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Biomedical subjects

G Hunsmann

Publications and source records attributed to G Hunsmann.

At least 217 records · Page 12Linked to original sources

Structural and epidemiological features of primate lymphotropic retroviruses.

Primate lymphotropic retroviruses (PLRV) have been isolated from man and various species of Old World monkeys. The human isolates adult T-cell leukemia virus (ATLV) or human T-cell leukemia virus (HTLV-I) are endemic among Japanese in the south-west of the country as well as Africans in both Africa and America. In these populations ATL and its lymphoma variant are also endemic and all these patients harbor the virus and have serum antibodies to viral env gene polypeptides gp68, gp46, p21 as well as core polypeptides p24, p19, and p15. ATLV and respective viral antibodies are very rare outside the endemic areas. Retroviruses structurally and serologically related have been found in various species of Old World monkeys especially in macaques. The human lymphadenopathy associated virus (LAV) later also known as HTLV-III most probably is the etiological agent of the acquired immunodeficiency syndrome (AIDS) in man and related syndromes. LAV/HTLV-III is highly endemic in certain parts of the population of America and Europe and apparently also in Africa. Human antisera react with viral envelope related gp120, gp100, gp46, and core related p24, p21, and p15 polypeptides. In our assays LAV/HTLV-III does not crossreact serologically with ATLV/HTLV-I and its major structural polypeptides may be related distantly to primate lymphotropic retroviruses (PLRVs) isolated from macaques but not to ATLV/HTLV-I.

Acquired Immunodeficiency Syndrome↗

Adult T-cell leukemia (ATL) virus-specific antibodies in ATL patients and healthy virus carriers.

The adult T-cell leukemia (ATL)-associated antigen complex (ATLA) is recognized by serum antibodies of carriers of ATL virus (ATLV). ATLA consists mainly of ATLV polypeptides and their precursors. The sera from 22 ATL patients, 21 healthy carriers and 9 healthy individuals were examined quantitatively by immunofluorescence assay (IF) for ATLA and by a newly developed radioimmunoprecipitation test with purified 125I-gp68, the putative env gene product of ATLV. More qualitative results were obtained by analysis on polyacrylamide gel (PAGE) of immunoprecipitates from lysates of 35S-cysteine-labelled cells producing ATLV, pelleted ATLV and cell-free culture supernatant. The two quantitative assays gave negative results with sera from all normal subjects and a few patients, but detected ATLA antibodies in all the healthy ATLV carriers. An important finding was that sera of patients that gave negative results in one assay gave positive results in the other, and vice versa. In contrast, all sera from ATL patients and healthy carriers, but not normal donors, precipitated ATLV-specific glycopolypeptides, gp68 and gp46 from 35S-labelled materials. But core polypeptides p28, p24, p19 and p15 were precipitated only by sera with IF titers of over 80. Thus, anti-ATLA antibodies in seropositive sera are predominantly directed against glycopolypeptides of ATLV, and the antibody reactivity to ATLA antigens does not differentiate between ATL patients at various stages of the disease and healthy ATLV carriers.

Antibodies, Neoplasm↗

Detection of serum antibodies to adult T-cell leukemia virus in non-human primates and in people from Africa.

The distribution of serum antibodies to adult T-cell leukemia virus (ATLV) was examined as a marker for virus infection among non-human primates as well as people from Africa and Germany. The virus is present in Africa in certain primate species including man. Altogether, 468 sera from 27 monkey species were examined. Only African green monkeys, less frequently also chimpanzees and crab-eating monkeys, were found to be infected. About 1-2% of people from Kenya have antibodies, while ATLV-antibodies may be present in well below 0.1% of the German population.

Animals↗

Sequence analysis of Gardner-Arnstein feline leukaemia virus envelope gene reveals common structural properties of mammalian retroviral envelope genes.

We have sequenced the envelope (env) gene and most of the adjacent 3' long terminal repeat (LTR) of Gardner-Arnstein feline leukaemia virus subtype B. The LTR of this virus contains, at corresponding positions, all signal sequence elements known from other retroviral LTRs. The deduced amino acid sequence of the longest open reading frame was compared with env polypeptide sequences of several murine leukaemia viruses. This allowed us to predict the positions of both p12/15env and gp70 polypeptides as well as a hydrophobic leader polypeptide. The env polypeptides of the different viruses show long stretches of homology and similar hydrophilicity profiles in the p12env region and in the carboxy-terminal half of gp70 (constant region). The most extensive variations are confined to certain parts of the amino-terminal half of gp70 (differential region). In this region, however, feline leukaemia virus and murine mink cell focus forming viruses are still closely related. A correspondingly spaced pattern of identical, short amino acid sequences appears in three different parts of the env polyprotein, suggesting its evolution from a primordial env-related precursor by tandem duplications.

Amino Acid Sequence↗

Active immunization with feline leukemia virus envelope glycoprotein suppresses growth of virus-induced feline sarcoma.

The efficiency was examined of immunization with feline leukemia virus glycoprotein complexes (gp85 rosettes) to protect cats against tumors induced by feline sarcoma virus (FeSV). The glycoprotein was isolated from feline leukemia virus (FeLV). Young cats were vaccinated with the purified viral glycoprotein and challenged with FeSV (FeLV). FeLV gp85 antibody levels were measured by enzyme-linked immunosorbent assay and tumor volumes were determined. In immunized animals tumor development was reduced. Gp85 antibody levels before challenge were correlated inversely with tumor size (r2 = 0.79). This method appears to be suitable for fast and efficient testing of future FeLV vaccines.

Animals↗

Virosomes constructed from lipid and purified Friend leukaemia virus glycoprotein.

Liposomes were loaded by a dialysis technique with purified envelope glycopolypeptide, gp85, of the Friend murine leukaemia virus (F-MuLV). The gp85 liposomes prepared from cellular lipid had a buoyant density of 1.05 g/ml and an apparent diameter of 50 to 300 nm. The gp85 was not simply entrapped by liposomes nor adsorbed non-specifically to their outer surface. Experiments with radioactively labelled protein, electron microscopic examinations, protease treatment and concanavalin A binding showed that gp85 is anchored in the liposomal membrane and oriented asymmetrically as in the virus envelope. Moreover, gp85-covered liposomes displayed some functions of the intact F-MuLV envelope, such as absorption of antibodies to gp70 and haemagglutination after enzyme treatment. To some extent, these lipid vesicles appeared to be reconstituted F-MuLV envelopes and thus, by analogy to other systems, were named retrovirosomes.

Dialysis↗

Nucleotide sequence of the envelope gene of Friend murine leukemia virus.

The envelope gene of the helper-independent, highly leukemogenic virus Friend murine leukemia virus was sequenced by using a molecular clone of a Friend murine leukemia provirus. The deduced amino acid sequences of the envelope proteins gp70 and p15env were homologous to the sequences of Moloney murine leukemia virus (86%) and Akv (76%). However, a stretch of about 40 amino acid residues near the middle of gp70 was dissimilar in Friend and Moloney murine leukemia viruses and Akv. In this type-specific region the gp70s of all three viruses contained more than 30% proline residues, giving this sequence a very rigid conformation. We suggest that this rigid and highly variable region of gp70 participates in infection by recognition of cell surface receptors and, in addition, might contribute to the different oncogenic spectra of murine leukemia viruses.

Amino Acid Sequence↗

Separation procedure and sugar composition of oligosaccharides in the surface glycoprotein of Friend murine leukemia virus.

The sugar composition of the surface glycoprotein from Friend murine leukemia virus was determined by gas-liquid chromatography of the alditol acetates and by the thiobarbituric acid method, respectively. N-Acetylglucosamine, mannose, galactose, sialic acid and fucose were found in a molar ratio around 15.2:11.6:7.4:3.3:1.0. Ten oligosaccharide fractions were obtained from glycoprotein preparations by a suitable sequence of degradation (with pronase, endo-beta-N-acetylglucosaminidase H, neuraminidase, and by hydrazinolysis) and separation procedures (concanavalin A-affinity chromatography and gel filtration). The qualitative sugar composition of these fractions was analyzed by in vivo labelling with D-[6-(3)H]glucosamine, D-[2-(3)H]mannose, D-[6-(3)H]galactose, or L-[6-(3)H]fucose, and their molecular weights were estimated from the gel elution volumina. Four fractions of N-glycosidically linked oligosaccharides of the oligomannosidic ('high mannose') type oligomannosidic7-oligomannosidic10, about seven to ten sugar residues), two of the mixed (M11 and M12), and four of the N-acetyllactosaminic ('complex') type (N-acetyllactosaminic9, probably nine sugar residues; N-acetyllactosaminica-N-acetyllactosaminic c, size unknown) were thus identified.

Chromatography, Affinity↗

Diverse effects: augmentation, inhibition, and non-efficacy of 12-O-tetradecanoylphorbol-13-acetate (TPA) on retrovirus genome expression in vivo and in vitro.

The action of 12-O-tetradecanoylphorbol-13-acetate (TPA) on several retrovirus-related functions was investigated in four virus-host cell systems. The following effects were recorded: (i) in STU-mice, infected with the Friend virus complex (Friend) murine leukaemia virus/Friend spleen focus forming virus) and treated with TPA (50 ng/g) for one week prior to infection, the number of spleen foci increased 5-fold over the control. (ii) Addition of TPA (0.04 to 40 ng/ml) to virus-producing cell systems resulted in a 2-fold increase of extracellular reverse transcriptase activity. The maximum response was observed in Friend leukemia virus-producing mouse cells at 0.1 to 0.4 ng TPA/ml and in simian sarcoma virus-producing rat cells at 4 ng/ml. (iii) The efficiency of transformation of BalbC 3T3 cells by Moloney murine sarcoma virus, tested in a focus formation assay, was slightly enhanced by TPA. (iv) TPA inhibited the induction of endogenous virus formation in B cell mitogen-stimulated spleen cell cultures from BalbC mice.

Animals↗

Glycoproteins of friend murine leukemia virus: separation and NH2-terminal amino acid sequences of gp69 and gp71.

The NH2-terminal amino acid sequences (initial 23 residues) of Friend murine leukemia virus gp71 and gp69 were determined and found to be different but highly related. Friend murine leukemia virus gp71 differed from Rauscher murine leukemia virus gp70 in only one position. Friend murine leukemia virus gp69 showed approximately 41% homology to these glycoproteins but lacked the glycosylation site (sequon) occurring at position 12 in Rauscher murine leukemia virus gp70.

Amino Acid Sequence↗

Antibody directed against Friend leukemia virus stimulates DNA synthesis in a subpopulation of mouse B lymphocytes.

Goat and rat antisera directed against Friend leukemia virus (anti-FLV) were found to be B-lymphocyte mitogens stimulating DNA synthesis in these cells but not in T lymphocytes. Membrane fluorescence microscopy showed that anti-FLV reacts with a subset of B lymphocytes of which the majority express immunoglobulin mu chains. The mitogenic effect was found with all mouse strains tested including 129 and AKR. Absorption experiments with purified viruses indicated that the mitogenic effect is specific for an antigen present in murine leukemia viruses of the FMR subgroup. In absorption experiments with viable cells, the antigen involved in mitogenicity was found to be expressed on Friend erythroleukemia cell lines (4/4) and on myelomas (2/2) but not on normal thymus T lymphomas (0/2) or on rabbit or mink cells infected with BALB/c xenotropic virus. Preincubation of spleen cells with anti-gp70 antiserum inhibited the mitogenic effect of anti-FLV but not of lipopolysaccharide.

Animals↗

A non-virion surface antigen on Moloney sarcoma virus-transformed non-producer and producer cells.

Sera from STU mice bearing sarcomas induced by cells producing the Moloney sarcoma-helper-virus (M-MSV/MLV) complex were cytotoxic for these cells as well as for M-MSV non-producer and M-MLV producer cells. Analysis by polyacrylamide gel electrophoresis of 125I-labelled surface antigens immunoprecipitated with such sera revealed the virus envelope glycoprotein gp71 on the producer cells and an additional antigen of mol. wt 55 K on the M-MSV-transformed producer and non-producer cells. This antigen was not found on non-transformed M-MLV-producing cells and was neither related serologically to structural polypeptides of murine C-type viruses nor to components of embryonal STU mouse fibroblasts and foetal bovine serum.

Animals↗

Envelope polypeptides of Friend leukemia virus: purification and structural analysis.

Roughly 10% of surface glycoproteins in the envelope of mature Friend murine leukemia virus are coupled to membrane polypeptides by disulfide bridges. The remaining 90% of these glycoproteins are associated noncovalently. However, they could also be linked to membrane polypeptides by the treatment of purified Friend murine leukemia virus with 2,2'dithiobis(m-nitropyridine). These amphiphilic heterodimer polypeptides, gp84/86, were recovered almost quantitatively in the form of aggregates, termed rosettes, when prepared by solubilization of the viral membrane with Triton X-100 and subsequent velocity sedimentation. gp69/71 and p12(E)/15(E) were purified from these protein micelles after reduction of the disulfide bonds by gel chromatography. Electron micrographs of rosettes, as well as of purified p12(E)/15(E), showed structures different from native viral knobs. Isolated gp84/86 could be reassociated and then displayed more similarity to these viral surface projections. As shown by peptide mapping, the primary structures of the glycoproteins gp69/71 are highly related as are those of the membrane polypeptides p12(E) and p15(E). Furthermore, it was shown by two-dimensional polyacrylamide gel electrophoresis and re-electrophoresis of purified gp84/86 that the larger component, gp86, was composed of gp71 associated with p15(E) and p12(E), whereas the smaller component, gp84, was formed by gp69 bound only to p12(E).

Animals↗

Possible cell surface receptor for Friend murine leukemia virus isolated with viral envelope glycoprotein complexes.

Water-soluble multimeric complexes of Friend leukemia virus envelope glycoprotein gp85 bind specifically to C57BL/6 mouse spleen leukocytes. Such complexes were used to isolate cell surface receptors for the virus, using an immunoprecipitation technique. The putative rceptor has a molecular weight of 14,000. Mouse H-2 histocompatibility antigens, which are receptors for Semliki Forest virus, are not receptors for Friend leukemia virus.

Animals↗