RÉSUMÉ
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.
Sujet(s)
Virus de l'arthrite-encéphalite caprine/génétique , ADN viral/analyse , Système génital de l'homme/virologie , Capra/virologie , Sperme/virologie , Animaux , Virus de l'arthrite-encéphalite caprine/isolement et purification , Hybridation in situ , Mâle , Réaction de polymérisation en chaîne , RT-PCRRÉSUMÉ
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.
Sujet(s)
Virus de l'arthrite-encéphalite caprine/croissance et développement , Infections à lentivirus/virologie , Animaux , Virus de l'arthrite-encéphalite caprine/composition chimique , Virus de l'arthrite-encéphalite caprine/génétique , Protéines de capside/métabolisme , Cellules cultivées , Techniques de coculture , ADN viral/génétique , Embryon de mammifère/virologie , Génome viral/génétique , Capra , Immunohistochimie , Réaction de polymérisation en chaîne , Provirus/génétique , Provirus/isolement et purification , Sensibilité et spécificité , Réplication viraleRÉSUMÉ
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.
Sujet(s)
Infections à lentivirus/médecine vétérinaire , Lentivirus ovins-caprins/isolement et purification , Ovocytes/cytologie , ARN viral/analyse , Maladies des ovins/transmission , Animaux , Séquence nucléotidique , Fragmentation de l'ADN , Femelle , Liquide folliculaire/virologie , Infections à lentivirus/génétique , Infections à lentivirus/transmission , Ovocytes/virologie , RT-PCR/médecine vétérinaire , Ovis , Maladies des ovins/génétique , Visna/génétique , Visna/transmission , Virus maedi-visna/isolement et purificationRÉSUMÉ
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.
Sujet(s)
Virus de l'arthrite-encéphalite caprine/pathogénicité , Leucocytes/physiologie , Glandes mammaires animales/virologie , Animaux , Virus de l'arthrite-encéphalite caprine/isolement et purification , Adhérence cellulaire , Cellules cultivées , Femelle , Capra , Glandes mammaires animales/immunologie , Réaction de polymérisation en chaîne , Provirus/isolement et purificationRÉSUMÉ
Primary milk epithelial cells were isolated from CAEV-uninfected goats and three cell lines designated TIGMEC-1, TIGMEC-2 and TIGMEC-3 were established. The three cell lines retained their morphological characteristics of epithelial cells and expressed specific epithelial cytokeratin marker as well as the immortalizing SV40 large T antigen. The kinetics of growth of TIGMEC1, TIGMEC2 and TIGMEC3 cell lines showed a doubling time of 24-48 hours while the parental cell lines became senescent after the passage 6 in cell culture. Like the parental primary cells, the three cell lines were found to be highly sensitive to CAEV-pBSCA, an infectious molecular clone of CAEV-CO strain, and to a French isolate CAEV-3112. TIGMEC cell lines infected with CAEV-pBSCA became chronically infected producing high virus titers in absence of cytopathic effects. These cell lines may be useful for study of the possible physiological alterations in mammary epithelial cells infected with small ruminant lentiviruses and their consequences on milk quality. On an other hand, these cell lines can be used to study their role in virus transmission and pathogenesis.
Sujet(s)
Virus de l'arthrite-encéphalite caprine/physiologie , Cellules épithéliales/virologie , Glandes mammaires animales/cytologie , Lait/cytologie , Animaux , Division cellulaire , Lignée cellulaire , Femelle , Maladies des chèvres/virologie , Capra , Immunohistochimie/médecine vétérinaire , Cinétique , Infections à lentivirus/médecine vétérinaire , Infections à lentivirus/virologie , Glandes mammaires animales/virologie , Mastite/médecine vétérinaire , Mastite/virologie , Lait/virologie , Transfection/médecine vétérinaire , Réplication virale/physiologieRÉSUMÉ
Barriers to replication of viruses in potential host cells may occur at several levels. Lack of suitable and functional receptors on the host cell surface, thereby precluding entry of the virus, is a frequent reason for noninfectivity, as long as no alternative way of entry (e.g., pinocytosis, antibody-dependent adsorption) can be exploited by the virus. Other barriers can intervene at later stages of the virus life cycle, with restrictions on transcription of the viral genome, incorrect translation and posttranslational processing of viral proteins, inefficient viral assembly, and release or efficient early induction of apoptosis in the infected cell. The data we present here demonstrate that replication of caprine arthritis-encephalitis virus (CAEV) is restricted in a variety of human cell lines and primary tissue cultures. This barrier was efficiently overcome by transfection of a novel infectious complete-proviral CAEV construct into the same cells. The successful infection of human cells with a vesicular stomatitis virus (VSV) G-pseudotyped Env-defective CAEV confirmed that viral entry is the major obstacle to CAEV infection of human cells. The fully efficient productive infection obtained with the VSV-G-protein-pseudotyped infectious CAEV strengthened the evidence that lack of viral entry is the only practical barrier to CAEV replication in human cells. The virus thus produced retained its original host cell specificity and acquired no propensity to propagate further in human cultures.
Sujet(s)
Virus de l'arthrite-encéphalite caprine/pathogénicité , Glycoprotéines membranaires , Récepteurs viraux/métabolisme , Réplication virale , Animaux , Virus de l'arthrite-encéphalite caprine/physiologie , Cellules cultivées , Capra , Humains , Tests aux précipitines , Récepteurs viraux/génétique , RT-PCR , Spécificité d'espèce , Transfection , Virus de la stomatite vésiculeuse de type Indiana/génétique , Protéines de l'enveloppe virale/génétique , Protéines de l'enveloppe virale/métabolismeRÉSUMÉ
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.
Sujet(s)
Virus de l'arthrite-encéphalite caprine , Infections à lentivirus/médecine vétérinaire , Maladies des ovins/virologie , Animaux , Virus de l'arthrite-encéphalite caprine/génétique , Virus de l'arthrite-encéphalite caprine/physiologie , Clonage moléculaire , Électrophorèse sur gel de polyacrylamide , Infections à lentivirus/virologie , Macrophages/virologie , Ovis , Réplication viraleRÉSUMÉ
The main route of small ruminant lentivirus dissemination is the ingestion of infected cells present in colostrum and milk from infected animals. However, whether only macrophages or other cell subtypes are involved in this transmission is unknown. We derived epithelial cell cultures, 100% cytokeratin positive, from milk of naturally infected and noninfected goats. One such culture, derived from a naturally infected goat, constitutively produced a high titer of virus in the absence of any cytopathic effect. The other cultures, negative for natural lentivirus infection, were tested for their susceptibility to infection with the CAEV-CO strain and a French field isolate CAEV-3112. We showed that milk epithelial cells are easily infected by either virus and produce viruses at titers as high as those obtained in permissive goat synovial membrane cells. The CAEV-CO strain replicated in milk epithelial cells in absence of any cytopathic effect, whereas the CAEV-3112 field isolate induced both cell fusion and cell lysis. Our results suggest that CAEV-infected milk epithelial cells of small ruminants may play an important role in virus transmission and pathogenesis.
Sujet(s)
Virus de l'arthrite-encéphalite caprine/physiologie , Cellules épithéliales/virologie , Infections à lentivirus/virologie , Lait/virologie , Animaux , Cellules cultivées , Femelle , Capra , Infections à lentivirus/anatomopathologie , Lait/cytologieRÉSUMÉ
We previously reported that infection of goats with caprine arthritis encephalitis virus (CAEV) tat- proviral DNA or virus results in persistent infection, since the animals seroconverted and direct virus isolation from cultures of blood-derived macrophages was positive. In this study we wanted to determine whether goats injected with CAEV tat- proviral DNA or virus were protected against challenge with the pathogenic homologous virus and to investigate whether CAEV tat- was still pathogenic. All animals injected with CAEV tat- became infected as indicated by seroconversion and virus isolation. Challenge at 8 or 9 months postinfection demonstrated protection in four of four animals injected with CAEV tat- but did not in three of three mock-inoculated challenged goats. Challenge virus was undetectable in the blood macrophages of protected animals during a period of 6 or 10 months postchallenge. In two of four protected animals, however, we were able to detect the challenge wild-type virus by reverse transcriptase PCR on RNA directly extracted from synovial membrane cells surrounding the inoculation site. This result suggests that protection was achieved without complete sterilizing immunity. Animals injected with CAEV tat- and mock challenged developed inflammatory lesions in the joints, although these lesions were not as severe as those in CAEV wild-type-injected goats. These results confirm the dispensable role of Tat in CAEV replication in vivo for the establishment of infection and pathogenesis and demonstrate in another lentivirus infection model the efficacy of live attenuated viruses to induce resistance to superinfection.
Sujet(s)
Virus de l'arthrite-encéphalite caprine/immunologie , ADN viral/immunologie , Protéines du gène tat/physiologie , Infections à lentivirus/prévention et contrôle , Vaccins à ADN/immunologie , Vaccins antiviraux/immunologie , Animaux , Anticorps antiviraux/immunologie , Virus de l'arthrite-encéphalite caprine/génétique , Délétion de gène , Protéines du gène tat/génétique , Protéines du gène tat/immunologie , Capra , Infections à lentivirus/immunologie , Infections à lentivirus/anatomopathologie , Provirus/génétique , ARN viralRÉSUMÉ
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.
Sujet(s)
Virus de l'arthrite-encéphalite caprine/immunologie , Maladies des chèvres/immunologie , Infections à lentivirus/immunologie , Infections à lentivirus/médecine vétérinaire , Sous-populations de lymphocytes T/immunologie , Animaux , Test ELISA/médecine vétérinaire , Femelle , Capra , Immunophénotypage/médecine vétérinaire , Lactation/immunologie , Numération des lymphocytes/médecine vétérinaire , Mâle , Mastite/immunologie , Mastite/médecine vétérinaireRÉSUMÉ
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.
Sujet(s)
Virus de l'arthrite-encéphalite caprine/enzymologie , Infections à lentivirus/microbiologie , Pyrophosphatases/déficit , Animaux , Virus de l'arthrite-encéphalite caprine/pathogénicité , Séquence nucléotidique , ADN viral/génétique , Gènes viraux , Maladies des chèvres/microbiologie , Maladies des chèvres/anatomopathologie , Capra/microbiologie , Infections à lentivirus/anatomopathologie , Données de séquences moléculaires , Monocytes/microbiologie , Mutation ponctuelle , Provirus/génétique , Membrane synoviale/ultrastructure , Distribution tissulaire , Protéines virales structurales/génétique , Réplication viraleRÉSUMÉ
Replication of vif-caprine arthritis encephalitis virus (CAEV) is highly attenuated in primary goat synovial membrane cells and blood-derived macrophages compared to the wild-type (wt) virus. We investigated the requirement for CAEV Vif for in vivo replication and pathogenicity in goats by intra-articular injection of either infectious proviral DNA or viral supernatants. Wild-type CAEV DNA or virus inoculation induced persistent infection resulting in severe inflammatory arthritic lesions in the joints. We were unable to detect any sign of virus replication in vif- CAEV DNA inoculated goats, while vif- CAEV virus inoculation resulted in the seroconversion of the goats. However, virus isolation and RT-PCR analyses on blood-derived macrophage cultures remained negative throughout the experiment as well as in joint or lymphoid tissues taken at necropsy. No pathologic lesions could be observed in joint tissue sections examined at necropsy. Goats inoculated with the vif- virus demonstrated no protection against a pathogenic virus challenge. These results demonstrate that CAEV Vif is absolutely required for efficient in vivo virus replication and pathogenicity and provide additional evidence that live attenuated lentiviruses have to establish a persistent infection to induce efficient protective immunity.
Sujet(s)
Virus de l'arthrite-encéphalite caprine/génétique , Gènes vif , Infections à lentivirus/virologie , Réplication virale/génétique , Animaux , Virus de l'arthrite-encéphalite caprine/immunologie , Virus de l'arthrite-encéphalite caprine/physiologie , Lignée cellulaire , Capra , Infections à lentivirus/immunologie , Infections à lentivirus/anatomopathologie , Infections à lentivirus/prévention et contrôle , Provirus/génétique , Latence viraleRÉSUMÉ
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.
Sujet(s)
Molécules d'adhérence cellulaire/biosynthèse , Capra/immunologie , Sous-populations de lymphocytes/immunologie , Sous-populations de lymphocytes/métabolisme , Lait/immunologie , Animaux , Cytométrie en fluxRÉSUMÉ
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.
Sujet(s)
Régulation de l'expression des gènes , Interleukine-8/génétique , Macrophages alvéolaires/immunologie , Pneumonie interstitielle progressive du mouton/immunologie , Animaux , Séquence nucléotidique , Technique de Northern , Cellules cultivées , ADN complémentaire , Poumon/immunologie , Données de séquences moléculaires , Pneumonie interstitielle progressive du mouton/anatomopathologie , Réaction de polymérisation en chaîne , ARN messager/biosynthèse , OvisSujet(s)
Virus de l'arthrite-encéphalite caprine/immunologie , Délétion de gène , Gènes tat , Gènes vif , Infections à lentivirus/prévention et contrôle , Vaccins atténués/immunologie , Vaccins antiviraux/immunologie , Animaux , Anticorps antiviraux/immunologie , Virus de l'arthrite-encéphalite caprine/génétique , Virus de l'arthrite-encéphalite caprine/physiologie , ADN viral/biosynthèse , Capra , Infections à lentivirus/immunologie , Infections à lentivirus/virologie , Réplication viraleRÉSUMÉ
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.
Sujet(s)
Virus de l'arthrite-encéphalite caprine/physiologie , Pyrophosphatases/métabolisme , Réplication virale/physiologie , Virus maedi-visna/physiologie , Animaux , Virus de l'arthrite-encéphalite caprine/enzymologie , Séquence nucléotidique , Cellules cultivées , ADN viral , Humains , Macrophages/cytologie , Données de séquences moléculaires , Mutation , Pyrophosphatases/génétique , Similitude de séquences d'acides aminés , Nucléotides thymidyliques/métabolisme , Virus maedi-visna/enzymologieRÉSUMÉ
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.
Sujet(s)
Poumon/physiopathologie , Pneumonie interstitielle progressive du mouton/anatomopathologie , Pneumonie interstitielle progressive du mouton/physiopathologie , Virus maedi-visna , Animaux , Infections à lentivirus/anatomopathologie , Infections à lentivirus/physiopathologie , Infections à lentivirus/médecine vétérinaire , Poumon/anatomopathologie , Ovis , Maladies des ovinsRÉSUMÉ
To link ovine lentivirus infection to lung tissue damage, we studied the procoagulant response in alveolar macrophages from experimentally infected lambs and in in vitro infected alveolar macrophages. We cloned ovine tissue factor cDNA and analysed its in vitro expression by Northern blotting. Visna-maedi virus induced tissue factor mRNA. In order to correlate this mRNA induction with its cellular function, we analysed macrophage procoagulant activity after in vitro and in vivo infection. The procoagulant activity was increased by interaction with the virus in both cases. Thus, visna-maedi virus-induced expression of tissue factor mRNA was associated with enhanced macrophage procoagulant activity. These findings indicate an active role of alveolar macrophages in the pathogenesis of these inflammatory lung lesions.
Sujet(s)
Facteurs de la coagulation sanguine/métabolisme , Pneumonie interstitielle progressive du mouton/étiologie , ARN messager/génétique , Thromboplastine/génétique , Virus maedi-visna/pathogénicité , Animaux , Séquence nucléotidique , Amorces ADN/génétique , ADN complémentaire/génétique , Expression des gènes , Techniques in vitro , Macrophages alvéolaires/métabolisme , Données de séquences moléculaires , Pneumonie interstitielle progressive du mouton/génétique , Pneumonie interstitielle progressive du mouton/métabolisme , ARN messager/métabolisme , OvisRÉSUMÉ
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.
Sujet(s)
Pneumonie interstitielle progressive du mouton/immunologie , Virus maedi-visna/immunologie , Animaux , Animaux nouveau-nés , Liquide de lavage bronchoalvéolaire/cytologie , Liquide de lavage bronchoalvéolaire/immunologie , Cellules cultivées , Femelle , Fibroblastes/microbiologie , Numération des leucocytes , Mâle , Pneumonie interstitielle progressive du mouton/imagerie diagnostique , Pneumonie interstitielle progressive du mouton/anatomopathologie , Radiographie , Ovis , Virus maedi-visna/croissance et développementRÉSUMÉ
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.