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(1) Autoimmune thyroiditis (AIT) is the most common cause of primary hypothyroidism and one of the most frequent organ-specific autoimmune diseases. Its pathogenesis is polygenic and still requires further research. The aim of the study was to assess, for the first time in the Caucasian population, the role of selected TPO gene promoter polymorphisms (rs2071399 G/A, rs2071400C/T, rs2071402 A/G, and rs2071403 A/G) in the development of AIT. A total of 237 patients diagnosed with AIT and 130 healthy controls were genotyped for four TPO gene polymorphisms, and the results were statistically analyzed to check for the role of these polymorphisms. There were no significant differences in the genotype and allele frequencies of the studied TPO gene promoter polymorphisms between patients and controls ( p > 0.05). The haplotype distribution (rs2071400-rs2071402-rs2071403) between the two studied groups was similar for the most common variants (CGA, CAG, TGG). Only a rare haplotype (CGG) occurred more frequently among patients compared to controls ( p = 0.04). The studied TPO gene promoter polymorphisms did not show an association with susceptibility to AIT in the Caucasian Polish population, contrary to the results in Japanese patients.

Aim: The association between the interleukin IL1 beta gene polymorphisms SNP-511 and SNP+3953 and susceptibility to the development of Hashimoto's thyroiditis among adult Caucasian-Polish population were analyzed., Patients and Methods: The group studied comprised of 115 unrelated patients with Hashimoto's thyroiditis (112 women and 3 men, mean age 53.3 years). All patients were euthyroid on thyroid replacement therapy, had extremely high serum anti-TPO levels and in 53 patients anti-TG levels were also increased. The control group consisted of 103 healthy blood donors without raised anti-TPO antibodies, in whom a personal and familial history of thyroid, autoimmune and inflammatory diseases was excluded. No goiter or thyroid dysfunction was found.2 polymorphisms of the IL1 beta were studied by PCR-RFLP analysis. To confirm the accuracy of the method used, randomly selected patients were analyzed by direct sequencing., Results: In both groups allele frequencies were in Hardy-Weinberg equilibrium. The significant statistical differences between the frequency of C and T allele for both SNPs (C-511T and C+3953T) in the group studied and in the controls were found (p=0.0081; OR=1.846; 95% CI: 1.183-2.878 and p=0.0099; OR=1.953; 95% CI: 1.183-3.224).The frequencies of the genotype C-511C compared to C-511T and T-511T as well as C+3953C compared to C+3953T and T+3953T also differed significantly (p=0.0057; OR=2.248; 95% CI: 1.292-3.912 and p=0.0043; OR=2.338; 95% CI: 1.305-4.191) between patients and controls., Conclusions: An association between the SNPs of the IL1 beta and susceptibility to Hashimoto's thyroiditis among the group of Caucasian-Polish population studied was found., (© Georg Thieme Verlag KG Stuttgart · New York.)

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.

Systemic lupus erythematosus (SLE) is a complex autoimmune disease of unknown etiology and limited available therapeutic options frustrating both clinicians and patients. However, recent advances in the understanding of disease mechanisms have given rise to numerous studies on specific approaches to SLE treatment. The purpose of this review is to explain the rationale for new treatments and results of the first clinical studies. We will focus on agents which deplete B cells (anti-CD20, anti-CD22), block cytokines (TNF α, Il 6), inhibit B/T cells interaction (CTLA-4Ig, anti-CD40L), or are even expected to reconstruct physiologic immunotolerance. Although preliminary results seemed promising, two randomized clinical trials with rituximab (EXPLORER and LUNAR study) failed to prove efficacy. Data analysis continues to explain the reasons. Trial design, subject population, limitations of the outcome measure instrument and site qualification have been questioned. Future studies are likely to focus on specific organ involvement or treatment combinations with other immunosuppressive agents.

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.

Objective: CD4+ T cells from patients with systemic lupus erythematosus (SLE) display defective function that contributes to abnormal activation of B cells and autoantibody production., Methods: We compared the transcript and protein levels of Fyn and CD70 in CD4+ T cells from patients with SLE (n = 41) and healthy individuals (n = 34). The CD4+ T cells were isolated by positive biomagnetic separation technique. The quantitative analysis of messenger RNA was performed by reverse transcription and real-time quantitative PCR. The protein contents in the CD4+ T cells were determined by Western blotting analysis., Results: We observed significantly higher levels of Fyn (p = 0.03) and CD70 (p = 0.029) transcripts in SLE CD4+ T cells than in controls. There was a significant increase in CD70 protein levels (p < 0.0001), but not Fyn protein levels (p = 0.081) in CD4+ T cells from patients with SLE compared to healthy individuals. In the group with high disease activity [SLE Disease Activity Index (SLEDAI) >/= 9], we observed a significantly higher Fyn protein content than in controls (p = 0.030). There was no correlation between Fyn and CD70 protein levels in SLE CD4+ T cells and disease activity as expressed in the SLEDAI scale., Conclusion: We confirmed previous observations of higher expression of CD70 in CD4+ T cells from patients with SLE. Our findings suggest that increased Fyn protein content in CD4+ T cells can be associated with high SLE disease activity.

Systemic lupus erythematosus (SLE) is a multisystem autoimmune connective tissue disorder characterized by various aberrations including increased production of IL-18. As IL-18 105 A>C polymorphic variants have been linked to increased production of this cytokine, we investigated the prevalence of IL-18 105 A>C (rs549908) polymorphic variants in SLE patients (n = 111) and controls (n = 152). There were no significant differences in the distribution of IL-18 105 A>C polymorphic variants in SLE patients and controls. However, there was a significant association between the IL-18 105 AA genotype (recessive model) and renal manifestations OR = 3.360 (1.523–7.415, P = 0.0039) and the P value remained statistically significant after Bonferroni correction (P corr = 0.0351).Our findings indicate that the IL-18 105 AA genotype variant can contribute to renal manifestations in patients with SLE.

Recently, a family-based association analysis showed that the haplotype carrying a low expression of the variant CD3Z 844 T>A (rs1052231) polymorphism located in the 3'-untranslated region of CD3Z predisposes to systemic lupus erythematosus (SLE) incidence. We analyzed the prevalence of the CD3Z 844 T>A polymorphism in SLE patients (n = 152) and controls (n = 304) in Poland. We observed that women with the CD3Z AA and CD3Z AT genotypes exhibited a 1.845-fold increased risk of SLE [95% confidence intervals (95% CI) = 1.222-2.787, P = 0.0038]. However, we did not find an increased risk for the homozygous CD3Z AA genotype (odds ratio = 1.204, 95% CI = 0.2838-5.108, P = 1.0000). This observation confers that genetic factors causing a decreased level of CD3-zeta in T cells may predispose to SLE incidence.

Objective: The aim of this study was to evaluate whether the IL-1beta gene could be a genetic marker of the thyroid-associated ophthalmopathy (TAO) development., Materials and Methods: The IL-1beta gene polymorphisms at -511 and +3953 regions in 117 TAO patients of Polish origin (ATA/NOSPECS class III or greater) and in 106 controls were studied., Results: We found no significant differences in the frequencies of genotypes and allelic variants for SNP -511 and SNP +3953 between the controls and the studied groups., Conclusions: No association between the IL-1beta polymorphisms and the TAO existed, so those polymorphisms are not suitable genetic markers for TAO.

The protein tyrosine phosphatase non-receptor 22 (PTPN22) 1858 C>T poly-morphic variant gene (rs2476601) displays an association with systemic lupus erythematosus (SLE) and other autoimmune diseases. However, its contribution to SLE has been found to be disputable. We therefore examined the association of PTPN22 1858 C>T polymorphism with susceptibility to SLE in the Polish population, among patients with SLE (n=150) and controls (n=300). We found a contribution of the PTPN22 1858 C>T polymorphism to the incidence of SLE. Women with the PTPN22 TT and PTPN22 CT genotypes displayed a 2.016-fold increased risk of SLE (95% CI=1.324 - 3.070, P=0.0014). However, we did not observe an increased risk for the homozygous PTPN22 TT genotype OR= 2.552 (95% CI=0.6748-9.64, p=0.1675). Our results confirm an association of the 1858 C>T polymorphism of the PTPN22 gene with SLE, which was previously observed in other populations.

Systemic lupus erythematosus (SLE) is a chronic and progressive autoimmune disease in which reactive oxygen species contribute to pathogenesis. We analysed the distribution of manganese superoxide dismutase (MnSOD2) 47C>T (Ala-9Val) functional polymorphic variants within the mitochondrial targeting sequence in SLE patients (n = 102) and controls (n = 199). We did not find significant differences in the distribution of MnSOD2 47C>T polymorphic variants in SLE patients and controls. However, we found that MnSOD2 Val/Val genotype (recessive model) showed a significant association with Raynaud's phenomenon, odds ratio (OR) = 12.000 [95% confidence interval (CI) = 2.315-62.193], p = 0.0015. We also found that the MnSOD2 Val/Val genotype contributes to immunologic manifestations, OR = 2.957 (95% CI = 1.207-7.243), p = 0.0222, and anti-dsDNA antibody presence OR = 3.365 (95% CI = 1.364-8.304), p = 0.0107, in patients. Our observations indicate that MnSOD2 Val/Val variant can be linked to some clinical manifestations in patients with SLE.

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.

Controversies regarding the safety and efficacy of immunization in patients with SLE have persisted for over 60 years. However infections are the main threat in these patients. There are concerns that immunization may cause SLE exacerbation. Evidence from prospective trials suggests that inactivated vaccines are probably safe in patients with stable or inactive disease. Live vaccines are contraindicated in immunocompromised hosts and patients on high dose steroids. Data regarding efficacy of vaccinations are scarce, mainly concerning influenza and pneumoccocal vaccine. Majority of patients have appropriate immunological response, but there are group without serological response after immunization. Risk factors for impaired response are not clear. In same cases it seems reasonable to assess protective antibody titer post vaccination. In this review we would like to present data concerning safety and efficacy of vaccines in patients with SLE to help doctor to decide when vaccination is advisable and when should be avoided.

Objective: Inherited thyroxine-binding globulin (TBG) deficiency is caused by mutations in the TBG gene (locus: Xq22.2), which result in defective synthesis or changes in the physical properties or biological function of a protein., Design: We report a novel mutation of the TBG gene causing a complete TBG deficiency in three brothers of Polish origin. DNA was extracted from all of the family members and subjected to sequence analysis. We analyzed the family with a heterozygous mother, a normal father, their three hemizygous affected sons, and their two normal sons., Main Outcome: Our studies revealed a novel mutation, a single nucleotide deletion (guanine) at position 1711, codon 201 (Asp) in exon 2 (GAC --> AC). This mutation led to a frame shift and premature termination at codon 206, causing a short TBG protein of 205 amino acids (AA) compared to 395 AA of the normal TBG. This new TBG-CD variant was found in the mother and her three affected sons., Conclusion: This is a new variant of TBG-CD (TBG-CD-PL Poland) containing 205 AA.

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.

The etiology of Graves ophthalmopathy (GO), representing the most common extrathyroidal manifestation of Graves disease, is multifactorial. Among multiple genetic, environmental, and endogenous factors, cytokines play a critical role in its etiopathogenesis. We studied an effect of glucocorticoid therapy on the serum IL-6, IL-4, and IL-13 levels in 18 GO patients. All the patients presented euthyroid GO with over 4 points according to the CAS classification (range 4-6; mean 4.94). The patients were treated with methylprednisolone (1 g every second day for three times) followed by 6 months oral prednisone (60 mg/day, with gradual reduction). The clinical examination (Clinical Activity Score and the GO severity by modified NOSPECS classification) and measurement of anti-TPO, anti-TG, anti-TSHR (TRAK), IL-6, IL-4, as well as IL-13 serum levels were performed before, after 2 weeks, and after 6 months of the glucocorticoid therapy. Significant serum IL-6 increases (p < 0.001) and moderate serum IL-4 and IL-13 increases (p < 0.05) were found in GO patients compared with healthy controls. After 2 weeks of the therapy, the serum IL-6 levels decreased in majority of the patients, however after 6-month observation, lower serum IL-6 levels were only in 8 patients who seemed to respond clinically to the therapy (mean value of the Clinical Activity Score decreased from 4.5 before the therapy initiation to 1.25 after 6 months of the glucocorticoid therapy). No changes in IL-4 and IL-13 serum levels during the therapy were observed. Statistical analysis revealed a good correlation between serum IL-6 level and the Clinical Activity Score (p < 0.01). Based on the obtained data, we conclude that IL-6 plays an important role in GO. It seems that IL-6 may serve as a useful factor in the inflammatory events of GO.

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.

Rheumatoid arthritis (RA) is a chronic, inflammatory, autoimmune disease leading to joint destruction. It is the most common cause of potentially treatable disabilities. The outcome of the disease varies from very mild to a refractory, rapidly progressive type with a high mortality rate. In recent years, profound changes in the traditional paradigms of RA therapy have been introduced. Instead of a therapeutically progressive approach, aggressive therapy is recommended for aggressive forms of RA. It has forced us to remodel the traditional treatment pyramid, and to start new strategies such as saw-tooth or step-down-bridge schedules. The last 10 years have seen wide acceptance of immunosuppressive therapy. These agents hold much promise for the further treatment of RA. A few years ago it seemed that we would be unable to influence the long term outcome in RA, but today the development of new drugs and techniques has increased our chances of fighting RA, and prospects for the future are even more promising.

The present study was undertaken in order to evaluate role of HLA DR Bl shared motif in prognosis of development of erosions in rheumatoid arthritis (RA). HLA genotyping was carried out in a retrospective analysis of 73 RA patients and 87 healthy controls using polymerase chain reaction. The assessment of disease activity was performed according to Mallya-Mace Index, whereas radiographs of hands were assessed according to the Larsen Index. HLA-DR4 and DR10 were significantly increased among RA patients. Relative risk was 7.540 and 4.646, respectively. Interestingly, the presence of DRl and DR14 did not enhance the relative risk in our group of patients. Determination of HLA-DR B1 alleles showed that among RA patients the most frequent was HLA-DR Bl&#x002A;0401, &#x002A;0404, and &#x002A;0408. They gave a rise to a relative risk of 4.010, 7.540, and 3.686 respectively. For the purpose of analysis the patients were divided into three groups. The first group comprised 14 patients with two high-risk alleles (HLA DR B1 &#x002A;01, &#x002A;0401, &#x002A;0404, &#x002A;0408, &#x002A;14). The second group gathered 35 patients with one high-risk allele. Twenty-four patients with no high-risk alleles were designated to the third group. We did not notice any differences in damage score and progression of damage score in rheumatoid arthritis patients with different number of high risk motifs. In conclusion, HLA-DR B1 shared motif was found to be significantly more frequent among analyzed erosive rheumatoid arthritis as compared to matched healthy controls. We did not observe any relation between the presence of shared motifs and outcome of the disease. Therefore, it seems that HLA DR B1 determination may very helpful in diagnosing or in establishing groups of risk but it is not likely to have a role in predicting development aggressive forms of RA.

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.

In the present study we investigated the influence of methotrexate (MTX) and azathioprine (AZA) on the serum levels of the IgA-alpha 1-antitrypsin (IgA-AT) complex in patients with the systemic form of juvenile chronic arthritis (JCA). Fifty-six JCA patients (22 treated with MTX, 18 treated with AZA, and 16 not treated with any immunosuppressive agent) were enrolled in the study. MTX dosage ranged from 0.3 to 0.5 mg/kg-1 week-1, while AZA was given daily at an average dose of 1 mg/kg. MTX was given for 13 months (SD = 7 months) whereas AZA for 11 months (SD = 6 months). The average value of the complex was higher in JCA patients than in both control groups (0.74 +/- 0.73 U vs 0.37 +/- 0.13 U (control children), P < 0.001 and vs 0.23 +/- 0.12 U (control adults), P < 0.001). Values exceeding the normal range were found in twenty-two JCA patients (39.4%). Serum IgA-AT level was lowest in the MTX group compared to AZA and non-treated patients (0.56 +/- 0.24 U, 0.76 = 0.43 U, 0.95 +/- 0.52 U, respectively, P < 0.05). IgA values exceeding normal levels for age were found in 14% of the patients. A correlation between the levels of the IgA-AT complex and C-reactive protein (r = 0.43, P < 0.01), alpha 1-acid-glycoprotein (r = 0.45, P < 0.01), alpha 1-antichymotrypsin (r = 0.52, P < 0.01), alpha 1-antitrypsin (r = 0.40, P < 0.01) and IgA (r = 0.56, P < 0.01) was established.

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.

In order to determine the role of memory/naive T cells in atopic allergy patients we analyzed peripheral blood mononuclear cells before and during the grass pollen season. The study comprised 28 patients with seasonal symptoms of atopic allergy and 18 with perennial symptoms. Flow cytometry was employed to detect the expression of CD3, CD4, CD4CD45RA, CD4CD45RO, CD8, CD16, and CD19 molecules on peripheral blood lymphocytes. Allergic patients showed a decreased proportion of memory (CD4+CD45RO+) T cells compared with healthy subject (p < 0.05). The proportion of naive (CD4+CD45RA+) helper T cells did not differ between allergic patients and controls. The percentage of CD4+CD45RO+ cells increased during natural antigen exposure (grass pollen season) in allergic patients with seasonal symptoms. The results show at least two important observations. A potential homing tendency to nasal, bronchial and conjunctival mucosa of memory T cells (CD45RO) in atopic allergy patients may explain their deficiency in peripheral blood. Secondly, the grass pollen season may switch their phenotype from naive into memory T cells causing the increase of CD45RO cells. These events do not occur in non-allergic individuals and may thus constitute new insight into the basic mechanism of atopic allergy.

In the present report, we serially measured the levels of interleukin-6 (IL-6) and some acute-phase proteins (APP) in 61 lung cancer patients undergoing radiotherapy in order to investigate the relationship between the response to the treatment and the changes in parameters of systemic inflammatory response. The patients were divided into two groups depending on the response to the treatment. The first group (referred to as responders) comprised 32 patients with stable disease, partial remission or total remission. Twenty-nine patients with progression of the disease were included to the second group (referred to as non-responders). Six patients died due to the lung cancer during the study. We showed a decrease in IL-6 serum level and C-reactive protein (CRP) level in responders but not in non-responders. However, the most interesting results were obtained after retrospective analysis of the data of six deceased patients. In these patients we observed an elevation of IL-6 and CRP before the patients' deaths. Following the changes in acute-phase response and interleukin-6 serum levels in lung cancer patients seems to be helpful in prognosis of the outcome of the disease. Based on our data, we conclude that an elevation in IL-6 and/or CRP level in patients with lung cancer may serve as an adverse prognostic factor.

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 order to get a better insight into cytokine network regulation in systemic lupus erythematosus (SLE), we analyzed levels of interleukin-10 (IL-10), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), and interferon-gamma (IFN-gamma) in the sera from 36 SLE patients. Moreover, C-reactive protein (CRP), alpha-1-acid-glycoprotein (AGP), and alpha-1-antichymotripsin (ACT) serum levels were evaluated. Serum levels of IL-10 and IL-6 were significantly increased when compared with healthy controls. TNF-alpha and IFN-gamma did not differ from normal values. We established the relationship between IL-10 and IL-6 as well as between IL-10 and TNF-alpha. None of the analyzed cytokines correlated with the acute phase protein levels. Based on the obtained data, we conclude that IL-10 may play the superior regulating role in SLE. A lack of correlation between the cytokines and acute phase proteins suggests their independence from cytokine regulation.

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.

In the present study we investigated the long-term effect of intravenous pulse cyclophosphamide (CY) on lymphocyte surface antigens in systemic lupus erythematosus (SLE) patients. Blood samples derived from 17 lupus erythematosus patients were analysed using two- and three-colour flow cytometry. During the CY therapy, the total number of T lymphocytes (CD3+) was reduced by 31.4%, B lymphocytes (CD19+) by 67.4% and NK cells (CD16+) by 27.4%. Six months after the end of the CY regimen, these values recovered to entry levels. At the onset of the study we observed increased percentages of CD3+ CD25+, CD3+ CD4- CD8-, CD4+ CD29+, CD19+ and CD19+ CD5+ cells. The CY treatment regimen decreased the CD3+ CD25+, CD3+ CD4- CD8-, CD19+ and CD19+ CD5+ cells, but increased the CD3+ CD8+ subpopulation. Taken together, a deficiency of CD8+ T cells associated with CD4+ CD29+ predominance may imply an immune regulatory imbalance leading to abnormal CD4+ cell activation and in consequence to autoimmunity. Depletion of CD19+ cells combined with an enlargement of CD8 cells as a result of CY therapy may reduce the enhanced immune response in SLE patients.

Immunoglobulin A-alpha-1-antitrypsin complex (IgA-AT) is a nonimmune complex formed by disulphide bonding between an active thiol group available on the cysteine residue of alpha heavy chains of IgA and a cysteine in position 232 of alpha l-antitrypsin in single polypeptide chain. The level of the complex can easy be determined using the ELISA method and findings are expressed in arbitrary units. In the healthy adults' sera the IgA-AT complex level is lower than 0.4 arbitrary unit. The elevated levels of the complex were found in a number of rheumatic diseases. In 50% of SLE patients, its levels are increased, particularly in those with current central nervous system involvement. Similarly, in approximately 50% sera derived from RA patients they are also found to be higher. Their presence correlates with anatomical progression of the disease. IGA-AT complex is found in RA (in 90% of cases) but not in the osteoarthritis synovial fluid. Our findings can be applied in clinical praxis in differential diagnosis of early rheumatoid arthritis and osteoarthritis. The IGA-AT complex can be also found in ankylosing spondylitis. The complex has been determined in a relatively large number of IgA myeloma sera. In 30% of the cases its levels were 10-fold higher than the upper limit for healthy adults.

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.