Les lentivirus présentent une évolution rapide du gène env, notamment dans la région codant la glycoprotéine de surface (SU). Un fait remarquable est que les mutations de la SU sont localisées dans des zones spécifiques, appelées régions variables (V), séparées par des régions dites constantes (C). Afin de déterminer s'il existe des signatures spécifiques des régions C et V, nous avons développé des modèles de Markov cachés, ou HMM (Hidden Markov Models), basés sur la composition en oligonucléotides de chaque type de région, capables de différencier les régions C et V des lentivirus. Nous avons entraîné des modèles de Markov cachés sur des séquences des SU des lentivirus équins, humains, simiens et des petits ruminants. Nous avons obtenu une délimitation claire des régions C et V de tous ces lentivirus ainsi que des lentivirus bovins et félins qui n'avaient pas été utilisés pour définir les modèles. Nos résultats suggèrent que les régions C et V des lentivirus ont des compositions statistiques en mots de nucléotides et d'acides aminés différentes. Des signatures caractéristiques des régions C et V ont été extraites à partir des modèles définis.

Lentiviruses can infect non-dividing cells, and various cellular nuclear transport proteins provide crucial functions for lentiviral nuclear entry and integration. Genome-wide small interfering RNA screens previously identified nuclear pore complex component nucleoporin 153 (NUP153) as being important for infection by human immunodeficiency virus type 1 (HIV-1). We found that HIV-1 infection of NUP153 depleted cells resulted in normal levels of reverse transcription, a moderate reduction of 2-long terminal repeat circles, and a relatively large reduction in integrated proviruses, consistent with a role for NUP153 during nuclear entry of the HIV-1 pre-integration complex. We ascertained the capsid (CA) to be the major viral determinant for NUP153 dependence during infection, and accordingly observed a direct interaction between the CA N-terminal domain and the phenylalanine-glycine (FG)-repeat enriched NUP153 C-terminal domain (NUP153C). NUP153C fused to the effector domains of the rhesus Trim5alpha restriction factor (Trim-NUP153C) potently restricted HIV-1, providing an intracellular readout for the NUP153C-CA interaction during retroviral infection. Primate lentiviruses and equine infectious anemia virus (EIAV) bound NUP153C under these conditions, results that correlated with direct binding between purified recombinant proteins in vitro. These binding phenotypes moreover correlated with the requirement for endogenous NUP153 function during infection. Mutagenesis experiments identified NUP153C and CA residues important for binding, and different FG motifs within NUP153C mediated binding to HIV-1 versus EIAV CA proteins. HIV-1 CA binding mapped to residues that line a common alpha helix 3/4 hydrophobic pocket that also mediates binding to the small molecule PF-3450074 (PF74) inhibitor and cleavage and polyadenylation specific factor 6 (CPSF6) protein, with Asn57 (Asp58 in EIAV) playing a particularly important role. PF74 and CPSF6 each competed with NUP153C for binding to HIV-1 CA, and significantly higher concentrations of PF74 were needed to inhibit HIV-1 infection in the face of Trim-NUP153C expression or NUP153 knockdown. Correlation between CA mutant viral cell cycle and NUP153 dependencies moreover indicated that the NUP153C-CA interaction underlies the ability of HIV-1 to infect non-dividing cells. We propose that HIV-1 CA binds NUP153 FG motifs to affect viral nuclear import, serving as a novel example of viral hijacking of a fundamental cellular process.

SerpinB2 (aka plasminogen activator type 2) is well described as an extracellular inhibitor of urokinase-type plasminogen activator (uPA). However, the majority of SerpinB2 is retained intracellularly, and many uPA-independent activities have been reported for SerpinB2 suggesting an alternate function. This thesis explores the role of SerpinB2 in epithelial tumour cell lines, highlights the problems associated with various expression systems and argues that SerpinB2 has no role in growth or apoptosis of tumour cells. A potential role for immune modulation and angiogenesis is suggested in in vivo models. Previous research using SerpinB2 transfected, clonally selected tumour cell lines suggested that SerpinB2 regulates the retinoblastoma tumour suppressor protein (Rb) by binding and protecting Rb from degradation. Despite the use of two techniques under numerous conditions and positive controls, no significant interaction between SerpinB2 and Rb was found. SerpinB2 was reported to bind Rb through a PENF homology motif located within the SerpinB2 C-D interhelical loop region. The PENF homology motif was postulated to represent the motif responsible for binding to the C-pocket of Rb. Epstein Barr Virus nuclear antigen 6 (EBNA6) is a known Rb binding protein, which contains two predicted PENF homology motifs. However, mutation of the two PENF homology motifs within EBNA6 did not reduce Rb binding. Furthermore, the SerpinB2 PENF homology motif is actually not well conserved between SerpinB2 proteins from multiple species, whereas other regions of the SerpinB2 C-D loop show a high level of conservation. These data do not support a role for SerpinB2 and the PENF homology motif in Rb binding. SerpinB2 has been proposed to have a role in regulating growth and apoptosis. To further investigate this proposed phenotype of SerpinB2, SerpinB2 was expressed in a range of epithelial tumour lines using transient transfection. No change in growth, apoptosis or Rb levels were found. After ≈2-3 month antibiotic selection for the SerpinB2-expressing plasmid, SerpinB2 protein was lost without the loss of the transgene, indicating selective pressure against long-term SerpinB2 protein expression. To further investigate long-term SerpinB2 expression adenovirus and lentivirus vectors were used. Infection of tumour cell lines with adenovirus vectors expressing SerpinB2 resulted in reduced cell growth, characterised by increased p53 (but not Rb) levels and G2 arrest or apoptosis. When SerpinB2 expressing lentivirus vectors were used to transduce the same tumour cell lines, high levels of long-term expression of functional SerpinB2 was achieved. However, SerpinB2-expressing cell lines showed no differences in growth, proliferation, Rb levels, or apoptosis induced by a range of agents. Growth and apoptosis observed with adenovirus SerpinB2 had all the characteristics of adenovirus-associated toxicity, which has been reported previously for specific proteins. These experiments highlighted the problems associated with SerpinB2 expression systems and suggest that SerpinB2 expression per se is not toxic nor has a role in regulating Rb, growth and apoptosis. Screening of a number of tumour cell lines identified the HPV16 transformed cervical cancer line as expressing high levels of SerpinB2. SerpinB2 was located both extracellularly and intracellularly with a cytoplasmic and nuclear distribution. A high molecular weight SerpinB2 species was identified in CaSki cells and was shown to be the N-linked glycosylated species. Sequencing showed the protein to be Type A SerpinB2 and the protein was shown to form an inhibitory complex with uPA. An abundant low molecular weight SerpinB2 species was also identified in CaSki cell supernatants and appeared to be a proteolytic fragment of SerpinB2. Treatment of CaSki with PMA, TNFα and IFNγ increased SerpinB2 levels. Lentiviral based shRNA failed to significantly down regulate SerpinB2 expression and increasing SerpinB2 levels with lentiviral expression did not change growth, apoptosis, Rb levels or E7 transcription. Lentiviral expression of SerpinB2 in (normally SerpinB2 negative) HPV16 transformed SiHa cells, also failed to show changes in Rb levels or E7 transcription. CaSki thus express wild-type and functional SerpinB2, but no evidence could found that SerpinB2 effects HPV16 E7 transcription or Rb levels. The data presented identifies CaSki as valuable source of biologically functional SerpinB2. SerpinB2 expression in breast cancer cells has been associated with positive prognosis. Tubo, a SerpinB2-negative murine breast carcinoma cell line, was transduced with lentivirus expressing SerpinB2 and grown subcutaneously in BALB/c mice. SerpinB2 expressing tumours appeared red and were larger than control tumours. Furthermore, SerpinB2 expressing tumours had a ≈2 fold higher density of blood vessels when compared to Tubo and Tubo expressing EGFP. Mice carrying tumours expressing SerpinB2 also showed reduced anti-tumour IgG2 responses. These data suggest that a role for SerpinB2 in regulating angiogenesis and antitumour immunity. In conclusion, this thesis challenges the notion that SerpinB2 regulates Rb, cell cycle, and apoptosis and suggests a potential role for SerpinB2 in tumour angiogenesis and immunity.

SerpinB2 (aka plasminogen activator type 2) is well described as an extracellular inhibitor of urokinase-type plasminogen activator (uPA). However, the majority of SerpinB2 is retained intracellularly, and many uPA-independent activities have been reported for SerpinB2 suggesting an alternate function. This thesis explores the role of SerpinB2 in epithelial tumour cell lines, highlights the problems associated with various expression systems and argues that SerpinB2 has no role in growth or apoptosis of tumour cells. A potential role for immune modulation and angiogenesis is suggested in in vivo models. Previous research using SerpinB2 transfected, clonally selected tumour cell lines suggested that SerpinB2 regulates the retinoblastoma tumour suppressor protein (Rb) by binding and protecting Rb from degradation. Despite the use of two techniques under numerous conditions and positive controls, no significant interaction between SerpinB2 and Rb was found. SerpinB2 was reported to bind Rb through a PENF homology motif located within the SerpinB2 C-D interhelical loop region. The PENF homology motif was postulated to represent the motif responsible for binding to the C-pocket of Rb. Epstein Barr Virus nuclear antigen 6 (EBNA6) is a known Rb binding protein, which contains two predicted PENF homology motifs. However, mutation of the two PENF homology motifs within EBNA6 did not reduce Rb binding. Furthermore, the SerpinB2 PENF homology motif is actually not well conserved between SerpinB2 proteins from multiple species, whereas other regions of the SerpinB2 C-D loop show a high level of conservation. These data do not support a role for SerpinB2 and the PENF homology motif in Rb binding. SerpinB2 has been proposed to have a role in regulating growth and apoptosis. To further investigate this proposed phenotype of SerpinB2, SerpinB2 was expressed in a range of epithelial tumour lines using transient transfection. No change in growth, apoptosis or Rb levels were found. After ≈2-3 month antibiotic selection for the SerpinB2-expressing plasmid, SerpinB2 protein was lost without the loss of the transgene, indicating selective pressure against long-term SerpinB2 protein expression. To further investigate long-term SerpinB2 expression adenovirus and lentivirus vectors were used. Infection of tumour cell lines with adenovirus vectors expressing SerpinB2 resulted in reduced cell growth, characterised by increased p53 (but not Rb) levels and G2 arrest or apoptosis. When SerpinB2 expressing lentivirus vectors were used to transduce the same tumour cell lines, high levels of long-term expression of functional SerpinB2 was achieved. However, SerpinB2-expressing cell lines showed no differences in growth, proliferation, Rb levels, or apoptosis induced by a range of agents. Growth and apoptosis observed with adenovirus SerpinB2 had all the characteristics of adenovirus-associated toxicity, which has been reported previously for specific proteins. These experiments highlighted the problems associated with SerpinB2 expression systems and suggest that SerpinB2 expression per se is not toxic nor has a role in regulating Rb, growth and apoptosis. Screening of a number of tumour cell lines identified the HPV16 transformed cervical cancer line as expressing high levels of SerpinB2. SerpinB2 was located both extracellularly and intracellularly with a cytoplasmic and nuclear distribution. A high molecular weight SerpinB2 species was identified in CaSki cells and was shown to be the N-linked glycosylated species. Sequencing showed the protein to be Type A SerpinB2 and the protein was shown to form an inhibitory complex with uPA. An abundant low molecular weight SerpinB2 species was also identified in CaSki cell supernatants and appeared to be a proteolytic fragment of SerpinB2. Treatment of CaSki with PMA, TNFα and IFNγ increased SerpinB2 levels. Lentiviral based shRNA failed to significantly down regulate SerpinB2 expression and increasing SerpinB2 levels with lentiviral expression did not change growth, apoptosis, Rb levels or E7 transcription. Lentiviral expression of SerpinB2 in (normally SerpinB2 negative) HPV16 transformed SiHa cells, also failed to show changes in Rb levels or E7 transcription. CaSki thus express wild-type and functional SerpinB2, but no evidence could found that SerpinB2 effects HPV16 E7 transcription or Rb levels. The data presented identifies CaSki as valuable source of biologically functional SerpinB2. SerpinB2 expression in breast cancer cells has been associated with positive prognosis. Tubo, a SerpinB2-negative murine breast carcinoma cell line, was transduced with lentivirus expressing SerpinB2 and grown subcutaneously in BALB/c mice. SerpinB2 expressing tumours appeared red and were larger than control tumours. Furthermore, SerpinB2 expressing tumours had a ≈2 fold higher density of blood vessels when compared to Tubo and Tubo expressing EGFP. Mice carrying tumours expressing SerpinB2 also showed reduced anti-tumour IgG2 responses. These data suggest that a role for SerpinB2 in regulating angiogenesis and antitumour immunity. In conclusion, this thesis challenges the notion that SerpinB2 regulates Rb, cell cycle, and apoptosis and suggests a potential role for SerpinB2 in tumour angiogenesis and immunity.

SerpinB2 (aka plasminogen activator type 2) is well described as an extracellular inhibitor of urokinase-type plasminogen activator (uPA). However, the majority of SerpinB2 is retained intracellularly, and many uPA-independent activities have been reported for SerpinB2 suggesting an alternate function. This thesis explores the role of SerpinB2 in epithelial tumour cell lines, highlights the problems associated with various expression systems and argues that SerpinB2 has no role in growth or apoptosis of tumour cells. A potential role for immune modulation and angiogenesis is suggested in in vivo models. Previous research using SerpinB2 transfected, clonally selected tumour cell lines suggested that SerpinB2 regulates the retinoblastoma tumour suppressor protein (Rb) by binding and protecting Rb from degradation. Despite the use of two techniques under numerous conditions and positive controls, no significant interaction between SerpinB2 and Rb was found. SerpinB2 was reported to bind Rb through a PENF homology motif located within the SerpinB2 C-D interhelical loop region. The PENF homology motif was postulated to represent the motif responsible for binding to the C-pocket of Rb. Epstein Barr Virus nuclear antigen 6 (EBNA6) is a known Rb binding protein, which contains two predicted PENF homology motifs. However, mutation of the two PENF homology motifs within EBNA6 did not reduce Rb binding. Furthermore, the SerpinB2 PENF homology motif is actually not well conserved between SerpinB2 proteins from multiple species, whereas other regions of the SerpinB2 C-D loop show a high level of conservation. These data do not support a role for SerpinB2 and the PENF homology motif in Rb binding. SerpinB2 has been proposed to have a role in regulating growth and apoptosis. To further investigate this proposed phenotype of SerpinB2, SerpinB2 was expressed in a range of epithelial tumour lines using transient transfection. No change in growth, apoptosis or Rb levels were found. After ≈2-3 month antibiotic selection for the SerpinB2-expressing plasmid, SerpinB2 protein was lost without the loss of the transgene, indicating selective pressure against long-term SerpinB2 protein expression. To further investigate long-term SerpinB2 expression adenovirus and lentivirus vectors were used. Infection of tumour cell lines with adenovirus vectors expressing SerpinB2 resulted in reduced cell growth, characterised by increased p53 (but not Rb) levels and G2 arrest or apoptosis. When SerpinB2 expressing lentivirus vectors were used to transduce the same tumour cell lines, high levels of long-term expression of functional SerpinB2 was achieved. However, SerpinB2-expressing cell lines showed no differences in growth, proliferation, Rb levels, or apoptosis induced by a range of agents. Growth and apoptosis observed with adenovirus SerpinB2 had all the characteristics of adenovirus-associated toxicity, which has been reported previously for specific proteins. These experiments highlighted the problems associated with SerpinB2 expression systems and suggest that SerpinB2 expression per se is not toxic nor has a role in regulating Rb, growth and apoptosis. Screening of a number of tumour cell lines identified the HPV16 transformed cervical cancer line as expressing high levels of SerpinB2. SerpinB2 was located both extracellularly and intracellularly with a cytoplasmic and nuclear distribution. A high molecular weight SerpinB2 species was identified in CaSki cells and was shown to be the N-linked glycosylated species. Sequencing showed the protein to be Type A SerpinB2 and the protein was shown to form an inhibitory complex with uPA. An abundant low molecular weight SerpinB2 species was also identified in CaSki cell supernatants and appeared to be a proteolytic fragment of SerpinB2. Treatment of CaSki with PMA, TNFα and IFNγ increased SerpinB2 levels. Lentiviral based shRNA failed to significantly down regulate SerpinB2 expression and increasing SerpinB2 levels with lentiviral expression did not change growth, apoptosis, Rb levels or E7 transcription. Lentiviral expression of SerpinB2 in (normally SerpinB2 negative) HPV16 transformed SiHa cells, also failed to show changes in Rb levels or E7 transcription. CaSki thus express wild-type and functional SerpinB2, but no evidence could found that SerpinB2 effects HPV16 E7 transcription or Rb levels. The data presented identifies CaSki as valuable source of biologically functional SerpinB2. SerpinB2 expression in breast cancer cells has been associated with positive prognosis. Tubo, a SerpinB2-negative murine breast carcinoma cell line, was transduced with lentivirus expressing SerpinB2 and grown subcutaneously in BALB/c mice. SerpinB2 expressing tumours appeared red and were larger than control tumours. Furthermore, SerpinB2 expressing tumours had a ≈2 fold higher density of blood vessels when compared to Tubo and Tubo expressing EGFP. Mice carrying tumours expressing SerpinB2 also showed reduced anti-tumour IgG2 responses. These data suggest that a role for SerpinB2 in regulating angiogenesis and antitumour immunity. In conclusion, this thesis challenges the notion that SerpinB2 regulates Rb, cell cycle, and apoptosis and suggests a potential role for SerpinB2 in tumour angiogenesis and immunity.

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 4 uppsatser.

Thesis (Ph.D.)--University of Alabama at Birmingham, 2007. / Title from PDF title page (viewed on Sept. 16, 2009). Includes bibliographical references.

Gene therapy is the introduction or alteration of genetic material with the intention to treat disease. To support this aim, viruses have been modified, with elements linked to viral pathogenicity removed from their genome and replaced by the genetic material to be delivered. Gene therapy vectors based on lentiviruses have many advantages, such as the ability to transduce non-dividing cells and to target specific cell types via pseudotyping. They have been successfully used in ex vivo clinical trials for several haematopoietic stem cell disorders. Lentiviral vectors, however, suffer from substantially lower titres than the more popular adeno-associated virus (AAV)-based vectors and therefore have limited applicability for in vivo gene therapy which requires much greater quantities of virus. The main aim of this thesis was to investigate strategies to improve lentiviral vector productivity during manufacture, in order to increase the likelihood of lentiviruses being adopted for disease treatment. Initial experiments were based on the lentiviral vector manufacturing process currently being developed by the United Kingdom Cystic Fibrosis Gene Therapy Consortium for the generation of highly concentrated, purified lentivirus for clinical use. Supplementation of FreeStyle 293 Expression Medium used during upstream processing was attempted, but none of the assessed supplements led to significant increases in lentiviral vector production. Investigation into intrinsic immunity to viral infection indicated that over-expression of the protein kinase RNA-activated (PKR) led to lower production titres, but over-expression of its inhibitors was not successful at increasing titres. The focus then shifted to reducing, or 'knocking-down', inhibitory factors present in the host cells, which could adversely affect viral titres. Investigation of the published HIV-1 literature revealed a possible 152 candidate inhibitory factors described as having a negative impact on HIV-1 replication in the late stages of the life cycle of the virus. A novel siRNA screen was developed to assess the effect of ‘knock-down' of inhibitory factors on lentiviral vector titre. Application of the screen to 89 candidate inhibitory factors identified nine genes which, when knocked-down, resulted in increased lentiviral vector production by more than 40%. Further work will be necessary to understand the role of the inhibitory factors in lentiviral vector production, but novel cell lines in which genes encoding these factors have been permanently deleted from producer cells could lead to higher titres, reducing costs in the manufacture of lentiviral vectors and making in vivo gene therapy more feasible from a health economics perspective.

Background: Pulmonary arterial hypertension (PAH) is a lethal disease for which the fundamental molecular mechanisms are only partially understood. Existing therapies, which primarily focus on endothelial dysfunction, have limited effects on improving outcomes. Increases in pulmonary vascular resistance in PAH may be attributed to complex lung arterial remodeling which result in obliterative “plexiform” lesions, a pathological hallmark of this disease. Recent studies have shown that endothelial cell (ECs) apoptosis may be a central trigger for PAH, resulting in the emergence of growth-dysregulated and apoptosis-resistant ECs that contribute to the formation of complex neoplastic-like vascular lesions. However, the mechanism which links ECs apoptosis to dysregulated growth is not yet known. Previous studies in our lab have identified increased expression of translationally controlled tumor protein (TCTP) and its gene (TPT1), previously implicated in the transformation of neoplastic cells in cancer, and in blood outgrowth ECs from patients with PAH. Moreover, TCTP expression was found to be elevated in the lungs of patients with PAH, and tightly localized to complex arterial lesions. In addition, it was detected in obliterative intimal lesions of an experimental rat model of severe PAH. Hypothesis: TCTP represents a central molecular mechanism linking ECs apoptosis to the emergence of growth-dysregulated lung vascular cells and occlusive, complex arterial remodelling in PAH. Specific Hypotheses: - Lentiviral overexpression of TCTP in human umbilical vein endothelial cells (HUVECs) and pulmonary artery smooth muscle cells (PASMCs) leads to a hyperproliferative and apoptosis-resistant phenotype. - Overexpression of TCTP will increase its export into apoptotic extracellular vesicles, thereby augmenting cell-cell signalling between ECs and neighbouring SMCs. Purpose: My objective was to examine the effects TCTP overexpression on ECs and SMCs survival in terms of proliferation and apoptosis, and TCTP release on the survival of nontransduced ECs and SMCs. Methods and Results: The effect of TCTP overexpression on ECs growth and survival was studied using in vitro models. TCTP was overexpressed via a lentivirus vector in HUVECs and PASMCs. Compared to non-transfected or null transfected cells, TCTP overexpression led to increases in BrdU incorporation, consistent with hyper-proliferation, and decreases in caspase activity, consistent with apoptosis resistance. As well, TCTP was selectively exported into the conditioned media of apoptotic ECs, but not SMCs, despite similar levels of overexpression. In addition, the level of release was greater in serum starved conditioned media in comparison to the exosome fraction. Finally, our data demonstrates a selective effect of conditioned media (CM) from serum-starved ECs on PASMCs, but not ECs, in terms of an increase in proliferation and a decrease in apoptosis. Conclusions: These support the idea that TCTP overexpression confers an increase in the survival of SMCs and HUVECs. Moreover, TCTP released from apoptotic ECs leads to a growth-dysregulated phenotype within SMCs (but not ECs) and may contribute to the formation of complex lung arterial lesions, leading to arteriolar obliteration in PAH. Finally, an increase in the level of TCTP expression via lentiviral transduction led to an increased TCTP export into the media, but this appeared to be mostly in the soluble portion, and less was associated with exosomes.

Submitted by (edna.saturno@ufrpe.br) on 2016-10-21T12:41:27Z No. of bitstreams: 1 Pedro Alves de Moura Sobrinho.pdf: 655310 bytes, checksum: cf5f69ddf5c19bb603c77fd7a7012e4e (MD5) / Made available in DSpace on 2016-10-21T12:41:27Z (GMT). No. of bitstreams: 1 Pedro Alves de Moura Sobrinho.pdf: 655310 bytes, checksum: cf5f69ddf5c19bb603c77fd7a7012e4e (MD5) Previous issue date: 2008-02-04 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The aim of the present study was to characterize the goat and sheep production systems, estimate the prevalence of small-ruminant lentiviruses (SRLV) and study predisposing factors to infection in herds in the state of Tocantins (Brazil). Investigative questionnaires were administered addressing data on 29 goat herds and 28 sheep herds distributed among 21 towns in the state of Tocantins. To estimate the prevalence of SRLV, 838 samples of sheep blood and 843 samples of goat blood were analyzed using the immunodiffusion test in agarose gel. There was a mean number of 79 and 340 animals for the goat and sheep herds, respectively. According to the survey, 79.3 and 71.4% of the goat and sheep farms, respectively, were implanted after 2001. The animals making up the herds originated in the states of Bahia and Sergipe. The Santa Inês (61.0%) and SRD (38.7%) were the most frequent breeds among the sheep, whereas the Anglo-Nubian (55.4%), SRD (36.6%) and Saanen (7.1%) were the most frequent goat breeds. 55.1% of the 49 properties surveyed bred cattle + goats or sheep. The semi-extensive system was adopted in 51.7 and 46.4% of the goat and sheep farms, respectively. The most frequently adopted hygiene practices among the breeders were de-worming, cuts and disinfection of the navel and burying dead animals. Vaccination is practiced by 24.1% of goat breeders, most frequently for clostridiosis (20.7%), aphthous fever (6.8%) and rabies (3.4%). The clinical signs and diseases most often cited as having considerable importance were caseous lymphadenitis (34.5%) and pododermatitis (17%) by goat breeders and pododermatitis (42.3%) and diarrhea (30.8%) by sheep breeders. 51.7 and 28.6% of the goat and sheep breeders, respectively, knew about lentiviruses (CAE and Maedi-Visna). The frequency of ovine blood reagents was 0.9 ±0.3% (8/838). The results regarding the micro-regions of the state were distributed in the following manner: 3.3 (2/60), 0.61 (1/178), 1.3 (2/150) and 2.0% (3/150) for Bico Papagaio, North, Miracema and South, respectively. No animals tested positive in the Jalapão, Porto Nacional, Rio Formoso and Southeast micro-regions. The Santa Inês was the breed with the highest percentage of blood reagent animals (3.9%; 6/511), followed by Undefined Breed (0.6%; 2/324). Regarding age, 0.6% (2/238) and 1.2% (96/510)of animals under 24 and over months of age, respectively, tested positive. 1.2% (2/161) of males and 0.9% (6/677) of females tested positive. There were no statistically significant differences in the prevalence of blood reagent regarding breed,age and gender (p > 0.05). The frequency of blood reagent goats was 2.7% (23/843). The results were distributed according to micro-region in the following manner: 10.0 (20/200) and 1.4% (3/207) for the North and Miracema micro-regions, respectively. The Saanen was the breed with the highest percentage of blood reagent animals (11.7%; 7/60), followed by the SRD (0.6%; 2/310) and Anglo-Nubian (3.0%; 14/466). Regarding age, 1.9% (6/314) and 3.2% (17/529) of animals under 24 and over months of age, respectively, tested positive. 2.4% (4/166) of males and 2.8% (19/677) of females tested positive. Analysis of the data revealed a predominance of the semiextensive breeding system with good installations, but little use of reproduction biotechniques and important hygiene management practices. The activity has been expanding in the state, with a tendency toward professionalization. SRLV infection occurs at a low prevalence among sheep and goats in the state of Tocantins and is evenly distributed according to micro-region, breeding system, breed, age and origin of the base herds. Control measures should be implanted to avoid the dissemination of the disease among the herds. / Objetivou-se com esta pesquisa caracterizar o sistema de produção de caprino e ovino, estimar a prevalência e estudar os fatores predisponentes à infecção por Lentiviroses de Pequenos Ruminantes (LVPR), em rebanhos no Estado do Tocantins. Foram aplicados questionários investigativos em 29 rebanhos de caprinos e 28 de ovinos, distribuídos em 21 municípios do Estado do Tocantins, sendo 15 para cada espécie. Para estimar a prevalência de LVPR, foram analisadas 838 amostras de soros ovinos e 843 amostras de caprinos, utilizando o teste de imunodifusão em gel de agarose – IDGA. Identificou-se um número médio de animais de 79 e 340 para os rebanhos de caprinos e ovinos, respectivamente. Conforme a pesquisa, 79,3 e 71,4% dos criatórios de caprinos e ovinos, respectivamente, foram implantados após 2001. Os animais para formação dos rebanhos bases tem como origem os Estados da Bahia e Sergipe. Os tipos raciais Santa Inês (61,0%) e o SRD (38,7%) são os mais encontrados entre os ovinos e na espécie caprina o Anglo-nubiano (55,4%), SRD (36,6%) e Saanen (7,1%). O estudo mostrou que 55,1% das 49 propriedades pesquisadas criam bovinos + caprinos ou ovinos. O sistema semi-extensivo é adotado em 51,7 e 46,4% dos criatórios de caprinos e ovinos, respectivamente. As práticas sanitárias adotadas com maior freqüência pelos criadores são vermifugação, cortes e desinfecção do umbigo e enterrar os animais mortos. A vacinação é prática adotada por 24,1% dos criadores de caprinos, sendo as mais freqüentes contra clostridioses (20,7%), febre aftosa (6,8%) e raiva (3,4%). Os sinais clínicos e doenças mais citados como de grande importância foram linfadenite caseosa (34,5%) e pododermatite (17%) pelos criadores de caprinos e Pododermatite (42,3%) e diarréias (30,8%) para os ovinos. Dos produtores de caprinos e ovinos, 51,7 e 28,6%, respectivamente, conhecem as lentiviroses (CAE e a Maedi-Visna). A frequência de ovinos sororreagentes encontrada foi de 0,9 ±0,3% (8/838). De acordo com a microrregião do Estado, os resultados foram assim distribuídos: 3,3 (2/60), 0,61 (1/178), 1,3 (2/150) e 2,0 % (3/150) para Bico Papagaio, Norte, Miracema e Sul e Sudeste, respectivamente. As microrregiões Jalapão, Porto Nacional, Rio Formoso e Sudeste não tiveram animais positivos. Entre as raças, a Santa Inês foi a que apresentou numericamente o maior percentual de animais sororreagentes, 3,9% (6/511), seguida da Sem Raça Definida, 0,6% (2/324). De acordo com a idade, os animais com idade inferior e superior a 24 meses apresentaram 0,6% (2/328) 1,2% (6/510), respectivamente. Os machos apresentaram 1,2% (2/161) de positivos e as fêmeas 0,9% (6/677). Não houve diferença significativa (p > 0,05) na prevalência de sororreagentes com relação às variáveis raça, idade e sexo. A frequência de caprinos sororreagentes encontrada foi de 2,7% (23/843). De acordo com a região, os resultados foram assim distribuídos: 10,0 (20/200) e 1,4% (3/207) para as microrregiões Norte e Miracema, respectivamente. Entre as raças, a Saanen foi a que apresentou numericamente o maior percentual de animais sororreagentes, 11,7% (7/60), a SRD 0,6% (2/310) e a anglonubiana 3,0% (14/466). De acordo com a idade, os animais com idade inferior e superior a 24 meses apresentaram 1,9% (6/314) e 3,2% (17/529)), respectivamente. Os machos apresentaram 2,4% (4/166) de positivos e as fêmeas 2,8% (19/677). A análise das informações mostrou uma atividade onde predomina o sistema de criação semiextensivo com boas instalações, mas pouco uso de biotécnicas da reprodução e uso de importantes práticas de manejo sanitário. Observou-se que a atividade vem se expandindo no Estado com uma tendência de profissionalização. A infecção por LVPR ocorre em ovinos e caprinos do Estado de Tocantins com baixa prevalência, homogeneamente distribuída de acordo com a microrregião, sistema de criação, raça, sexo, idade e origem dos rebanhos base. Medidas de controle devem ser implantadas no sentido de evitar a disseminação da doença entre os rebanhos.