OBJECTIVE: The adhesion molecule that mediate cell-cell and cell-extracellular matrix adhes.ion provides very important role in growth and differentiation of cells and tissue. VLA integrin is a prototype of adhesion molecule which participate in cell-cell and cell-extracellular matrix interacton, especially for collagen, laminin and fibronectin. The biologic functions of VLA-integrin are very diverse. Cartilage is the target tissue of various arthritides including rheumatoid arthritis and osteoarthritis and the process of homeostasis in cartilage matrix may be very important in preservation of cartilage. Although VLA integrin may participate in the process of cartilage degeneration and repair mechanism, tissue.distribution and exact role of VLA integtin in cartilage was not yet clearly defined. METHODS:Immunohistochemical analysis of VLA-integrin in cryostat section of articular cartilage was conducted using monoclonal antibody and avidin-biotin complex method. Analysis was performed in 10 rheumatoid arthritis specimens, 7 osteoarthritis specimens and 1 normal control. RESULTS: 1) Normal cartilage showed strong and diffuse stain with CD29, CD51 and moderate stain of VLA-5. Staining pattern of VLA-1 and 3 was inconstant and weak in intensity. 2) The intensity of VLA expression in articular cartilage of osteoarthritis was upregulated chiefly in CD29, CD51 and slightly in VLA-5. The positive rate of VLA-1 and 3 was similar to that of normal cartilage though the intensity was increased especially at cluster of chondrocytes. 3) VLA-integrin expression of rheumatoid arthritis cartilage was similar to that of osteoarthritis cartilage. CONCLUSION: VLA integrins functioning as fibronectin receptor such as VLA-5 and alpha, beta1 were upregulated in osteoarthritis and rheumatoid arthritis. Intensity was increased at clusters of chondrocytes. It was able to presume from above findings that VLA molecule has some role in the maintenance and repair of articular cartilage. The regulation of expression by cytokines and growth factors and exact function of VLA molecule in cartilage have to be elucidated.
Arthritis ; Arthritis, Rheumatoid ; Cartilage ; Cartilage, Articular* ; Chondrocytes ; Collagen ; Cytokines ; Fibronectins ; Homeostasis ; Integrin alpha1beta1 ; Integrin alpha5beta1 ; Integrins ; Intercellular Signaling Peptides and Proteins ; Laminin ; Osteoarthritis

BACKGROUND: It has been found that Helper T cells in the peripheral blood are decreased in the cell mediated immunity in the pulmonary tuberculosis But it has not been confirmed yet that only decrease in number of cells which has phenotype in the peripheral blood is defined to decrease in cell mediated immunity. The immunocytochemical study was performed to observe the change of the percentage of T-lymphocytes with their subsets and activated T cells in the peripheral blood of pulmonary tuberculosis and to know how many T cells would be activated, relative to resting cells in the peripheral blood. METHODS: The peripheral blood obtained from twenty two patients and ten healthy controls were smeared on the gelatin coated slide glass prepared for of mononuclear cells. The double bridge technique of alkaline phosphatase-antialkaline phosphatase(APAAP) method was used. As the primary antibodies, T1(anti-human T cell), T4(anti-human helper/inducer T cells) and T8(anti-human supressor/cytotoxic T cell) antibodies and interleukin-2 receptor (for early activated T cell),very late activation antigen (for activated cytotoxic T cell), T cell lineage specific activation antigen monoclonal actibodies were used. RESULTS: 1) There were significantly decrease in the absolute number of T4(+) cells but significantly increase of T8(+) cells in the peripheral blood of pulmonary tuberculosis (p<0.05). 2) The percentage of T4(+) cells showed significantly decrease in pulmonary tuberculosis but T8 (+)cells significantly increase(p<0.05). T4(+)/T8 (+) ratio showed significantly decrease in the peripheral blood of the pulmonary tuberculosis(p <0.05) 3) There were significantly increase in the absolute number of variable stages of activated T cells in the peripheral blood of the pulmonary tuberculosis(p<0.05). 4) The percentage of IL-2R, VLA-1, TLiSA were 6.45+1.56%, 7.64+1.34*, 10.45+1.16% in order which showed significantly increase in the peripheral blood of the pulmonary tuberculosis(p <0.05). CONCLUSION: We speculate that only a few percentage of T lymphocyte is activated in cell mediated immunity in pulmonary tuberculosis.
Antibodies ; Cell Lineage ; Gelatin ; Glass ; Humans ; Immunity, Cellular ; Integrin alpha1beta1 ; Interleukin-2 ; Lymphocytes ; Phenotype ; T-Lymphocytes* ; T-Lymphocytes, Helper-Inducer ; Tuberculosis, Pulmonary*

Animals ; Arginine Vasopressin ; physiology ; Connective Tissue Growth Factor ; physiology ; Glomerular Mesangium ; pathology ; Humans ; Integrin alpha1beta1 ; physiology ; Kidney Diseases ; etiology ; Proto-Oncogene Protein c-ets-1 ; physiology

In order to help define the possible role of adhesion molecules in corneal inflammation, we investigated the distribution of adhesion molecules in normal and Aspergillus fumigatus keratitis-induced corneas of rabbits in process if time. Each 4 corneas were resected at 3, 12, 24, and 72 hours after intracorneal injection with A. fumigatus. Normal corneas (4 eyes) were used as a control. Monoclonal antibodies to beta 1 subunit of VLAs, alpha 1 subunit of VLA-1, LFA-1, ICAM-1,and ELAM-1 were used for immunohistochemical staining of 20 corneas.The results were as follows: In normal cornea, beta 1 subunit of VLAs was expressed on all parts of the cornea, and ICAM-1 was expressed on corneal stroma and endothelium. Vascular endothelium showed expression of beta 1 subunit of VLAs and ICAM-1 after 12 hours of intracorneal injection, and alpha1 subunit of VLA-1 and ELAM-1 at 72 hours after injection. In inflamed cornea, beta 1 subunit of VLAs was expressed strongly at 72 hours after injection. Alpha1 subunit of VLA-1 was detected on corneal stroma after 12 hours of injection, and on corneal endothelium at 72hours after injection. Expression of beta 2 subunit of LFA-1 was weak on corneal stroma after 3 hours injection, and on corneal endothelium at 72 hours after injection. ICAM-1 expression was detected weakly on corneal epithelium, and increased on corneal stroma and endothelium at 72 hours after injection. ELAM-1 was expressed weakly on corneal stroma after 3 hours of injection, and on corneal endothelium at 72 hours after injection.In this study, it was found that the invasion of A. fumigatusinto the cornea causes localized inflammatory reaction that results in activation of corneal cells (keratocytes, corneal endothelial cells and epithelial cells), and subsequent expression of adhesion molecules in the cornea. Expression of adhesion molecules facilitates the inflammatory cells to be migrated into the cornea with inflammmation.
Antibodies, Monoclonal ; Aspergillus fumigatus* ; Aspergillus* ; Cornea* ; Corneal Stroma ; E-Selectin ; Endothelial Cells ; Endothelium ; Endothelium, Corneal ; Endothelium, Vascular ; Epithelium, Corneal ; Inflammation ; Integrin alpha1beta1 ; Intercellular Adhesion Molecule-1 ; Keratitis* ; Lymphocyte Function-Associated Antigen-1 ; Rabbits

We produced beta1 gene which is about 2400 bp by reverse transcription polymerase chain reaction (RT-PCR) from bovine trachea, reclaimed and purified, then cloned the amplified fragment to pGEM-T easy vector, confirmed by sequencing. The immune-dominant epitope of beta1 gene was chosen by computer analysis and then syncretized ligand-binding domain from 346 bp to 843 bp of ecytoplasm with six histidine, expressed LBD protein massly in E. coli BL21 (DE3), and identified by SDS-PAGE. The fusion protein was purified with Ni-NTA affinity chromatography and immunized New Zealand rabbits preparing of its polyclonal antibody, the specific antibody titer was above 1:12,800 detected by indirect ELISA, the result of Western blot showed that this antibody could be recognized by LBD fusion protein.
Animals ; Antibodies, Monoclonal ; biosynthesis ; Cattle ; Escherichia coli ; genetics ; metabolism ; Foot-and-Mouth Disease Virus ; physiology ; Integrin alpha1beta1 ; biosynthesis ; genetics ; immunology ; Ligands ; Protein Binding ; Protein Interaction Domains and Motifs ; Rabbits ; Receptors, Virus ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology