Defective polymorphonuclear leucocyte chemotaxis in rheumatoid arthritis associated with a serum inhibitor.

Cellular and/or serum components of polymorphonuclear leucocyte chemotaxis were assessed in 21 patients with rheumatoid arthritis. No difference in the chemotactic migration of control and patient cells in response to a number of chemotactic solutions could be detected (P = 0.38). Deficient generation of chemotactic activity in patient sera (P = 0.58) as compared to control sera (P = 0.014) after incubation of the sera with Escherichia coli lipopolysaccharide, resulted in a significant difference in the chemotactic activity of the control and rheumatoid serum preparations for polymorphonuclear leucocytes (P = 0.0012). This defect was associated with the presence of a serum inhibitor of chemotaxis, the potency of which was inversely correlated with the level of chemotactic activity generated in the rheumatoid sera (r = -0.941, P less than 0.001).     

Griseofulvin inhibition of polymorphonuclear leucocyte chemotaxis in Boyden chambers.

Griseofulvin inhibited the chemotaxis of polymorphonuclear leucocytes (PMN's) in vitro in the concentration range 0.1-1.0 mug/ml, i.e. at concentrations comparable to those obtained in serum during peroral treatment with griseofulvin. It is suggested that PMN chemotaxis is inhibited by griseofulvin interference with the redistribution of cytoplasmic microtubules, which is thought to be essential in the direction-finding of PMNs during chemotaxis. Furthermore, it is suggested that the griseofulvin inhibition of PMN chemotaxis - together with the previously known pharmacodynamic properties of griseofulvin - may provide the rationale for griseofulvin therapy in PMN-mediated tissue injury of the gut.     

Abnormal polymorphonuclear leucocyte chemotaxis in Beh?et's syndrome.

Experiments both in vitro and in vivo have been performed to study the chemotactic response of polymorphonuclear leucocytes (PMNs) in Behçet''s syndrome. The experimental results were apparently contradictory. Using modified Boyden chambers we found that the PMNs from patients with Behçet''s syndrome responded to a greater extent in vitro than normal cells, but with skin chambers placed over abrasions the in vivo response was less than normal. The significance of these findings is discussed and related to the histological appearances that may be seen in this condition.     

Evidence of impaired polymorphonuclear leucocyte phagocytic functions and chemiluminescence response in rheumatoid arthritis

Summary In this study, the phagocytic uptake of ³H-thymidine-labelled Staphylococcus aureus and bacterial killing (Bk) by polymorphonuclear leucocytes (PMN) from patients with rheumatoids arthritis (RA) were investigated and compared to the luminol-enhanced chemiluminescence response (LCL) to phorbolmyristic acetate (PMA), a receptor and second message-independent activator of respiratory burst activity in definite or classical rheumatoid arthritis (RA), with regard to the inflammatory activity of the disease process, and compared to patients with osteoarthritis (OA) and healthy controls. PMN of RA patients showed a significant reduction in uptake (57.8±4.4%) and less Bk capacity (27.3±4.2%) compared to OA patients (uptake 71.4±4.3%, Bk 20.6±2.9%; P<0.001) and controls (uptake 73.2±5.2%, Bk19.3±4.2%;P<0.001). In contrast, PMN LCL response was markedly enhanced in RA patients compared to OA patients (P<0.001) and controls (P<0.001). There was no significant influence of inflammatory activity on various PMN functions in RA and no difference was found between OA and control subjects. These data clearly demonstrated impaired PMN phagocytic functions (uptake, Bk) and enhanced LCL in RA, suggesting priming and/or activated peripheral blood PMN, which might be of clinical importance concerning altered host defence in these patients.     

Degradation in vivo of articular cartilage in rheumatoid arthritis by leucocyte elastase from polymorphonuclear leucocytes

Summary Using a specific substrate, no leucocyte elastase activity could be detected in 55 synovial fluids, including 29 from patients with rheumatoid arthritis (RA). However, a high percentage of samples contained phagocytic inclusions of elastase, ₁-proteinase inhibitor (₁-PI) and ₂-macroglobulin (₂-MG) in both the polymorphonuclear (PMN) and mononuclear phagocytes. Immunofluorescence and indirect peroxidase-antiperoxidase staining of articular cartilage (ACA) from 52% of 21 patients with RA and one with juvenile RA (JRA) showed presence of elastase in the superficial layer of microscopically intact but proteoglycan depleted pannus-free ACA. In histologically altered pannus-free RA-ACA superficial elastase deposits were found in 24% of the cases. Adjacent ACA sections contained IgG, C 3, ₁-PI and rarely ₂-MG. RA-ACA below or surrounded by pannus showed close contact with intact and decaying PMN in 62% and 48% of the cases, respectively. ACA specimens from patients with degenerative disease and systemic lupus were negative. These findings strongly suggest that PMN leucocyte elastase is operative in the degradation of RA-ACA and JRA-ACA, and that this activity is largely dependent upon the presence of entrapped immune complexes in such cartilage.     

Circulating human leucocyte elastase in rheumatoid arthritis

Summary Elastase-inhibitor complex (EIC) levels were determined in EDTA-plasma samples of 40 patients with connective tissue disease by a double antibody immunoassay technique. In active rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE), EIC levels were significantly higher than in the normal controls (P<10^–8) and fell on remission. The mean EIC level in active RA was significantly greater than in inactive disease (P=0.0001) but there was no statistically significant difference between the EIC levels in the acute and inactive disease states in SLE (P=0.49). In active RA, there was a positive correlation between EIC levels and white blood cell count (WBC) but not with erythrocyte sedimentation rate (ESR). In SLE there was no significant correlation between EIC levels and ESR or WBC. EIC measurement may be useful in the objective assessment of activity in RA.     

Inhibition of neutrophil chemotaxis in methotrexate-treated rheumatoid arthritis patients

Summary Polymorphonuclear cell (PMN) chemotaxis was assessed using the in vitro under agarose assay in ten rheumatoid arthritis patients prior to and following a single 10-mg dose of methotrexate (MTX). PMNs obtained from patients after MTX showed a decreased chemotactic migration response to both zymosan activated serum (P<0.005) and N-formyl-L-methionyl-L-phenylalanine (P<0.01). In similar conditions, no significant difference in chemotactic migration could be detected in six rheumatoid arthritis patients not on MTX. In contrast to the in vivo effects of MTX, there was no inhibition of normal PMN chemotactic migration following a 30-min in vitro incubation of the cells with MTX (P<0.99).     

Ketoprofen-induced reduction of polymorphonuclear cell activation in rheumatoid arthritis

Ten early rheumatoid arthritis (RA) patients and 10 healthy volunteers were given 200 mg of ketoprofen daily for 8 days. Polymorphonuclear cell functional studies were performed just before and immediately after this treatment. In RA patients, increased chemotactic index and adherence returned to normal after this short-term treatment whilst reduced phagocytosis and bacterial function were not significantly modified by the drug. In healthy volunteers, no significant effect was shown. In addition, ketoprofen diminished the chemotactic activity induced by zymosan in RA patients and also in healthy volunteer plasma.     

Drug induced impairment of polymorphonuclear cell bactericidal ability in rheumatoid arthritis.

Thirty three patients with rheumatoid arthritis (RA) treated only with non-steroidal anti-inflammatory drugs were divided into two groups according to the drug received (group A: diclofenac, group B: indomethacin or ketoprofen), and their polymorphonuclear (PMN) cell functions were investigated. We found that bactericidal ability was the only function significantly impaired in group A as compared with group B patients and normal controls. This modification correlated well with the reduction of control PMN bactericidal ability when this test was carried out in patient plasma. These differences between drug effects might explain some of the discrepancies between reports of PMN cell functions in RA.     

Scanning electron microscopy of rheumatoid arthritis peripheral blood polymorphonuclear leucocytes.

Peripheral blood polymorphonuclear leucocytes (PMNs) were isolated from six normal individuals and from 27 patients with rheumatoid arthritis (RA) by the Ficoll-Hypaque rapid single step centrifugation technique, fixed in suspension, and examined by scanning electron microscopy (SEM). In addition, four of the preparations from normal individuals and eight from patients with RA were examined by transmission electron microscopy (TEM). Most PMNs in preparations from normal subjects were spherical, unpolarised, and had their surface membrane elaborated into irregular ridges and small ruffles; they contained few phagocytic vacuoles and large numbers of electron dense primary and secondary granules. A minority of the cells were non-spherical, polarised, and had portions of their surface membrane elaborated into ruffled pseudopodia. In contrast, preparations of RA PMNs frequently contained fewer unpolarised PMNs and a higher number of polarised PMNs than did preparations of normal PMNs. Some preparations of RA PMNs also contained substantial numbers of spherical cells whose surface was covered mainly by bulges and blebs. Concurrent examination by TEM showed that RA PMNs frequently contained more phagocytic vacuoles and fewer electron dense primary and secondary granules than normal PMNs. The morphological and ultrastructural changes seen in RA PMNs resembled those which normal PMNs are known to undergo on exposure to C5a in vitro, during adherence to endothelial cells in vivo, or during phagocytosis in vivo or in vitro. Our observations, therefore, provide a useful morphological correlation to those in vitro studies in which differences in the functional activity of RA and normal PMNs have been shown. The possibility that the difference seen between RA and normal PMNs is artefactual and does not represent the genuine in vivo states of these cells is discussed.     

Impaired diastolic function in active rheumatoid arthritis. Relationship with disease duration.

OBJECTIVE: Using digitized M-mode and Doppler echocardiography, we evaluated left ventricular (LV) function in 54 patients (43 women and 11 men; mean age 50 years) suffering from active rheumatoid arthritis (RA) without obvious cardiovascular disease, and compared them with 54 age- and sex-matched normal subjects. RESULTS: No differences were found in LV end-diastolic diameter, systolic function and parietal thickness between the patients and controls. However, a significant reduction in various indexes of LV diastolic function was found, including E/A (ratio of early to late filling waves of mitral inflow Doppler) and the peak lengthening rate of the LV diameter (an index of LV relaxation evaluated by M-mode echocardiography). The former was correlated with patient age and was independent of disease duration, while the latter was more markedly correlated with disease duration than with patient age. CONCLUSION: The relationship between diastolic impairment and disease duration in active RA may open new perspectives in the study of RA-associated cardiovascular disease.     

Impaired delayed type cutaneous hypersensitivity in rheumatoid arthritis reversed by chrysotherapy.

A prospective 24 week study of 31 patients with active rheumatoid arthritis (18 women, 13 men) was undertaken to determine whether weekly intramuscular sodium aurothiomalate (gold) would influence delayed type cutaneous hypersensitivity (DTH) and other indices of cell mediated immunity. DTH to seven recall antigens was measured by Multitest on three occasions during the study. Twenty five patients completed the study. At entry 13 patients (12 female) were anergic, and no significant correlations were found between DTH and other clinical and immunological indices. Women showed a significantly greater depression of DTH than men. At week 24 only three of the patients were anergic with significant increase in mean DTH score being noted particularly to tuberculin, candida, and streptococcus. Improvement in DTH was observed in both gold responders and non-responders. In conclusion, patients with active rheumatoid arthritis show impairment of DTH, which is reversed by chrysotherapy. This effect is most apparent in women and appears to be relatively independent of the clinical response.     

An inhibitor of leucocyte movement in the plasma of patients with rheumatoid arthritis.

Plasma from patients with rheumatoid arthritis shows inhibitory activity towards the random and stimulated movement of human polymorphonuclear leucocytes and peripheral phagocytic monocytes in vitro. The inhibitory activity is heat-labile, not present in plasma from patients with noninflammatory diseases, and appears to exert its inhibitory effects by occupying sites on the cell cell membrane from where it may be removed by washing. It is suggested that this factor(s) is responsible for the diminished accumulation of polymorphonuclear leucocytes into skin chambers with occurs in patients with rheumatoid arthritis.     

Impaired antigen-specific suppressor cell activity in patients with rheumatoid arthritis

Antigen-specific suppressor cell activity of peripheral blood mononuclear cells was investigated in 20 patients with rheumatoid arthritis (RA) and 16 age- and sex-matched healthy controls. Suppressor cell activity was generated by priming peripheral blood mononuclear cells with high dose antigen (ovalbumin) and adding the washed primed or control (unprimed) cells to autologous, optimally stimulated, target plaque forming cell (PFC) cultures. The ability of the primed cells to interfere with an optimal ovalbumin specific PFC response in the target culture was used as a measure of antigen-specific suppressor cell activity. The results demonstrated that the mean (+/- SE) PFC response of the rheumatoid patients (669 +/- 76 PFC/10(6) cells) was not statistically different from that of the normal controls (722 +/- 83 PFC/10(6) cells), P = 0.1. However, reduced suppressor cell activity was observed in the rheumatoid patients relative to controls (46.4 +/- 4.2% versus 64.6 +/- 2.7% suppression, respectively; P < 0.001). No correlation was demonstrated between suppressor cell activity in rheumatoid patients and disease activity or therapy.     

Antibody-mediated leucocyte cytotoxicity to Chang human liver cells in rheumatoid arthritis and other diseases.

The incidence of an IgG-antibody which induces lymphocyte cytotoxicity to Chang human liver cells in culture was estimated in the sera of patients with rheumatoid arthritis, systemic lupus erythematosus, ankylosing spondylitis, psoriatic arthritis, Crohn's disease, ulcerative colitis, and in healthy controls. It was found in 4.1% of control subjects and in 31% of patients with rheumatoid arthritis. None of the other patient groups differed from the control group. This may be the first demonstration of an antibody response to an antigen or antigens which is almost entirely confined to patients with rheumatoid arthritis. The possibility that an antigenic similarity exists between the rheumatoid synovial membrane and Chang cells is currently under investigation.