RESUMEN
Porcine endogenous retroviruses (PERVs) represent a risk for xenotransplantation using pig cells or organs since they are integrated in the genome of all pigs and infect human cells in vitro. Recombinants between PERV-A and PERV-C have been described in pigs in vivo and found de novo integrated in the genome of somatic cells, but not in the germ line. To study whether PERV-A/C can infect and have a pathogenic effect in normal pigs, German landrace pigs were inoculated with high-titre PERV-A/C. No provirus integration was found in blood cells or in various tissues, and no antibody production was observed, indicating the absence of infection.
Asunto(s)
Retrovirus Endógenos/genética , Retrovirus Endógenos/patogenicidad , Recombinación Genética , Infecciones por Retroviridae/veterinaria , Enfermedades de los Porcinos/virología , Animales , Provirus/genética , Provirus/aislamiento & purificación , Infecciones por Retroviridae/virología , Porcinos , Enfermedades de los Porcinos/patología , Integración ViralRESUMEN
Herpes B virus (BV) infection of macaques persists in the natural host, but is mainly asymptomatic. However, BV can cause fatal disease in humans and in several non-macaque species such as capuchin monkeys (Cebus apella). The BV infection described here in a colony of capuchin monkeys was persistent but asymptomatic. Initially the infection was detected serologically in five out of seven animals. However, using polymerase chain reaction (PCR) developed specifically for BV, we found the virus in all seven clinically healthy animals. It is probable that the infection was transferred from BV-infected macaques housed in different cages but in the same room for several years. We have no evidence to indicate that similar asymptomatic infections may occur in other New World species but the possibility should not be discounted. We recommend that the housing of capuchin monkeys in close proximity to macaques should be avoided and that greater caution should be used when handling capuchin monkeys and possibly other New World species that have been in contact with macaques. All may act as a source of BV infection in humans, hence routine, repeated testing of all primates is essential.
Asunto(s)
Cebus , Infecciones por Herpesviridae/veterinaria , Herpesvirus Cercopitecino 1/crecimiento & desarrollo , Macaca , Enfermedades de los Monos/virología , Técnicos de Animales , Animales , Anticuerpos Antivirales/sangre , ADN Viral/química , ADN Viral/genética , Femenino , Infecciones por Herpesviridae/transmisión , Infecciones por Herpesviridae/virología , Herpesvirus Cercopitecino 1/genética , Inmunohistoquímica/veterinaria , Masculino , Enfermedades de los Monos/transmisión , Reacción en Cadena de la Polimerasa/veterinaria , Análisis de Secuencia de ADNRESUMEN
In primates, little has been reported about malignant mesenchymal uterine tumours. A case of a spontaneous metastasising uterine sarcoma in a 17-year-old rhesus monkey is presented. Clinically, transient abdominal pain, spasms, nausea, anaemia, a firm uterus and bloody vaginal discharge were noted. In a diagnostic laparoscopy, both massive adhesions in the lesser pelvis and 10 ml of ascites fluid were detected. In necropsy, in addition to peritonitis with massive adhesions, a cauliflower-shaped, irregular, tough, greyish-white uterine tumour was seen. Two cherry-sized tumour metastases were noticed in the greater omentum. In histology, both in the uterus and the metastases, a sarcoma with a low amount of connective tissue and well-differentiated cell nuclei was identified.
Asunto(s)
Macaca mulatta , Enfermedades de los Monos/diagnóstico , Epiplón/patología , Neoplasias Peritoneales/veterinaria , Sarcoma/veterinaria , Neoplasias Uterinas/veterinaria , Animales , Diagnóstico Diferencial , Femenino , Inmunohistoquímica/veterinaria , Enfermedades de los Monos/patología , Neoplasias Peritoneales/diagnóstico , Neoplasias Peritoneales/secundario , Sarcoma/diagnóstico , Sarcoma/secundario , Sarcoma/ultraestructura , Neoplasias Uterinas/diagnóstico , Neoplasias Uterinas/patología , Neoplasias Uterinas/ultraestructuraRESUMEN
For xenotransplantation, the transplantation of animal cells, tissues and organs into human recipients, to date, pigs are favored as potential donors. Beside ethical, immunological, physiological and technical problems, the microbiological safety of the xenograft has to be guaranteed. It will be possible to eliminate all of the known porcine microorgansims in the nearby future by vaccinating or specified pathogen-free breeding. Thus, the main risk will come from the porcine endogenous retroviruses (PERVs) which are present in the pig genome as proviruses of different subtypes. PERVs will therefore be transmitted, with the xenograft, to the human recipient. PERVs can infect numerous different types of human primary cells and cell lines in vitro and were shown to adapt to these cells by serial passaging on uninfected cells. Furthermore, PERVs have high homology to other retroviruses, such as feline leukemia virus (FeLV) or murine leukemia virus (MuLV), which are known to induce tumors or immunodeficiencies in the infected host. To evaluate the potential risk of a trans-species transmission of PERV in vivo, naive and immunosuppressed rats, guinea pigs and minks were inoculated with PERV and screened over a period of 3 months for an antibody reaction against PERV proteins or for the integration of proviral DNA into the genomic DNA of the host's cells. Furthermore, we inoculated three different species of non-human primates, rhesus monkey (Macaca mulatta), pig-tailed monkey (Macaca nemestrina) and baboon (Papio hamadryas) with high titers of a human-adapted PERV. To simulate a situation in xenotransplantation, the animals received a daily triple immunosuppression using cyclosporine A, methylprednisolone and RAD, a rapamycin derivative, presently under development by Novartis. None of the small laboratory animals or the non-human primates showed production of antibodies against PERV or evidence of integration of proviral DNA in blood cells or cells of several organs, 3 months after virus inoculation, despite the observation that cells of the animals used in the experiment were infectible in vitro. This apparent difference in the outcome of the in vitro and the in vivo data might be explained by an efficient elimination of the virus by the innate or adaptive immunity of the animals.
Asunto(s)
Retrovirus Endógenos/patogenicidad , Porcinos/virología , Trasplante Heterólogo/efectos adversos , Animales , Línea Celular , Femenino , Cobayas , Humanos , Macaca mulatta , Masculino , Visón , Modelos Animales , Ratas , Seguridad , Trasplante Heterólogo/inmunología , Integración ViralRESUMEN
Porcine endogenous retroviruses (PERVs) are considered a special risk for xenotransplantation because they are an integral part of the porcine genome and are able to infect cells of numerous species including humans in vitro. Among these cells, the mink lung epithelial cell line Mv1Lu could be productively infected with PERV. Provirus integration was detected by PCR, expression of viral proteins was shown by immunostaining and reverse transcriptase was detected in cell supernatants. PERV produced from mink cells could infect both, uninfected mink Mv1Lu cells and uninfected human 293 cells, with considerably higher virus production by human cells. Typical type C retroviruses were observed in PERV-infected mink cells using electron microscopy together with numerous multivesicular body (MVB)-like structures containing small virus-like particles, not present in uninfected mink cells. These MVBs could be stained with PERV-specific serum. In an attempt to establish a small animal model, PERV grown on mink cells was inoculated into adult and newborn American minks. Neither antibody production against PERV nor integration of viral DNA or production of viral proteins in tissues of different organs could be detected 12 weeks post virus inoculation, indicating that PERV infection had not occurred.
Asunto(s)
Retrovirus Endógenos/fisiología , Retrovirus Endógenos/patogenicidad , Visón/virología , Infecciones por Retroviridae/transmisión , Porcinos/virología , Animales , Western Blotting , Línea Celular , Humanos , Riñón/citología , Pulmón/citología , Microscopía Electrónica , Infecciones por Retroviridae/virologíaRESUMEN
Several cases of hydatid echinococcosis were diagnosed in a laboratory colony of 19 pig-tailed macaques (Macaca nemestrina) at the Paul Ehrlich Institute, Germany. Three hydatid cysts were found in the liver of an euthanized animal. The diagnosis of an Echinococcus granulosus infection was confirmed by histopathology and the results of a specific polymerase chain reaction. The serum of five of 14 other monkeys tested for Echinococcus antibodies using a genus-specific enzyme-linked immunosorbent assay (ELISA) was positive or weakly positive; none of the animals, however, showed specific reactions in a E. multilocularis-specific ELISA. On ultrasonographic examination, alterations in the liver were found in four of the serologically positive monkeys, and two animals showed clinical signs such as progressive anorexia, apathy and icterus. The monkeys had most probably acquired the E. granulosus infection in their breeding colony in Slovenia.
Asunto(s)
Equinococosis Hepática/veterinaria , Echinococcus/aislamiento & purificación , Macaca nemestrina/parasitología , Enfermedades de los Monos/diagnóstico , Animales , Anticuerpos Antihelmínticos/sangre , ADN de Helmintos/análisis , Equinococosis Hepática/diagnóstico , Equinococosis Hepática/inmunología , Echinococcus/genética , Echinococcus/inmunología , Ensayo de Inmunoadsorción Enzimática/veterinaria , Femenino , Hígado/parasitología , Hígado/patología , Masculino , Enfermedades de los Monos/inmunología , Enfermedades de los Monos/parasitología , Reacción en Cadena de la Polimerasa/veterinariaRESUMEN
We used multidirectional chromosome painting with probes derived by bivariate fluorescence-activated flow sorting of chromosomes from human, black lemur (Eulemur macaco macaco) and tree shrew (Tupaia belangeri, order Scandentia) to better define the karyological relationship of tree shrews and primates. An assumed close relationship between tree shrews and primates also assists in the reconstruction of the ancestral primate karyotype taking the tree shrew as an "outgroup" species. The results indicate that T. belangeri has a highly derived karyotype. Tandem fusions or fissions of chromosomal segments seem to be the predominant mechanism in the evolution of this tree shrew karyotype. The 22 human autosomal painting probes delineated 40 different segments, which is in the range found in most mammals analyzed by chromosome painting up to now. There were no reciprocal translocations that would distinguish the karyotype of the tree shrew from an assumed primitive primate karyotype. This karyotype would have included the chromosomal forms 1a, 1b, 2a, 2b, 3/21, 4-11, 12a/22a, 12b/22b, 13, 14/15, 16a, 16b, 17, 18, 19a, 19b, 20 and X and Y and had a diploid chromosome number of 2n=50. Of these forms, chromosomes 1a, 1b, 4, 8, 12a/22a, and 12b/22b may be common derived characters that would link the tree shrew with primates. To define the exact phylogenetic relationships of the tree shrews and the genomic rearrangements that gave rise to the primates and eventually to humans further chromosome painting in Rodentia, Lagomorpha, Dermoptera and Chiroptera is needed, but many of the landmarks of genomic evolution are now known.
Asunto(s)
Pintura Cromosómica/métodos , Cromosomas Humanos/genética , Lemur/genética , Musarañas/genética , Animales , Sondas de ADN , Femenino , Humanos , Procesamiento de Imagen Asistido por Computador , Hibridación in Situ/métodos , Cariotipificación , Masculino , Metafase/genética , Primates/genética , Especificidad de la Especie , Cromosoma YRESUMEN
We used reciprocal chromosome painting with both African green monkey (C. aethiops) and human chromosome specific DNA probes to delineate homologous regions in the two species. Probes were derived by fluorescence-activated chromosome flow sorting and then were reciprocally hybridized to metaphase spreads of each species. Segments in the size range of a single chromosome band were identified, demonstrating the sensitivity of the approach when comparing species that diverged more than 20 million years ago. Outgroup analysis shows that the great difference in diploid numbers between the African green monkey (2n = 60) and humans (2n = 46) is mainly owing to fissions, and the direction of change is towards increasing diploid numbers. However, most break points apparently lie outside of the centromere regions, suggesting that the changes were not solely Robertsonian as has been previously assumed. No reciprocal translocations have occurred in the phylogenetic lines leading to humans or African green monkeys. The primate paints established here are a valuable tool to establish interspecies homology, to define rearrangements, and to determine the mechanisms of chromosomal evolution in primate species.
Asunto(s)
Chlorocebus aethiops/genética , Pintura Cromosómica/métodos , Diploidia , Animales , Secuencia de Bases , Mapeo Cromosómico , Cartilla de ADN , Sondas de ADN , Ligamiento Genético , Humanos , Cariotipificación , FilogeniaRESUMEN
Several studies have demonstrated that newborn animals are more susceptible to disease development following infection with retroviruses than adults. Adult African green monkeys (AGMs) infected with SIVagm do not develop AIDS-like disease and the objective of the study was to determine whether experimental infection of newborn AGMs with SIVagm would result in pathogenesis. Neonatal AGMs were found to have a higher percentage of circulating CD4+ lymphocytes than adults (62% versus 14%) and therefore a higher potential pool of target cells for SIVagm infection. However, no differences in the in vitro replication kinetics of SIVagm in peripheral blood mononuclear cells of adult or neonatal AGMs could be observed. In vivo, the neonatal AGMs became viremic at the earliest two months after inoculation whereas the adult AGMs had evidence of virus replication already 2 to 6 weeks after infection. None of the animals developed AIDS-like symptoms upon infection. In the heterologous cynomolgus macaque host, a newborn infected with SIVagm developed early high virus loads and died two months after birth with AIDS-like histopathologic features. It would therefore appear that in contrast to the situation with many other retroviruses, newborn AGMs are no more permissive to SIVagm infection than are adults.
Asunto(s)
Animales Recién Nacidos , Chlorocebus aethiops , Síndrome de Inmunodeficiencia Adquirida del Simio/inmunología , Virus de la Inmunodeficiencia de los Simios/patogenicidad , Viremia/inmunología , Envejecimiento/inmunología , Animales , Anticuerpos Antivirales/biosíntesis , Linfocitos B/citología , Recuento de Linfocito CD4 , Relación CD4-CD8 , Linfocitos T CD8-positivos/citología , Línea Celular , Células Cultivadas , Progresión de la Enfermedad , Citometría de Flujo , Inmunidad Innata , Hibridación in Situ , Leucocitos Mononucleares/virología , Recuento de Linfocitos , Tejido Linfoide/patología , Tejido Linfoide/virología , Síndrome de Inmunodeficiencia Adquirida del Simio/virología , Virus de la Inmunodeficiencia de los Simios/inmunología , Virus de la Inmunodeficiencia de los Simios/fisiología , Viremia/virología , Replicación ViralRESUMEN
Two chimeric proviruses comprising the U3 promoter and the nef gene of simian immunodeficiency virus (SIV) smmPBj1.9 in addition to other genomic regions of SIVagm3mc from African green monkeys (Cercopithecus aethiops) were constructed. The derived chimeric viruses (SIVagm3mc/SIVsmmPBj1.9) were both able to replicate in nonstimulated peripheral blood leukocytes from pig-tailed macaques (Macaca nemestrina), a biological property often correlated with acute pathogenicity. However, only one of the chimeric viruses was acutely pathogenic, inducing a rapid depletion of the peripheral CD4+ T cells in two infected pig-tailed macaques within 10 days after infection in a manner similar to infection with SIVsmmPBj1.9 itself. The other chimeric virus actively replicated during the first 8 weeks after experimental infection of two pig-tailed macaques but induced neither acute disease nor CD4+ T-cell depletion for 113 weeks after infection. Thus, the U3 promoter and the nef gene of SIVsmmPBj1.9 alone appear to be insufficient to confer acute pathogenicity to SIVagm3mc.
Asunto(s)
Genes nef , Regiones Promotoras Genéticas , Síndrome de Inmunodeficiencia Adquirida del Simio/virología , Virus de la Inmunodeficiencia de los Simios/genética , Virus de la Inmunodeficiencia de los Simios/patogenicidad , Animales , Linfocitos T CD4-Positivos/citología , Linfocitos T CD8-positivos/citología , Chlorocebus aethiops/virología , Depleción Linfocítica , Macaca nemestrina , Mutagénesis , Síndrome de Inmunodeficiencia Adquirida del Simio/inmunología , Virus de la Inmunodeficiencia de los Simios/inmunologíaRESUMEN
Microglia are the major target for human immunodeficiency virus (HIV) infection within the central nervous system. Because only a few cells are productively infected, it has been suggested that an aberrant cytokine production by this cell population may be an indirect mechanism leading to the development of neurological disorders in HIV-infected patients. Therefore we decided to study the secretion pattern of several interleukins (IL) by microglial cells and peripheral blood macrophages isolated from uninfected and simian immunodeficiency virus (SIV)-infected Rhesus monkeys. We found that uninfected, unstimulated primate microglia produce more IL-6 and less TNF alpha than peripheral blood macrophages, but generate comparable levels of IL-1 beta and IL-8. After infection with SIV in vitro, synthesis of all cytokines tested is increased compared to uninfected cultures and to peripheral blood macrophages. Microglia isolated from infected animals produce more IL-8 and TNF alpha than the uninfected cultures and display a strongly increased capacity to secrete TNF alpha upon stimulation with lipopolysaccharide. In addition, production of IL-6 by in vivo-infected microglia increases with time in culture to very high levels despite the fact that only a few cells contained replicating virus. These findings clearly show that the cytokine production of microglia is impaired after SIV infection both in vitro and in vivo and that a low level of viral replication is sufficient for these alterations to occur. In conclusion, the results of this study further support a possible role of cytokines in the pathogenesis of neuro-AIDS.
Asunto(s)
Citocinas/metabolismo , Macrófagos/inmunología , Microglía/inmunología , Síndrome de Inmunodeficiencia Adquirida del Simio/inmunología , Virus de la Inmunodeficiencia de los Simios/inmunología , Animales , Células Cultivadas , Citocinas/biosíntesis , Humanos , Interleucina-1/biosíntesis , Interleucina-1/metabolismo , Interleucina-6/biosíntesis , Interleucina-6/metabolismo , Interleucina-8/biosíntesis , Interleucina-8/metabolismo , Macaca mulatta , Macrófagos/metabolismo , Microglía/metabolismo , Factor de Necrosis Tumoral alfa/biosíntesis , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
To clarify the pathogenesis of SIV-induced thymus atrophy, the presence of SIV within thymus stromal cell cultures (epithelial cells, IDC, macrophages or fibroblasts) was investigated. The material studied consisted of 15 thymus specimens of rhesus macaques infected with SIVmac251 (2-4 months postinoculation). No viral antigen was detected, either in the cultures, by immunohistochemistry, or in cell culture supernatants, by ELISA (p17 antigen), and no viral RNA was detected by in situ hybridization. Only after coculture with the C8166 cell line, was virus detected in 2 out of 15 stroma cultures. The fact that the virus could only be detected after several passages of coculture with the C8166 cell line indicates that the virus exists in the thymus stroma cells in the form of proviral DNA. The infection of thymus stromal cells may contribute to the destruction of the thymus microenvironment and to the SIV-induced thymus atrophy.
Asunto(s)
Síndrome de Inmunodeficiencia Adquirida del Simio/virología , Virus de la Inmunodeficiencia de los Simios/aislamiento & purificación , Timo/virología , Animales , Atrofia , Muerte Celular , Línea Celular , Células Cultivadas , Macaca mulatta , Provirus/aislamiento & purificación , Síndrome de Inmunodeficiencia Adquirida del Simio/etiología , Síndrome de Inmunodeficiencia Adquirida del Simio/patología , Virus de la Inmunodeficiencia de los Simios/patogenicidad , Linfocitos T/patología , Linfocitos T/virología , Timo/patologíaRESUMEN
The absence of AIDS-like symptoms in HIV-infected chimpanzees and SIV-infected African Green monkeys (AGMs) may provide important clues about the pathogenic mechanism of AIDS and about mechanisms of resistance. HIV-infected persons and SIV-infected rhesus macaques have, on the average, markedly decreased cysteine, cystine, and glutathione levels and elevated plasma glutamate concentrations. Glutamate inhibits the membrane transport of cystine and a combination of low plasma glutamate and high cystine levels was found to be correlated with high CD4+ T cell numbers even in HIV-negative healthy human individuals. We have now found that glutamate and cystine levels are also correlated with CD4+ T cell numbers in chimpanzees. But infection of chimpanzees, AGMs, and goats with HIV-1, SIV, and caprine arthritis encephalitis virus (CAEV), respectively, does not induce significant changes in plasma cystine or glutamate levels, although infected AGMs and goats have, on the average, significantly elevated plasma levels of the biochemically related amino acid proline.
Asunto(s)
Aminoácidos/sangre , Infecciones por Lentivirus/sangre , Animales , Virus de la Artritis-Encefalitis Caprina , Chlorocebus aethiops , Cisteína/sangre , Cistina/sangre , Glutamatos/sangre , Ácido Glutámico , Enfermedades de las Cabras/sangre , Cabras , Infecciones por VIH/sangre , VIH-1 , Hepatitis B/sangre , Humanos , Infecciones por Lentivirus/veterinaria , Macaca mulatta , Pan troglodytes , Prolina/sangre , Síndrome de Inmunodeficiencia Adquirida del Simio/sangreRESUMEN
A spontaneous Yersinia pseudotuberculosis (Y.p.) infection in one African Green Monkey (Cercopithecus aethiops), and nine Squirrel Monkeys (Saimiri sciureus), is reported. The clinical findings, laboratory results and pathological findings are presented. The digestive system was the most affected organ in the acute phase of the Y.p. infection, while in the subacute and chronic phases, the alterations of the lymphatic tissues, spleen or liver were severe.
Asunto(s)
Chlorocebus aethiops , Enfermedades de los Monos/patología , Saimiri , Infecciones por Yersinia pseudotuberculosis/veterinaria , Animales , Sistema Digestivo/patología , Femenino , Ganglios Linfáticos/patología , Masculino , Bazo/patología , Infecciones por Yersinia pseudotuberculosis/patologíaRESUMEN
Eight rhesus macaques were immunized intramuscularly four times (0, 1, 2, and 4 months) over a period of 4 months with a formalin-inactivated whole Simian immunodeficiency virus (SIV) vaccine in the presence of muramyl dipeptide (MDP) as adjuvant. Four animals received 0.5 mg and the other four received 0.1 mg immunogen per injection. Three weeks after the final immunization, the vaccinated monkeys along with two control monkeys were challenged intravenously with 50 MID50 of SIVmac251-32H. At the time of challenge, one monkey from the high dose group had a high titer (1:761) of antibody able to neutralize in vitro the homologous 32H strain of SIV mac. All other animals had lower but measurable titers (1:57-1:453), although there was no correlation between the levels of neutralizing antibody and subsequent protection. Upon challenge, three of four animals from the low dose group (plus the nonvaccinated control animals) became infected as demonstrated by reisolation of virus from peripheral blood mononuclear cells, by SIVmac-specific polymerase chain reaction, and by the development of a strong anamnestic response. In the sera from these animals the titer of neutralizing antibody rose to over 1:5,100. All other animals (one from low dose group and all four of the high dose group) remained negative by all parameters at 4 months post challenge. These data indicate that when used in conjunction with MDP, the amount of immunogen required per immunization for protection against challenge is between 0.1 and 0.5 mg.