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Purpose: Our aim was to determine the effects of infliximab on bone mineral metabolism in rheumatoid arthritis (RA) patients and analyze the relationship between inflammatory markers of acute phase thought to play a major role in bone remodeling., Materials and Methods: 36 patients with established RA were investigated. All patients underwent physical examination and blood and urinary analysis at baseline, 2 weeks, 14 weeks, 6 months and 12 months after the initiation of treatment. The serum levels of: tumor necrosis factor alpha (TNF-alpha), tumor necrosis factor alpha receptor 1 (TNFR1), TNFR2, interleukin 6 (IL-6), IL-17, IL-23 and markers of bone remodeling such as osteocalcin (BGP), deoxypyridynoline (Dpd), and N-telopeptide of type I collagen (NTx) were measured by ELISA., Results: The results showed significant decrease of all the above cytokines levels in RA patients in comparison with those after 2 weeks of treatment. After 6 months, the markers of bone formation and resorption decreased compared to baseline values. We found positive correlation between the levels of NTx and the levels of IL-6, IL-17 and TNFR1, and between the levels of Dpd and IL-6 and Dpd and TNFR2, whereas negative correlation between BGP and IL-23. After 12 months the positive association was found at the BGP level and IL-6 as well as Dpd and the level of IL-6. We also observed a positive relation between Dpd and TNF-alpha and negative between BGP and TNFR1., Conclusion: We suggest that infliximab treatment may limit the risk of osteoporosis in RA patients.

Interleukin-17F (IL-17F) is a novel proinflammatory cytokine. IL-17F gene is an excellent candidate for chronic inflammatory disease. We investigated the association between rheumatoid arthritis (RA) and His161Arg (7488A/G; rs763780) and Glu126Gly (7383A/G; rs2397084) polymorphism of IL-17F gene. The gene polymorphisms in 220 Polish patients with RA and 106 healthy subjects were amplified by polymerase chain reaction with restriction endonuclease mapping. Overall, the polymorphisms of the IL-17F gene were not correlated with susceptibility to RA in Polish population. However, the IL-17F His161Arg variant was associated with parameters of disease activity, such as number of tender joints, HAQ score or DAS-28-CRP. Moreover, our findings have shown that Glu126Gly IL-17F gene polymorphism may be correlated with longer disease duration in patients with RA. Our results for the first time showed the relationship between IL-17F gene polymorphisms and severity of RA.

Background and Objectives: In order to gain a better insight into the pathogenesis of the anemia of chronic disease (ACD) accompanying rheumatoid arthritis, we analyzed the density of the integrins very late antigen (VLA) 4 and VLA-5 on the surface of erythroblasts from bone marrow in patients with rheumatoid arthritis. We also measured the concentration of interleukin (IL) 3 and tumor necrosis factor (TNF) alpha in bone marrow. Finally, we analyzed the relationship between integrin expression on hematopoietic cells and the degree of anemia and concentration of cytokines in bone marrow in patients with rheumatoid arthritis., Results: Patients with rheumatoid arthritis who also had ACD were found to have lower hemoglobin levels and higher C-reactive protein and erythrocyte sedimentation rate compared to patients who had rheumatoid arthritis without ACD or osteoarthritis of the hip. The mean bone marrow concentration of IL-3 was elevated in patients with rheumatoid arthritis and ACD compared to those without ACD or patients with osteoarthritis. IL-3 concentration in bone marrow showed a significant negative correlation with VLA-4 and VLA-5 expression on erythroblasts, but only in patients with rheumatoid arthritis and ACD., Conclusion: Patients with rheumatoid arthritis and ACD have abnormal erythroblasts (decreased VLA density), possibly through an effect on early stages of erythroblast development. Increased levels of IL-3 and the negative correlation between IL-3 concentration in bone marrow and expression of the integrins VLA-4 and VLA-5 may suggest positive feedback between erythroblasts and IL-3, probably associated with decreased sensitivity of bone marrow erythroblasts to IL-3.

Objective: Glucocorticoids (GCs) are among the most frequently used drugs for the treatment of rheumatoid arthritis (RA). Unfortunately, up to 30% of patients with RA fail to respond to the treatment. We investigated the hypothesis that patients with RA who did not respond to GC treatment have steroid-resistant peripheral blood mononuclear cells (PBMCs)., Methods: Forty-four patients with RA were enrolled in the study. PBMCs were isolated from blood samples. The effect of methylprednisolone (MP) on the proliferation of stimulated cells was measured. After taking the blood samples, 10 days of MP therapy (20 mg i.v.) was started, in order to classify the patients into either a GC-sensitive (RA/GCS) or a GC-resistant (RA/GCR) group., Results: A quarter of our patients did not show any improvement after short-term GC therapy and were assigned to the RA/GCR group. The inhibition of PBMC proliferation after MP treatment was significantly lower in the RA/GCR as compared to the RA/GCS group., Conclusion: Based on the close relationship between clinically observed GC resistance and a diminished response of PBMCs to MP treatment, we conclude that measurement of the steroid sensitivity of PBMCs may be a useful tool in predicting the therapeutic effect of GC in patients with RA.

Osteoporosis associated with rheumatoid arthritis (RA) is induced by chronic inflammation. Glucocorticosteriods (GCS) applied in the treatment of RA chronically reduce production of proinflammatory cytokines (IL-1, IL-6, and TNF) which are potent stimulators of bone resorption. On the other hand they directly reduce bone mass by inhibition of osteoblast. In order to assess bone turnover the following parameters have been measured: Alkaline phosphatase (AP), alkaline phosphatase-bone formation (AP-B), deoxypirydynoline (Dpd) and carboxyterminal telopeptides of type I collagen (CTx). Based on the obtained findings we conclude that: 1. Decrease in level of AP-B and CTx may suggest reduction of bone turnover, 2. Short-term low dose GCS therapy dramatically reduce inflammation which temporarily may reduce the loss of bone mass.

A study was made to evaluate bone turn-over in rheumatoid arthritis (RA) patients treated with infliximab. Twenty-two patients with established RA were included. In all patients, biochemical markers of osteoporosis: osteocalcin (BGP), alkaline phosphatase (bone isoenzyme), deoxypyridinoline (Dpd), acute phase proteins (CRP, AGP, ACT, AGP-RC), and interleukin 6 (IL-6) were determined before treatment, at week 30, and at week 46. Two markers (BGP, Dpd) were significantly decreased at both weeks 30 and 46. Moreover, a fall in serum levels of acute phase proteins and IL-6 was seen. The results suggest that anti-TNF treatment with infliximab not only decreases activity of inflammation but also may slow down bone turn-over. Further research is needed to assess its potential in reducing risk of osteoporosis in RA.

In the present study we investigated the relation between cyclophosphamide and methotrexate toxicity and the presence of HLA- DR B1 alleles in rheumatoid arthritis patients. Seventy-eight such patients (67 women and 11 men) were observed for 12 months. Eighteen were treated with intravenous cyclophosphamide, 28 with oral methotrexate, and 32 with intramuscular gold salts. The prevalence of this shared motif was higher in the study population than in the healthy controls. However, detailed observations did not demonstrate a relation between particular genotype and drug intolerance. Based on the obtained findings we concluded that HLA-DR B1 typing cannot affect cyclophosphamide or methotrexate tolerance in rheumatoid arthritis patients. However, taking into account the relatively small number of patients expressing single genotype, further studies are recommended.

Unlabelled: The aim of this study was to assess of bone mineral content (BMC) and bone mineral density (BMD) of the forearm using DXA technique (DTX-200) and to evaluate broadband ultrasound attenuation (BUA) and speed of sound (SOS) of the heel using QUS technique (DTU-ONE). We examined 83 RA patients: 73 women and 10 men, at average age (55.0 +/- 12.2 yrs), ranging from 29 to 85 yrs. Average disease duration was 112.6 +/- 98.1 months. Disease activity was assessed according to Mallya and Mace index and radiological stage of the disease according to Steinbrocker index. We found significant correlation between BMC, BMD and BUA (r = 0.6572, r = 0.6081, respectively) and between BMC, BMD and SOS (r = 0.4704, r = 0.4723, respectively)., In Conclusion: quantitative ultrasound parameters (BUA and SOS) significant correlate with BMC and BMD values of the forearm assessed by DXA technique in rheumatoid arthritis patients.

The present study was undertaken in order to investigate the relationship between tumor necrosis factor-alpha (TNF-alpha) gene polymorphism and the radiological progression of rheumatoid arthritis (RA) within the first 3-years of the disease. Sixty-eight RA patients (59 women and nine men) were observed for 3-years. TNF-alpha polymorphism analysis was performed in all patients. Radiographs of the hands were taken at the onset of study and after 3-years of follow-up. Radiographs were assessed according to the Larsen index (damage score and progression of damage score). We did not observe any correlation between TNF gene polymorphism and damage score or progression of damage score. The obtained data suggests that TNF-308 polymorphism cannot serve as an indicator of the disease course in RA patients.

Glucocorticosteroids have been recognized as a well known risk factor for drug induced osteoporosis. Many studies have shown a decrease in bone mass, bone quality disorders and an increase in the risk of fractures in patients with long-term corticosteroid therapy. Rheumatic patients, particular with rheumatoid arthritis, who are usually chronic steroid users are at the highest risk. On the other hand uncontrolled active inflammatory process is also a main factor for rapid bone loss. Some studies suggest that patients with low dose corticosteroid therapy (prednisone 5 to 7.5 mg per day) are not at increased risk of osteoporosis. Our study of 36 rheumatoid arthritis women treated with daily prednisone doses between 5 to 7.5 mg in comparison with non-steroid control group confirmed the above suggestion.

In order to better understand the immunological mechanisms involved in the pathogenesis of rheumatoid arthritis (RA), the level of various lymphocyte subsets in the peripheral blood of 29 patients with erosive RA was determined. All the patients were treated with methotrexate for 2 years. The total number and the proportion of CD3 cells, CD3+CD4+ and CD3+CD8+ cells did not change during the study. The initially increased level of CD19+ B-cells and CD19+CD5+ cells decreased during the treatment. The percentage of CD3-CD16+ natural killer cells was not affected by the treatment. At the inception of the study, we observed a deficiency of CD4+ CD45RA+ cells and the level of CD3+CD29+ cells was slightly increased. During the treatment we noticed significant elevation of CD45RA cells. Consequently, the CD29/CD45RA ratio significantly decreased. We showed significant correlation between changes in disease activity and changes in the level of CD19+ cells and CD4+CD29+ cells. Our results suggest that low-dose methotrexate may affect immunocompetent cells. The lowering in the CD29+ subset population associated with depletion of CD19 B-cells after methotrexate therapy may limit abnormal CD4+ cell activation and reduce the migration of lymphocytes into inflamed synovium.

Rheumatoid arthritis (RA) is associated with HLA-DR4 and DR1 antigens. HLA-DRB1 gene sequences analysis demonstrated that only a limited set of alleles is positively associated with RA. Third hypervariable region sequences (70-74, 86) Q(R)R(K)RAA, G(V) are found in up to 95% of erosive RA. The presence of disease-associated allels may predict severe outcome of the disease. Therefore, their presence may allow us to start aggressive therapy in early stage of the disease.

Glycosylation is the main posttranslational modification of protein molecules. Human immunoglobulin G (IgG) is unique among serum glycoproteins because it is associated with at least 30 different biantennary oligosaccharide moieties. Elevated ratio of circulating IgG molecules lacking galactose from the oligosaccharides on the CH2 domain correlates with severity of the disease in a number of different autoimmune illnesses. Moreover, it may be helpful in differential diagnosis of rheumatoid arthritis (RA). By that we may try to predict outcome of RA. Its seems that agalactosyl IgG is distinctively connected with rheumatoid arthritis and may be involved in its immunopathogenesis.

We sought to investigate the influence of interleukin-10 (IL-10) and IL-6 on the acute phase proteins (APP) in systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). IL-10, IL-6, Creactive protein (CRP), alpha-1-acid glycoprotein (AGP), and alpha1 antichymotrypsin (ACT) serum levels were determined in one hundred-eight patients (71 with SLE, 37 with RA). Quantification of the serum IL-10 level showed increased levels in SLE and RA patients as compared to healthy controls. Serum IL-6 level was found to be elevated in SLE and RA patients. A correlation between IL-10 and IL-6 serum level was found only in SLE. CRP and AGP serum levels were increased in RA as compared to controls, whereas in SLE only AGP was found elevated. A statistically significant correlation between IL-6 serum level and CRP, AGP and ACT was found only in RA. No correlation between IL-10 and serum level of CRP, AGP and ACT was established. Since IL-10 has a potent immunosuppressive activity, we expected it to be negatively correlated with APP levels. Surprisingly, IL-10 did not correlate with APP either in SLE or RA patients. However, the elevation of IL-10 serum levels in SLE and RA and the correlation between IL-10 and IL-6 in SLE may suggest that IL-10 may play a central role in inflammatory connective tissue diseases.

Synovial fluid samples from 21 rheumatoid arthritis patients were analyzed for lymphocyte subsets using flow cytometry and antibodies to the lymphocyte surface antigens CD3, CD4, CD8, CD16, CD19, CD29, and CD45RA. Synovial fluid levels of interleukin-6, rheumatoid factors and acute phase proteins were also measured. Depletion of CD45RA+ cells and predominance of CD29+ cells were found. Interleukin-6 levels were markedly elevated (1155 pg/ml; range, 24-3875 pg/ml), as were levels of IgM, IgG and IgA rheumatoid factors and of acute phase proteins. CD4 + CD29+ counts were significantly correlated with interleukin-6 levels. Interleukin-6 levels were significantly correlated with levels of all three rheumatoid factor classes but not with levels of acute phase proteins. Significant correlations were also found between CD19+ B counts and levels of all three rheumatoid factor classes. These data suggest that synovial fluid CD4 + CD29+ cells may be involved in the immune dysregulation characteristic of rheumatoid arthritis. The correlation between CD4 + CD29+ counts and interleukin-6 levels is consistent with the recent hypothesis that, together with monocytes, CD4 + CD29+ cells are an important source of the elevated levels of interleukin-6 seen in rheumatoid synovial fluid.

One hundred and fifty patients with rheumatoid arthritis (RA) were treated with immunosuppressive drugs for 12 months. Fifty nine patients were treated with methotrexate (MTX), 15 with cyclophosphamide (CTX), 30 with prednisone (PRE) and 46 with non steroidal antiinflammatory drugs (NSAID). The cell surface phenotype of lymphocyte was analyzed using immunofluorescence methods ans a variety of monoclonal antibodies. The studies were performed using fluorescence activated cell scanner (FACScan) and epifluorescence microscope. No changes in the percentage of CD3, CD4, CD8 were observed. However, in the MTX treated group the percentage of CD19+ (15.1% before treatment vs 10.2% after 12 months MTX treatment (p < 0.05) and CD5+CD19+ B-cells was decreased. In CTX treated group the percentage of both B cells (17.4% vs 11.0% (p < 0.5)) and activated T cells was decreased (CD25+: 2.8% vs 1.1%, p < 0.05 and HLA-DR+: 22.1% vs 12.7%, p < 0.01). Patients treated with prednisone expressed several changes in lymphocyte phenotype i.e. decreasing of activated T cells (CD3+CD25+: 6.9% vs 3.6%, p < 0.01), B cells (CD5+CD19+: 3.4% vs 0.8%, p < 0.001), and NK cells (CD16+: 14.8% vs 8.5, p < 0.01 and CD56+: 16.3% vs 10.7%, p < 0.01). Analyzing immunosuppressive drugs appeared to be very sufficient in the RA treatment. Moreover, the correlation between percentage of both B lymphocytes and activated T cells, and activity of disease were shown.

In the present study, in order to get a better insight into the mechanism of action of cyclophosphamide (CY) in rheumatoid arthritis (RA), we monitored the changes in lymphocytes' expression of leukocyte function associated antigen 1 (LFA-1). A group of 28 patients with refractory severe RA were treated with CY and methylprednisolone (MO) intravenously. Using flow cytometry we evaluated the changes in LFA-1 molecule expression on peripheral lymphocytes. In the analyzed group of patients the proportion of LFA-1 "dim" cells was reduced. After the treatment the ratio was partly normalized. Twelve months after cessation of the therapy high proportion of LFA-1 "dim" was observed only among CY/MP treated patients. The changes were related to clinical improvement. Based on the obtained data, it seems, that the treatment affecting the expression of LFA-1 may slow down lymphocyte migration and by that limit chronic inflammation within the synovium.

Ninety-six patients with refractory rheumatoid arthritis were treated with methotrexate for 48 months. We analyzed the effect of low doses of methotrexate (7.5-15 mg weekly) on the activity of the disease and on the progression in damage score. Clinical improvement was observed few weeks after the study was introduced. Following 48 months observation only 32 (33%) remained in the study. Seven of them needed additionally prednisone administration. In 64 (67%) patients the drug was withdrawn: in 18 (19%) due to the side effects, in 24 (25%) due to the lack of effectiveness, in 22 (23%) due to other reasons. Methotrexate did not affect the progression of damage score in our patients. Methotrexate appeared to be usefull drug in the treatment of rheumatoid arthritis, however clinical improvement was observed only during drug administration. After discontinuous of the therapy a flare of the disease was observed.

The mode of action of methylprednisolone and cyclophosphamide in the treatment of rheumatoid arthritis still remains unclear. We sought to determine whether methylprednisolone and/or cyclophosphamide affect surface antigens on peripheral blood lymphocytes. Twenty-eight patients with severe refractory rheumatoid arthritis were observed for 12 months. Thirteen patients were treated with an intravenous pulse of methylprednisolone, and fifteen with methylprednisolone combined with cyclophosphamide. The surface antigens of lymphocytes and natural killer (NK) cells isolated from peripheral blood were determined using flow cytometry. The clinical improvement was observed in 16 (57%) patients (8 treated with methylprednisolone and 8 with methylprednisolone/cyclophosphamide). However, after cessation of the treatment in 9 patients, a flare up of the disease was observed. A striking decrease in total lymphocyte count was observed. The percentage of CD3+ and CD3+CD4+ cells remained unchanged. We observed a decrease in the percentage of CD3+CD8+ in patients treated with methylprednisolone/cyclophosphamide. Moreover, the percentage of activated T cells (CD25+ cells and HLA-DR+ cells) was reduced. The depletion of CD8+CD25+ cells was observed after combined treatment. The percentage of CD19+CD5+ was reduced due to the treatment. We also observed a decrease in CD16+CD56+ NK cells. Amelioration of the course of the refractory rheumatoid arthritis was observed in patients treated with both methylprednisolone and methylprednisolone/cyclophosphamide. A stronger effect on lymphocyte phenotype was observed in those given methylprednisolone/cyclophosphamide, but it was not followed by further benefit. On the other hand, the clinical improvement was more stable in methylprednisolone/cyclophosphamide-treated patients. The use of cyclophosphamide should be reserved for patients with rapidly progressing rheumatoid arthritis or life-threatening complications.

Immunoglobulin G-lacking galactose (Gal[0]) appears to be helpful in differential diagnosis of early synovitis, and correlates with disease activity in rheumatoid arthritis (RA). Its utility for RA monitoring and prognosis has been evaluated in the present study. Forty-eight patients with early RA were observed for 3 years. Hand radiographs were assessed according to Larsen and results were expressed as damage score (DS) and progression of damage score (PDS). Gal[0], DS, and PDS were evaluated at the onset of the study and after 1 and 3 years. The average values of Gal[0] in RA patients at the onset of the observation were significantly higher as compared to healthy controls (0.43 +/- 0.22 vs. -0.03 +/- 0.09, P < 0.05). The findings of Gal[0] after a 3-year follow-up were also higher as compared to healthy controls (0.37 +/- 0.21 vs. -0.03 +/- 0.09, P < 0.05). Radiological progression (PDS > 15) was observed in 16 patients. This group was characterized by a constantly high level of Gal[0]. The level of Gal[0] in patients without or with moderate radiological progression (PDS < 15) was significantly lower at the onset of the study and remained low during the observation. The relationship between Gal[0] and radiological progression was shown. The data thus far obtained suggest that Gal[0] may serve as an indicator for the disease course in patients with RA. Secondly, we cannot exclude the possibility that the constantly elevated level of Gal[0] causes erosions.

We sought to investigate an influence of immunosuppressive drugs on acute phase response (APR) in rheumatoid arthritis (RA). Ninety-six patients (pts) were treated with methotrexate (MTX), or with cyclophosphamide (CTX) (9-intravenously, 19-orally), or with cyclosporin A (CSA). C-reactive protein (CRP), alpha-1-acid glycoprotein (AGP), and alpha-1 antichymotrypsin (ACT) serum levels were measured by rocket immunoelectrophoresis. AGP and ACT microheterogenities evaluated using immunoelectrophoresis were expressed as reactivity coefficient (RC). Clinical improvement was observed in 71.4% MTX pts, 77.8% CTX intravenously pts, 36.8% CTX orally pts, 60.0% CSA pts. The number of side effects was the highest in CTX oral group (57.9% left the study). CRP, AGP, and ACT serum levels were increased in all groups of RA pts as compared to healthy controls. CRP level decreased only after MTX and CTX intravenous treatment. Moreover, a decrease in ACT was observed in CTX intravenously treated pts. AGP-RC was lower in the initial population of RA pts as compared to healthy control. After 6 months of treatment RC became significantly higher in MTX pts only. In opposite ACT-RC in RA pts was found to be elevated as compared to controls. After the treatment it fell down. The decrease was found to be significant only in pts treated with MTX. From our study we can conclude that MTX is the safest and the most effective agent among immunosuppressive drugs applied in RA. CTX given orally causes a number of adverse reactions, which frequently make continuous and effective treatment impossible. CTX intravenously and CSA are attractive in the treatment of the patients with severe and refractory RA. A lack of clinical benefit is reflected in the absence of acute markers changes.

We followed the levels of serum IgA-alpha-1-antitrypsin (IgA-AT) complex in 37 patients with early rheumatoid arthritis (RA) during the first 3 years of the disease. The changes in IgA-AT were correlated with a radiological damage score (DS) of the hands assessed according to Larsen. At the onset of the disease, the IgA-AT serum concentration was significantly higher as compared to the control group (0.72 +/- 0.22 U vs 0.29 +/- 0.14 U, P < 0.01). The level significantly decreased during the 3-year observation period. The DS was significantly higher after 3 years. However, this difference was due to changes in only 11 patients; in 26 patients the DS was almost unchanged. In the group of 11 patients with radiological progression, the level of IgA-AT either remained high or increased significantly (0.95 +/- 0.18 U at the onset, 0.97 +/- 0.25 U after 3 years), whereas we observed a decrease in IgA-AT in 26 patients without radiological progression (0.63 +/- 0.16 U at the onset of the disease, 0.45 +/- 0.10 U after 3 years, P < 0.01). Moreover, a relationship between changes in IgA-AT serum level and radiological progression was shown (r = 0.60, P < 0.01). Our studies suggested that the relationship between IgA-AT level and radiological progression of the disease should be considered. We cannot exclude the possibility that the constant high level of IgA-AT may cause worsening in bone erosions.

We sought to investigate whether clinical improvement after immunosuppressive treatment reflects changes in acute phase response (APR) in rheumatoid arthritis (RA). Fifty-eight patients (pts) were treated with methotrexate (MTX), nineteen with intravenous cyclophosphamide (CTX), and fifteen with cyclosporin A (CSA). C-reactive protein (CRP), alpha-1-acid glycoprotein (AGP), alpha-1 antichymotrypsin (ACT), and alpha-1 antitrypsin (AT) serum levels were measured by nephelometry or rocket immunoelectrophoresis. Clinical improvement was observed in 67% MTX pts, 53% CTX pts, and 47% CSA pts. Baseline serum levels of CRP, AGP, ACT, and AT were significantly higher as compared to healthy controls. After MTX and CTX therapy CRP level significantly decreased. The decrease in serum level of ACT and AT in CTX treated patients was also observed. All analyzed acute phase proteins remained substantially elevated after CSA therapy despite a clear reduction in disease activity. We established a correlation between changes in disease activity and all acute phase proteins (APP) in MTX and CTX pts. From our study we can conclude that clinical improvement after immunosuppressive treatment correlated with quantitative changes in all APR markers in MTX and CTX treated pts, and none in CSA pts. Although measurement of APP remains the best marker for monitoring RA pts, not always they properly reflect changes in disease activity.

In order to evaluate the relationship between serum concentrations of interleukin-10 (IL-10), IL-6, and acute phase proteins in rheumatoid arthritis (RA) patients treated with methotrexate (MTX) or intramuscular gold (IMG) we determined IL-10, IL-6, C-reactive protein (CRP), alpha-1-acid glycoprotein (AGP) and alpha-1-antichymotrypsin (ACT) in the sera of 35 RA patients. IL-10 and IL-6 levels were evaluated using an enzyme-linked immunoassay (ELISA). AGP and ACT level were measured using rocket immunoelectrophoresis. IL-10 serum level was not increased in RA patients as compared to controls (58.7 +/- 18.1 pg/ml vs. 57.2 +/- 11.9 pg/ml). IL-6 level was significantly elevated (91.6 +/- 46.9 pg/ml vs. 45 +/- 19 pg/ml, p < 0.05). CRP was significantly increased as compared to healthy controls (35 +/- 19 mg/l vs. 3 +/- 2 mg/l, p < 0.05). Patients treated with MTX or IMG presented an increased level of IL-10 and decreased amounts of IL-6, as compared to those treated with NSAID only. However, only changes between patients treated with IMG and NSAID were found to be statistically significant. A good negative correlation between IL-10 and IL-6 serum level was found (r = -0.75, p < 0.05). A positive significant correlation between IL-6 serum level and CRP (r = 0.62, p < 0.05), AGP (r = 0.78, p < 0.05) and ACT (r = 0.45, p < 0.05) was established.(ABSTRACT TRUNCATED AT 250 WORDS)

To evaluate the effect of interleukin 10 (IL-10) on acute phase response, we determined serum levels of IL-10, interleukin 6 (IL-6), C-reactive protein (CRP), alpha-1-acid glycoprotein (AGP), alpha-1-anti-chymotrypsin (ACT) in 34 rheumatoid arthritis (RA) patients. IL-10 and IL-6 levels were evaluated using an enzyme-linked immunoassay (ELISA). CRP, AGP and ACT levels were measured using rocket immunoelectrophoresis. The results showed that IL-10 serum level was increased in RA patients as compared to controls (60.0 +/- 17.5 pg/ml vs. 7.2 +/- 1.9 pg/ml). IL-6 level was significantly elevated (87.3 +/- 32.7 pg/ml vs. 45 +/- 19 pg/ml, p < 0.05). CRP was significantly increased as compared to healthy controls (34 +/- 19 mg/1 vs. 3 +/- 2 mg/1, p < 0.05). AGP and ACT serum levels were increased in RA patients, but we did not find these changes to be statistically significant. A good negative correlation between IL-10 and IL-6 serum level was found (r = -0.73, p < 0.05). A positive significant correlation between IL-6 serum level and CRP (r = 0.62, p < 0.05), AGP (r = 0.74, p < 0.05) and ACT (r = 0.45, p < 0.05) was established. Moreover, a negative correlation between IL-10 and serum level of CRP (r = -0.76, p < 0.05), AGP (r = -0.60, p < 0.05) and ACT (r = -0.37, p < 0.05) was also shown. According to the data thus far obtained it seems that IL-10 decreases IL-6 production, and by that indirectly affects acute phase response, decreasing CRP, AGP and ACT synthesis.

Microheterogeneity of alpha-1-acid glycoprotein was studied using affinity immunoelectrophoresis with concanavalin A as a ligand in the samples of serum and synovial fluid obtained at the same time from 22 patients with rheumatoid arthritis. Individuals with intercurrent infection or other illnesses were excluded from the study. The results were expressed as reactivity coefficient (RC). Disease activity was evaluated by Mallya-Mace Activity Score, Lansbury Joint Index and laboratory tests. In most of the studied samples the glycosylation pattern was similar, composed of a nonreactive variant and 2 reactive (the first and the second) with Con A variants. In seven samples of synovial fluid an extra third peak representative of the strongly reactive one with Con A fraction was observed. It was the cause of the remarkable elevation of AGP-RC. Moreover, the level of IgM and IgA RF was higher in the synovial fluid derived from these patients--with the presence of the third peak in AFF-EP with Con A--than in those without the considered fraction.

In order to investigate the role of the acute-phase response in early rheumatoid arthritis (RA), we followed the changes of alpha 1-acid glycoprotein (AGP), alpha 1-antitrypsin (AT) and alpha 1-antichymotrypsin (ACT) in the sera of 25 patients with refractory rheumatoid arthritis (RA) during the first 3 years of the disease. Serum levels of AGP, ACT and AT were measured using rocket immunoelectrophoresis, and AGP, ACT and AT microheterogeneities were performed using two-dimensional immunoelectrophoresis with concanavalin A (Con A) as ligand. On average, serum levels of AGP, ACT and AT proteins were higher at the onset of the disease as compared with healthy controls. After 3 years, a significant decrease in serum levels of all three acute-phase proteins (APPs) was observed, but only in patients without anatomical progression was this decrease statistically significant. At the beginning of the study, only the AT reactivity coefficient (RC) was decreased in comparison with healthy subjects. However, after 3 years of disease, AGP, ACT and AT RCs all decreased. This investigation provided two new observations. The first is that only AT RC is significantly lower in comparison with normal values at the beginning of RA, whereas AGP RC and ACT RC remain within the normal range of values; a decrease in AGP RC and ACT RC appeared later. The second observation is that the highly elevated levels of APPs at the onset of RA decrease during the course of the disease. Moreover, this decrease does not depend on the disease activity, but a relationship with radiological progression was shown.

This study was undertaken to determine whether the rheumatoid factor (RF) isotypes affect the level of IgA-AT complex in early rheumatoid arthritis. IgA-AT complex and IgM, IgG, and IgA RF were evaluated using a double antibody enzyme-linked immunosorbent assay (ELISA) in sera of 83 (61 women and 22 men) patients. The mean level of the complex was higher in RA patients as compared with the control group (0.60 +/- 0.41 U vs 0.19 +/- 0.11 U, p < 0.05). The IgA-AT complex values exceeding the norm were found in 19 of 83 RA cases (23%), mainly in patients non-treated with DMARDs. IgM RF values judged as positive were present in 53 (64%), IgG RF in 55 (66%), and IgA RF in 56 (67%) patients. Higher RF values of IgM and IgG, but not those of IgA class, were found more frequently in patients with elevated IgA-AT levels. A good correlation was established between IgA and IgM RF (r = 0.61, p < 0.01), but not between IgA and IgG, or IgM and IgG RF in the group of patients under study. However, we did not find any significant correlation between isotypes of RF and parameters of the disease activity.

One-hundred-and-two-patients (pts) with rheumatoid arthritis (RA) were observed for 12 months. Forty-eight pts were treated with a weekly low-dose of methotrexate (MTX), 23 pts with cyclophosphamide (CTX) (eight pts with one single intravenous dose, and 15 pts orally with a single daily dose), and 31 pts with nonsteroidal antiinflammatory drugs (NSAID) only. In all individuals acute phase response, i.e., C-reactive protein (CRP) and alpha-1-acid glycoprotein (AGP) serum levels, and AGP microheterogeneity using affinoimmunoelectrophoresis with concanavalin A were evaluated. The phenotype of lymphocytes isolated from peripheral blood was characterized using immunofluorescence technique. Following treatment the increased level of CRP significantly decreased whereas AGP serum level remained unchanged. Among the patients, microheterogeneity of AGP expressed as reactivity coefficient (RC) was lower before treatment when compared with 17 controls (0.95 +/- 0.23 vs. 1.35 +/- 0.15, p < 0.01). After 12 months of MTX therapy AGP-RC rose significantly (1.19 +/- 0.13, p < 0.01). No changes were observed in AGP-RC levels in CTX and NSAID treated individuals. No significant differences were observed in the percentage of CD3+, CD4+, and CD8+ cells in all the patient groups, except in CTX intravenously treated patients. In this group of patients a decrease of CD4+ cells was noticed (60.1 +/- 11.5% and 43.8 +/- 12.5% before and after treatment respectively -p < 0.01). The percentage of CD19 positive cells decreased significantly during 12 months of treatment with MTX and CTX. The percentage of activated T cells (CD25+ cells and HLA-DR+ cells) remained unchanged in MTX treated patients and was reduced in both CTX groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Twenty three rheumatoid arthritis (RA) patients treated with cyclophosphamide (CTX) were observed for 6 months. Eight patients received CTX in a single intravenous dose (group I) and 15 orally in a single daily dose (group II). The surface antigens of lymphocytes, isolated from peripheral blood, were determined using immunofluorescence method. The clinical improvement was observed both in group I and in group II of patients. However, in patients receiving CTX intravenously amelioration of the disease appeared quicker. The percentage of CD3+ and CD8+ T cells remained unchanged. In group I we observed decrease in the number of CD4+ T cells (60.1 +/- 11.5% and 43.8 +/- 12.5%, before and after treatment respectively, p < 0.01), in group II this level remained unchanged. In both groups the percentage of CD19+ B cells decreased (14.7 +/- 9.2% and 8.0 +/- 6.1%, before and after treatment respectively in group I, p < 0.01; 17.4 +/- 12.3% and 11.0 +/- 7.1%, before and after treatment respectively in group II, p < 0.01). Moreover, the percentage of activated T cells (CD25+ cells and HLA-DR+ cells) was reduced in both groups.

Seventy nine patients with rheumatoid arthritis (RA) were monitored for 12 months. Forty eight individuals were treated weekly with a low dose of methotrexate (MTX), and 31 received nonsteroid antiinflammatory drugs (NSAID) only. The mean total dose of MTX was 413 mg with a range from 250 to 735 mg. The phenotype of lymphocytes was analyzed using double immunofluorescence and a panel of monoclonal antibodies. The clinical status of 33 MTX-treated patients improved, in 9 neither clinical nor laboratory improvement was noticed. Six patients were withdrawn from the study because of adverse reactions. We observed no changes in the percentage of CD3, CD4, CD8 and CD25 positive lymphocytes in both MTX and NSAID-treated groups. However, the percentage of CD19 positive cells significantly decreased during 12 month observation (15.1 +/- 6.5% v.s. 10.2 +/- 5.0, p < 0.01) in MTX-treated subjects. Moreover, this percentage increased (9.4 +/- 6.7% v.s. 17.1 +/- 1.1%, p < 0.01) in NSAID-treated patients.

The etiology of rheumatoid arthritis (RA) is still unknown. Among the pathogenic mechanisms being discussed, the antigen recognition seems to be of great importance. Probably the "abnormal" interaction between the presented peptide of undefined nature with CD4 T-cell, triggers the pathological events. Immunosuppressive drugs, affecting antigen presentation, are in common use in the treatment of RA. We discuss the effects of alkylating agents, antimetabolites and other immunomodulatory procedures on the course of the disease.

Sixty-two patients with rheumatoid arthritis (RA) were treated with methotrexate (MTX) for 3 months. Methotrexate was given orally in a single dose of 7.5-15.0 mg/week. At the onset of the treatment increased activity of aminotransferases has been observed. Eighteen patients (29%) displayed higher enzyme activity in the first 3 weeks. Despite the continuous drug administration the activity of aminotransferase became normal at week 12 in all but one patient. In this patient MTX had to be withdrawn.

Thirty-four patients with rheumatoid arthritis were treated with oral methotrexate for 6 months. Methotrexate was given as a single weekly dose of 7.5-10 mg. A correlation between serum concentration of CRP and proportion of CD8+ lymphocytes in peripheral blood was found in this group of patients. This correlation can be described by a linear regression equation. According to the results obtained, CRP suppresses lymphocyte function in patients with RA. This is the consequence of the stimulating effect on suppressor mechanisms.