Search

Hamophiliepatienten besitzen ein hohes Risiko, an AIDS zu erkranken. Bei einem von uns untersuchten Patientenkollektiv waren 60% der Hamophiliepatienten HTLV III-Antikorper-positiv [1]. Man schatzt, das 5–15% aller HTLV III-Antikorperpositiven Patienten an AIDS erkranken werden. Eine Abschatzung des individuellen Risikos der Patienten, an AIDS zu erkranken, ware wunschenswert. Wir versuchen, mit Hilfe immunologischer Profile Risikogruppen innerhalb des Hamophiliepatientenkollektivs zu definieren, die ein erhohtes Risiko besitzen, an AIDS zu erkranken.

The study provides information on the epidemiology of HTLV-III infection and the lymphadenopathy syndrome (LAP) in 374 German homosexual men. Sexual contacts in the USA and rectal enemas before receptive anal intercourse are the main risk factors associated with virus transmission. HTLV-III seropositivity is significantly correlated with LAP. Prominent clinical signs are infreqquent. Immunological and haematological abnormalities are prevalent, and the retrovirus infection is frequently associated with serological markers of other viruses (hepatitis B, herpes group viruses). Lymphadenopathy as a manifestation of HTLV-III infection is discussed within the context of AIDS-related disorders.

A technique originally described for the isolation of Friend leukaemia virus envelope polypeptides [1] yields equivalent structures from Moloney leukaemia, AKR and BALB/c xenotropic virus as well as feline leukaemia virus. The envelope polypeptides are obtained as micellar protein complexes, named rosettes. Rosettes of the five m am malian type-C viruses examined are indistinguishable by electron microscopy. Separation of these aggregates in polyacrylamide gel electrophoresis under nonreducing conditions reveals a glycoprotein of about 85 000 d as their major component. Tryptic peptide analyses identify the viral origin of these polypeptides and emphasize strain specific differences in their primary structure

Ninety analogues of suramin have been examined for their ability to inhibit the exogenous reverse transcriptase (RT) of human immunodeficiency virus type I (HIV-I). Of these compounds, 57 inhibited the poly(rC).oligo(dG)-dependent RT activity. Three classes of dose-response curves could be discriminated. Allocation of a compound to one class did not correspond with obvious structural features. Twenty-four substances were superior to suramin in our RT inhibition assay. The RT-inhibitory activity of these compounds did not correlate with their effect against filariae or trypanosomes. Preliminary antiviral evaluation in susceptible human T cells inoculated with HIV-I demonstrated in vitro therapeutic efficacy for some compounds with lower drug-related cellular toxicity than suramin. Certain structural features relevant for the RT-inhibitory effect of these compounds were recognized. Predictions are made for the design of more effective RT inhibitors. Such compounds will help to understand the molecular mechanism of reverse transcription and might be useful in the therapy of retroviral infections.

Summary We have compared the structural polypeptides of an adult T-cell leukaemia (ATL) virus (ATLV) isolate from a Japanese patient with ATL with those of a similar virus derived from a Japanese macaque monkey. Both are distinct but related entities. Their core polypeptides p19 could not be distinguished, but p24, another core polypeptide, and their envelope glycopolypeptides differ. The human virus directs the synthesis of a single intracellular glycopolypeptide, gp68, while the macaque virus specifies two such glycopolypeptides, gp57 and gp50. Furthermore, the glycopolypeptides of both viruses are serologically distinct. Thus, these viruses represent subtypes of the ATLV family and the macaque virus is apparently not involved in human ATL.

Cells producing endogenous and exogenous type-C retroviruses of murine, feline and primate origin were evaluated for expression of those virus-specific cell-surface antigens which cross-react with antibodies to interspecies determinants of mammalian type-C viral polypeptides. Surface polypeptides of cells replicating endogenous and exogenous type-C retroviruses were iodinated by the lactoperoxidase method. Labelled antigens were immunoprecipitated and analyzed in polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. This method detected gp71 and less frequently p15E/p12E on the surface of virus-producing cells; in addition, p27 was found on F422 cells replicating feline leukemia virus. Antibodies to the membrane protein p15E displayed the broadest cross-reactivity but only antibodies to gp71 mediated complement-dependent interspecies cell lysis. The pattern of cross-reactivity reflected known genetic relationships among these mammalian viruses. Although antiserum to the simian sarcoma virus complex (SSV) was strongly cytotoxic to some human tumor cell lines, this reactivity could not be attributed to antibodies cross-reacting with SSV gp71.

The adult T-cell leukemia (ATL)-associated antigen complex (ATLA) was first discovered with indirect immunofluorescence by Hinuma et al. (1981). Biochemical analysis with MT-2 cells revealed that ATLA consisted mainly of human T-cell leukemia virus (HTLV) structural polypeptides and their precursors (Yamamoto and Hinuma 1982a; Schneider et al. 1984). In this study, we have investigated the molecular nature of the ATLA antigen complex in various HTLV-positive human cell lines established by different methods including independently established HTLV-infected HUT 102 cells. We found that HTLVs infecting these cell lines have similar core polypeptides, p24 and p19, as well as an envelope glycopolypeptide, gp46, in all these cells. The intracellular gp61 and p53 appear to be precursors of the viral envelope and core polypeptides, respectively. Interestingly, MT-2 and MT-2 related T-cell lines contain two different species of envelope proteins, gp68 and gp61, whereas cell lines not related to MT-2 express only gp61.

Antibodies to HTLV-I have been detected in sera from 15 (2.0%) of 736 adult blood-donors in Nigeria, in 4 (20.0%) of 20 patients with chronic lymphatic leukaemia, 3 (10.0%) of 30 with non-Hodgkin's lymphoma, one of 12 with Burkitt's lymphoma and one of 7 with acute lymphoblastic leukaemia. The frequency of positivily was higher (3.6%) in the blood-donors from the guinea and wooded savanna of northern Nigeria than in those from the rain-forest and mangrove swamps of southern Nigeria (1.8% in Lagos and 0.7% in Calabar). Two of the 3 seropositive patients with lymphoma had clinical presentation and courses similar to those of Japanese and Caribbean patients with adult T-cell leukaemia/lymphoma.

Summary Sera of individuals infected with adult T-cell leukaemia virus (ATLV) react predominantly with the polypeptides gp68, p24 and p19. These polypeptides were isolated from ATLV-infected MT-2 cells and virus. The radioiodinated polypeptides were used to quantify respective antibodies in individual ATLV carrier sera. Heteroantisera prepared in rabbits against isolated polypeptides facilitated studies on the biosynthesis of the core and envelope polypeptides of ATLV. Pulse-chase experiments revealed a polypeptide of mol. wt. 48000 (48K) as the precursor to the core polypeptides p24 and p19. A 28K polypeptide related to p19 appeared to be an early side-product of the gag gene or a translate of a defective viral message. Antiserum to the putative env gene product gp68 recognized gp68, gp66 and small amounts of gp62. In tunicamycin-treated cells gp68, gp66 and gp62 were no longer synthesized, but a 54K polypeptide reacted with antiserum to gp68. Polypeptide p54 is structurally related to gp68 and therefore apparently represents the unglycosylated form of gp68. Moreover, the apparent mol. wt. of p54 and p48 agree with those predicted for respective env and gag precursors from the nucleotide sequence of an ATLV provirus.

In 1984 10,281 sera were collected in the FRG and examined for antibodies to HTLV-III (LAV) with an enzyme-linked immunosorbent assay and confirmative tests. Of the German AIDS patients 81% have antibodies. Individuals belonging to AIDS risk groups, homosexuals, haemophiliacs and i.v. drug abusers, have antibody frequencies between 25%–72%. The detection of HTLV-III antibodies in blood donours indicates that the virus is being transmitted by blood transfusions.

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.

A 240-bp DNA fragment encoding a peptide, designated ENV(80), homologous to a conserved part of the gp41 transmembrane glycoprotein of human immunodeficiency virus (HIV) was chemically synthesized and inserted into different plasmid expression vectors. Escherichia coli transformants containing these plasmid constructs produced upon induction high amounts of either an ENV(80) peptide of relative molecular mass (Mr) of 10,000 or the same ENV(80) peptide N-terminally fused to E. coli chloramphenicol acetyltransferase (CAT) or to mouse dihydrofolate reductase (DHFR) having Mr of 36,000 and 31,000 respectively. All polypeptides containing the ENV(80) sequences were strongly reactive with antibodies present in sera from AIDS virus-infected individuals, but not with control sera. The strategy of gene assembly allowed the expression of ENV(80) subfragments fused to DHFR. The serodiagnosis of 15 positive sera by Western blot analysis using these bacterially synthesized ENV(80) subfragments revealed the presence of several immunoreactive epitopes on the 80-amino acid polypeptide which were recognized differently by the various patients.

Eight lymphoblastoid cell lines were established from the peripheral blood of individual African green monkeys (AGM). The AGM-2206 line grew out spontaneously. The others - AGM-6, 7, 8, 10, 12, 13, and 16 - were obtained after infection of peripheral AGM lymphocytes with cell-free culture supernatant of AGM-2206. All lines contained, and were probably transformed by, AGM-EBV. Moreover, they expressed immunoglobulins but lacked the Leu l T-cell marker. Thus they were B cells. Since a high percentage of AGMs are naturally infected with a virus similar to adult T-cell leukemia virus (ATLV), we examined these cell lines for ATLV. With immunofluorescence tests we detected ATLV-related antigens (ATLA) in three of the eight cell lines. EBV membrane antigen was present in three out of four. The highest percentage (40%) of ATLA-positive cells was found in the AGM-13 line. After metabolic labelling of these cells, ATLV-specific polypeptides p24, p19, p15, and p10 were detected. Hybridization experiments showed that both AGM-2206 and AGM-13 cell lines contained ATLV-proviral DNA. Electron micrographs of AGM-13 revealed a few type-C particles morphologically similar to the MT-2 virus. By cocultivation this AGM virus was able to infect and immortalize human peripheral blood lymphocytes. One such human cell line, NA-13, expressed polypeptides closely related to ATLV core antigens but a 68,000 mol.wt. glycopolypeptide was serologically distinct from MT-2 ATLV gp68.

Die humanpathogenen Retroviren HTLV-I und HTLV-III sind wie eine Reihe von Viren, die bisher mit MS in Verbindung gebracht wurden, sowohl lymphotrop wie neurotrop, sie persistieren zeitlebens. Der vor kurzem erhobene Verdacht, Verwandte dieser Viren konnten mit MS assoziiert sein, basierte auf den Nachweisen kreuzreagierender Serumantikorper sowie mit HTLV-I verwandter RNS in Lymphozyten von MS-Patienten (5). Um diesem Verdacht nachzugehen, haben wir Seren und Liquores von MS-Patienten im „enzyme linked immunosorbent assay“ (ELISA), in der indirekten Immunfluoreszenz (IF) und der Immunprazipitation (IP) auf Antikorper gegen HTLV-I gepruft. Dieses mehrstufige Vorgehen schliest falsch positive Resultate mit hoher Wahrscheinlichkeit aus.

To establish an animal model for AIDS, 6 juvenile rhesus monkeys were infected intravenously with cell-free SIVagmTYO-1, 5, or 7. One animal was infected with SIV of known pathogenicity isolated from a sooty mangabey (SIVsmm). The 2 animals infected with TYO-1, 1 of 2 infected with TYO-7 and the 1 infected with SIVsmm seroconverted within 4-8 weeks after infection and infectious virus could be recovered 8-10 or 44 weeks after infection. Three of the 4 seroconverted monkeys developed a persistent lymphadenopathy 12 weeks after infection. Rhesus monkeys infected with SIVagm could serve as a model to study HIV infection and for vaccine trials.

Sera from five Japanese patients with adult T-cell leukemia (ATL) showed in the immunofluorescence test for ATL-associated antigen (ATLA) titers ranging from 320 to 1280. Control sera from three healthy adults were negative. These eight sera were used to immunoprecipitate radiolabeled polypeptides from three cell lines infected with adult T-cell leukemia virus (ATLV) and two noninfected human cell lines. Cells were labeled either metabolically with [35S]cysteine, [35S]methionine, or [3H]glucosamine, or chemically with 125-iodine. Immunoprecipitates from cells, virus, and concanavalin A-enriched supernatants were analyzed by polyacrylamide gel electrophoresis. Cells producing ATLV contain gp68, the putative precursor to ATLV envelope polypeptides, gp46, and possibly p15, in addition to several nonglycosylated polypeptides between 40 to 70 kDa. Gp46 is shed into the culture medium and appears to be loosely attached to the viral and cellular surface. After purification on density gradients viral particles contain immunoreactive p24, p19, p15, and small amounts of gp46. Kinetics of synthesis, distribution, size, biochemical characteristics, and immunoreactivity of these polypeptides strongly suggest that most of them are structural components of ATLV or their precursors. Apparently, the intracellular ATLA complex predominantly represents precursors of viral structural polypeptides and gp46 is the viral envelope glycopolypeptide.

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 35-S-cysteine-la-belled 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.

Objective To study the diagnostic sensitivity and specificity of polymerase chain reaction (PCR) for the non-invasive diagnosis of progressive multifocal leukoencephalopathy (PML) in HIV-1-infected individuals. Design Retrospective analysis of stored cerebrospinal fluid (CSF) samples by PCR of HIV-1-infected patients. Methods Results of the PCR analysis of the CSF of three AIDS patients with autopsy-proven PML were compared with the results in 15 neurologically asymptomatic HIV-1-infected patients and with 15 AIDS patients with other opportunistic infections of the central nervous system (CNS). A polyclonal antiserum to simian virus 40 (SV40) cross-reacting with JC virus (JCV) late antigens was used for immunocytochemical confirmation of the diagnosis. Two different primer pairs, one taken from the VP1/large T gene and the other from the large T gene, were used to amplify JCV-specific DNA sequences from CSF. Results Five CSF samples were analysed and JCV-specific DNA found in three patients with autopsy-proven PML. No JCV-specific DNA was detected in 47 CSF samples, including serial samples from 14 of the 30 non-PML patients. The diagnosis of PML was confirmed in all three cases by immunocytochemistry. Conclusion: PML can be diagnosed by PCR analysis of CSF. The sensitivity and specificity of the method depends on the sensitivity of the primers used for amplification. Using a primer pair from the large T gene, JCV-specific DNA was amplified in three cases with PML as early as the day of presentation with the first neurological symptom of PML.

Progressive multifocal leukoencephalopathy (PML) is a subacute viral infection of oligodendrocytes by JC virus occurring almost exclusively in immunocompromised patients. By use of partially purified recombinant VP1 as antigen, the IgG response was analyzed by a quantitative ELISA of paired cerebrospinal fluid (CSF) and serum samples. An intrathecal immune response to VP1, defined as an antibody-specificity index of CSF to serum antibody titers greater than or equal to 1.5, was found in 76% of PML patients (47/62) but in only 3.2% of controls (5/155) (P < .001). Intra-blood-brain barrier synthesis of VP1-specific IgG antibodies is 76% sensitive and 96.8% specific for the diagnosis of PML. Furthermore, the excellent correlation (r = .985) between the plasma cell count in brain tissue and the humoral intrathecal immune response to VP1 in PML patients suggests a role for B cells in this disorder.

A suspension tissue culture producing large amounts of Friend leukemia virus was developed. Properties of the cells and the virus are described.

A glycoprotein of an apparent molecular mass of 46000, gp 46, was enriched by affinity chromatography from the virus-and cell-free culture medium of adult T-cell leukemia virus (ATLV) infected cells, gp 46 was specifically precipi­tated with sera from patients with adult T-cell leukemia known to react with the adult T-cell leukemia associated antigen (ATLA). Thus, gp 46 is a novel component of the ATLA antigen complex.

Mice could be protected against Friend leukemia virus infection by inoculation with highly purified viral glycoprotein GP71 or its specific antiserum