RESUMO
The aim of this study was to determine the infectious status of semen and genital tract tissues from male goat naturally infected with the caprine lentivirus. Firstly, polymerase chain reaction (PCR) was used to detect the presence of CAEV proviral-DNA in the circulating mononuclear cells, semen (spermatozoa and non-spermatic cells), and genital tract tissues (testis, epididymis, vas deferens, and vesicular gland) of nine bucks. RT-PCR was used to detect the presence of CAEV viral RNA in seminal plasma. Secondly, in situ hybridization was performed on PCR-positive samples from the head, body, and tail of the epididymis. CAEV proviral-DNA was identified by PCR in the blood cells of 7/9 bucks and in non-spermatic cells of the seminal plasma of 3/9 bucks. No CAEV proviral-DNA was identified in the spermatozoa fraction. The presence of CAEV proviral-DNA in non-spermatic cells and the presence of CAEV in the seminal plasma was significantly higher (p<0.01) in bucks with PCR-positive blood. Two of the three bucks with positive seminal plasma cells presented with at least one PCR-positive genital tract tissue. Proviral-DNA was found in the head (3/9), body (3/9), and tail (2/9) of the epididymis. In situ hybridization confirmed the presence of viral mRNA in at least one of each of these tissues, in the periphery of the epididymal epithelium. This study clearly demonstrates the presence of viral mRNA and proviral-DNA in naturally infected male goat semen and in various tissues of the male genital tract.
Assuntos
Vírus da Artrite-Encefalite Caprina/genética , DNA Viral/análise , Genitália Masculina/virologia , Cabras/virologia , Sêmen/virologia , Animais , Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Hibridização In Situ , Masculino , Reação em Cadeia da Polimerase , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
The aim of this study was to examine the Maedi-Visna virus (MVV) infection status of oocytes, cumulus cells, and follicular fluid taken from 140 ewes from breeding flocks. MVV proviral-DNA and MVV RNA were detected using nested-PCR and RT-PCR MVV gene amplification, respectively in the gag gene. Nested-PCR analysis for MVV proviral-DNA was positive in peripheral blood mononuclear cells in 37.1% (52/140) of ewes and in 44.6% (125/280) of ovarian cortex samples. The examination of samples taken from ovarian follicles demonstrated that 8/280 batches of cumulus cells contained MVV proviral-DNA, whereas none of the 280 batches of oocytes taken from the same ovaries and whose cumulus cells has been removed, was found to be PCR positive. This was confirmed by RT-PCR analysis showing no MVV-viral RNA detection in all batches of oocytes without cumulus cells (0/280) and follicular fluid samples taken from the last 88 ovaries (0/88). The purity of the oocyte fraction and the efficacy of cumulus cell removal from oocytes was proved by absence of granulosa cell-specific mRNA in all batches of oocytes lacking the cumulus cells, using RT-PCR. This is the first demonstration that ewe cumulus cells harbor MVV genome and despite being in contact with these infected-cumulus cells, the oocytes and follicular fluid remain free from infection. In addition, the enzymatic and mechanical procedures we used to remove infected-cumulus cells surrounding the oocytes, are effective to generate MVV free-oocytes from MVV-infected ewes.
Assuntos
Infecções por Lentivirus/veterinária , Lentivirus Ovinos-Caprinos/isolamento & purificação , Oócitos/citologia , RNA Viral/análise , Doenças dos Ovinos/transmissão , Animais , Sequência de Bases , Fragmentação do DNA , Feminino , Líquido Folicular/virologia , Infecções por Lentivirus/genética , Infecções por Lentivirus/transmissão , Oócitos/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Ovinos , Doenças dos Ovinos/genética , Visna/genética , Visna/transmissão , Vírus Visna-Maedi/isolamento & purificaçãoRESUMO
A characteristic lesion in goats infected by the lentivirus CAEV is mastitis with lymphoid hyperplasia. In order to investigate the mechanism of lesion formation, cultures highly enriched in microvascular endothelial cells, mature and immature luminal epithelial cells, fibroblasts and myoepithelial cells were established from goat mammary gland biopsies. Their susceptibility to in vitro infection with two distinct types of CAEV was investigated by PCR, antigen expression and cytopathy. The capacity of infected mammary gland cells to bind uninfected caprine leukocytes was determined by flow cytometry. All cell types tested were susceptible to CAEV infection in vitro, with different levels of sensitivity according to cell phenotype. Our results suggest that the limited extent of natural infection of mammary gland cells reflects a protective local immune response, and that the myoepithelial cell could act as a reservoir cell. After infection, the mature luminal cell acquires the capacity to bind leukocytes in vitro, which could indicate a facilitation of cellular interactions. The distinct reactions of the different cell types to CAEV infection may be correlated with events leading to progressive lesion development during the natural infection.
Assuntos
Vírus da Artrite-Encefalite Caprina/patogenicidade , Leucócitos/fisiologia , Glândulas Mamárias Animais/virologia , Animais , Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Adesão Celular , Células Cultivadas , Feminino , Cabras , Glândulas Mamárias Animais/imunologia , Reação em Cadeia da Polimerase , Provírus/isolamento & purificaçãoRESUMO
Visna-maedi virus is a lentivirus closely related to the human immunodeficiency virus type I (HIV-I). During spontaneous infection of sheep by Visna-maedi virus an interstitial lung disease is observed. It is characterized by an alveolitis, peribronchovascular lymphoid nodules, alveolar wall thickening and myomatosis. In order to decipher the pathology of this lentiviral infection we have induced this disease in colostrum-deprived newborn lambs.
Assuntos
Pneumonia Intersticial Progressiva dos Ovinos/imunologia , Vírus Visna-Maedi/imunologia , Animais , Animais Recém-Nascidos , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Células Cultivadas , Feminino , Fibroblastos/microbiologia , Contagem de Leucócitos , Masculino , Pneumonia Intersticial Progressiva dos Ovinos/diagnóstico por imagem , Pneumonia Intersticial Progressiva dos Ovinos/patologia , Radiografia , Ovinos , Vírus Visna-Maedi/crescimento & desenvolvimentoRESUMO
The effect of natural infection by Caprine Arthritis Encephalitis Virus on the phenotypic pattern of T lymphocytes in peripheral blood was studied in a herd of 127 milking goats by flow cytometry. Total leukocyte and T-lymphocyte numbers tend to decrease with age, with only small changes in the CD4/CD8 ratio. The lymphocyte phenotypes show no strong correlation with seropositivity to CAEV or the presence of clinical symptoms, suggesting that this macrophagetropic lentivirus does not greatly effect the lymphocyte population.
Assuntos
Vírus da Artrite-Encefalite Caprina/imunologia , Doenças das Cabras/imunologia , Infecções por Lentivirus/imunologia , Infecções por Lentivirus/veterinária , Subpopulações de Linfócitos T/imunologia , Animais , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Cabras , Imunofenotipagem/veterinária , Lactação/imunologia , Contagem de Linfócitos/veterinária , Masculino , Mastite/imunologia , Mastite/veterináriaRESUMO
Cytometric analysis of cells in milk from healthy goats was achieved after elimination of interfering signals from debris and dead cells by irreversible staining with ethidium monoazide. Some 61% of milk lymphocytes are CD8+ T cells, 17% are CD4+ and about 20% of these express class II antigens: less than 4% are B cells. Compared with blood, the CD4/CD8 ratio was inverted and fewer lymphocytes expressed CD4SR or L-selectin. Monoclonal antibodies to ovine integrins recognised the caprine alpha 4 chain on most lymphocytes from milk or blood, while beta 1 was more frequent on milk cells.
Assuntos
Moléculas de Adesão Celular/biossíntese , Cabras/imunologia , Subpopulações de Linfócitos/imunologia , Subpopulações de Linfócitos/metabolismo , Leite/imunologia , Animais , Citometria de FluxoRESUMO
OBJECTIVE: To determine whether monocyte-derived macrophages from Mouflon-domestic sheep hybrids (Ovis musimon X Ovis spp) were susceptible to productive infection with caprine arthritis-encephalitis virus (CAEV) in vitro and whether experimental inoculation of Mouflon-domestic sheep hybrids with a molecularly cloned CAEV would result in persistent infection. ANIMALS: 5 Mouflon hybrids. PROCEDURE: Macrophage monolayers were inoculated with virus in vitro. Three animals were inoculated with virus intratracheally. RESULTS: Productive replication of CAEV was demonstrated in monocyte-derived macrophages following in vitro and in vivo inoculation. Titer of infectious cytopathic CAEV produced by macrophages from the Mouflon hybrids was similar to titers produced by macrophages from an infected goat or by synovial membrane cells. Isolation of virus from monocyte-derived macrophages and use of a semiquantitative polymerase chain reaction assay to amplify a portion of the viral genome demonstrated persistent virus replication in all 3 inoculated animals. Two weeks after inoculation of sheep, approximately 1 of 5,000 monocytes was harboring the virus. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that Mouflon-domestic sheep hybrids are susceptible to infection with isolates of CAEV that cause infection in domestic small ruminants.
Assuntos
Vírus da Artrite-Encefalite Caprina , Infecções por Lentivirus/veterinária , Doenças dos Ovinos/virologia , Animais , Vírus da Artrite-Encefalite Caprina/genética , Vírus da Artrite-Encefalite Caprina/fisiologia , Clonagem Molecular , Eletroforese em Gel de Poliacrilamida , Infecções por Lentivirus/virologia , Macrófagos/virologia , Ovinos , Replicação ViralRESUMO
Lentiviruses belong to the retroviruses family (ie RNA viruses with reverse transcriptase activity); they induce inflammatory and/or degenerative slowly progressive diseases, affecting various organs. Some lentiviruses preferentially infect lymphocytes (HIV-1 and HIV-2, SIV and FIV) and are associated with infectious and tumoral disorders. Most lentiviruses induce a pulmonary disease, typically diffuse interstitial pneumonia. The visna/maedi-virus of sheep infects monocyte macrophage cells and the pulmonary lesions are macrophagic and neutrophilic alveolitis, lymphoid infiltration, myomatosis and interstitial fibrosis. Such pulmonary lesions are also induced by the goat and equine lentiviruses. In humans infected by HIV-1 or HIV-2, a diffuse interstitial lung disease also occurs; the histological findings are of alveolitis associated with lymphoid peribronchovascular infiltrates. The mechanism of formation of the lesions involves complex cellular interactions (especially between macrophage and lymphocyte, via cytokine production). These interactions are well modelled by small ruminant lentivirus induction of interstitial pneumonia.
Assuntos
Síndrome da Imunodeficiência Adquirida/complicações , Pneumonia Intersticial Progressiva dos Ovinos/etiologia , Fibrose Pulmonar/etiologia , Síndrome da Imunodeficiência Adquirida/patologia , Adulto , Animais , Criança , Infecções por Deltaretrovirus/complicações , Humanos , Infecções por Lentivirus/complicações , Pulmão/patologia , Pneumonia Intersticial Progressiva dos Ovinos/patologia , Fibrose Pulmonar/patologia , OvinosAssuntos
Vírus da Artrite-Encefalite Caprina/imunologia , Deleção de Genes , Genes tat , Genes vif , Infecções por Lentivirus/prevenção & controle , Vacinas Atenuadas/imunologia , Vacinas Virais/imunologia , Animais , Anticorpos Antivirais/imunologia , Vírus da Artrite-Encefalite Caprina/genética , Vírus da Artrite-Encefalite Caprina/fisiologia , DNA Viral/biossíntese , Cabras , Infecções por Lentivirus/imunologia , Infecções por Lentivirus/virologia , Replicação ViralRESUMO
Zona-pellucida-free embryos at 8-16 cell stage were co-cultured for 6 days in an insert over a mixed cell monolayer infected with CAEV-pBSCA. Embryos were washed and transferred to an insert on CAEV indicator goat synovial membrane cells for 6 h, then they were washed and cultivated in B2 Ménézo for 24 h, finally, embryo cells were dissociated and cultivated on a feeder monolayer for 8 days. After 5 weeks, multinucleated giant cells typical of CAEV infection were observed in indicator GSM cell monolayers. In the acellular medium, the early embryonic cells produced at least 10(3.25) TCID50/ml over 24 h. The monolayer of cultivated embryonic cells developed cytopathic lesions within 8 days, and CAEV RNA, CAEV proviral DNA and protein p28 of the capsid were detected. All of these results clearly demonstrate that caprine early embryonic cells are susceptible to infection with CAEV and that infection with this virus is productive.
Assuntos
Vírus da Artrite-Encefalite Caprina/crescimento & desenvolvimento , Infecções por Lentivirus/virologia , Animais , Vírus da Artrite-Encefalite Caprina/química , Vírus da Artrite-Encefalite Caprina/genética , Proteínas do Capsídeo/metabolismo , Células Cultivadas , Técnicas de Cocultura , DNA Viral/genética , Embrião de Mamíferos/virologia , Genoma Viral/genética , Cabras , Imuno-Histoquímica , Reação em Cadeia da Polimerase , Provírus/genética , Provírus/isolamento & purificação , Sensibilidade e Especificidade , Replicação ViralRESUMO
Retroviruses from small ruminants are spread between susceptible animals by mononuclear phagocytes which are also virus targets. Young seronegative goats were inoculated with in vitro infected monocytes producing caprine arthritis encephalitis virus (10(5) cells/animal) by intravenous route corresponding to 10(5) syncytia forming units. After 3 months the same goats received dexamethasone treatment (5 mg/animal each 2 days over a 10-day period). The observed immune response differed if the animal was infected with its own cells or with homologous cells. Before dexamethasone treatment, antibody production evaluated by gel precipitation, ELISA or western blot was delayed in autologous by comparison with homologous conditions. After dexamethasone treatment, the appearance of infectious monocytes in blood and subsequent arthritis were observed in all animals in homologous conditions and only in 1 animal out of 3 in autologous conditions. Host reaction to the infected cells determines virus expression at the time of contamination.
Assuntos
Artrite/veterinária , Encefalite/veterinária , Doenças das Cabras/etiologia , Monócitos/microbiologia , Infecções por Retroviridae/veterinária , Animais , Anticorpos Antivirais/biossíntese , Artrite/etiologia , Dexametasona , Encefalite/etiologia , Cabras , Retroviridae/imunologia , Retroviridae/fisiologia , Infecções por Retroviridae/etiologiaRESUMO
Lambing in small ruminants is a time of high lentivirus expression; infected mononuclear phagocytes are frequent in colostrum and milk. We have studied mammary secretions in 5 multiparous ewes and shown that infected macrophages in milk are accompanied by an augmentation of leucocyte number. The lymphocyte CD8 subpopulation increased in size simultaneously with the onset of infected cell excretion. The udder infection by coagulase negative staphylococci did not modulate milk lymphocyte content. Although infected cell excretion was restricted to one half of the udder, virus-specific lesions were found in both udder halves. Milk leukocytes changes are a marker of infected macrophage presence; they do not control lentivirus spread.
Assuntos
Infecções por Lentivirus/veterinária , Leucócitos/imunologia , Complicações do Trabalho de Parto/veterinária , Complicações Infecciosas na Gravidez/veterinária , Doenças dos Ovinos/imunologia , Animais , Colostro/microbiologia , Feminino , Infecções por Lentivirus/imunologia , Contagem de Leucócitos/veterinária , Macrófagos/microbiologia , Glândulas Mamárias Animais/microbiologia , Glândulas Mamárias Animais/patologia , Leite/citologia , Leite/microbiologia , Complicações do Trabalho de Parto/imunologia , Gravidez , Complicações Infecciosas na Gravidez/imunologia , OvinosRESUMO
A very small number of goat positive for caprine arthritis encephalitis virus are also capable of in vitro inhibition of specific tissue culture lesions. This neutralizing activity for apparition of multinucleated cells is present in sera at low dilutions. We found this seric activity under natural conditions in young goats and in culled animals. Under experimental conditions the inhibitory activity was shown in the serum in a goat infected during gestation and in two animals infected when treated with repeated cortisone injection. The presence of syncitia formation inhibitory activity in sera seems to be related with virus reactivation.
Assuntos
Artrite Infecciosa/veterinária , Encefalite/veterinária , Cabras/imunologia , Soros Imunes/imunologia , Infecções por Retroviridae/veterinária , Animais , Artrite Infecciosa/imunologia , Encefalite/imunologia , Infecções por Retroviridae/imunologiaRESUMO
Milk is the most important route of lentivirus spread in sheep and goat. Blood and milk cell populations were characterised with specific monoclonal antibody at lambing time in 4 primiparous seropositive ewes. Two of the ewes (3/4 half udders) produced milk with infected cells. The cell number/ml was always higher in milk and blood from virus-producing animals. In ewes which spread infected macrophages CD8 lymphocyte number was increased in blood and milk. Serological tests are able to detect infected animals but virus production is reflected immediately by an increase of CD8 lymphocytes in milk.
Assuntos
Trabalho de Parto/imunologia , Subpopulações de Linfócitos , Leite/microbiologia , Pneumonia Intersticial Progressiva dos Ovinos/imunologia , Vírus Visna-Maedi/imunologia , Animais , Anticorpos Antivirais/sangue , Feminino , Contagem de Leucócitos/veterinária , Leite/citologia , Pneumonia Intersticial Progressiva dos Ovinos/microbiologia , Gravidez , Ovinos , Linfócitos T Reguladores , Vírus Visna-Maedi/isolamento & purificaçãoRESUMO
The major characteristic lesion observed following spontaneous infection of sheep by the prototype lentivirus, maedi-visna virus (MVV), is a lymphocytic intestitial pneumonia. Similar lesions may be observed with variable frequency following infection of other species by pathogenic lentiviruses, for example in children infected by HIV-1. Further, lentivirus-induced lesions involving organs other than the lungs frequently involve a comparable cellular infiltration. The cellular composition of bronchoalveolar lavage specimens from naturally- or experimentally-infected sheep has been examined with a view to describing the pathological progression of lentivirus-induced lung lesions. The naturally-infected sheep presented advanced lesions typical of 'maedi', while the experimentally-infected newborn lambs permitted the study of early lesions which we refer to as 'pre-maedi'. In both cases there was a considerable infiltration of lymphocytes, predominantly CD8+ in maedi, but with nearly equal numbers of CD4+ cells in pre-maedi. A large proportion of the alveolar lymphocytes in spontaneous maedi, but not in experimentally-infected lambs, express high levels of MHC class II antigen, suggesting an activated phenotype. Activated macrophages, the chief target cells for MVV infection, are also present at this advanced stage of the disease suggesting the involvement of mediators such as IL-8 in the cellular interactions leading to the localization of particular lymphocyte sub-populations in the pulmonary parenchyma during lentiviral disease.
Assuntos
Pulmão/fisiopatologia , Pneumonia Intersticial Progressiva dos Ovinos/patologia , Pneumonia Intersticial Progressiva dos Ovinos/fisiopatologia , Vírus Visna-Maedi , Animais , Infecções por Lentivirus/patologia , Infecções por Lentivirus/fisiopatologia , Infecções por Lentivirus/veterinária , Pulmão/patologia , Ovinos , Doenças dos OvinosRESUMO
The importance of the virally encoded dUTPase for CAEV replication, invasiveness, pathogenesis, and genetic stability was investigated in goats infected by viruses with single point (DU-G) and deletion (DU-1) mutations of the dUTPase gene (DU gene). The DU gene was found to be dispensable for CAEV replication in vivo as judged by times taken to seroconvert, frequencies of viral isolation, and tissue distribution of viral RNAs. DU- reversion at week 34 in one of three goats infected with the single point mutant DU-G, however, suggested that the viral dUTPase confers some advantages for replication in vivo. Moreover, we show that dUTPase is necessary for the timely development of bilateral arthritic lesions of the carpus. Finally, dUTPase was shown to efficiently prevent accumulation of G-to-A transitions in the viral genome.
Assuntos
Vírus da Artrite-Encefalite Caprina/enzimologia , Infecções por Lentivirus/microbiologia , Pirofosfatases/deficiência , Animais , Vírus da Artrite-Encefalite Caprina/patogenicidade , Sequência de Bases , DNA Viral/genética , Genes Virais , Doenças das Cabras/microbiologia , Doenças das Cabras/patologia , Cabras/microbiologia , Infecções por Lentivirus/patologia , Dados de Sequência Molecular , Monócitos/microbiologia , Mutação Puntual , Provírus/genética , Membrana Sinovial/ultraestrutura , Distribuição Tecidual , Proteínas Estruturais Virais/genética , Replicação ViralRESUMO
The virion-associated dUTPase activities of caprine arthritis-encephalitis virus (CAEV) and visna virus were determined by using an assay which measure the actual ability of the dUTPase to prevent the dUTP misincorporations into cDNA during reverse transcription. We showed that the CAEV molecular clone from the Cork isolate was dUTPase defective as a result of a single amino acid substitution. Using this point mutant and deletion mutants of CAEV as well as a deletion mutant of visna virus, we demonstrated that dUTPase-deficient viruses replicate similarly to wild-type viruses in dividing cells but show delayed replication in nondividing primary macrophages.
Assuntos
Vírus da Artrite-Encefalite Caprina/fisiologia , Pirofosfatases/metabolismo , Replicação Viral/fisiologia , Vírus Visna-Maedi/fisiologia , Animais , Vírus da Artrite-Encefalite Caprina/enzimologia , Sequência de Bases , Células Cultivadas , DNA Viral , Humanos , Macrófagos/citologia , Dados de Sequência Molecular , Mutação , Pirofosfatases/genética , Homologia de Sequência de Aminoácidos , Nucleotídeos de Timina/metabolismo , Vírus Visna-Maedi/enzimologiaRESUMO
Sheep infected by visna-maedi virus, a lentivirus related to the human immunodeficiency virus, develop a chronic interstitial lung disease. Since monocyte/macrophages are known to be specifically infected by visna-maedi virus, we investigated the role of macrophages in the appearance of pulmonary lesions in animals with naturally occurring disease. Alveolitis in maedi leads to a doubling in bronchoalveolar lavage total cell counts and of macrophages as compared to normal sheep. A significant increase in the relative percentage of neutrophils was also observed, accompanied by an increased spontaneous release of neutrophil chemotactic activity by alveolar macrophages of diseased animals, suggesting that they may be activated. Macrophage activation is also demonstrated by the observation of a significant (x3) increase of spontaneous fibronectin release by alveolar macrophages from maedi lungs, and furthermore by the high level expression of major histocompatibility complex class II antigens on most of these cells. Thus viral infection, although restricted to a small population of macrophages, is able to modulate extensive activation of macrophages in the lung. Activated macrophages release mediators likely to play a role in the development of the alveolitis and the parenchymal desorganization. These findings may be relevant to our understanding of the mechanisms by which human immunodeficiency virus infection leads to pulmonary disease other than that caused by opportunistic infections.
Assuntos
Ativação de Macrófagos/imunologia , Pneumonia Intersticial Progressiva dos Ovinos/imunologia , Alvéolos Pulmonares/imunologia , Animais , Líquido da Lavagem Broncoalveolar/citologia , Fatores Quimiotáticos/biossíntese , Fibronectinas/metabolismo , Antígenos de Histocompatibilidade Classe II/análise , Interleucina-8 , Macrófagos/imunologia , Ovinos , Vírus Visna-Maedi/isolamento & purificaçãoRESUMO
In order to investigate the contribution of lymphocytes to interstitial lung disease in animals with visna-maedi infection, we studied in parallel bronchoalveolar cells and lung tissue from slaughter-house animals (n = 29) and from colostrum-deprived lambs transtracheally inoculated with field isolates of visna-maedi virus (n = 9) or saline (n = 6). Lymphocyte subpopulations were identified in bronchoalveolar lavage by immunofluorescence and flow cytometry analysis and in lung tissue using indirect immunohistochemistry. In infected animals a lymphocytic alveolitis containing CD4 and CD8 lymphocytes was observed. Peribronchovascular lymphoid nodules comprise mostly CD4 lymphocytes. Alveolar lymphocytes of both subsets displayed increased expression of MHC class II antigens in animals with naturally occurring maedi but not in experimentally infected ones. A sequential process of lymphocyte attraction and activation is likely to occur in vivo as part of the alveolitis.
Assuntos
Subpopulações de Linfócitos/imunologia , Visna/imunologia , Animais , Antígenos de Histocompatibilidade Classe II/análise , Subpopulações de Linfócitos/microbiologia , Alvéolos Pulmonares/imunologia , Alvéolos Pulmonares/patologia , Ovinos , Visna/patologia , Vírus Visna-MaediRESUMO
Visna-maedi virus is a lentivirus which causes inflammatory disorders in sheep, including a chronic interstitial lung disease resembling that observed in human immunodeficiency virus type 1 (HIV 1) infection. In view of our previous demonstration of the production of neutrophil chemotactic activity by alveolar macrophages, and given the lymphocytic and neutrophilic nature of the alveolar cell infiltrate in both naturally and experimentally infected animals, we hypothesized that interleukin-8 (IL8) could be a candidate for at least part of the chemotactic activity we described. In this study, we investigated IL8 mRNA expression following visna-maedi virus infection. Northern analysis of total RNA using an ovine IL8-specific probe demonstrated that the IL8 gene is upregulated in alveolar macrophages as a consequence of in vitro infection and in alveolar cells from experimentally infected animals. Using a semi-quantitative RT-PCR method, we showed that various levels of IL8 mRNA are expressed by alveolar cells from infected animals and that they correlate with the intensity of the lesions. In conclusion, visna-maedi virus is able to induce IL8 mRNA expression in sheep alveolar cells. Results from in vivo infected animals suggest that IL8 could play a role in the early build-up of visna-maedi virus-induced lesions.