RESUMEN
The comparative analysis of expression level of FCRL1 gene encoding human B-cell surface receptor in healthy individuals and patients with autoimmune diseases was carried out. For the expression estimation we used results of DNA dot-hybridization on the membranes, containing cDNA samples from subpopulations of blood cells of patients with autoimmune diseases. The quantitative estimation of hybridization signals showed that expression level of FCRL1 gene in peripheral blood B-lymphocytes was significantly higher in patients with a multiple sclerosis, lupus anticoagulans, Takayasu's arteritis and also in von Willebrand disease than in healthy individuals. FCRL1-specific monoclonal and polyclonal antibodies were raised. They were proven to detect FCRL1 in Western blotting, immunohistochemistry and flow cytometry. It was found that FCRL1 is expressed on the surface of CD19+ mature B-cells. In tonsil FCRL1-positive cells were located in crypt area: in mantle zone of secondary lymphoid follicles and among cells of lymphoepithelium. FCRL1-positive cells were also found in B-cell follicles of the spleen.
Asunto(s)
Linfocitos B/metabolismo , Inhibidor de Coagulación del Lupus/genética , Proteínas de la Membrana/genética , Esclerosis Múltiple/genética , Arteritis de Takayasu/genética , Enfermedades de von Willebrand/genética , Anticuerpos/inmunología , Antígenos CD19/inmunología , Autoinmunidad , Linfocitos B/inmunología , Western Blotting , Estudios de Casos y Controles , ADN Complementario/análisis , Expresión Génica , Humanos , Inmunohistoquímica , Inhibidor de Coagulación del Lupus/inmunología , Esclerosis Múltiple/inmunología , Esclerosis Múltiple/patología , Tonsila Palatina/inmunología , Tonsila Palatina/metabolismo , Bazo/inmunología , Bazo/metabolismo , Arteritis de Takayasu/inmunología , Arteritis de Takayasu/patología , Enfermedades de von Willebrand/inmunología , Enfermedades de von Willebrand/patologíaRESUMEN
The IFGP family is a recently identified group of human and mouse genes structurally related to the genes of leukocyte Fc-receptors. In this study we compared expression patterns of six human and four mouse IFGP genes. With the exception of mouse IFGP2, the genes of the family were found to be predominantly expressed in haemopoietic cells. Expression of human IFGP1-IFGP5 and mouse IFGP3 was B cell-specific. Mouse IFGP1 transcripts were mainly found in B cells, but this gene may be either expressed by nonlymphoid cells. Expression of the human IFGP6 was detected in CD8+ T cells and NK cells. We further demonstrated that alternative splicing of human IFGP1 and IFGP6 mRNA may generate transcripts coding for the previously unknown isoforms. The novel IFGP1 isoform lacks transmembrane domain, whereas the IFGP6 isoform has altered cytoplasmic tail. The data obtained indicate that the receptors of family may contribute to the regulation of development and/or functions of three effector types of lymphocytes, namely B cells, CD8 T cells and NK cells.
Asunto(s)
Empalme Alternativo , Linfocitos B/inmunología , Células de la Médula Ósea/inmunología , Receptores Inmunológicos/genética , Animales , Expresión Génica , Humanos , Ratones , ARN Mensajero/metabolismoRESUMEN
cDNA library in the lambda gt11 phage was constructed using poly (A+)-mRNA from mink spleen as a template. Immunoscreening of the library allowed the identification of 2 lambda-related clones containing 370 and 803 bp insertion (lambda IGL-1 and lambda IGL-2). Analysis of the primary structure of lambda IGL-2 demonstrated that it contains a large portion of V lambda-segment, J lambda-segment, C lambda-gene and its 3'-untranslated part. The nucleotide sequences known for the immunoglobulin genes were compared to the sequence of the lambda IGL-2 clone. The highest degree of homology was established for the rabbit lambda-genes, this being 63, 94 and 72% for the RF3 region of V lambda-segment, J lambda-segment and C lambda-region, respectively.
