http://repub.eur.nl/res/aut/2032/ List of Publications en http://repub.eur.nl/static-eur/img/logo.png http://repub.eur.nl http://repub.eur.nl/res/pub/3839/ 2001-05-14T00:00:00Z Rinderpest virus (RPV), a member of genus Morbillivirus in the family Paramyxoviridae, causes an acute and often fatal disease in cattle and other large ruminants. A subunit rinderpest vaccine consisting of an immune-stimulating complex (ISCOM) incorporating the RPV haemaggulutinin (H) protein, was examined for its ability to induce protective immunity in cattle, the natural host of RPV. All of four cattle vaccinated with the ISCOM vaccine survived challenge with virulent virus. Three were solidly protected, showing no clinical signs of infection, while the fourth animal developed only mild and transient symptoms. Virus neutralizing antibodies were produced at a significant level in all vaccinated cattle. These results indicate that this ISCOM vaccine is effective in producing protective immunity in cattle and should be a suitable means of delivering glycoprotein antigens from other morbilliviruses. http://repub.eur.nl/res/pub/3695/ 1999-12-01T00:00:00Z The evolution of simian immunodeficiency virus (SIV)–specific cytotoxic T lymphocyte precursors (CTLps) and their relationship with virus replication were studied in SIV‐infected macaques. After primary viremia, 3 of 8 macaques lost culturable virus and polymerase chain reaction–detectable provirus in peripheral blood. Although proviral DNA persisted in the spleen and lymph nodes, virus loads were below or barely above detection levels. Throughout the study, the 3 macaques remained asymptomatic, with stable CD4+ cell counts. These findings were associated with the detection of CTLps directed against both structural and regulatory SIV proteins. The response peaked during the first 7 months of infection but waned subsequently. CTLps increased after rechallenge of 1 macaque, suggesting that limited antigenic stimulation contributed to their disappearance from circulation. Transient viremia with increasing CTLp frequencies and antibody titers also suggested at least partial susceptibility to reinfection. These findings bear implications for vaccination strategies aimed at inducing protective CTLs against lentiviruses. http://repub.eur.nl/res/pub/3681/ 1999-06-04T00:00:00Z Intro: More than 15 years after the discovery of HIV-1 as the causative agent of AIDS, and numerous attempts to develop a vaccine, it has become clear that the efficacy of the currently considered HIV-1 vaccine candidates will generally be limited. This is at least in part due to the relative resistance of so-called primary HIV strains to neutralization by HIV-1 envelope specific antibodies: even the most potent HIV-1 neutralizing antibodies failed to provide protection in in vivo models, at concentrations that can be maintained for longer periods in human vaccinees. http://repub.eur.nl/res/pub/3641/ 1998-05-01T00:00:00Z The development of a safe and effective vaccine for the prevention of AIDS has thus far proven to be extremely difficult, at least in part due to complexities associated with HIV-1 and its pathogenesis. The recent description of individuals transiently infected with HIV-1, as well as persons who survived HIV-1 infection for more than 15 years, indicates the ability of the immune response of certain individuals to control HIV-1 infection. Moreover, vaccination-challenge experiments in macaques infected with simian immunodeficiency virus have shown that protection against infection or development of disease may be achieved in the absence of sterilizing immunity, suggesting that the goals for AIDS vaccine development may have to be redefined. In addition, evaluation of new lentivirus vaccine strategies may largely benefit from the use of the newly developed chimeric simian-human immunodeficiency viruses, allowing the testing of HIV-1 antigen based vaccines in macaques. http://repub.eur.nl/res/pub/8467/ 1998-01-01T00:00:00Z http://repub.eur.nl/res/pub/3619/ 1997-12-01T00:00:00Z The efficiency of paraformaldehyde (PFA) and binary ethylenimine (BEI) in inactivating recombinant vaccinia virus (rVV), present in baby hamster kidney cells expressing simian immunodeficiency virus envelope glycoproteins (SIV-Env), was measured in a series of inactivation studies. Both compounds were shown to be effective in reducing rVV titres. The use of standard 3-day titration assays proved to be inadequate to measure PFA inactivation, since upon prolonged incubation, residual rVV infectivity was detected in cultures negative at 3 days. Different procedures using PFA or BEI were selected to assess their influence on the antigenicity and immunogenicity or rVV expressed SIV-Env. Antigenicity, as defined by the ability to react with a panel of monoclonal antibodies recognizing major antigenic sites, and immunogenicity, as defined by the ability to induce SIV envelope specific and virus neutralizing serum antibodies in rats, proved to be preserved after either inactivation procedure. These data show that both protocols using PFA or BEI can be used successfully as part of the procedures to remove residual rVV infectivity. http://repub.eur.nl/res/pub/18557/ 1997-11-05T00:00:00Z Since its discovery about 15 years ago, human immunodeficiency virus type 1 (HtV-1) continues·to spread at an alarming rate. It is estimated that by the year 2000 between 30 and 40 million people will be infected with this virus worldwide'. Of these, about 90 % live in developing countries. To date, more than six million people have already developed to the fatal acquired immunodeficiency syndrome (AIDS). Despite numerous research efforts no vaccination or generally accessible therapeutic approach has become available yet. In 1981, the first case of AIDS was recognized'. Epidemiologic studies implicated an infectious agent and in 1983, the isolation of a previously unknown human retrovirus called lymphadenopathy-associated virus (LA V) from an AIDS patient was reported by Barre-Sinoussi et al.'. In 1984, Gallo et aI' and Levy et a/s also reported the isolation of retroviruses from AIDS patients which they called human T-Iymphotropic virus type III (HTLV-III) and AIDS-associated retrovirus (ARV), respectively. Soon thereafter, these retroviruses were recognized as members of the Lentivirinae, a subfamily of the Retroviridae, and were indeed identified as the etiological agents of AIDS. In 1986, these retroviruses were named human immunodeficiency virus (HIV). Today, two types of HIV are recognized, HIV-1 and HIV-2. Of these, HIV-1 is the primary etiologic agent of the current AIDS pandemic. The search for an HIV-1 counterpart in other primate species led to the identification of several simian immunodeficiency viruses (SIV), which may cause AIDS like syndromes in monkeys'. Currently, macaque lentivirus infections are considered the most appropriate animal models to study HIV pathogenesis and possible intervention strategies. In the following section, a concise overview is given of the biology of primate lentiviruses, including the host immune response and correlates of immune-mediated protection. Aspects important for vaccine development are highlighted. http://repub.eur.nl/res/pub/3595/ 1997-01-01T00:00:00Z A detailed analysis of simian immunodeficiency virus (SIV)-specific cytotoxic T lymphocyte (CTL) responses and the identification of the proteins and epitopes they target may improve the design of immunotherapeutic interventions and provide insights into AIDS pathogenesis. Here, we identified a new CTL epitope in the SIV Gag protein, recognized by CD8+ and MHC class I-restricted CTL clones from a long-term asymptomatic cynomolgus macaque (Macaca fascicularis) infected with SIVmac32H-J5. Using overlapping synthetic peptides, the optimal minimal epitope was characterized as a nine amino acid peptide representing amino acids 242-250 of p26 (SVDEQIQWM). CTL recognition was shown to be abolished by amino acid substitutions observed within homologous human immunodeficiency virus (HIV)-1 and HIV-2 sequences. http://repub.eur.nl/res/pub/3607/ 1997-01-01T00:00:00Z Detailed analyses of simian immunodeficiency virus (SIV)-specific cytotoxic T lymphocyte (CTL) responses in vaccinated and infected macaques may help to clarify the role of CTL immunity in protection against lentiviruses. Here, the optimal conditions for the measurement of SIV Gag-specific CTL were investigated by bulk and limiting dilution assays of peripheral blood mononuclear cells (PBMC) from naive and vaccinated cynomolgus macaques (Macaca fascicularis) infected with SIVmac32H(J5). In vitro restimulation was generally required for CTL detection. Selective activation of CD8+ and MHC-restricted SIV Gag-specific CTL was induced by stimulation with autologous para-formaldehyde-fixed B-lymphoblastoid cell lines infected with a recombinant vaccinia virus expressing SIV Gag. Applied to limiting dilution assays, antigenic stimulation reproducibly demonstrated SIV Gag-specific CTL precursors (CTLp) in PBMC of all animals studied, including those lacking significant responses in standard bulk CTL assays. http://repub.eur.nl/res/pub/3547/ 1995-01-01T00:00:00Z Shortly after infection of two rhesus monkeys (Macaca mulatta) either with a SIVmac32H challenge stock or with the same virus that had been passaged in another rhesus monkey for 11 months, SIV-envelope genes were cloned from their peripheral blood mononuclear cells and subsequently expressed by recombinant vaccinia viruses. The molecular weights and antigenicities of the thus produced envelope glycoproteins were largely identical to those of the native SIV. The envelope glycoprotein derived from the in vivo passaged virus proved to be poorly recognized by virus neutralizing monoclonal antibodies directed against one of the seven antigenic sites for which monoclonal antibodies were available. Immunization studies in rats showed that this protein was also less efficient in inducing antibodies against this antigenic site, and that it induced significantly lower levels of virus neutralizing antibodies than the other SIV-envelope glycoprotein. The immunogenicity of the SIV-envelope glycoprotein incorporated into immune stimulating complexes (iscoms) was compared to that of the same protein presented with Quil A or MDP-tsl. http://repub.eur.nl/res/pub/3552/ 1995-01-01T00:00:00Z To gain further insight into the ability of subunit vaccines to protect monkeys from experimental infection with simian immunodeficiency virus (SIV), two groups of cynomolgus macaques were immunized with either recombinant SIVmac32H-derived envelope glycoproteins (Env) incorporated into immune-stimulating complexes (iscoms) (group A) or with these SIV Env iscoms in combination with p27gag iscoms and three Nef lipopeptides (group B). Four monkeys immunized with recombinant feline immunodeficiency virus Env iscoms served as controls (group C). Animals were immunized intramuscularly at weeks 0, 4, 10, and 16. Two weeks after the last immunization, monkeys were challenged intravenously with 50 monkey 50% infectious doses of virus derived from the J5 molecular clone of SIVmac32H propagated in monkey peripheral blood mononuclear cells. High titers of SIV-neutralizing antibodies were induced in the monkeys of groups A and B. In addition, p27gag-specific antibodies were detected in the monkeys of group B. Vaccine-induced cytotoxic-T-lymphocyte precursors against Env, Gag, and Nef were detected on the day of challenge in the monkeys of group B. Env-specific cytotoxic-T-lymphocyte precursors were detected in one monkey from group A. In spite of the observed antibody and T-cell responses, none of the monkeys was protected from experimental infection. In addition, longitudinal determination of cell-associated virus loads at weeks 2, 4, 6, 9, and 12 postchallenge revealed no significant differences between vaccinated and control monkeys. These findings illustrate the need to clarify the roles of the different arms of the immune system in conferring protection against primate lentivirus infections. http://repub.eur.nl/res/pub/3510/ 1994-11-11T00:00:00Z The immunogenicity and efficacy of an inactivated whole SIVmac (32H) preparation adjuvanted with muramyl dipeptide (SIV-MDP) and a gp120-enriched SIVmac (32H) ISCOM preparation (SIV-ISCOM), were compared by immunizing four rhesus macaques (Macaca mulatta) four times with SIV-MDP and four others in the same way with SIV-ISCOM. Two monkeys immunized with whole inactivated measles virus (MV) adjuvanted with MDP (MV-MDP) and two monkeys immunized with MV-ISCOM served as controls. In the SIV-ISCOM-immunized monkeys higher SIV-specific serum antibody titres were found than in the SIV-MDP-immunized monkeys. In contrast to the MV-immunized monkeys all SIV-MDP- and SIV-ISCOM-immunized monkeys were protected against intravenous challenge 2 weeks after the last immunization with 10 median monkey infectious doses (MID50) of a cell-free SIVmac (32H) challenge stock propagated in the human T-cell line C8166. After 43 weeks the protected monkeys were reboosted and 2 weeks later rechallenged with 10 MID50 of the same virus produced in peripheral blood mononuclear cells (PBMC) from a rhesus macaque. None of these animals proved to be protected against this challenge. In a parallel experiment in which the same numbers of monkeys were immunized in the same way, the animals were challenged intravenously with 10 MID50 of PBMC from an SIVmac (32H)-infected rhesus macaque. Two out of four SIV-MDP- and two out of four SIV-ISCOM-immunized monkeys proved to be protected from SIV infection.