lewis-x-antigen and Arthritis--Rheumatoid

To investigate if granulocyte and monocyte apheresis mitigates the symptoms of rheumatoid arthritis (RA), and influences production of panmyelocytes (CD15+ CD16- cells) at the bone marrow level, 27 RA patients who had elevated granulocyte counts were recruited. The granulocyte and monocyte apheresis column (G-1 column) is an extracorporeal type device packed with 220 g cellulose acetate beads to which granulocytes and monocytes specifically adhere. Patients received apheresis of 1 hr duration twice per week, 8 times over a period of 4 weeks. To prepare CD15+CD16- cells, iliac bone marrow aspirate was obtained at baseline and at 2 weeks after completion of the apheresis course. Ex-vivo proliferation of bone marrow low density cells and production of IgM-RF were also investigated. Following granulocyte and monocyte apheresis, there was a suppressed tendency in the number of CD15+CD16- cells in patients with high bone marrow CD15+CD16- cell counts at baseline. Clinical assessments 2 weeks after the completion of apheresis therapy showed improvements in swollen joint count (P < 0.001), tender joint count (P < 0.001) and duration of morning stiffness (P < 0.005). The results suggest that granulocytes and monocytes/macrophages have a pathological role in RA and apheresis treatment to reduce or suppress these cells should benefit patients with RA.

Topics: Adult; Aged; Arthritis, Rheumatoid; Blood Transfusion, Autologous; Cytokines; Female; Flow Cytometry; Granulocytes; Humans; Leukapheresis; Leukocyte Count; Leukocyte Transfusion; Lewis X Antigen; Male; Middle Aged; Monocytes; Pilot Projects; Receptors, IgG; Rheumatoid Factor; Treatment Outcome

As reported by us, a new myeloid cell population with an oncofetal membrane marker, dimeric Lex (di-Lex; III3FucV3 FucnLc6), was found in the epiphyseal bone marrow adjacent to the involved joints of patients with severe rheumatoid arthritis (RA). Patients with RA received intradermal (id) injections of di-Lex incorporated in liposome or of high molecular weight glycoprotein, or tumor associated carbohydrate antigen (TCA), containing the same carbohydrate epitope as di-Lex. The epiphyseal myeloid cells were reduced or sometimes eliminated during id injection. In random trials of id injection, observation under clinical and laboratory conditions showed improvement in 63% (17/27) of the patients treated for 6 months with appropriate doses of di-Lex (III3FucnLc4), and in 72% (31/43) of those treated with an identical protocol for TCA. However, id injection with monomeric Lex had no effect.

Topics: Adult; Aged; Antigens, Tumor-Associated, Carbohydrate; Arthritis, Rheumatoid; Bone Marrow; Carbohydrate Sequence; Dose-Response Relationship, Drug; Drug Carriers; Female; Fluorescent Antibody Technique; Glycoproteins; Humans; Injections, Subcutaneous; Lewis X Antigen; Liposomes; Male; Middle Aged; Molecular Sequence Data; Time Factors

The purpose of this study was to investigate the relationship between clinical disease activity in patients with advanced stage rheumatoid arthritis (RA) on treatment with Disease Modifying Antirheumatic Drugs (DMARDs) and histopathological scores of synovial inflammation. To this end, synovial biopsies of 62 RA patients who underwent surgery for either synovectomy or total joint arthroplasty were assessed by a general synovitis score (GSS) and an immunologic synovitis score (IMSYC). The clinical disease activity index (CDAI) was significantly correlated with both the GSS and the IMSYC (r = 0.65, p = <0.001, r = 0.68, p = <0.001). Compared to patients with moderate and high disease activity, there was a significantly lower expression of T cell (CD3), B cell (CD20) and neutrophil (CD15) markers in synovial tissue of patients with low activity, but similar expression of the macrophage marker CD68. Subgroup analyses revealed no differences between small and large joints, seropositive and seronegative RA and patients with or without prednisolone treatment. However, we found a significantly stronger correlation of CDAI with IMSYC in patients undergoing arthroplasty (r = 0.82) than in patients undergoing synovectomy (r = 0.55). In addition, there was a stronger correlation of CDAI with GSS in patients treated with methotrexate (r = 0.86) than in patients with TNFα blockade (r = 0.55). In summary, the present study demonstrates that the histopathological scores GSS and IMSYC in general reflect clinical disease activity in patients with advanced stage rheumatoid arthritis, but that there is some heterogeneity between subgroups of patients within the cohort. In the future, molecular characterization of synovial inflammatory cell populations, including plasma cell infiltrates, will help to further defined clinically important subtypes of RA and treatment response.

Topics: Adult; Aged; Arthritis, Rheumatoid; Biopsy; CD3 Complex; Female; Gene Expression Regulation; Humans; Inflammation; Lewis X Antigen; Macrophages; Male; Middle Aged; Severity of Illness Index; Synovial Fluid; Synovitis

Immunohistochemical synovial tissue biomarkers are used increasingly to classify arthropathies, study their pathogenesis, and to measure disease activity in clinical trials. We have used receiver operating characteristic (ROC) analysis to quantify the discriminatory abilities of markers for common inflammatory cells (subintimal CD15, CD68, CD3, CD20, CD38, and lining CD68), proliferating cells (Ki-67) and blood vessels (von Willebrand factor, vWF) among inflammatory (chronic septic arthritis, early arthritis and rheumatoid arthritis (RA)) and degenerative arthropathies (osteoarthritis (OA) and orthopedic arthropathies) and normal synovium. Six of the eight markers distinguished accurately between RA and the degenerative arthropathies (area under the curve (AUC) 0.91-0.97), whereas subintimal CD68 (AUC 0.92) and Ki-67 (AUC 0.87) distinguished best between OA and normal synovium. Fold differences in mean expression correlated only modestly with AUCs (r(2) = 0.44). Multicategory ROC analysis ranked Ki-67, subintimal CD68, and CD15 as discriminating best among all six sample groups, and thus identified them as the most broadly applicable markers.

Topics: Antigens, CD; Antigens, CD20; Antigens, Differentiation, Myelomonocytic; Area Under Curve; Arthritis, Infectious; Arthritis, Rheumatoid; Biomarkers; CD3 Complex; Humans; Ki-67 Antigen; Lewis X Antigen; Osteoarthritis; ROC Curve; Synovial Membrane; Synovitis; von Willebrand Factor

To investigate enhanced granulopoiesis in bone marrow of patients with rheumatoid arthritis (RA), and the role of neutrophils in RA pathogenesis.. Aspirated bone marrow cells and peripheral blood leukocytes from patients with RA and non-RA patient controls were analyzed morphologically and by 2 color flow cytometry. Thirteen iliac bones (8 RA, 5 non-RA) were examined by light and transmission electron microscope (TEM).. The percentage of CD15+CD16- cells (immature neutrophils) in RA bone marrow (64.3+/-13.4%, mean +/- SD) increased significantly compared to that of non-RA controls (43.2+/-14.3%), whereas the fraction of CD15+CD]6+ cells (mature neutrophils)was greatly decreased (RA 21.8+/-10.1%; non-RA 38.1+/-8.9%). The absolute number of CD15+CD16- cells also increased markedly in RA bone marrow. The ratio of immature cells to the total granulocytes (% CD15+CD16- to % CD15+) correlated with the Lansbury Index score (R = 0.76, p<0.0001). TEM observations revealed that abundant immature neutrophils adhered closely to the trabeculae of the iliac bone. Margins of trabeculae were mostly irregular, especially in severe RA, and collagenous fibers frequently disappeared in those trabeculae with ragged margins.. In RA bone marrow, immature neutrophils (CD15+CD16-) were markedly increased in number; by contrast, no changes were found for mature cells. Augmented production of immature neutrophils (at the promyelocyte-to-myelocyte stage) might lead to the destruction of collagenous fibers in RA bone trabeculae, as revealed by TEM. Generalized bone destruction in RA might, at least in part, be caused by enhanced production of immature neutrophils.

Topics: Adult; Aged; Aged, 80 and over; Arthritis, Rheumatoid; Bone Marrow Cells; Cell Division; Cellular Senescence; Disease Progression; Female; Flow Cytometry; Humans; Ilium; Leukocyte Count; Leukopoiesis; Lewis X Antigen; Male; Microscopy, Electron; Middle Aged; Neutrophils; Receptors, IgG

The adhesion molecules CD11b (a beta2-integrin component) and CD54 (ICAM-1) on blood leukocytes were studied by flow cytometry in patients with rheumatoid arthritis (RA). The fractions of CD4+ cells co-expressing CD11b were elevated in 16 patients with active RA compared with those in 16 RA patients who improved during therapy and 8 healthy controls: 0.8+/-0.12% (mean+/-SEM) versus 0.3+/-0.06% (p<0.002) and 0.3+/-0.06% (p<0.005), respectively. Increased levels of CD11b+CD45R0+ cells were observed in patients with active RA compared to those with improved RA and controls: 12.6+/-3.9% versus 4.8+/-2.7% (p<0.002) and 6.1+/-1.2% (p<0.003), respectively. Disease activity, determined by C-reactive protein, correlated with the numbers of CD11b+CD45R0+ cells: r=0.62 (p<0.001). Seven patients were followed during induction of remission with methotrexate and glucocorticoids. The numbers of CD11b+CD4+ and CD11b+CD45R0+ cells fell significantly after clinical improvement. The levels of CD11b+CD14+ cells (monocytes) did not differ between the groups. The number of CD11b+CD15+ cells (neutrophils) was elevated in patients with RA irrespective of disease activity. The levels of CD54+ cells were not different between the RA and control groups. We conclude that the increased numbers of CD11b+ memory T cells may arise from exposure to stimuli outside the synovial compartment.

Topics: Adult; Aged; Aged, 80 and over; Arthritis, Rheumatoid; Case-Control Studies; CD4-Positive T-Lymphocytes; Female; Humans; Immunologic Memory; Intercellular Adhesion Molecule-1; Leukocyte Common Antigens; Lewis X Antigen; Lipopolysaccharide Receptors; Macrophage-1 Antigen; Male; Middle Aged; Monocytes; Neutrophils; Up-Regulation

We previously reported the accumulation of abnormal myeloid cell populations reacting with CD14 (MY4) monoclonal antibody in the iliac and epiphyseal bone marrow of patients with severe rheumatoid arthritis (RA). Therefore, we investigated in vitro production and modulation of CD14+ myeloid cells from iliac bone marrow cells.. Mononuclear cells were prepared from iliac bone marrow aspirates from patients with RA. The presence of unusual myeloid cells was assessed by 2 color flow cytometry of cells cultured under various conditions.. Cultured iliac bone marrow cells of patients with severe RA produced 14.7% of CD14+ CD15+ cells on average. Cultures derived from healthy donors and from patients with a milder form of RA produced fewer CD14+ CD15+ cells (< 10%). The production of CD14+ CD15+ cells was enhanced by granulocyte macrophage colony stimulating factor and interleukin 1beta, but inhibited by T lymphocytes.. Production and modulation of CD14+ myeloid cells were observed in iliac bone marrow of patients with severe RA.

Topics: Adult; Arthritis, Rheumatoid; Bone Marrow Cells; Cell Line; Cell Separation; Cytokines; Female; Humans; Ilium; Interleukin-1; Lewis X Antigen; Lipopolysaccharide Receptors; Male; Middle Aged; T-Lymphocytes

The membrane carbohydrate antigen, sialyl Lewis x (sLe(x)), is involved in cellular adhesive interactions in many diseases, such as cancer, inflammation and thrombosis. This antigen is also found on soluble macromolecules, such as serum glycoproteins, but the precise role of soluble sLe(x) in modifying disease processes, or reflecting the pathological changes is still unclear. Although methods were previously reported for the measurement of soluble sLe(x), many of these were not well characterised, measurements were mainly made on mixtures of molecules, and the anti-sLe(x) antibodies were used at concentrations that made the assay expensive. In this study an ELISA has been devised that detects sLe(x) in purified soluble glycoconjugates using the anti-sLe(x) antibody, CSLEX I. Commercially-available haptoglobin (Hp) and synthetic complexes of Lewis antigens with polyacrylamide were used as model substances in developing the procedure. Key steps were washing the antibody/antigen complex with ten times diluted salt solution to prevent dissociation of the complex and the use of bovine serum albumin for blocking non-specific interactions. The assay was shown to be very specific, its precision was in the range 6-12%, and it could detect less than a pmol of sLe(x). It could also distinguish between different densities of sLe(x) on the same amount of glycoconjugate. Determination of sLe(x) in Hp isolated from small groups of healthy individuals, cancer patients, and rheumatoid arthritis sufferers suggested that the antigen expression is increased in disease. This method, which is an improvement on those previously described will be useful for determining sLe(x) in many different types of soluble glycoconjugate, and used in combination with synthetic carbohydrate polyacrylamide complexes, will help to standardize measurements of soluble sLe(x) in the future.

Topics: Adult; Aged; Antibodies, Monoclonal; Arthritis, Rheumatoid; Enzyme-Linked Immunosorbent Assay; Female; Glycoconjugates; Haptoglobins; Humans; Lewis X Antigen; Male; Middle Aged; Ovarian Neoplasms; Reproducibility of Results; Sensitivity and Specificity

Endothelial adhesion molecules facilitate the entry of leukocytes into inflamed tissues. This in turn promotes neovascularization, a process central to the progression of rheumatoid arthritis, tumor growth and wound repair. Here we test the hypothesis that soluble endothelial adhesion molecules promote angiogenesis. Human recombinant soluble E-selectin and soluble vascular cell adhesion molecule-1 induced chemotaxis of human endothelial cells in vitro and were angiogenic in rat cornea. Soluble E-selectin acted on endothelial cells in part through a sialyl Lewis-X-dependent mechanism, while soluble vascular cell adhesion molecule-1 acted on endothelial cells in part through a very late antigen (VLA)-4 dependent mechanism. The chemotactic activity of rheumatoid synovial fluid for endothelial cells, and also its angiogenic activity, were blocked by antibodies to either soluble E-selectin or soluble vascular cell adhesion molecule-1. These results suggest a novel function for soluble endothelial adhesion molecules as mediators of angiogenesis.

Topics: Animals; Arthritis, Rheumatoid; Cell Adhesion; Cell Adhesion Molecules; Chemotaxis; Cornea; E-Selectin; Endothelium, Vascular; Humans; Lewis X Antigen; Neovascularization, Pathologic; Rats; Recombinant Proteins; Solubility; Synovial Fluid; Vascular Cell Adhesion Molecule-1

To analyze the differentiation stages of myeloids statistically, we adopted a two-color FACS system and used appropriate monoclonal antibodies belonging to CD15, CD16 and CD11b. By using HL60 treated with DMSO or human bone marrow MNCs from patients with rheumatoid arthritis, it was proved that with this system, myeloids could be clearly separated according to differentiation stages. Furthermore, the number of myeloids at certain stages of differentiation in the epiphyseal bone marrow of patients with RA or OA was measured. Nine of 15 samples from RA patients showed immature and relatively mature myeloids, while none of the 8 OA samples did. When the proportions of myeloids in epiphyseal bone marrow MNCs were compared with the clinical features, disease subsets in RA and the degree of synovitis, seemed to be important factors for abnormal myelopoiesis.

Topics: Adult; Aged; Antibodies, Monoclonal; Antigens, Differentiation; Arthritis, Rheumatoid; Bone Marrow; CD11 Antigens; Cell Differentiation; Dimethyl Sulfoxide; Female; Flow Cytometry; Growth Plate; Humans; Joints; Leukemia, Myeloid, Acute; Lewis X Antigen; Male; Middle Aged; Osteoarthritis; Receptors, Fc; Receptors, IgG; Tumor Cells, Cultured

The purpose of this study is to show that anti-Leu M1 antibody (anti-CD15), which has different staining characteristics in lymphoid and non-lymphoid cells, reacted against the surface antigen of a defined monoclonal B cell line. This antibody recognizes the sugar moiety, lacto-N-fucopentaose (LNF-III), which is linked to the cell membrane protein in several kinds of cells, but not in B cells. However, a human monoclonal B-cell line (TKS-1) which was established from the peripheral blood of a patient with rheumatoid arthritis, expressed the Leu M1 antigen spontaneously. The analysis of surface markers using a fluorescence-activated cell sorter (FACS) has revealed that the surface markers of TKS-1 were anti-mu, delta, kappa, HLA-DR, DQ, Leu 12 (CD19) and Leu M1 (CD15). TKS-1 cells were not reactive with any of the following antibodies: anti-OK M1 (CD11b), Leu M2, Leu M3 (CD14), Leu M4, Leu 1 (CD5), Leu 2 (CD8), Leu 3 (CD4), Leu 4 (CD3), Leu 7 and Leu 11 (CD16). In addition, TKS-1 was positive to Epstein-Barr nuclear antigen, weakly positive to non-specific esterase without staining inhibition by NaF, and negative to peroxidase. TKS-1 cells produced IgM in the culture supernatant and have kappa-light chain rearrangement in its DNA. As shown in other studies, distribution of Leu M1 is very wide. This antigen is not a specific immunodiagnostic marker to distinguish the cell type. We conclude that it is possible to express Leu M1 antigen on the membrane of a B-cell lineage cell.

Topics: Antibodies, Monoclonal; Antigens, Differentiation, Myelomonocytic; Arthritis, Rheumatoid; B-Lymphocytes; Clone Cells; Gene Rearrangement, B-Lymphocyte, Light Chain; Humans; Lewis X Antigen