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Biomedical subjects

R Thorstensson

Publications and source records attributed to R Thorstensson.

18 recordsLinked to original sources
PubMed1992

Prevention of HIV-2 and SIVSM infection in cynomolgus monkeys by active or passive immunization.

We have established experimental infection with HIV-2 and SIVsm in cynomolgus monkeys and successfully used these models for vaccine experiments. Protection against homologous HIV-2 infection was demonstrated in two of two monkeys immunized with a Triton-X100-treated whole HIV-2SBL-6669 vaccine in incomplete Freund's adjuvant and in 2 of 4 monkeys immunized with a formalin-inactivated whole HIV-2 vaccine in RIBI adjuvant. Monkeys preinfected with a live poorly replicating HIV-2 strain were shown to develop cross-protection against SIV-induced disease. We have shown also that HIV-2 and SIVsm infection in cynomolgus monkeys can be prevented by passive immunization. These results raise hope for effective immunization against HIV infections in humans.

AIDS Vaccines↗
PubMed1991

Prevention of HIV-2 and SIVsm infection by passive immunization in cynomolgus monkeys.

Infection of macaques with simian immunodeficiency virus (SIV) and human immunodeficiency virus type 2 (HIV-2) are useful models for studies of immunotherapy and vaccination against HIV as well as for testing of antiviral drugs. Vaccine research showing protective immunity in immunized monkeys has indicated that it will be possible to develop a vaccine for prevention of human HIV infection, although many hurdles remain. The design of an HIV vaccine would be helped if the basis of the protective immunity could be elucidated. Passive immune prophylaxis offers a means to determine the relative role of antibodies in protection against infection. We have studied whether a transfer of antibodies can prevent HIV-2 and SIVsm (SIV of sooty mangabey origin) infection in cynomolgus monkeys. Sera with high antibody titres were collected, heat-treated and injected into naive animals 6 h before challenge with 10-100 monkey-infectious doses of live homologous virus. All control animals treated with normal monkey serum (n = 6) or no serum (n = 39) became infected by the challenge virus, whereas five out of seven animals pretreated with antibody-containing serum at a dose of 9 ml kg-1 resisted infection. Thus passively transferred antibodies can protect against a low-dose lentivirus challenge in a nonhuman primate.

Animals↗
PubMed1991

Hyperimmune antisera against synthetic peptides representing the glycoprotein of human immunodeficiency virus type 2 can mediate neutralization and antibody-dependent cytotoxic activity.

Twenty-five 13- to 35-amino-acid-long peptides representing regions of human immunodeficiency virus type 2 (HIV-2), strain SBL6669, envelope proteins were evaluated for their immunogenic activity in guinea pigs. The peptides were selected to provide homologous representation of sites in the HIV-1 envelope proteins that were previously documented to have a particular immunogenic importance. A number of the HIV-2 peptides were found to be capable of inducing strain SBL6669 neutralizing and antibody-dependent cellular cytotoxicity (ADCC) antibodies. Two overlapping peptides covering amino acids 311-337 representing the central and C-terminal part of the variable third (V3) region, terminology according to Modrow et al. [Modrow, S., Hahn, B., Shaw, G. M., Gallo, R. C., Wong-Staal, F. & Wolf, H. (1987) J. Virol. 61, 570-578], showed the most pronounced capacity to induce neutralizing antibodies. One of the peptides (amino acids 318-337) also induced antibodies mediating ADCC. Two additional regions in the large glycoprotein, gp125, containing linear sites reacting with neutralizing antibodies were identified (amino acids, 119-137 and 472-509). The transmembrane protein, gp36, of HIV-2 harbored two regions of importance for induction of neutralizing antibodies (amino acids 595-614 and 714-729). ADCC activity was induced by two additional gp125-specific peptides (amino acids 291-311 and 446-461). Thus, except for the single V3-specific site there was no correlation between linear immunogenic sites stimulating neutralizing antibody and ADCC activity. These findings pave the way for development of synthetic vaccines against HIV-2 and possibly also simian immunodeficiency virus infections. The capacity of such a product to induce protective immunity can be evaluated in macaque monkeys.

Amino Acid Sequence↗
PubMed1991

HIV reverse transcriptase inhibiting antibodies detected by a new technique: relation to p24 and gp41 antibodies, HIV antigenemia and clinical variables.

A new assay for HIV reverse transcriptase activity inhibiting antibodies (RTI-ab) was used for the analysis of a large collection of sera sampled before and after confirmation of HIV infection. In this assay HIV-RT was preincubated with diluted serum, after which residual RT activity was determined by a technique using a template coupled to macrobeads and 125I-lodo-deoxyuridine-triphosphate as the tracer-substrate. Of the 936 sera analysed, 818 were found positive for RTI-ab, and 824 were positive in Western blot (Wb). The prevalence of RTI-ab compared to Wb was therefore 99.3%. The corresponding figure for 930 sera analysed for envelope-ab, i.e., gp41-ab, was 823 positive, and of these 930 sera 815 were Wb positive, giving a comparative prevalence of 101%. In contrast, only 678 samples of 993 analyzed for core ab, i.e., p24, were positive, giving a prevalence of 77.0% as 880 of these samples were Wb positive. Thus, RTI-ab was as prevalent as gp41-ab, and although the analyses of RTI-ab amounts in different stages showed decreasing levels in stage IV compared to stages II or III, all of the sera except 1 were found positive in stages III and IV. Further, it was found that both the few RTI-ab negative samples in stage II and the few RTI-ab positive samples among Wb negative sera were sampled in connection with seroconversion. The specificity of the RTI-ab assay was 100% in a test of 200 serum samples from HIV negative blood donors. It was concluded that RTI-ab analyses can be made highly sensitive and specific and useful for studies of HIV infection.

AIDS Serodiagnosis↗
PubMed1991

Vaccine protection against HIV-2 infection in cynomolgus monkeys.

The aim of this study was to determine if protection against an infectious human immunodeficiency virus type 2 (HIV-2) challenge could be obtained in cynomolgus macaques by active immunization using whole killed virus vaccine. Four monkeys were immunized with killed HIV-2SBL-6669, two of them with five intramuscular (im) injections of viral preparation containing 100 or 300 micrograms protein emulsified in incomplete Freund's adjuvant (IFA) and the two remaining received four im injections of 25-50 micrograms viral protein in iscoms. Each of the four vaccinated cynomolgus monkeys, along with four unvaccinated controls, were challenged intravenously two weeks after the last booster with approximately 100 animal infectious doses (ID50) of live HIV-2SBL-6669. All four immunized monkeys developed antibodies to HIV-2 envelope and core proteins before challenge exposure to HIV-2, but only the two animals vaccinated with virus in IFA developed detectable neutralizing antibodies. The two monkeys immunized with killed virus in IFA have shown no evidence of infection nine months after challenge with live virus. When blood and lymph node cells from these animals were transfused into naive cynomolgus monkeys, the recipients remained free of infection. In contrast, virus was recovered repeatedly in all nonimmunized animals and in the two animals immunized with iscom-associated viral antigens, which had a low content of envelope gp125 antigen. The demonstration of vaccine-induced protection against HIV-2 in a nonhuman primate raises hope for effective immunization against HIV infections in humans as well.

Animals↗
PubMed1991

A capture enzyme immunoassay for detection of HIV-2/SIV antigen.

An antigen capture enzyme immunoassay was developed for the demonstration of human immunodeficiency virus type 2 (HIV-2) and simian immunodeficiency virus (SIV). The assay (HIV-2 CE) has a sensitivity of approximately 250 pg/ml of HIV-2/SIV antigen. The HIV-2 CE was 4-16 times more sensitive than the Abbott HIV-1 antigen assay for detection of HIV-2/SIV antigen in cell culture supernatant. The sensitivity for detection of HIV-2/SIV antigen in serum or plasma was 98.5% in the HIV-2 CE and 95.5% in the Abbott HIV-1 antigen assay. The specificity of the HIV-2 CE was 99.2% (one false positive among 119 negative sera) whereas the Abbott assay was 100% specific. The HIV-2 CE detected antigen in supernatants from cultures of peripheral blood mononuclear cells of macaque monkeys infected with HIV-2 or SIV about 1 week before a reverse transcriptase (RT) microassay was positive and remained positive for at least 1 week after the RT assay had become negative. Three cultures from persistently infected monkeys were positive only in the HIV-2 CE, reflecting a higher sensitivity compared to the RT microassay. Thus, the HIV-2 CE was more sensitive than the Abbott HIV-1 antigen assay for detection of HIV-2/SIV antigen in culture supernatants as well as in serum/plasma. Furthermore, the HIV-2 CE showed a higher sensitivity than the RT microassay for detection of HIV-2/SIV in cell culture supernatants.

Animals↗
PubMed1990

Infection of cynomolgus monkeys with HIV-2 protects against pathogenic consequences of a subsequent simian immunodeficiency virus infection.

Simian immunodeficiency virus (SIV) infection in cynomolgus macaques leads to severe immunodeficiency with a fatal outcome. In contrast, HIV-2 infects these primates without apparently causing any immunological abnormalities. In this study three cynomolgus monkeys were experimentally infected with HIV-2 strain SBL-K135 and 168 days later challenged with 10-100 animal infectious doses of the closely related SIV strain SM to study protective immunity. At the time of SIV challenge the HIV-2-infected monkeys had neutralizing antibodies against HIV-2, but virus could no longer be recovered from their peripheral blood mononuclear cells (PBMCs) and no clinical symptoms or decrease in CD4+ lymphocytes were observed. Follow-up for 9 months after challenge with SIV showed that the HIV-2-infected monkeys were protected against SIV-induced immunodeficiency (no decrease of CD4+ lymphocytes) and lymphadenopathy. However, they were not resistant to SIV infection since virus could be recovered from their PBMCs and they developed anamnestic antibody responses. Four naive control monkeys which were inoculated with the same dose of SIV became persistently infected and developed a decrease of the absolute numbers of CD4+ cells and showed a marked lymphadenopathy. Two out of four control animals died 58-265 days postinfection with an immunosuppressive disease. Immunohistochemical examination showed abundant viral antigen in lymph-node biopsies from the SIV-infected control monkeys but absence of SIV or HIV-2 antigens in the biopsies from the three HIV-2-preinfected and SIV-superinfected monkeys. The present study demonstrates possibilities for induction of immunity against immunodeficiency induced by a primate lentivirus, a concept with application also to HIV infection and AIDS in man.

Acquired Immunodeficiency Syndrome↗
PubMed1989

Experimental infection of cynomolgus monkeys (Macaca fascicularis) with HIV-2.

Ten healthy adult cynomolgus monkeys (Macaca fascicularis) were inoculated with two different isolates of human immunodeficiency virus type 2 (HIV-2), SBL-6669 and SBL-K135, to establish an animal model for HIV infection. HIV-2SBL-6669 had been propagated for a long time in continuous human cell lines whereas HIV-2SBL-K135 had been grown only in fresh human and monkey lymphocyte cultures or previously for a short time in a continuous cell line. Virus was isolated from three or four animals inoculated with HIV-2SBL-K135 but in none of six monkeys inoculated with HIV-2SBL-6669. All animals seroconverted although the antibody response was higher in SBL-K135 virus-infected monkeys. Varying degrees of lymphadenopathy were observed but there were no significant changes in the numbers of CD4+ and CD8+ T cells. The infection was transferred to two monkeys inoculated with blood from a previously SBL-K135 virus-infected monkey. Four animals inoculated with HIV-2SBL-K135, which had never been propagated in continuous human cell lines, showed strong antibody responses against both gag- and env- encoded proteins of HIV-2. None of the SBL-6669 infected monkeys showed antibodies to core proteins. HIV-2 infection of cynomolgus monkeys represents a useful experimental model for HIV vaccine trials and antiviral testing.

Acquired Immunodeficiency Syndrome↗
PubMed1989

Experimental infection of cynomolgus monkeys (Macaca fascicularis) with simian immunodeficiency virus (SIVsm).

Five healthy cynomolgus monkeys were inoculated intravenously with simian immunodeficiency virus (SIVsm) propagated in human lymphocytes. All five animals became infected. Virus was recovered from blood mononuclear cells and viral antigen was detected in serum 12 days postinoculation (PI) in all inoculated animals. Virus was also isolated in all five animals tested 74 to 226 days PI. Antibodies to different structural proteins of SIV and HIV-2 were demonstrated by ELISA, Western blot, and radioimmunoprecipitation assay from day 31 PI concomitantly with a reduction of viral proteins in the serum. Reappearance of antigen accompanied by a fall in antibody to gag products (p26) was observed in two monkeys 69 days PI. All SIV-infected monkeys showed a pronounced decrease in CD4+ lymphocytes demonstrable already 12 days PI. They also developed persistent lymphadenopathy. Thus, infection of cynomolgus monkeys with SIVsm mimics events in human immunodeficiency virus infection in humans but the course of evolution of pathogenic events in the monkey is markedly compressed. This experimental model will be useful for evaluation of HIV vaccines and antiviral testing.

Acquired Immunodeficiency Syndrome↗
PubMed1989

HIV-2-associated AIDS and HIV-2 seroprevalence in Bissau, Guinea-Bissau.

During a 6-month period in 1987, we examined patients clinically suspected of the acquired immune deficiency syndrome (AIDS) at the national hospital in Bissau, the capital of Guinea-Bissau, and found 20 cases that fulfilled the criteria for AIDS. The two most prevalent major symptoms were weight loss and diarrhea, and the most common minor symptoms generalized lymphadenopathy and generalized dermatitis. Six of 20 patients died within a couple of months. Nineteen of 20 patients were tested for human immunodeficiency virus type 1 (HIV-1) and type 2 (HIV-2) antibodies and were shown to be HIV-2 seropositive. During the same time period, a seroprevalence study of HIV-2 and HIV-1 was carried out, including 2,122 patients or healthy persons in Bissau. Antibodies to HIV-2 were demonstrated by enzyme-linked immunosorbent assay and verified by Western blot analysis in 8.6% (46/535) of prenatal women, in 7.9% (9/114) of women attending a family-planning clinic, in 4.4% (19/427) of applicants for scholarships, in 17.6% (16/91) of blood donors tested during the first 2 months and 5.3% (10/189) of blood donors tested during the following months, in 5.7% (2/35) of police officers, in 36.7% (11/30) of female prostitutes, in 15.8% (97/614) of outpatients suspected of having tuberculosis, and in 55.2% (48/87) of patients clinically suspected of AIDS or AIDS-related disease. One of 2,001 subjects tested had antibodies specific for HIV-1. Another subject had an antibody pattern compatible with both HIV-1 and HIV-2 infections.(ABSTRACT TRUNCATED AT 250 WORDS)

Acquired Immunodeficiency Syndrome↗
PubMed1988

Enzyme immunoassays for the demonstration of antibodies to HIV-2SBL-6669 and HTLV-IV (SIVmac).

Enzyme-linked immunosorbent assays (ELISA) were developed for the demonstration of antibodies to HIV-2 using disrupted virions of the SBL-6669 isolate of HIV-2 and the so-called human T-lymphotropic virus type IV (HTLV-IV), recently found to be identical with the simian immunodeficiency virus (SIVmac), as antigens. Three hundred sera from West African subjects, attending an outward clinic in Bissau for examination of suspected tuberculosis, were tested by these two assays as well as by a commercially available anti-HIV-2 ELISA (ELAVIA II). Fifty of these sera were positive in all three ELISAs as well as in Western blot tests against HTLV-IV. Thirty-eight of these positive sera were also tested by an anti-HIV-2 Western blot kit (LAV-Blot II) with positive results. The ELISAs based on SBL-6669 and HTLV-IV antigens had a specificity of 99.6% (one false positive among 250 negative sera) whereas the specificity of ELAVIA II was 94.6% using the recommended cut-off value and 98.4% using a higher cut-off value. Another 58 sera from West African patients, clinically suspected of having AIDS or HIV-related disease, were tested for HIV-2/HTLV-IV antibodies by Western blot and by ELISA against SBL-6669 and HTLV-IV antigens; all of the 30 sera which were positive by Western blot were found to be positive in both ELISAs.(ABSTRACT TRUNCATED AT 250 WORDS)

Africa, Eastern↗
PubMed1987

Anticardiolipin antibodies in patients with systemic lupus erythematosus.

We studied a group of 59 unselected patients with systemic lupus erythematosus (SLE); these patients were from a defined population who lived in southern Sweden. We found that serum concentrations of anticardiolipin antibodies were increased in 32 SLE patients (54.2%). No significant correlation between increased amounts of anticardiolipin antibodies and clinical symptoms, such as thrombocytopenia or thrombosis, was found. Serial serum samples from 28 patients (12 patients were from the epidemiologic cohort) were analyzed. Sixteen of these 28 patients (57.1%) had increased levels of anticardiolipin antibodies; in most cases, there was no variation in these values with regard to clinical disease flares or treatment. Increased concentrations of anticardiolipin were observed in 4 patients with cerebral infarction. However, very high concentrations of anticardiolipin antibodies were observed in several patients with inactive SLE who had no history of thrombosis or thrombocytopenia. Our results underscore the importance of studying unselected patient groups when correlating laboratory data with clinical manifestations of disease.

Adult↗
PubMed1985

Preparation of highly purified F-actin-depolymerizing factor of human serum.

F-actin depolymerizing factor (ADF) of human serum was purified 250-400-fold to more than 98% purity with high reproducibility. The purification included (1) 30-50% (NH4)2SO4 precipitation, (2) ion-exchange chromatography on DEAE-Sepharose, (3) chromatofocusing on Polybuffer exchanger 94 and (4) affinity chromatography on ConA-Sepharose. The recovery of ADF was estimated to be 20-30% whereas the ADF activity yield was 5-17%. The lower activity yield was thought to be due-partly to proteolysis and partly to destabilization of highly purified ADF.

Actin Depolymerizing Factors↗
PubMed1979

F-Actin-depolymerizing activity of human serum.

Non-heated human and animal sera contain a factor which exhibited an inhibiting activity on the staining of actin-containing structures by anti-actin antibodies in indirect immunofluorescence experiments. The presence of this factor lowered the viscosity of F-actin preparations and caused, as studied by electron-microscopy, a depolymerization of F-actin filaments as well as inhibition of filament formation of G-actin. The factor was, after its reaction with F-actin, liberated seemingly unaffected, indicating an enzymatic activity. The factor tentatively termed 'F-actin depolymerizing factor' was heat-sensitive and trypsin sensitive but resisted reduction. It was Ca2+ dependent and the staining inhibiting reaction was faster at 30 degrees C and 37 degrees C than at lower temperatures. Gel filtration experiments on Sephadex G-200 suggested a molecular size of the actin depolymerizing factor slightly higher than that of albumin. The electrophoretic mobility was that of gamma 2 globulin. The physiological role of the factor might be to prevent the presence of F-actin filaments within the circulation.

Actins↗
PubMed1977

The reaction of cells with anti-actin sera in relation to the amount of cellular actin.

The staining pattern of anti-actin sera on various cells smeared on glass was compared to the relative amount of cellular actin estimated by SDS-polyacrylamide-gel electrophoresis with subsequent scanning of the gel. Although the cells showed a varying stainability the actin content was fairly constant. Thus, the staining differences reflected changes in the organization of cellular actin rather than actual differences in the amount of actin.

Actins↗
PubMedDate not supplied

A formalin inactivated whole SIVmac vaccine in Ribi adjuvant protects against homologous and heterologous SIV challenge.

Eight monkeys were immunized at 0, 4, 9, and 18 weeks with a total of 2 mg of formalin inactivated SIVmac vaccine with Ribi adjuvant. Two weeks after the last booster four immunized monkeys and two controls were challenged with 10 MID50 of live homologous virus SIVmac, and the remaining four vaccinated animals along with two controls were challenged with the heterologous SIVsm strain. All eight vaccinated monkeys resisted the virus challenge, whereas all controls became infected. Three months after the first challenge the monkeys were rechallenged with the same virus strain, without further boosting. Two of four vaccinated monkeys were still resistant to the homologous SIV strain, and three of four monkeys were resistant to the heterologous SIVsm strain. This study demonstrates vaccine induced cross-protection between SIV strains.

Adjuvants, Immunologic↗