RESUMO
OBJECTIVES: Nurse-like stromal cells (NLC) in synovia and bone marrow of patients with rheumatoid arthritis (RA) can support pseudoemperipolesis, protect from apoptosis and enhance immunoglobulin production of peripheral blood B cells isolated from healthy individuals, suggesting the profound contribution of hyperactivation of B cells in RA. In the course of establishing RA-NLC from RA patients, we observed the growth of B cells in the presence of RA-NLC. METHODS: We cloned B cells from the synovium or bone marrow of RA patients using the limiting dilution technique. For established clones, nucleotide sequences of immunoglobulin and surface antigens were investigated. To investigate the dependence of these clones on NLC, differences in the proliferation and the amount of immunoglobulin produced in the presence or absence of NLC were compared. Immunocytochemical staining of various cells was performed using the antibody these clones produced. RESULTS: Nine B-cell clones established from RA patients showed RA-NLC-dependent growth. These B-cell clones expressed CD19, CD20, CD38, CD39 and CD40, suggesting that the cloned cells were mature and activated. All clones secreted immunoglobulins in culture media, which were specific for intracellular components of various cell lines, including RA-NLC. Interestingly, we found limited usage of immunoglobulin heavy-chain variable regions (VH) among B-cell clones from RA patients. These repertoires were reported to be detected preferentially in fetal livers. CONCLUSION: The present study provides a novel insight into the involvement of RA-NLC in the immunopathogenesis of RA via an autoreactive B cell development and/or activation mechanism.
Assuntos
Artrite Reumatoide/imunologia , Linfócitos B/imunologia , Genes de Imunoglobulinas , Cadeias Pesadas de Imunoglobulinas/genética , Região Variável de Imunoglobulina/genética , Antígenos CD/metabolismo , Artrite Reumatoide/genética , Autoanticorpos/biossíntese , Autoantígenos/imunologia , Comunicação Celular/imunologia , Proliferação de Células , Células Clonais/imunologia , Humanos , Imunoglobulinas/biossíntese , Imunofenotipagem , Ativação Linfocitária/imunologia , Células Estromais/imunologia , Membrana Sinovial/imunologia , Células Tumorais CultivadasRESUMO
The nucleotide sequence of the matrixprotein (M) gene of the lapinized rinderpest virus (RPV-L) was determined. The full-length cDNA of the RPV-L M gene is composed of 1460 base pairs and is supposed to contain an open reading frame of 1005 nucleotides encoding on M protein of 335 amino acids. The homology of the predicted amino acid among congeneric morbilliviruses such as RPV Kabete 'O' strain (wild strain of RPV), RPV RBOK strain (vaccine strain of RPV for cattle), measles virus (MV), and canine distemper virus (CDV), is approximately 94%, 93%, 87% and 77%, respectively. In the present study, all coding regions of the RPV-L strain have been determined.
Assuntos
Vírus da Peste Bovina/genética , Proteínas da Matriz Viral/genética , Sequência de Aminoácidos , Sequência de Bases , DNA Viral/química , DNA Viral/genética , Genes Virais , Dados de Sequência Molecular , Filogenia , RNA Viral/química , RNA Viral/genética , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Peste Bovina/virologia , Vírus da Peste Bovina/química , Homologia de Sequência de Aminoácidos , Proteínas da Matriz Viral/químicaRESUMO
Forty Caspian seals were surveyed seroepidemiologically between 1993 and 1998 around the times of mass mortality that occurred in 1997 in the Caspian Sea and seven Baikal seals were also surveyed in 1998. Virus neutralizing tests and ELISA clearly suggested that distemper virus epidemic was caused in Caspian seals before the spring of 1997 and that CDV infection continued to occur in Lake Baikal in recent years.
Assuntos
Anticorpos Antivirais/sangue , Surtos de Doenças/veterinária , Vírus da Cinomose Focina/imunologia , Infecções por Morbillivirus/veterinária , Focas Verdadeiras/virologia , Animais , Ensaio de Imunoadsorção Enzimática/métodos , Ensaio de Imunoadsorção Enzimática/veterinária , Infecções por Morbillivirus/epidemiologia , Infecções por Morbillivirus/virologia , Federação Russa/epidemiologia , Estudos Soroepidemiológicos , Sibéria/epidemiologiaAssuntos
Animais Selvagens/virologia , Carnívoros/virologia , Surtos de Doenças/veterinária , Vírus da Cinomose Canina/isolamento & purificação , Cinomose/virologia , Sequência de Aminoácidos , Animais , Anticorpos Antivirais/sangue , Cinomose/epidemiologia , Vírus da Cinomose Canina/classificação , Japão/epidemiologia , Filogenia , Estudos SoroepidemiológicosRESUMO
Antigenic properties between new Japanese field isolates and vaccine strains of canine distemper virus (CDV) have been compared using four monoclonal antibodies (MAbs) (JD-5, JD-7, JD-11 and d-7) against the hemagglutinin (H) proteins of CDV. JD-5, JD-7 and JD-11 are newly established antibodies. Three MAbs, namely d-7, JD-5 and JD-11, reacted similarly to all the CDV strains examined. However, JD-7 reacted much more strongly with the vaccine strains and an old field isolate than the recent field isolates in immunofluorescence, radio immunoprecipitation and virus neutralization assays. These results indicate that an antigenic region in the H protein, concerned with neutralization and recognized by JD-7, has been altered in the recent field isolates.
Assuntos
Vírus da Cinomose Canina/imunologia , Cinomose/imunologia , Hemaglutininas Virais/imunologia , Animais , Anticorpos Monoclonais/imunologia , Antígenos Virais/classificação , Antígenos Virais/imunologia , Chlorocebus aethiops , Efeito Citopatogênico Viral , Cinomose/virologia , Vírus da Cinomose Canina/classificação , Cães , Ensaio de Imunoadsorção Enzimática/veterinária , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Hemaglutininas Virais/classificação , Japão , Camundongos , Camundongos Endogâmicos BALB C , Testes de Neutralização/veterinária , Ensaio de Radioimunoprecipitação/veterinária , Células Vero , Vacinas Virais/imunologiaRESUMO
The nucleotide sequences of the phosphoprotein (P) of canine distemper virus (CDV) strains isolated between 1992 and 1996 in Japan were determined. This is the first report of the complete sequences of the P genes of recently prevalent CDV strains. The deduced amino acid sequences of the P, C and V proteins showed that in the new Japanese isolates, these proteins have approximately 93%, 90-91% and 92% identities with those of the Onderstepoort vaccine strain, respectively. The predicted functional regions were conserved. RNA editing resulting in a shift to the open reading frame (ORF) of the V protein was shown to occur with the same efficiency in both the field isolates and vaccine strain.
