Thyrocytes isolated from autoimmune-diseased thyroids secrete soluble tumor necrosis factor-R1 that is related to their elevated protein kinase C activity.

Soluble tumor necrosis factor (TNF)-alpha receptors have the potential to modulate TNF-alpha activity during autoimmune thyroiditis. In this study we examined cell-surface TNF-alpha receptors and soluble TNF-alpha receptor production by thyrocytes from normal and MRL-lpr(-/-) (diseased) mice, which spontaneously develop autoimmune thyroiditis. We found that murine thyrocytes possess the 55-kd receptor (TNF-R1). Examination of soluble TNF-R1 production revealed that diseased thyrocytes produced sevenfold more soluble TNF-R1 than normal thyrocytes. Furthermore, basal protein kinase C (pKC) activity in diseased thyrocytes was 67% higher than that found in normal murine thyrocytes. The elevated basal pKC activity in diseased thyrocytes was related to their enhanced production of soluble TNF-R1 because inhibition of pKC activity with calphostin C caused soluble TNF-R1 production to decrease significantly. Additionally, soluble TNF-R1 production by murine thyrocytes was not a result of cell-surface receptor shedding but through secretion of a truncated version of TNF-R1. This was evident when cell-surface TNF-R1 levels were unchanged after treatment of diseased thyrocytes with calphostin C. Also, the 28-kd form of TNF-R1, which corresponds to the soluble receptor, was present in the intracellular membranes of the diseased thyrocytes.

Circulating antibodies to guanosine in systemic lupus erythematosus: correlation with nephritis and polyserositis by acute and longitudinal analyses.

Systemic lupus erythematosus (SLE) is characterized by autoantibodies, including antibodies to the nucleosides of DNA. Guanosine is the most immunogenic nucleoside. In this study serum antiguanosine antibody levels were compared with disease activity, determined by their SLEDI score, in 86 patients with SLE. Sera from these patients were tested, by ELISA, for autoantibodies to guanosine, single-stranded DNA (ssDNA), and double-stranded DNA (dsDNA). Anti-double-stranded DNA levels were also measured by RIA. Resultant values from these assays were correlated with SLE disease activity, and compared with specific features of SLE. The strongest correlation was higher levels of antiguanosine antibodies in patients with active lupus nephritis and polyserositis compared to patients with inactive disease (P < 0.0001). Antiguanosine levels also correlated with arthritis (P < 0.006), CNS lupus (P < 0.005), and hematologic manifestations of SLE (P < 0.002). To test the validity of this association in chronic SLE, serum antiguanosine antibodies were measured in patients with SLE at various phases of disease activity. Twelve patients with SLE had serum samples drawn at active, active-improved, and inactive phases over a 3-7 y period. Differences were significant for serum antiguanosine antibodies in the active group compared to the inactive group (P < 0.05) and the active vs the active-improved group (P < 0.02), unlike those for dsDNA and ssDNA by ELISA or RIA. Antiguanosine antibodies correlated more closely with disease activity in SLE patients in this longitudinal study than either anti-dsDNA or ssDNA antibodies. Thus, antibodies to guanosine correlated as well or better with disease activity than the other anti-DNA antibodies measured and should be considered to contribute to the pathology of SLE, especially lupus nephritis.

The effectiveness of the clinical self-assessment (MHAQ) compared with other clinical and laboratory tests used to monitor the activity of rheumatoid arthritis by consensus analysis.

Disease activity in rheumatoid arthritis (RA) is difficult to measure objectively. Both clinical and laboratory measures were evaluated by the statistical method of consensus analysis. In the literature, laboratory tests as a group have proved more effective; however, studies did not include self-assessment questionnaires. We evaluated the effectiveness of a Modified Health Assessment Questionnaire (MHAQ) relative to other clinical and laboratory tests measuring disease activity in 100 patients with RA. Hemoglobin, hematocrit, mean corpuscular volume, fibrinogen level, Wes-tergren erythrocyte sedimentation rate and C-reactive protein tests, along with tests of morning stiffness, visual analog pain scale, Ritchie index, total joint count, and MHAQ, were evaluated simultaneously. The erythrocyte sedimentation rate was confirmed as a highly effective test to follow disease activity. The MHAQ was the best clinical test; in the methotrexate-treated subgroup it ranked as highly as the most effective laboratory assays. MHAQ is a more effective tool than many laboratory tests currently ordered for monitoring RA disease activity and should be a part of the record of each visit of an arthritis patient.

Characterization of autoimmune thyroiditis in MRL-lpr/lpr mice.

MRL-lpr/lpr mice are genetically predisposed to develop a systemic lupus erythematosus-like syndrome that is clinically very similar to the human disease. The results presented here demonstrate, for the first time to our knowledge, that MRL-lpr/lpr mice also develop thyroiditis as part of their systemic autoimmune disorder. The thyroid gland was infiltrated by immunocomponent cells with defined lymphoid follicular centers and extensive interstitial lymphocytes dispersed throughout the thyroid epithelium. All the diseased mice were hypothyroid with reduced, relative levels of thyroid hormone (free T4) and elevated levels of thyroid-stimulating hormone (TSH). They also had high concentrations of circulating IgG class autoantibodies directed against thyroglobulin, thyroperoxidase and double-stranded DNA. The MRL-+/+ age-matched allelic counterpart mice had relatively few lymphocytes in their thyroid tissue, and normal levels of thyroxine and TSH. The non-diseased mice also had undetectable levels of thyroid reactive autoantibodies tested for by enzyme-linked immunosorbent assays. Collectively these findings document that the MRL-lpr/lpr mice spontaneously develop autoimmune thyroiditis and can be used as a model for the study of thyroid-specific autoimmunity.

Increased serum levels of anti-elastin antibodies in patients with Peyronie's disease.

The cause of Peyronie's disease is unknown. Immunological mechanisms in the pathogenesis have been previously suggested. Antibodies to elastin are present in all individuals. However, abnormal serum levels of anti-tropoelastin (reflecting elastin synthesis) and anti-alpha-elastin (reflecting elastin destruction) are seen in a variety of autoimmune diseases. We show that patients with Peyronie's disease have higher levels of antibodies to tropoelastin (p < 0.047) and alpha-elastin (p < 0.012) than age-matched controls, suggesting an increase in elastin synthesis and breakdown, respectively. These findings suggest the presence of autoimmune mechanisms in the pathogenesis of Peyronie's disease, which may have future diagnostic and therapeutic implications.

Eosinophilic fasciitis in association with chronic vasculitic-like leg ulcerations.

Vasculitic lesions are not generally associated with eosinophilic fasciitis. Eosinophilic fasciitis is reported to be a syndrome distinct from progressive systemic sclerosis (PSS). More recent studies, however, note overlapping features in the clinical, pathologic, and laboratory findings of eosinophilic fasciitis and scleroderma. We report a typical presentation of eosinophilic fasciitis that developed vasculitic-like leg ulcerations as seen in scleroderma.

Serum anti-tropo:anti-alpha-elastin antibody ratio assessing elastin turnover in scleroderma.

Serum antibodies to native (tropo) and denatured (alpha) elastins appear to correlate with the production and breakdown respectively of elastic tissue. Elastin may be degraded as a part of autoimmune diseases. This possibility was tested by measuring IgG antibodies to tropo- and alpha-elastins by ELISA in the sera of 111 patients with a variety of connective tissue diseases compared with 18 healthy individuals. Anti-alpha-elastin antibodies were significantly higher in sera from 18 scleroderma patients than from healthy controls (p less than 0.008). Conversely, anti-tropoelastin antibody levels for scleroderma patients (p less than 0.03) and for patients with a variety of other connective tissue diseases (p less than 0.02) were lower than in healthy controls. Low antibody levels to native elastin and high levels of antibodies to denatured elastin suggest a low synthesis: degradation ratio for elastin in scleroderma. Scleroderma may be a unique model for elastin turnover because of its heretofore unrecognized accelerated elastolysis.

Apheresis enhances the selective removal of antinuclear antibodies in systemic lupus erythematosus.

Apheresis suppresses clinical manifestations of lupus and reduces levels of antinuclear antibodies implicated in the pathogenesis of systemic lupus erythematosus (SLE). It is not known, however, if reduced levels of antinuclear antibodies are due to nonspecific removal, or specific mechanisms associated with decreased production, or enhanced clearance from the circulation. In order to distinguish between specific and nonspecific effects of apheresis on antinuclear antibodies in SLE, we compared plasma levels of IgG antibodies to DNA and IgG antibodies to microbial antigens in 13 SLE patients before and after apheresis. Although apheresis lowered plasma levels of IgG (21% mean reduction), there was a disproportionate reduction in IgG antibodies to DNA (42% mean reduction, p less than 0.13). In marked contrast, reduction in antibodies to microbial antigens did not exceed those of plasma IgG. A rapid rebound of serum anti-DNA antibodies following apheresis in certain SLE patients suggests that the selective reduction in anti-DNA antibodies is due to enhanced clearance from the circulation rather than decreased production. These results indicate that apheresis enhances selective removal of antinuclear antibodies in some patients with SLE.

Rheumatoid factors specific for active rheumatoid arthritis.

To measure rheumatoid factors specific for patients with rheumatoid arthritis an enzyme linked immunosorbent assay (ELISA) was developed to measure rheumatoid factors in human serum that bind a cross-reactive determinant shared on human and other mammalian IgG. Rheumatoid factors that cross link human IgG and sheep IgG in a double binding ELISA were almost completely specific (greater than 99%) for rheumatoid arthritis in assays of 108 sera from patients with rheumatoid arthritis compared with 231 sera from patients with other connective tissue diseases and 365 sera from healthy subjects and patients without these diseases. Moreover, positive tests occurred primarily in patients with active arthritis (r = 0.68). In contrast, these rheumatoid factor autoantibodies were not detected in sera from most of the patients with other autoimmune diseases, including patients with systemic lupus erythematosus. These results show that rheumatoid factors identified in human sera by the double binding test are specific for active rheumatoid arthritis.

A conserved anti-DNA antibody idiotype associated with nephritis in murine and human systemic lupus erythematosus.

In order to identify unique structural features of pathogenic autoantibodies to DNA in SLE, a murine anti-anti-DNA (anti-Id) mAb (mAb 1C7) was produced in response to immunization of lupus mice with a syngeneic anti-DNA mAb (mAb 3E10). Immunization of lupus mice with mAb 3E10 inhibited production of native anti-DNA antibodies, suppressed development of lupus kidney disease (nephritis), and induced production of anti-anti-DNA (anti-Id) antibodies. mAb 1C7 bound F(ab')2 fragments of mAb 3E10, and it bound other murine anti-DNA mAb, but not murine mAb or polyclonal serum antibodies unreactive with DNA. Moreover, binding of mAb 1C7 anti-Id to mAb 3E10 was inhibited by DNA, suggesting anti-Id binding within or near the binding site for DNA. Furthermore, mAb 1C7 bound serum IgG immunoglobulins from 9/12 patients with lupus nephritis and serum anti-DNA antibodies compared to only 3/12 SLE patients with comparable serum levels of anti-DNA antibodies, but without nephritis (p = 0.04), and only 1/53 SLE patients without serum anti-DNA antibodies, 0/49 patients with rheumatoid arthritis, and 1/47 healthy subjects (p less than 0.001). These results provide evidence that mAb 1C7 identifies a conserved Id associated with anti-DNA antibodies in murine and human SLE and may be useful as a structural probe to characterize pathogenic anti-DNA antibodies in SLE.

Human monoclonal IgG rheumatoid factor has structural homology with bacterial Fc receptor proteins.

A cloned lymphoblast cell line, hRF-1, that secreted human monoclonal IgG4 rheumatoid factor autoantibody was produced by Epstein-Barr virus transformation of lymphocytes from rheumatoid arthritis synovium. The binding of hRF-1 rheumatoid factor to IgG globulins of different mammalian species was similar to the binding specificity of Staphylococcus aureus protein A (SpA) and to antibodies found in the sera from patients with rheumatoid arthritis. hRF-1 also had the same binding pattern to human IgG subclasses as SpA. Direct competition was observed between SpA and hRF-1 in binding IgG Fc. These results provide evidence for structural homology between a bacterial Fc receptor protein (SpA) and the monoclonal IgG rheumatoid factor.

Thymoma associated with rheumatoid arthritis in a patient taking methotrexate.

Recently many patients with rheumatoid arthritis (RA) have been effectively treated with methotrexate. Until now, there have been no treatment related neoplasms reported in patients with RA taking methotrexate. Furthermore, there have been no reports of thymoma associated with isolated RA. We describe a patient with classical RA who developed a thymoma while being treated with methotrexate.

Antiguanosine antibodies: a new marker for procainamide-induced systemic lupus erythematosus.

Antinuclear antibodies are present in most patients receiving procainamide. To ascertain whether IgG antiguanosine antibodies are associated with the development of the symptoms of systemic lupus erythematosus, we compared the levels of these antibodies in the sera of 65 patients receiving procainamide: 18 with procainamide-induced symptoms and 47 asymptomatic patients. Antinuclear antibodies measured by immunofluorescence were present in the 18 patients with drug-induced symptoms but also in 24 asymptomatic patients. Similarly, elevated serum levels of antibodies to single-stranded DNA were found in 15 patients with symptoms and in 20 asymptomatic patients. In contrast, levels of IgG antiguanosine antibodies were elevated in 15 patients with drug-induced symptoms, but in only 3 asymptomatic patients. Antiguanosine antibodies binding to single-stranded DNA were found primarily in patients with arthritis, pleuritis, and pericarditis. These results suggest a strong association between IgG antiguanosine antibodies and major manifestations of procainamide-induced systemic lupus erythematosus.

Hodgkin's lymphoma in a patient treated for Wegener's granulomatosis with cyclophosphamide and azathioprine.

Only 5 neoplasms have been reported associated with the use of cytotoxic drugs in the treatment of Wegener's granulomatosis. Described here for the first time, to our knowledge, is a patient with Wegener's granulomatosis treated with cyclophosphamide and azathioprine who later developed Hodgkin's lymphoma.

Impaired response of neutrophils to a lymphokine by sera from patients with connective tissue disease.

The studies reported here were designed to determine whether sera from various patients could prevent neutrophils from responding to the lymphokine, neutrophil migration inhibition factor from T lymphocytes (NIF-T). Neutrophils from healthy donors were treated with sera from 84 subjects and assayed for responses to NIF-T. Serum from 7 of 37 patients (19%) with rheumatoid arthritis, systemic lupus erythematosus, and various forms of vasculitis showed blocking activity. In contrast, none of 47 subjects, including healthy individuals and patients with spondylarthropathies, cancer, and active infections had a serum factor that prevented neutrophils from responding to NIF-T (P less than 0.01). Serum blocking activity occurred transiently in association with infection by Staphylococcus aureus in one patient with rheumatoid arthritis. Moreover, autologous neutrophils from this same patient showed impaired responses to NIF-T. Blocking activity could be eluted from protein A-Sepharose in three of three patients studied. In three of seven patients, blocking activity was detected in serum cryoprecipitates, with a recovery of 46 to 78% of the blocking activity and overall enrichment (purification) of 137- to 281-fold. Analysis of cryoprecipitates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed the predominance of immunoglobulins M and G. In one patient, the serum blocking activity was not cryoprecipitable, and cryoprecipitates from a patient with essential cryoglobulinemia failed to prevent neutrophils from responding to NIF-T. Blocking activity was relatively specific for NIF-T, as there was no effect on F-met-leu-phe-induced chemotaxis of neutrophils. Serum blocking activity in patients with connective tissue disease showed some correlation (r = 0.50; P less than 0.01) with immune complexes detected by polyethylene glycol precipitation but not Clq binding. These studies suggest that the response of neutrophils to NIF-T may be blocked by serum, possibly as a result of immune complexes or autoantibodies found primarily in patients with connective tissue disease.

Effect of methylprednisolone on the production of neutrophil migration inhibition factor by T lymphocytes (NIF-T).

Glucocorticoids may suppress cell-mediated immunity by inhibiting lymphocyte mediator production or reducing the responsiveness of target cells to these mediators. Our laboratory recently described a newly recognized T-lymphocyte mediator, neutrophil migration inhibition factor from T-lymphocytes (NIF-T). In this report we assessed the effect of glucocorticoids on NIF-T activity. Methylprednisolone (MP) at concentrations as low as 10-7 M inhibited NIF-T activity from peripheral blood lymphocytes (PBL) in response to staphylococcal protein A (SPA) and concanavalin A (Con A). However, MP at concentrations as high as 10-4 M did not after the responsiveness of neutrophils to NIF-T. Therefore, the effect of MP on NIF-T activity was due to inhibition of mediator production. The effect of MP on NIF-T production was reversible in 24 hours. This finding is consistent with the clinical observation that alternate day therapy does not suppress cell-mediated immunity. Serum taken from a patient as early as one hour after oral administration of 100 mg of prednisone inhibited NIF-T production in vitro; serum obtained at 48 hr after prednisone had no measurable effect on NIF-T activity, In addition. MP inhibited NIF-T production by previously activated lymphocytes.

Effect of immunoglobulin G on the responsiveness of human neutrophils to soluble staphylococcal protein A-induced neutrophil migration inhibition factor from T-lymphocytes.

Staphylococcal protein A is a bacterial cell wall product that binds human immunoglobulin G and thereby interferes with opsonization and phagocytosis of Staphylococcus aureus by neutrophils. Phagocytic cells are also responsive to various non-immunoglobulin lymphocyte mediators. We utilized the detection of a newly recognized mediator, a neutrophil migration inhibition factor from T-lymphocytes (NIF-T), to show that aggregates of staphylococcal protein A and immunoglobulins G could inhibit the responsiveness of neutrophils to NIF-T. That such aggregates may alter the responsiveness of neutrophils to lymphocyte mediators that amplify or modulate phagocytic functions may have important pathogenetic implications in staphylococcal infection.