of syncytia at 33 degrees C but not at the restrictive temperature (39.5 degrees C) were selected for detailed study. No viral RNA synthesis was detected after infection at the restrictive temperature with six of the mutants (RNA-) whereas six others were RNA+, although they displayed RNA synthesis which was generally reduced. No differences have been detected in the size of the genome or the viral-intracellular RNA species found in wild type virus or ts mutant infected cells at permissive temperature. The pattern of virus-induced proteins analyzed after immunoprecipitation by SDS-PAGE was similar in wild type virus and RNA+ mutant infected cells at 39.5 degrees C. Complementation experiments between ts mutants enabled us to distinguish five groups. Three of the groups contained RNA- mutants and two of them RNA+. Plaques made by mutants in one group displayed characteristic features that distinguished them from the wild type.
Animals↗
Organization of RNA transcripts from a 7.8-kb region of the frog virus 3 genome.
The detailed organization of the RNAs transcribed from a region of the FV 3 genome (Sa/I-F fragment and adjacent sequences) has been determined. The information was derived from the cell-free translation of hybrid-selected RNA to locate the genes encoding specific polypeptides, RNA filter hybridization to size the transcripts, and S1 nuclease mapping to locate the 5'- and 3'-ends of the RNAs on the genome. Three genes are contiguous and are transcribed from the same strand: two immediate early genes encoding transcripts of about 1.3 kb that directed the in vitro synthesis of 42K and 46K polypeptides, separated by the late gene encoding the major capsid protein (48K). At an advanced stage in infection, transcripts derived from the immediate early genes are also present. A set of RNAs with different 5'-ends ranging from 1.7 to 0.58 kb is produced from the p46 gene region whereas RNAs, 0.98 and 0.6 kb in size, complementary to the 5'-end of the p42 message, are synthesized. This gene cluster is located between two genes transcribed in the opposite direction from the rightward-reading strand: a late gene whose message is 0.5 kb in size and encodes a 15K polypeptide and a gene transcribed at immediate early and late times of infection which encodes a protein of 70 kDa. The 5'-end of the late RNA maps downstream of the 5'-end of the early one, their sizes being 1.85 and 2 kb, respectively, but both of them can be translated in vitro into a 70K polypeptide. These observations suggest that transcription is not regulated by the organization of the genes; they suggest rather that specific DNA sequences are responsible for the promotion of immediate early and late transcriptions.
DNA, Viral↗
Protection of mice from fatal measles encephalitis by vaccination with vaccinia virus recombinants encoding either the hemagglutinin or the fusion protein.
Vaccinia virus recombinants encoding the hemagglutinin or fusion protein of measles virus have been constructed. Infection of cell cultures with the recombinants led to the synthesis of authentic measles proteins as judged by their electrophoretic mobility, recognition by antibodies, glycosylation, proteolytic cleavage, and presentation on the cell surface. Mice vaccinated with a single dose of the recombinant encoding the hemagglutinin protein developed antibodies capable of both inhibiting hemagglutination activity and neutralizing measles virus, whereas animals vaccinated with the recombinant encoding the fusion protein developed measles neutralizing antibodies. Mice vaccinated with either of the recombinants resisted a normally lethal intracerebral inoculation of a cell-associated measles virus subacute sclerosing panencephalitis strain.
Animals↗
Interaction of mouse hepatitis virus 3 with Kupffer cells explanted from susceptible and resistant mouse strains. Antiviral activity, interleukin-1 synthesis.
The genetic sensitivity of mouse strains to mouse hepatitis virus 3 (MHV 3) has been related in vitro to a delay of virus replication in liver sinusoidal cells. In vivo immuno-histochemical studies of the liver from infected mice have demonstrated that mechanisms other than direct viral injury are in operation. To examine potential mechanisms, the interaction of lipopolysaccharide (LPS)-stimulated Kupffer cells with MHV 3 was studied. We first observed a dramatic inhibition in viral replication in LPS-treated Kupffer cells explanted from A/J resistant mice. Second, we demonstrated that MHV 3 induced a dose-dependent interleukin 1 (IL-1) activity in the supernatants of infected Kupffer cells of both strains. These results led us finally to examine the antigen-processing function of the Kupffer cells of both strains of mice. No striking differences were observed in the ability of Kupffer cells from resistant or sensitive mice to collaborate with immunocompetent lymphocytes. Our data suggest that Kupffer cells play a double role which is crucial in the pathogenesis of MHV 3-induced hepatitis. First, they act directly as the genetically determined sensitivity of mice to MHV 3 infection is correlated with the efficiency of the antiviral activity induced in Kupffer cells by LPS. Second, they act indirectly through the synthesis of different amounts of IL-1 induced by MHV 3. This hypothesis is further borne out by the effects of indomethacin treatment on the course of MHV 3 infection in A/J resistant mice in vivo.
Animals↗
A cowpox virus gene required for multiplication in Chinese hamster ovary cells.
Cowpox virus, in contrast to vaccinia virus, can multiply in Chinese hamster ovary cells. To study the genetic basis for this difference in host range, recombinants between vaccinia and cowpox viruses were isolated and their DNA restriction patterns were examined. The ability to multiply in Chinese hamster ovary cells could be correlated with the conservation of cowpox virus sequences mapping at the left end of the genome. This was further demonstrated by marker rescue of the host range phenotype with restricted cowpox virus DNA. Marker rescue with cloned restriction fragments of decreasing size enabled the fine localization of the host range function to a 2.3-kilobase-pair fragment. Nucleotide sequencing revealed that the fragment encoded a single major polypeptide of approximately 77,000 daltons. It is suggested that the role of the host range gene from cowpox virus is to prevent the early and extensive shutoff of protein synthesis that normally occurs in Chinese hamster ovary cells infected by vaccinia virus.
Amino Acid Sequence↗