Influence of low socioeconomic status on mortality in granulomatosis with polyangiitis.

Objectives: To evaluate the influence of low socioeconomic status (SES) on mortality among patients with granulomatosis with polyangiitis (GPA).Methods: Using nationwide registers, we established a cohort of 827 patients diagnosed with GPA in the public hospital system of Denmark. For each patient, information regarding educational level, civil status, employment status, and comorbidities at time of GPA diagnosis was collected. We used Cox regression analyses to calculate hazard ratios (HRs) adjusted for age, gender, calendar period of GPA diagnosis, and Charlson Comorbidity Index score for preceding illnesses as a measure of relative risk of death. We assessed the risk of death associated with three measures of low SES: basic schooling only, civil status as single, and being unemployed or recipient of disability pension.Results: The median age of patients at GPA diagnosis was 61 (interquartile range 51-69) years, and 508 were 18-64 years old. During a total of 4337 person-years, 237 patients died. Among patients aged 18-64 years at GPA diagnosis, all three measures of low SES were identified as risk factors for death [basic schooling only: HR = 2.04, 95% confidence interval (CI) 1.30-3.19; civil status as single: HR = 1.95, 95% CI 1.24-3.05; being unemployed or recipient of disability pension: HR = 2.96, 95% CI 1.72-5.08]. The association between low SES and mortality was less pronounced among patients aged ≥ 65 years.Conclusions: Our observations indicate that low SES is associated with increased mortality in GPA, especially among patients of working age.

Long-term risk and outcome of infection-related hospitalization in granulomatosis with polyangiitis: a nationwide population-based cohort study.

OBJECTIVE: To assess the long-term risk and outcome of infection-related hospitalization (IH) among patients with granulomatosis with polyangiitis (GPA). METHOD: We used administrative databases to establish a GPA cohort (n = 398), construct a comparison cohort of population controls (n = 3980), and collect clinical data. Cox regression analyses were used to determine hazard ratios (HRs) as a measure of relative risk. Follow-up began at date of GPA diagnosis and continued for up to 10 years. RESULTS: GPA patients had a markedly increased long-term risk of IH compared to controls [HR (95% confidence interval) year 1: 9.5 (7.0-12.8); years 2-5: 3.2 (2.4-4.3); years 6-10: 2.6 (1.8-3.9)]. Increased long-term risks were found for hospital-treated pneumonia, urinary tract infection, sepsis, and skin infection. We did not observe a lower risk of IH for people diagnosed with GPA during 2005-2014 than for those diagnosed during 1995-2004. Mortality at 3 and 6 months after IH did not differ significantly between patients diagnosed with vasculitis during 2005-2014 and those diagnosed during 1995-2004. Charlson Comorbidity Index score ≥1 was identified as a predictor of pneumonia and urinary tract infection in the GPA cohort, but not of sepsis or skin infection. CONCLUSION: Patients with GPA have a high risk of IH, even after prolonged follow-up. The long-term risk of IH and mortality after IH did not decline across recent calendar periods among Danish GPA patients. These observations underscore the need for clinical strategies to reduce the burden of infectious complications in GPA.

Clinical and biochemical presentation of sarcoidosis with high and normal serum angiotensin-converting enzyme.

OBJECTIVES: The presentation of sarcoidosis can involve symptoms from all organs and the diagnosis is therefore often difficult. A raised serum level of serum angiotensin-converting enzyme (sACE) can be detected in 41-58% of patients. However, whether the sACE level per se reflects the severity of the sarcoid inflammation at the onset of the disease is not well described. The purpose of this study was to investigate the clinical and laboratory significance of high versus normal sACE levels in sarcoidosis. METHOD: Journal data were retrospectively extracted from 101 patients from our clinic. Clinical and biochemical data were compared between patients with high sACE levels (> 115 U/L) on at least one occasion and normal sACE levels (< 115 U/L). RESULTS: In total, 48% (n = 48) of the patients had high ACE and 52% (n = 53) had normal ACE. The most common extrapulmonary manifestation for both groups was arthritis, followed by skin and eye involvement, but none of these differed between the two groups. Serum ionized calcium was significantly higher in the high sACE group, with a correlation coefficient of 0.112 (p = 0.460). CONCLUSION: Our study demonstrates that serum ionized calcium is significantly higher in the high sACE group but there was no statistical correlation to sACE. No other clinical or biochemical differences were observed.

Incidence of granulomatosis with polyangiitis (Wegener's) in Greenland and the Faroe Islands: epidemiology of an ANCA-associated vasculitic syndrome in two ethnically distinct populations in the North Atlantic area.

OBJECTIVES: Previous studies suggest that the incidence of granulomatosis with polyangiitis (Wegener's; GPA) increases along a south-north gradient in the Northern Hemisphere with an incidence of 8.0/million/year reported for the population of Northern Norway. In the present study, we assessed the incidence of GPA in the predominantly Inuit population of Greenland and in the Caucasian population of the Faroe Islands. METHODS: Greenlandic and Faroese patients affected by severe rheumatic diseases are routinely referred to the National University Hospital in Denmark for treatment. By means of the Danish National Hospital register, we identified all Greenlandic and Faroese patients treated at the hospital under a diagnosis of GPA during 1992-2011. For each patient, the GPA diagnosis was validated by medical files review. RESULTS: One patient born and living in Greenland and 6 from the Faroe Islands were identified. The incidence of GPA was 1.0/million/year (95% CI 0.02-5.6) in Greenland and 6.4/million/year (95% 2.4-14.0) in the Faroe Islands. During the period of study, no cases of GPA occurred among Greenlanders aged 0-44 years, while an incidence of 4.1/million/year (95% CI: 0.1-22.9) was calculated for those aged ≥45 years. In the Faroese population, incidences of 1.7/million/year (95% CI 0.4-9.4) and 14.8/million/year (95% CI 4.8-34.6) were calculated for the age-groups 0-44 and ≥45 years, respectively. CONCLUSIONS: The occurrence of GPA is lower among Inuit in Greenland than among Caucasians living in the Faroe Islands. This observation demonstrates that the risk of GPA varies across ethnic groups populating the northernmost regions of the world.

Deoxyspergualin in relapsing and refractory Wegener's granulomatosis.

OBJECTIVES: Conventional therapy of Wegener's granulomatosis with cyclophosphamide and corticosteroids is limited by incomplete remissions and a high relapse rate. The efficacy and safety of an alternative immunosuppressive drug, deoxyspergualin, was evaluated in patients with relapsing or refractory disease. METHODS: A prospective, international, multicentre, single-limb, open-label study. Entry required active Wegener's granulomatosis with a Birmingham vasculitis activity score (BVAS) > or =4 and previous therapy with cyclophosphamide or methotrexate. Immunosuppressive drugs were withdrawn at entry and prednisolone doses adjusted according to clinical status. Deoxyspergualin, 0.5 mg/kg per day, was self-administered by subcutaneous injection in six cycles of 21 days with a 7-day washout between cycles. Cycles were stopped early for white blood count less than 4000 cells/mm(3). The primary endpoint was complete remission (BVAS 0 for at least 2 months) or partial remission (BVAS <50% of entry score). After the sixth cycle azathioprine was commenced and follow-up continued for 6 months. RESULTS: 42/44 patients (95%) achieved at least partial remission and 20/44 (45%) achieved complete remission. BVAS fell from 12 (4-25), median (range) at baseline to 2 (0-14) at the end of the study (p<0.001). Prednisolone doses were reduced from 20 to 8 mg/day (p<0.001). Relapses occurred in 18 (43%) patients after a median of 170 (44-316) days after achieving remission. Severe or life-threatening (> or = grade 3) treatment-related adverse events occurred in 24 (53%) patients mostly due to leucopaenias. CONCLUSIONS: Deoxyspergualin achieved a high rate of disease remission and permitted prednisolone reduction in refractory or relapsing Wegener's granulomatosis. Adverse events were common but rarely led to treatment discontinuation.

Reduced folate carrier polymorphism determines methotrexate uptake by B cells and CD4+ T cells.

OBJECTIVE: To examine if polymorphism 80G --> A in the Reduced Folate Carrier (RFC) affects uptake of MTX in B- and CD4+ T-cells. METHODS: Mononuclear cells were isolated from peripheral blood of healthy persons. Real-time PCR was used to detect the RFC80 variants. FITC-labelled MTX was added to cells stimulated with Candida albicans or tetanus toxoid, and the uptake of MTX was measured by flow cytometry. A FITC-conjugated monoclonal antibody against RFC was used to detect the cellular RFC expression. RESULTS: Antigen-stimulated CD4+ T cells and B cells from individuals with the GG variant (n = 9) exhibited lower uptake of MTX than individuals expressing the AA variant (n = 8), or the GA variant (n = 8). No difference could be demonstrated between the three groups with respect to the expression of RFC by CD4+ T cells and B cells, and CD4+ T cells from individuals homozygous for the G allele exhibited lower uptake of MTX per receptor than CD4+ T cells from individuals homozygous for the A allele. CONCLUSION: MTX is taken up more efficiently via the A allele than via the G allele. This difference between the variant forms of RFC suggests that genotyping could be relevant for determining the relevant dosage of MTX in the treatment of neoplastic and autoimmune disease.

A rapid and semi-quantitative method for the detection of autoantibodies by multiple spot immunoassay.

A method for rapid simultaneous determination of multiple autoantibodies in sera has been developed. 42 different antigens were coated onto a nitrocellulose membrane in a miniblotter apparatus with 42 lanes. The membrane was also coated with different concentrations of human IgG to create a standard curve. The blotting apparatus after coating, blocking and washing was turned 90 degrees so that all lanes crossed the antigen coated lanes. Thereafter, 42 sera were incubated with the antigens. After this stage the membrane was treated as in the usual dot-blot procedure. After incubation with alkaline phosphatase labelled anti-human IgG, staining and drying of the membrane, the staining intensity of individual lanes was scanned into a data file and analyzed by computer. By this method it was possible in 4 h to examine 42 sera for autoantibodies against 42 antigens, i.e., more than 1600 tests. Furthermore, the amount of antibody could be semi-quantified, using the IgG standards. The method should be useful for rapid screening of autoantibodies in a routine laboratory.

Clinical correlates and substrate specificities of antibodies exhibiting neutrophil nuclear reactivity--a methodological study.

In a serological laboratory with a routine service for determining autoantibodies to human neutrophils, antibodies giving a selective or preferential reaction with the nucleus or perinuclear area of neutrophils are not uncommon. The aim of this study was to look for clinical correlates with the presence of such neutrophil-reactive autoantibodies. The specificity of such antibodies for nuclear or cytoplasmic antigens was studied in 65 consecutive sera displaying nuclear/perinuclear reactivity at a titre of at least 80 using the indirect immunofluorescence technique (IIF) on ethanol-fixed leucocytes. The sera were also investigated by IIF on formalin-acetone fixed leucocytes and on HEp-2 cells. ELISA techniques were used to measure antibodies to azurophil granule constituents (ANCA), purified myeloperoxidase (MPO-ANCA), and lactoferrin (LF-ANCA). Furthermore a qualitative spot immunoassay was used for the detection of antibodies to alpha, beta, and gamma fractions, and the nuclear fraction of neutrophils, purified proteinase 3 (PR3), MPO, enolase, lysozyme, elastase, lactoferrin, and cathepsin G. The diagnoses linked to such GS-ANA/pANCA positivity were arthritides, vasculitides, inflammatory bowel disease and chronic hepatic conditions. MPO was the main antigen recognized in the vasculitis group, but apart from that, rather limited antigen reactivity was demonstrable by these techniques, lysozyme being the most frequently recognized autoantigen in patients with arthritides. Human lymphocytes served as a suitable control substrate when distinguishing between GS-ANA/pANCA and ANA, whereas HEp-2 cells usually could not be used if both classes of antibodies were present in a sample. Furthermore, formalin-acetone fixation is not recommended for routine use.

Measurements of proteinase 3 and its complexes with alpha 1-proteinase inhibitor and anti-neutrophil cytoplasm antibodies (ANCA) in plasma.

Wegener's granulomatosis (WG) is a systemic vasculitis which is diagnosed on clinicopathological findings. The diagnosis may be aided by the presence of anti-neutrophil cytoplasm antibodies (ANCA). In WG, ANCA are primarily directed to proteinase 3 (PR3), a serine protease of the azurophilic granules of the neutrophilic granulocyte. The main plasma inhibitor of PR3 is alpha 1-proteinase inhibitor (PI). To study if free PR3 or complexes between the enzyme and PI or PR3 and ANCA could be found in the plasma from patients with WG we have developed three ELISA systems for the detection of these complexes and free PR3. In all three assays monoclonal antibodies against PR3 were used as capture antibodies. After incubation with plasma, free PR3 was detected by affinity purified rabbit anti-PR3 followed by alkaline phosphatase-labelled swine anti-rabbit IgG. Serial dilutions of purified PR3 was used as standard. The detection limit was 3 ng/ml. PR3 complexed with PI was measured by rabbit anti-PI antibodies and alkaline phosphatase-labelled swine anti-rabbit IgG. Pre-formed in vitro complexes of PR3/PI in serial dilutions were used as standard. The detection limit of this assay was 1 ng/ml. PR3/IgG-ANCA complexes were detected by alkaline phosphatase labelled goat anti-human IgG. A positive plasma sample in serial dilutions was used as standard. Plasma samples from nine patients with WG, eight patients with fever of infectious origin without evidence of vasculitis and ten healthy donors were examined by these methods. Free PR3 could not be found in any of the plasma samples. PR3/PI complexes were detected in healthy donors at levels between 41-85 ng/ml. All WG patients, both active and inactive, had PR3/PI concentrations above this level, and so had all patients with fever. PR3/IgG-ANCA was found in three of the patients with WG, two being ANCA negative with inactive disease and one was ANCA positive with active disease. Thus, the developed methods can be useful for future studies of the clinical relevance of these complexes in patients with WG and possibly other vasculitides.

Anti-neutrophil cytoplasm autoantibodies (ANCA). The need for specific and sensitive assays.

Anti-neutrophil cytoplasm antibodies (ANCA) are a group of autoantibodies primarily associated with systemic vasculitis. Hitherto, the method of choice for ANCA detection has been indirect immunofluorescence (IIF). By this method two major patterns can be seen: a cytoplasmic pattern (cANCA) or a perinuclear pattern (pANCA). The cANCA pattern is most often caused by antibodies directed against proteinase-3 (PR3) and in rare cases it is caused by anti-myeloperoixdase (MPO) antibodies. The pANCA pattern can de caused by antibodies directed against a large group of proteins i.e. MPO, lactofenin and bactericidal/permeability-increasing protein. Often there is a discrepancy between the results obtained by IIF and those reported from the use of assays with purified antigens. This causes confusion. Until now only anti-PR3 and anti-MPO have been found of any clinical value. Therefore, it would be more proper to use assays with these highly purified antigens instead of an unspecific method like IIF.

Effect of 8 wk of bicycle training on the immune system of patients with rheumatoid arthritis.

The effect of 8 wk of progressive bicycle training on the immune system was evaluated in a controlled study on 18 patients with rheumatoid arthritis and moderate disease activity. Maximal O2 uptake increased significantly, whereas heart rate at stage 2 and rate of perceived exertion decreased significantly, in the training group compared with the controls. Resting levels of a number of immune parameters were measured before and after 4 and 8 wk of training. Training did not induce changes in blood mononuclear cell subpopulations, proliferative response, or natural killer cell activity. Furthermore the plasma concentrations of interleukin-1 alpha, interleukin-1 beta, and interleukin-6 did not change in response to training. It is concluded that 8 wk of bicycle training does not influence the immune system of patients with rheumatoid arthritis.

Vitamin D3 metabolism in patients with rheumatic diseases: low serum levels of 25-hydroxyvitamin D3 in patients with systemic lupus erythematosus.

1,25-dihydroxyvitamin D3 (1,25(OH)2 D3) has been shown to modulate lymphocyte activation in vitro. Through binding to specific receptors 1,25-(OH)2 D3 inhibits proliferation, immunoglobulin production and the release of cytokines. Moreover, 1,25-(OH)2 D3 is efficiently produced by activated monocytes. These findings suggest that 1,25-(OH)2 D3 may play a role as a regulator of immunological activation. Consequently, we found it of interest to study the serum levels of the two major metabolites of vitamin D3 in patients with systemic lupus erythematosus (SLE) (n = 21), rheumatoid arthritis (RA) (n = 29) and osteoarthritis (n = 12). In patients with SLE the levels of 25-OH D3 were below those of the healthy controls (p = 0.0008) and OA (p = 0.0168). The levels 1,25-(OH)2 D3 corresponded to normal levels. There were no significant correlations between 25-OH D3 levels and clinical or paraclinical disease manifestations. Further, the phenotypic distribution of Gc-globulin, which binds vitamin D3 metabolites in circulation, was normal. The serum concentrations of 1,25-(OH)2 D3 and 25-OH D3 in patients with RA and OA corresponded to those of the controls. Although the cause of the reduced 25-OH D3 levels in SLE patients is unclear, possible beneficial effects of administration of vitamin D to these patients should be considered.

Anticardiolipin antibodies and lipoprotein (a) levels in patients with chronic renal failure treated with dialysis: associations with atherothrombotic disease.

The present study evaluated the importance of anticardiolipin antibodies (ACA) and lipoprotein (a) (Lp(a)) as markers of atherothrombotic disease in a retrospective study of patients on maintenance hemodialysis (HD) or peritoneal dialysis (PD). ACA and Lp(a) were measured in 22 patients on PD, and 64 on HD. Three patients were ACA IgM seropositive, whereas none were ACA IgG seropositive. The mean number of previous atherothrombotic events was 2.0 (1-3) in ACA seropositive patients, as compared to 0.7 (0-4) in ACA seronegative patients. The mean Lp(a) level was 56.7 mg/dl (3.0-217.7) in PD patients and 38.8 mg/dl (2.0-255.6) in HD patients (n.s.). Levels of Lp(a) greater than 30 mg/dl were not significantly associated with a history of atherothrombotic events, but all patients who had suffered a myocardial infarction or cerebrovascular insult had Lp(a) levels above 30 mg/dl. We conclude that ACA seropositivity is rare. All ACA seropositive patients had suffered atherothrombotic disease in the current study, whereas all patients with myocardial infarction or cerebrovascular insult had Lp(a) levels above 30 mg/dl.

[Humero-scapular periarthrosis]. / Periarthrosis humeroscapularis.

Humero-scapular periarthrosis (PH) is defined as an idiopathic painful condition in the shoulder with simultaneous limitation of active and passive movements in all directions. The condition is characterized by three phases: a painful period, a period dominated by stiffness and a period of recovery. PH is commonest between the ages of 40 and 60 years and the cumulated risk is estimated to be 2%. The incidence is higher in diabetics. The cause of PH is unknown and knowledge of the pathogenesis is very limited. Histological examination shows inflammation and fibrosis. Association with other pathological conditions of the upper limb or medical conditions, except diabetes mellitus, has not been documented with certainty. The average duration is 30 months but recovery without functional defects is common. Treatment is symptomatic and consists primarily of analgesics and rest during the painful phase. Local injections of steroids may be of effect. Exercises may be employed once the pain is under control and these have the object of improving mobility and increasing muscular strength.

[Persons aged 65 years and over admitted to the medical department of a hospital in the Danish provinces with 4 specialized departments. An analysis of extra days of hospitalization related to the services for senior citizens in 4 communities]. / Indlagte på 65 år og derover på medicinsk afdeling på firdelt sygehus i provinsen. En analyse af ekstra indlaeggelsesdage relateret til fire kommuners aeldreservice.

In a retrospective investigation of year 1985, it was found that out of 438 medical patients greater than or equal to 65 years there were in all 42 patients who together were responsible for 1,543 bed-days after the time when they were considered to have concluded hospital treatment. The extra bed-days correspond to approximately 9% of the total medical bed capacity. The patients came from four communities whose coverage with nursing homes, home nursing services and home help arrangements were very different. It was found that increase in nursing home capacity was followed by decrease in the number of extra bed-days among the patients who had extra bed-days. When the number of home nurses increased, fewer elderly patients from the community concerned, were admitted who ended by having extra long hospitalization. These conditions are discussed and the results are compared with previously published investigations. It is concluded that improved cooperation between communities and hospitals, is required.

Methotrexate induces poly(ADP-ribose) polymerase-dependent, caspase 3-independent apoptosis in subsets of proliferating CD4+ T cells.

The mechanism of action of methotrexate (MTX) in autoimmune diseases (AID) is unclear. A pro-apoptotic effect has been demonstrated in mitogen-stimulated peripheral blood mononuclear cells (PBMC), but studies employing conventional antigens have disputed a pro-apoptotic effect. CD4+ T helper (Th) cells play a significant role in most AID. We therefore examined directly, by flow cytometry, the uptake of MTX by the T helper (Th) cells stimulated for 6 days with Candida albicans (CA) or tetanus toxoid (TT), and its consequences with respect to induction of apoptosis. While none of the resting Th cells took up MTX, nearly all the dividing Th cells did, and this abrogated further cell division. Among dividing Th cells, MTX induced an approximately sixfold increase over baseline levels in the proportion of apoptotic cells. This proportion could be reverted to baseline by the addition of folic acid. Exposure of CA-stimulated PBMC to MTX significantly increased their level of cleaved poly(ADP-ribose) polymerase (PARP), and a similar tendency was observed in TT-stimulated cells. Unlike CA and TT, the mitogen phytohaemagglutinin (PHA) induced proliferation of both CD4- and CD4+ T cells, and induced apoptosis in both undivided and divided Th cells. PHA-induced apoptosis involved activation of caspase-3 and the anti-apoptotic protein Bcl-2 in addition to PARP cleavage, suggesting that PHA induces apoptosis via different pathways than CA and TT. We suggest that the latter are more representative of stimulation with self-antigens in AID, and that a pro-apoptotic effect of MTX on self-antigen-stimulated Th cells contributes to the effect of MTX in the treatment of AID.

Insulin resistance in patients with rheumatoid arthritis: effect of anti-TNFalpha therapy.

OBJECTIVES: We undertook this study to test the hypotheses that patients with active rheumatoid arthritis (RA) are insulin resistant and that anti-tumour necrosis factor-alpha (TNFalpha) therapy improves not only the clinical state of these patients but also their glucose metabolism. METHODS: Nine RA patients with active disease and nine healthy subjects, matched for sex, age, and body mass index (BMI), underwent a hyperinsulinaemic euglycaemic clamp. The RA patients received anti-TNFalpha therapy with Humira(adalimumab) and had the insulin clamp re-evaluated after 8 weeks of treatment. RESULTS: Patients with RA had marked insulin resistance (glucose infusion rate (GIR) area under the curve (AUC) was 499+/-55 mg/kg in the RA group compared to 710+/-77 mg/kg in the control group; p<0.05). However, insulin sensitivity did not differ before and after 8 weeks of adalimumab therapy. The RA patients demonstrated a reduction in C-reactive protein (CRP) and interleukin-6 (IL-6) levels after the therapy as compared to pretreatment values, but there was no concomitant effect on plasma levels of TNFalpha. CONCLUSION: RA patients with active disease showed marked insulin resistance that was not influenced by anti-TNFalpha therapy despite a reduction in systemic inflammation during the treatment.

Magnetic resonance imaging for accelerated assessment of drug effect and prediction of subsequent radiographic progression in rheumatoid arthritis: a study of patients receiving combined anakinra and methotrexate treatment.

OBJECTIVES: By MRI to assess the efficacy of addition of anakinra for controlling synovitis and stopping erosive progression in patients with clinically active RA despite receiving methotrexate, and to determine the predictive value of MRI for subsequent radiographic erosive progression. METHODS: 100 mg anakinra subcutaneously/day was added to the treatment of 17 patients with clinically active RA despite methotrexate. MRI of the non-dominant wrist and 2nd-5th MCP joints (OMERACT evaluation) was performed at weeks 0, 12, and 36, and radiography of both hands and wrists (modified Sharp evaluation) at weeks 0 and 36. RESULTS: MRI synovitis scores were not significantly changed. Radiography of both hands and wrists after 36 weeks showed erosive progression in 11 patients, and MRI after 12 weeks in 10 patients. Nine of 10 patients with MRI progression at 12 weeks had radiographic progression at 36 weeks. Baseline MRI synovitis and erosion scores, but no clinical/biochemical parameters, correlated significantly with subsequent erosive progression. CONCLUSION: Addition of anakinra did not significantly reduce MRI signs of synovitis, and most patients had progressive joint destruction. Baseline MRI findings predicted subsequent radiographic erosive progression. Unilateral wrist and MCP joint MRI after 12 weeks had a similar sensitivity for detection of erosive progression as bilateral hand and wrist radiography after 36 weeks.

Some patients with anti-myeloperoxidase autoantibodies have a C-ANCA pattern.

Rapidly progressive glomerulonephritis with or without other signs of systemic vasculitis is often accompanied by antibodies to myeloperoxidase. Such antibodies normally produce a perinuclear pattern on ethanol-fixed neutrophils (perinuclear anti-neutrophil cytoplasm antibodies (P-ANCA)) at indirect immunofluorescence. We report here sera from three patients that are anti-myeloperoxidase-positive in ELISA that instead produce a cytoplasmic pattern (classical anti-neutrophil cytoplasmic antibodies (C-ANCA)), a pattern normally seen in conjunction with antibodies to proteinase 3. These sera did not react with proteinase 3. For two of the sera the specificity of the anti-myeloperoxidase reaction was confirmed with inhibition-ELISA experiments and with immunoblotting. A mouse anti-myeloperoxidase MoAb that produces a cytoplasmic pattern is also described. Competition ELISA experiments show that this antibody and anti-myeloperoxidase sera with cytoplasmic pattern recognize epitopes that are separate from epitopes recognized by another perinuclear pattern producing anti-myeloperoxidase MoAb. 'Cytoplasmic pattern' epitopes as well as 'perinuclear pattern' epitopes can be found on all three major myeloperoxidase isoforms, after separation by ion exchange chromatography. Affinity chromatography, using the cytoplasmic pattern producing anti-myeloperoxidase monoclonal antibody, shows that the epitope recognized by this MoAb is present on all myeloperoxidase molecules. This epitope is not confined to any special subpopulation. These findings indicate that all myeloperoxidase do not relocate after ethanol fixation, and that C-ANCA and P-ANCA epitopes exist simultaneously on the same myeloperoxidase molecule. We propose that the two immunofluorescence patterns arise due to different availabilities of the epitopes in the microenvironment where myeloperoxidase is present.

Frozen shoulder: current concepts.

The literature on frozen shoulder (FS) is reviewed. The etiology of FS is still not known and our understanding of the pathogenesis is limited. Studies on treatment programs under controlled conditions are few and incomplete. Further research is urgently needed.