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1.
Eur J Med Chem ; 130: 440-457, 2017 Apr 21.
Artigo em Inglês | MEDLINE | ID: mdl-28282613

RESUMO

A novel series of feruloyl-donepezil hybrid compounds were designed, synthesized and evaluated as multitarget drug candidates for the treatment of Alzheimer's Disease (AD). In vitro results revealed potent acetylcholinesterase (AChE) inhibitory activity for some of these compounds and all of them showed moderate antioxidant properties. Compounds 12a, 12b and 12c were the most potent AChE inhibitors, highlighting 12a with IC50 = 0.46 µM. In addition, these three most promising compounds exhibited significant in vivo anti-inflammatory activity in the mice paw edema, pleurisy and formalin-induced hyperalgesy models, in vitro metal chelator activity for Cu2+ and Fe2+, and neuroprotection of human neuronal cells against oxidative damage. Molecular docking studies corroborated the in vitro inhibitory mode of interaction of these active compounds on AChE. Based on these data, compound 12a was identified as a novel promising drug prototype candidate for the treatment of AD with innovative structural feature and multitarget effects.


Assuntos
Doença de Alzheimer/tratamento farmacológico , Indanos/farmacologia , Terapia de Alvo Molecular/métodos , Piperidinas/farmacologia , Acrilatos/química , Acrilatos/farmacologia , Animais , Anti-Inflamatórios , Antioxidantes , Linhagem Celular , Células Cultivadas , Inibidores da Colinesterase/química , Inibidores da Colinesterase/farmacologia , Donepezila , Desenho de Fármacos , Humanos , Indanos/química , Masculino , Camundongos , Simulação de Acoplamento Molecular , Neurônios/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Piperidinas/química , Relação Estrutura-Atividade
2.
Microsc Res Tech ; 79(9): 845-55, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27380767

RESUMO

This study describes the mucosa-associated lymphoid tissue (MALT) in odontocetes from the Brazilian coast and freshwater systems. Seven species were evaluated and tissue samples were analyzed by light, scanning and transmission electron microscopy, and immunohistochemistry. Laryngeal tonsil was a palpable oval mass located in the larynx, composed of a lymphoepithelial complex. Dense collections of lymphocytes were found in the skin of male fetus and calf. Clusters of lymphoid tissue were found in the uterine cervix of a reproductively active juvenile female and along the pulmonary artery of an adult female. Lymphoid tissues associated with the gastrointestinal tract were characterized by diffusely arranged or organized lymphocytes. The anal tonsil was composed of an aggregate of lymphoid tissue occurring exclusively in the anal canal, being composed of squamous epithelium branches. MALT was present in different tissues and organic systems of cetaceans, providing constant protection against mucosal pathogens present in their environment.


Assuntos
Tecido Linfoide , Tonsila Palatina , Baleias , Animais , Feminino , Tecido Linfoide/citologia , Tecido Linfoide/ultraestrutura , Masculino , Microscopia Eletrônica de Varredura , Mucosa/citologia , Mucosa/ultraestrutura , Tonsila Palatina/citologia , Tonsila Palatina/ultraestrutura , Baleias/anatomia & histologia , Baleias/imunologia
3.
An Acad Bras Cienc ; 87(3): 1727-36, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26221989

RESUMO

The reproductive morphology of cetaceans is poorly studied and, despite the large number of strandings, reports on this subject are scarce due to access to carcasses mostly in an advanced state of decomposition. The present study aimed to describe histological characteristics of the female genital tract of Sotalia guianensis, in order to assist in future studies on the reproductive biology of these animals. Females of different ages, from stranding events on beaches in northeastern Brazil, were used. Fragments of all organs were collected and processed for light and scanning electron microscopy. Histological analyses showed that these structures were similar to those found in terrestrial mammals, with some peculiarities, such as the presence of differentiated cells in the vulvar subepidermal layer, not described in the literature on cetaceans. Reproductive studies with a morphological description of the female genital organs are extremely important, since they would enable a better understanding of the species reproductive physiology and assist in the development of new strategies for the species conservation.


Assuntos
Golfinhos/anatomia & histologia , Genitália Feminina/anatomia & histologia , Animais , Brasil , Golfinhos/classificação , Feminino , Genitália Feminina/ultraestrutura , Microscopia Eletrônica de Varredura
4.
Microsc Res Tech ; 77(5): 348-55, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24644104

RESUMO

Aquatic mammals underwent morphological and physiological adaptations due to the transition from terrestrial to aquatic environment. One of the morphological changes regards their vision since cetaceans' eyes are able to withstand mechanical, chemical, osmotic, and optical water conditions. Due to insufficient information about these animals, especially regarding their sense organs, this study aimed to describe the morphology of the Humpback whale (Megaptera novaeangliae) eyeball. Three newborn females, stranded dead on the coast of Sergipe and Bahia, Brazil, were used. Samples were fixed in a 10% formalin solution, dissected, photographed, collected, and evaluated through light and electron microscopy techniques. The Humpback whale sclera was thick and had an irregular surface with mechanoreceptors in its lamina propria. Lens was dense, transparent, and ellipsoidal, consisting of three layers, and the vascularized choroid contains melanocytes, mechanoreceptors, and a fibrous tapetum lucidum. The Humpback whale eyeball is similar to other cetaceans and suggests an adaptation to diving and migration, contributing to the perception of differences in temperature, pressure, and lighting.


Assuntos
Olho/anatomia & histologia , Jubarte/anatomia & histologia , Animais , Câmara Anterior/anatomia & histologia , Câmara Anterior/ultraestrutura , Corioide/anatomia & histologia , Corioide/ultraestrutura , Corpo Ciliar/anatomia & histologia , Corpo Ciliar/ultraestrutura , Córnea/anatomia & histologia , Córnea/ultraestrutura , Olho/ultraestrutura , Feminino , Iris/anatomia & histologia , Iris/ultraestrutura , Cristalino/anatomia & histologia , Cristalino/ultraestrutura , Microscopia , Microscopia Eletrônica , Nervo Óptico/anatomia & histologia , Nervo Óptico/ultraestrutura , Retina/anatomia & histologia , Retina/ultraestrutura , Esclera/anatomia & histologia , Esclera/ultraestrutura
5.
Microsc Res Tech ; 76(8): 803-10, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23733492

RESUMO

The poultry industry is a sector of agribusiness which represents an important role in the country's agricultural exports. Therefore, the study about embryogenesis of the domestic chicken (Gallus gallus domesticus) has a great economic importance. The aim of this study was to evaluate embryonic development of the endoderm in chicken (Gallus gallus domesticus). Forty fertilized eggs of domestic chickens, starting from the 1st day of gestation and so on until the 19 days of the incubation were collected from the Granja São José (Amparo, SP, Brazil). Embryos and fetus were fixed in 10% formaldehyde solution, identified, weighed, measured, and subjected to light and scanning electron microscopy. The endoderm originates the internal lining epithelium of the digestive, immune, respiratory systems, and the organs can be visualized from the second day (48 h) when the liver is formed. The formation of the digestive system was complete in the 12th day. Respiratory system organs begin at the fourth day as a disorganized tissue and undifferentiated. Their complete differentiation was observed at the 10 days of incubation, however, until the 19 days the syrinx was not observed. The formation of immune system at 10th day was observed with observation of the spleen, thymus, and cloacal bursa. The study of the organogenesis of the chicken based on germ layers is very complex and underexplored, and the study of chicken embryology is very important due the economic importance and growth of the use of this animal model studies such as genetic studies.


Assuntos
Embrião de Galinha/embriologia , Desenvolvimento Embrionário , Endoderma/embriologia , Animais , Embrião de Galinha/anatomia & histologia , Embrião de Galinha/ultraestrutura , Galinhas/anatomia & histologia , Galinhas/crescimento & desenvolvimento , Endoderma/anatomia & histologia , Endoderma/ultraestrutura , Fígado/anatomia & histologia , Fígado/embriologia , Fígado/ultraestrutura , Microscopia Eletrônica de Varredura , Baço/anatomia & histologia , Baço/embriologia , Baço/ultraestrutura
6.
Infect Genet Evol ; 13: 18-26, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23000111

RESUMO

Infectious bursal disease is a highly contagious disease of young chickens caused by Infectious bursal disease virus (IBDV). Genome segment A encodes the capsid protein (VP2), while segment B encodes the RNA-dependent RNA polymerase (VP1). In the present study, we trace the molecular epidemiology of IBDV in Brazil by analyzing 29 isolates collected in the major regions of poultry production. To genetically characterize the isolates, phylogenetic and population dynamic analyses were conducted using 68 VP1 (2634 nt) and 102 VP2 (1356 nt) coding sequences from IBDV isolates from different regions of the world. Furthermore, the evolution of IBDV was analyzed by characterizing the selective forces that operated during the diversification of viral isolates. We show that IBDV isolates were introduced into Brazil mainly from the Netherlands and the USA. These introductions were associated with all Brazilian poultry production regions analyzed in this work. In addition, we show that the evolution of IBDV has been shaped by a combination of very low recombination rates and relatively high rates of nucleotide substitution (2.988×10(-4) for VP1 and 3.2937×10(-4) for VP2), which themselves are a function of purifying selection operating on VP1 and VP2. Furthermore, our extended Bayesian skyline plot suggests that the increase in the effective population size of isolates of IBDV is consistent with its epidemiological history, with a large increase during the emergence of acute outbreaks of IBD in the 1980s.


Assuntos
Infecções por Birnaviridae/epidemiologia , Vírus da Doença Infecciosa da Bursa/genética , Doenças das Aves Domésticas/epidemiologia , Animais , Brasil/epidemiologia , Recombinação Homóloga , Vírus da Doença Infecciosa da Bursa/classificação , Vírus da Doença Infecciosa da Bursa/isolamento & purificação , Dados de Sequência Molecular , Taxa de Mutação , Filogenia , Seleção Genética , Proteínas Estruturais Virais/genética , Virulência/genética
7.
Arq. neuropsiquiatr ; Arq. neuropsiquiatr;70(4): 271-272, Apr. 2012.
Artigo em Inglês | LILACS | ID: lil-622597

RESUMO

Paroxysmal dyskinesias (PD) are thought to be rare movement disorders. The overwhelming majority of reported cases are primary. Secondary PD has seen reported to occur in some conditions, mainly in multiple sclerosis and head trauma. The anatomic origin of the lesion is also rarely seen at the spinal cord. Our objective was to describe four patients with paroxysmal dystonia secondary to spinal lesions during the recovering phase of a neuromyelitis optica (NMO) bout. In the reviewed literature, we do not find any report of PD related to NMO.


Discinesias paroxísticas (DP) são distúrbios do movimento raros. A maioria dos casos relatados é de origem primária. DP secundárias têm sido relatadas em algumas condições, principalmente na esclerose múltipla e no trauma craniano. A origem anatômica da lesão também é raramente observada na medula. O objetivo deste trabalho foi descrever quatro pacientes com distonia paroxística secundária a lesões medulares, ocorrida durante a fase de recuperação do surto de neuromielite óptica (NMO). Na literatura consultada, não encontramos qualquer relato de DP secundárias à NMO.


Assuntos
Adulto , Idoso , Feminino , Humanos , Pessoa de Meia-Idade , Distonia/complicações , Neuromielite Óptica/complicações , Anticonvulsivantes/uso terapêutico , Carbamazepina/uso terapêutico , Distonia/diagnóstico , Distonia/tratamento farmacológico
8.
Arq Neuropsiquiatr ; 70(4): 271-2, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22358312

RESUMO

Paroxysmal dyskinesias (PD) are thought to be rare movement disorders. The overwhelming majority of reported cases are primary. Secondary PD has seen reported to occur in some conditions, mainly in multiple sclerosis and head trauma. The anatomic origin of the lesion is also rarely seen at the spinal cord. Our objective was to describe four patients with paroxysmal dystonia secondary to spinal lesions during the recovering phase of a neuromyelitis optica (NMO) bout. In the reviewed literature, we do not find any report of PD related to NMO.


Assuntos
Distonia/complicações , Neuromielite Óptica/complicações , Adulto , Idoso , Anticonvulsivantes/uso terapêutico , Carbamazepina/uso terapêutico , Distonia/diagnóstico , Distonia/tratamento farmacológico , Feminino , Humanos , Pessoa de Meia-Idade
9.
Virus Res ; 163(1): 320-7, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22056846

RESUMO

Porcine circovirus-2 (PCV-2) is an emerging virus associated with a number of different syndromes in pigs known as Porcine Circovirus Associated Diseases (PCVAD). Since its identification and characterization in the early 1990s, PCV-2 has achieved a worldwide distribution, becoming endemic in most pig-producing countries, and is currently considered as the main cause of losses on pig farms. In this study, we analyzed the main routes of the spread of PCV-2 between pig-producing countries using phylogenetic and phylogeographical approaches. A search for PCV-2 genome sequences in GenBank was performed, and the 420 PCV-2 sequences obtained were grouped into haplotypes (group of sequences that showed 100% identity), based on the infinite sites model of genome evolution. A phylogenetic hypothesis was inferred by Bayesian Inference for the classification of viral strains and a haplotype network was constructed by Median Joining to predict the geographical distribution of and genealogical relationships between haplotypes. In order to establish an epidemiological and economic context in these analyses, we considered all information about PCV-2 sequences available in GenBank, including papers published on viral isolation, and live pig trading statistics available on the UN Comtrade database (http://comtrade.un.org/). In these analyses, we identified a strong correlation between the means of PCV-2 dispersal predicted by the haplotype network and the statistics on the international trading of live pigs. This correlation provides a new perspective on the epidemiology of PCV-2, highlighting the importance of the movement of animals around the world in the emergence of new pathogens, and showing the need for effective sanitary barriers when trading live animals.


Assuntos
Infecções por Circoviridae/veterinária , Circovirus/classificação , Circovirus/genética , Filogeografia , Doenças dos Suínos/epidemiologia , Animais , Infecções por Circoviridae/virologia , Circovirus/isolamento & purificação , Biologia Computacional/métodos , DNA Viral/genética , Genótipo , Epidemiologia Molecular , Suínos , Doenças dos Suínos/virologia
10.
GED gastroenterol. endosc. dig ; GED gastroenterol. endosc. dig;30(Supl.1): 3-33, out.-dez. 2011. ilus
Artigo em Português | LILACS | ID: lil-621069

RESUMO

A encefalopatia hepática (EH) é um distúrbio funcional do sistema nervoso central (SNC) associado à insufi ciência hepática, de fisiopatologia multifatorial e complexa. Devido aos avanços no conhecimento sobre o manejo da EH na cirrose e na insuficiência hepática aguda (IHA), a diretoria da Sociedade Brasileira de Hepatologia (SBH) promoveu uma reunião monotemática acerca da fi siopatologia, diagnóstico e tratamento da EH, abordando aspectos controversos relacionados ao tema. Com a utilização de sistemática da medicina baseada em evidências, foram abordados o manejo da EH e da hipertensão intracraniana na IHA, o manejo da EH episódica na cirrose, as controvérsias no manejo da EH e a abordagem da EH mínima. O objetivo desta revisão é resumir os principais tópicos discutidos na reunião monotemática e apresentar recomendações sobre o manejo da síndrome votadas pelo painel de expertos da SBH.


Hepatic encephalopathy (HE) is a functional disorder of the central nervous system (CNS) associated with liver failure, either end-stage chronic liver disease or fulminant hepatic failure. Its pathogenesis remains complex and poorly understood. In view of recent advances in the management of HE, the Brazilian Society of Hepatology endorsed a monothematic meetingregarding HE in order to gather experts in the to discuss related data and to draw evidence-based recommendations concerning: management of HE and intracranial hypertension in FHF, treatment of episodic HE in cirrhosis, controversies in the management of EH including difficult to treat cases and diagnostic and treatment challenges for minimal HE. The purpose of this review is to summarize the lectures and recommendations made by the panel of experts of the Brazilian Society of Hepatology.


Assuntos
Humanos , Encefalopatia Hepática , Fibrose , Encefalopatia Hepática/diagnóstico , Encefalopatia Hepática/fisiopatologia , Transplante de Fígado , Falência Hepática Aguda , Hipertensão Intracraniana/prevenção & controle , Amônia , Hipertensão Portal
11.
Microb Drug Resist ; 17(2): 215-20, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21332364

RESUMO

In Brazil, the spread of an endemic clone of SPM-1-producing Pseudomonas aeruginosa has been reported. Recently, a higher genomic variety has been observed among the SPM-1-producing P. aeruginosa isolates. The principal aim of this study was to analyze through multilocus sequence typing (MLST) analysis whether the recently isolated SPM-1-producing P. aeruginosa descend or not from a common ancestor. A total of 50 SPM-1-producing P. aeruginosa exhibiting 11 distinct ribotyping genotypes collected from 11 different Brazilian cities were studied. Three IMP-1-producing P. aeruginosa and two non-metallo-beta-lactamase-producing P. aeruginosa isolates were included in the study as controls. For assignment of allelic numbers and subsequent determination of sequence type (ST), the obtained sequences were compared to existing sequences in the MLST database (www.pubmlst.org/paeruginosa). The eBURSTv3 software was used in this study for establishing the evolutionary relationship and phylogenetic analysis. A total of 5 different STs were identified among 55 P. aeruginosa isolates. All of the SPM-1-producing P. aeruginosa presented an identical allelic profile (ST277), except for one strain. The three IMP-1-producing P. aeruginosa strains were classified as belonging to the ST593, whereas the non-metallo-beta-lactamase-producing P. aeruginosa showed two new distinct STs, ST594 and ST595. Our study shows that SPM-1-producing P. aeruginosa isolates as well as the IMP producers evaluated in this study descend from a common ancestor.


Assuntos
DNA Bacteriano/genética , Filogenia , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/classificação , Pseudomonas aeruginosa/isolamento & purificação , beta-Lactamases , Alelos , Antibacterianos/farmacologia , Automação Laboratorial , Brasil/epidemiologia , Carbapenêmicos/farmacologia , DNA Bacteriano/classificação , DNA Bacteriano/isolamento & purificação , Bases de Dados Genéticas , Surtos de Doenças , Farmacorresistência Bacteriana/genética , Eletroforese em Gel de Campo Pulsado/métodos , Genótipo , Humanos , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus/métodos , Infecções por Pseudomonas/epidemiologia , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/genética , Ribotipagem/métodos , Software , beta-Lactamases/classificação , beta-Lactamases/genética , beta-Lactamases/isolamento & purificação
12.
Cephalalgia ; 31(1): 13-7, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20974599

RESUMO

UNLABELLED: The digiti quinti sign (DQS), described originally as a clinical indication of subtle motor deficit, consists of a relatively greater abduction of the fifth finger on the affected side when both arms are extended forwards. This sign was previously observed interictally in three consecutive hemiplegic migraine (HM) patients. PATIENTS AND METHODS: To verify whether the DQS specifically discriminates HM from non-hemiplegic migraine (nHM), the angle between the fourth and fifth fingers (ANG) was measured interictally in 10 HM patients, 44 migraine with aura and migraine without aura patients, and 45 healthy controls. RESULTS: The ANG was significantly wider at the symptomatic side in HM as compared with nHM and controls. The differences between the symptomatic and non-symptomatic (for HM) or between the right and left sides (absolute values for nHM and controls) were, respectively, 10.10° ± 9.58°, 4.15° ± 3.95° and 5.37° ± 4.74° (p = .007). The optimal cutoff point for ANG was 15° at the symptomatic side (sensitivity and specificity of 80.0% and 72.2%, respectively), 10.5° at the non-symptomatic side (sensitivity and specificity of 60.0% and 52.3%), and 3° for the difference between sides (sensitivity and specificity of 90.0% and 79.5%). CONCLUSION: Data show that the DQS discriminates HM from nHM and controls.


Assuntos
Dedos/fisiopatologia , Transtornos de Enxaqueca/diagnóstico , Adulto , Feminino , Humanos , Masculino , Transtornos de Enxaqueca/complicações , Transtornos de Enxaqueca/fisiopatologia , Debilidade Muscular/etiologia , Curva ROC , Sensibilidade e Especificidade
13.
Pesqui. vet. bras ; 29(1): 76-82, 2009. ilus, graf
Artigo em Inglês | VETINDEX | ID: vti-430

RESUMO

Porcine circovirus 2 (PCV2) is generally associated with the porcine circovirosis syndrome, which is considered an important disease of swine and has potentially serious economic impact on the swine industry worldwide. This article describes the construction of a recombinant plasmid expressing the PCV2 structural protein and the evaluation of cellular and humoral immune responses produced by this recombinant vaccine in BALB/c mice. The vaccine candidate was obtained and analyzed in vivo, in an effort to determine the ability to induce a specific immune response in mice. DNA was extracted from a Brazilian PCV2 isolate and the gene coding for Cap protein was amplified by PCR and inserted into an expression plasmid. Groups of BALB/c mice were inoculated intra-muscularly and intradermally in a 15-day interval, with 100 µg and 50 µg of the vaccine construct, respectively. Another group was inoculated intramuscularly with 100 µg of empty plasmid, corresponding to the control group. Seroconversion and cellular response in BALB/c mice were compared and used for vaccine evaluation. Seroconversion was analyzed by ELISA. After a series of 3 immunizations the spleen cells of the immunized animals were used to perform lymphocyte proliferation assays. Seroconversion to PCV2 was detected by ELISA in the animals inoculated with the vaccine construct when compared with control groups. Lymphocyte proliferation assays showed a stronger cell proliferation in the inoculated animals compared with the control group. Thus, the vaccine candidate construct demonstrated to be able to induce both humoral and cellular responses in inoculated mice.(AU)


O circovírus suíno 2 (PCV2) é geralmente associado à síndrome da circovirose suína, que é considerada uma importante doença de suínos e possui um sério impacto econômico na suinocultura mundial. Este trabalho descreve a construção de um plasmídeo recombinante que expressa a proteína estrutural do PCV2 e a avaliação das respostas imune humoral e celular por meio de vacinação em camundongos BALB/c. O candidato vacinal foi submetido a análises in vivo, determinando a capacidade de induzir resposta imune específica em camundongos. O DNA de um isolado brasileiro de PCV2 foi extraído e o gene que codifica para a proteína do capsídeo foi amplificado por PCR e inserido num plasmídeo de expressão. Grupos de camundongos BALB/c foram inoculados por via intramuscular e intradérmica a cada 15 dias, com 100µg e 50µg da construção vacinal, respectivamente. Outro grupo foi inoculado com 100µg do plasmídeo original, correspondente ao grupo controle. A soroconversão e a resposta celular dos grupos de camundongos BALB/c vacinados foram comparados como parâmetros de avaliação vacinal. A soroconversão foi avaliada por um teste de ELISA. Após 3 imunizações, as células esplênicas dos animais imunizados foram utilizadas nos ensaios de linfoproliferação. A soroconversão para o PCV2 foi detectada por ELISA nos animais inoculados com a construção vacinal quando comparados com o grupo controle. Nos ensaios de linfoproliferação foi observada uma grande proliferação celular nos animais inoculados comparados ao grupo controle. Portanto, o candidato vacinal demonstrou ser capaz de induzir tanto uma resposta humoral e celular nos camundongos inoculados.(AU)


Assuntos
Animais , Circovirus/isolamento & purificação , Vacinas de DNA/efeitos adversos , Ensaio de Imunoadsorção Enzimática/métodos , Camundongos
14.
Pesqui. vet. bras ; Pesqui. vet. bras;29(1): 76-82, jan. 2009. ilus, graf
Artigo em Inglês | LILACS | ID: lil-509259

RESUMO

Porcine circovirus 2 (PCV2) is generally associated with the porcine circovirosis syndrome, which is considered an important disease of swine and has potentially serious economic impact on the swine industry worldwide. This article describes the construction of a recombinant plasmid expressing the PCV2 structural protein and the evaluation of cellular and humoral immune responses produced by this recombinant vaccine in BALB/c mice. The vaccine candidate was obtained and analyzed in vivo, in an effort to determine the ability to induce a specific immune response in mice. DNA was extracted from a Brazilian PCV2 isolate and the gene coding for Cap protein was amplified by PCR and inserted into an expression plasmid. Groups of BALB/c mice were inoculated intra-muscularly and intradermally in a 15-day interval, with 100 µg and 50 µg of the vaccine construct, respectively. Another group was inoculated intramuscularly with 100 µg of empty plasmid, corresponding to the control group. Seroconversion and cellular response in BALB/c mice were compared and used for vaccine evaluation. Seroconversion was analyzed by ELISA. After a series of 3 immunizations the spleen cells of the immunized animals were used to perform lymphocyte proliferation assays. Seroconversion to PCV2 was detected by ELISA in the animals inoculated with the vaccine construct when compared with control groups. Lymphocyte proliferation assays showed a stronger cell proliferation in the inoculated animals compared with the control group. Thus, the vaccine candidate construct demonstrated to be able to induce both humoral and cellular responses in inoculated mice.


O circovírus suíno 2 (PCV2) é geralmente associado à síndrome da circovirose suína, que é considerada uma importante doença de suínos e possui um sério impacto econômico na suinocultura mundial. Este trabalho descreve a construção de um plasmídeo recombinante que expressa a proteína estrutural do PCV2 e a avaliação das respostas imune humoral e celular por meio de vacinação em camundongos BALB/c. O candidato vacinal foi submetido a análises in vivo, determinando a capacidade de induzir resposta imune específica em camundongos. O DNA de um isolado brasileiro de PCV2 foi extraído e o gene que codifica para a proteína do capsídeo foi amplificado por PCR e inserido num plasmídeo de expressão. Grupos de camundongos BALB/c foram inoculados por via intramuscular e intradérmica a cada 15 dias, com 100µg e 50µg da construção vacinal, respectivamente. Outro grupo foi inoculado com 100µg do plasmídeo original, correspondente ao grupo controle. A soroconversão e a resposta celular dos grupos de camundongos BALB/c vacinados foram comparados como parâmetros de avaliação vacinal. A soroconversão foi avaliada por um teste de ELISA. Após 3 imunizações, as células esplênicas dos animais imunizados foram utilizadas nos ensaios de linfoproliferação. A soroconversão para o PCV2 foi detectada por ELISA nos animais inoculados com a construção vacinal quando comparados com o grupo controle. Nos ensaios de linfoproliferação foi observada uma grande proliferação celular nos animais inoculados comparados ao grupo controle. Portanto, o candidato vacinal demonstrou ser capaz de induzir tanto uma resposta humoral e celular nos camundongos inoculados.


Assuntos
Animais , Circovirus/isolamento & purificação , Ensaio de Imunoadsorção Enzimática , Camundongos , Vacinas de DNA/efeitos adversos
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