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Biomedical subjects

J Pahle

Publications and source records attributed to J Pahle.

At least 19 recordsLinked to original sources

Responses of normal and rheumatic human articular chondrocytes cultured under various experimental conditions in agarose.

The purpose of the present study was to test if agarose could support the maintenance of normal and arthritic human chondrocytes in culture, and under which experimental conditions they could be successfully grown. Cultures of chondrocytes isolated from articular cartilage from patients with rheumatoid arthritis (RA), juvenile rheumatoid arthritis (JRA), and healthy controls were assessed by light microscopy, alcian blue staining, formazan uptake and incorporation of radiosulfate into the extracellular matrix. The results showed that both normal and arthritic chondrocytes proliferated, and synthesized proteoglycan (PG) in agarose in short term and long term culture. Proliferation and PG synthesis occurred at a slower rate in chondrocytes from adult rheumatic patients than from healthy controls. Supplements to the medium influenced chondrocyte proliferation, PG synthesis and release into the medium. Serum from RA patients stimulated chondrocyte responses more than normal human serum (NHS), and NHS promoted PG synthesis more than fetal calf serum (FCS). Exposure to inflammatory synovial fluid (SF) enhanced PG synthesis of healthy chondrocytes, but suppressed it in arthritic chondrocytes. We conclude that species-specific serum is optimal for chondrocyte cultures, and that disease related culture conditions change the chondrocyte response. As metabolic responses of human chondrocytes are maintained in agarose, this culture system appears as a suitable in vitro tool for further studies of human joint disease.

Adolescent↗

IgG and IgA subclass distribution of total immunoglobulin and rheumatoid factors in rheumatoid tissue plasma cells.

The subclass distribution of IgG and IgA plasma cells, and in IgG and IgA rheumatoid factor (RF) producing cells was studied in sections of synovial tissue from seropositive RA and various types of seronegative arthritis, including ankylosing spondylitis, psoriatic arthritis, and Reiter's syndrome. The study was performed with immunofluorescence technique and monoclonal IgG and IgA subclass specific antibodies. IgG RF producing cells were identified by their ability to bind and activate factors both in the early (C3) and late (C5b-9) part of the complement cascade. IgA RF cells were identified by double staining experiments with heat-aggregated IgG and monoclonal antibodies to IgA subclasses. In 23 tissues tested for total IgG, IgG1 cells were usually predominant, while the frequency of IgG3 cells was usually higher than that of IgG2. In 19 tissues also tested for IgA, both IgA subclasses were present in all tissues. IgA1 plasma cells were always predominant, with a mean ratio of IgA1 to IgA2 cells of approximately 10. In the 13 tissues tested for RF-producing cells, the highest frequency of IgG RF cells was found among the IgG3 cells, followed by IgG1 and IgG2. IgA RF cells were found in only one case, all cells being IgA1.

Antigen-Antibody Complex↗

Phenotypes and spontaneous cell cytotoxicity of mononuclear cells from patients with seronegative spondyloarthropathies: ankylosing spondylitis, psoriatic arthropathy and pauciarticular juvenile chronic arthritis--analysis of mononuclear cells from peripheral blood, synovial fluid and synovial membranes.

T-cell subpopulations and natural killer (NK) cells from peripheral blood, synovial fluid and synovial membranes from patients with seronegative spondyloarthropathies were investigated. Thirty-four patients with ankylosing spondylitis, sixteen patients with psoriatic arthropathy and six patients with pauciarticular juvenile chronic arthritis were studied. All the patient groups had normal proportions of T4+ and T8+ cells as well as normal T4/T8 ratios in peripheral blood. In the synovial fluids the T4/T8 ratios were reduced in ankylosing spondylitis and psoriatic arthropathy (p less than 0.05). Although both the T4 and T8 subpopulations were reduced, the T4/T8 ratios in the synovial membranes of patients with these two disorders tended to be within the normal range of that of peripheral blood. Increased numbers of T-cells in the synovial fluid from patients with ankylosing spondylitis expressed class II MHC antigens. The natural killer cell activity was normal in peripheral blood and synovial fluids of patients with ankylosing spondylitis and psoriatic arthropathy while it tended to be reduced, although not significantly, in pauciarticular juvenile chronic arthritis. Synovial membranes were almost devoid of NK cell activity. The number of Leu 7+ cells were reduced in synovial fluid of patients with psoriatic arthropathy (p less than 0.04), but not as significantly as in the two other patient groups.

Adult↗

Characteristics of human rheumatoid synovial and normal blood dendritic cells. Retention of class II major histocompatibility complex antigens and accessory function after short-term culture.

Dendritic cells (DC) were isolated from synovial tissue and synovial fluids of patients with rheumatoid arthritis and from peripheral blood of healthy donors. The cells were analysed for various surface antigens in indirect immunofluorescence by means of monoclonal antibodies. Surface antigen expression and accessory activity of the DC during short-term cultures were also investigated. Both the rheumatoid synovial and the normal blood DC were strongly positive for panleucocyte antigen and class II major histocompatibility complex (MHC) antigens (HLA-DP, HLA-DQ, and HLA-DR). The DC suspensions (purity approximately 80-85%) showed very low percentage of cells staining for various other cell membrane markers, including B cell, T cell, natural killer (NK) cell, and various monocyte/macrophage markers as well as markers specific for dendritic reticulum cells and Reed Sternberg cells. Moreover, neither rheumatoid nor normal DC reacted with the RFD1 monoclonal antibody, which is specific for interdigitating cells of human thymus. In contrast to Langerhans' cells, the DC lacked the thymocyte (T6) marker. The various DC expressed neither complement receptors (CR1, CR3), transferrin receptors, nor Fc receptors. They also lacked enzyme markers like peroxidase and nonspecific esterase. The DC formed clusters with autologous T cells. Cluster formation was readily inhibited by anti-HLA-DR and anti-CD2 (T11) monoclonal antibodies. After 3 to 5 days in culture the DC still expressed class II MHC antigens and were potent stimulators in allogeneic mixed lymphocyte reactions (MLR). Only a small number of cells in the DC suspensions from synovial tissue expressed fibroblast antigens before and after culture.

Antigen-Presenting Cells↗

Complement activation in rheumatoid arthritis evaluated by C3dg and the terminal complement complex.

Complement activation was evaluated in plasma and synovial fluid from patients with rheumatoid arthritis. The activation fragment C3dg and the fluid-phase terminal complement complex were used as indicators of initial and terminal activation, respectively. Considerable activation of the whole complement cascade was demonstrated in most synovial fluid samples and in one-third of the plasma samples. No correlation was found between the level of activation products in synovial fluid and the level in plasma. Therefore, both compartments must be examined in order to evaluate local and systemic complement activation in rheumatoid arthritis.

Arthritis, Rheumatoid↗

Dendritic cells from human rheumatoid synovial inflammatory tissue and peripheral blood as accessory cells in mitogen stimulation of T lymphocytes.

Dendritic cells (DC) were purified from the peripheral blood (PB) of normal individuals and from the synovial fluid (SF) and synovial tissue (ST) of patients with rheumatoid arthritis. These cells are strongly HLA-DR positive and lack B-cell, T-cell, and monocyte markers as well as Birbeck granules. The DC were compared with monocytes and non-T cells from PB for their ability to act as accessory cells for T-cell responses to concanavalin A (Con A) and phytohaemagglutinin (PHA). DC from PB, SF and ST were much more efficient accessory cells for the mitogenic responses than autologous monocytes from PB. The mean PHA responses in cpm obtained with DC from the various compartments were 4-20 times greater than the responses obtained with monocytes from PB. The Con A responses obtained when the various DC populations were used as accessory cells were 3-13 times greater than those obtained with monocytes from PB. The mitogenic responses seen with monocytes were very low. The non-T cells, which comprise a mixture of cells obtained after removal of T cells, also gave low T-cell responses to PHA and Con A compared with DC as accessory cells.

Antigen-Presenting Cells↗

In situ characterization of mononuclear cells in rheumatoid tissues, using monoclonal antibodies. No reduction of T8-positive cells or augmentation in T4-positive cells.

The reactivity of monoclonal mouse anti-human antibodies specific for mononuclear cell surface antigens were studied by the indirect immunofluorescence technique in frozen synovial tissue sections from patients with rheumatoid arthritis (RA). Most of the proliferating synovial lining cells were positive for HLA-DR antigens and monocyte-specific antigens, since they reacted with the OKIa1 and OKM1. Cells positive for HLA-DR and monocyte antigens were also seen scattered or in small nests in the synovial stroma, probably representing synovial cells or monocytes/macrophages. Some of the HLA-DR-positive cells may also be B lymphocytes or activated T cells. Endothelial cells were also HLA-DR antigen-positive. Monoclonal antibodies with specificity for all T cells (OKT3), for helper/inducer cells (OKT4), and for suppressor/cytotoxic cells (OKT8) reacted with cells often located in follicle-like structures around vessels. Cells with the T4 phenotype tended to be located in the centre of the follicles, whereas the T8 positive cells were more peripherally situated. In most instances fewer cells were positive for the T8 than for the T4 marker. In some instances there was as many T8-positive cells as T4-positive cells. Complete lack of T-lymphocyte subpopulations was not seen.

Animals↗

Physical activity in the treatment of inflammatory rheumatic disorders.

A search of the literature in this area shows that there is little national or international documentation available on the effect of physical activity on the development of disease in inflammatory rheumatic disorders. Low capacity for physical work has been demonstrated and is based on poor circulation and reduced muscle strength in rheumatism: several studies from Norway and elsewhere have documented the value of physical training, especially in improving the rheumatic patient's physical condition (1--2, 8). No investigations have been able to document any arrest of progress of the disease, but physical training seems to be able to maintain existing joint function and also to some extent prevent progressive deterioration in condition. There is therefore no disagreement about the significance of physiotherapy, but the intensity and duration of training are more controversial.

Exercise Therapy↗

Spontaneous release of leucocyte migration-inhibitory factor by mononuclear cells eluted from rheumatoid synovial tissue.

Spontaneous leucocyte migration-inhibitory factor (LIF) was assayed by an indirect system in which mononuclear cells (MNC) eluted from rheumatoid synovial tissues or isolated from peripheral blood of normal donors were cultured without antigen. The supernatants from these cultures and control supernatants, heated to 80 degrees C for 30 min to destroy the LIF activity, were used in the test. In seven out of eleven rheumatoid arthritis patients the eluted synovial MNC produced LIF spontaneously, whereas none of the twelve normal blood donors showed any such production. The ability to show spontaneous LIF production was primarily seen in the joint tissues from sero-negative patients.

Arthritis, Juvenile↗

Lymphocytes from synovial tissue of a boy with X-linked hypogammaglobulinemia and chronic polyarthritis.

Lymphocytes were eluted from synovial tissue of a boy with X-linked hypogammaglobulinemia and chronic polyarthritis. The cell suspension contained 53% lymphocytes and 28% peroxidase-positive, macrophagelike cells, No B lymphocytes, 83% T lymphocytes, and 10% Fc-receptor-bearing lymphocytes were detected. Lymphocyte transformation was induced by polyclonal mitogens (phytohemagglutinin and pokeweed mitogen) whereas no response to antigens (purified protein derivative and Candida albicans antigen) was obtained. The eluted cells displayed antibody-dependent cytotoxicity.

Agammaglobulinemia↗

Capillary column gas chromatography--mass spectrometry in studies on rheumatoid arthritis.

A method is described for determining the organic acid profile of synovial fluid. The method requires 50--100 microgram of sample and involves ion exchange for isolation of the acids followed by combined capillary column gas chromatography--mass spectrometry. Normal synovial fluid and synovial fluid from patients with rheumatoid arthritis show a similar qualitative pattern of acids (mainly fatty acids) resembling the pattern found in serum. The concentrations of the organic acids in the abnormal synovial fluids are 5--10 times higher than those in the fluid from the normal joints. In patients receiving treatment with gold thiomalate, free thiomalate was excreted in the urine. The release of this thiol upon gold therapy is of consequence for the further understanding of the mechanism of action of certain drugs against rheumatoid arthritis.

Arthritis, Rheumatoid↗

Lymphocytes eluted from synovial tissue of juvenile rheumatoid arthritis patients.

Synovial tissues from 11 patients with juvenile rheumatoid arthritis were investigated. The elution of lymphocytes was performed according to a procedure previously described for synovial tissue of adult rheumatoid arthritis patients (1). The T lymphocytes were pre dominant (mean: 71%) in all cell suspensions studied, whereas the average proportion of B lymphocytes was 4%. In addition, Fc-receptor-bearing lymphocytes were demonstrable (mean: 8%). Transformation of the lymphocytes was induced by the unspecific mitrogens phytohemagglutinin, pokeweed mitogen, and concanavallin A, whereas antigens such as ppd and candida albicans antigen were usually ineffective.

Adolescent↗

Antibody-dependent cytotoxicity mediated by cells eluted from synovial tissues of patients with rheumatoid arthritis and juvenile rheumatoid arthritis.

Cell suspensions containing an average of 78% lymphocytes were obtained from synovial tissues of 26 patients with rheumatoid arthritis and 10 patients with juvenile rheumatoid arthritis. These cells were shown to mediate cytotoxicity against 51Cr-labeled chicken erythrocytes sensitized with a rabbit anti-chicken erythrocyte antiserum. Nylon column filtration of the cells increased the proportion of lymphocytes and usually also the cytotoxicity, which suggested that at least some of the effector cells were lymphocytes. The cytotoxic activity of the cells obtained from rheumatoid synovial tissue was always lower than that obtained with the patients' peripheral blood lymphocytes. No significant change in cytotoxicity of normal peripheral blood lymphocytes was observed after these cells had been treated in the same manner as the rheumatoid synovial tissues.

Antibody-Dependent Cell Cytotoxicity↗

Antigen and unspecific mitogen stimulation of lymphocytes eluted from rheumatoid inflammatory tissue.

Lymphocytes were eluted from synovial tissues of 17 patients with classical rheumatoid arthritis, using a procedure previously reported. Stimulation was obtained with the unspecific mitogens phytohemagglutinin (PHA), pokeweed mitogen (PWM), and concanavalin A (Con A) as well as with purified protein derivative of tuberculin (PPD) and mitoycin-C-treated allogeneic lymphocytes, whereas candida antigen usually gave a low response. The pattern of reactivity to unspecific mitogens was similar to that obtained with lymphocytes from peripheral blood of rheumatoid arthritis patients. Two different PPD preparations usually gave transformation of the same magnitude as seen with PHA. This was in contrast to the reactivity of the peripheral blood lymphocytes. It could be demonstrated that the elution procedure initiated some degree of lymphocyte transformation, mainly potentiating the responses to PHA and Con A.

Antigens↗

Elution and characterization of lymphocytes from rheumatoid inflammatory tissue.

Lymphocytes were eluted from the synovial tissue of 19 patients with classical rheumatoid arthritis. The tissue was minced and dissociated by treatment with crude collagenase and DNase. The cell suspension obtained was filtered and incubated in plastic culture flasks overnight at 37 degrees C. The cells that did not adhere to the plastic surface were harvested and the lymphocytes further purified by the Ficoll-Isopaque gradient centrifugation technique. The lymphocyte yield varied from 0.64 to 32 times 10(6) cells. Differential counts showed on the average 85% lymphocytes, 12% mocrophage-like cells, and variable proportions of polymorphonuclear granulocytes, unclassified cells, and dead cells. An average of 77% of the cells were viable as assessed by the trypan blue exclusion test. This cell suspension was investigated for lymphocyte populations. T lymphocytes were predominant in all experiments (mean, 73.6%). The mean percentage of B lymphocytes was 9.7%, whereas the proportion of Fc-receptor-bearing lymphocytes was on the average 6.0%.

Adult↗