Polymorphism of FCGR2A, FCGR2C, and FCGR3B Genes in the Pathogenesis of Sarcoidosis.

We have previously presented evidence that the polymorphism of the FCGR3A gene, encoding the receptor for Fc fragment of immunoglobulin G IIIa (FcγRIIIa) plays a role in the enhancement of circulating immune complexes (CIs) with the occurrence of Mycobacterium tuberculosis heat shock proteins in patients with sarcoidosis (SA). The immunocomplexemia might be caused by decreased affinity of CIs to Fcγ receptors, with the subsequently decreased receptor clearance by immune cells. In the present study we examined whether the polymorphisms of other related genes (FCGR2A, FCGR2C, FCGR3B) encoding other activatory Fcγ receptors, could have a similar effect. To this end, we genotyped 124 patients with sarcoidosis and 148 healthy volunteers using polymerase chain reaction with sequence-specific primers. We revealed a significant decrease in the percentage of the FCGR2A and FCGR2C variants that ensure effective CIs clearance, with a concomitant increase of less functional variants of these genes in Stages I/II, compared with Stages III/IV of SA. There was no aberration in FCGR3B allele/genotype frequencies. We conclude that the FCGR2A and FCGR2C polymorphisms may also contribute to immunocomplexemia present in SA. The assessment of FCGR genes could become a tool in presaging a clinical course of sarcoidosis and in its personalized therapy.

Peroxynitrite in Sarcoidosis: Relation to Mycobacterium Stationary Phase.

There is evidence that the same mycobacterial heat shock proteins (Mtb-HSPs), especially HSP16, the main marker of mycobacteria dormant stage, occur in sarcoid tissues and in circulated immune complexes and prompt the immune responses against the different genetic background, leading to the development of acute sarcoidosis (SA)/Löfgren syndrome, chronic SA, latent tuberculosis (TB), or active TB. In SA there is increased monocytes phagocytic activity, decreased clearance of antigens/immune complexes by monocytes, which are resistant to apoptosis, and decreased serum microbicidal/degradable nitrate/nitrite (NOx) concentration. Reduction in NOx may result from the reaction of NOx with superoxide with subsequent production of peroxynitrite (ONOO-). In this study, therefore, we evaluated NOx and ONOO- levels in supernatants of peripheral blood mononuclear cells cultures treated with Mtb-HSPs from 20 SA patients, 19 TB patients, and 21 healthy volunteers using Griess and rhodamine fluorescence methods. We found significantly greater NOx and ONOO- concentrations with/without Mtb-HSPs stimulation in SA and TB patients than in controls. However, there were significantly lower NOx and higher ONOO- levels after Mtb-HSPs induction in SA than TB patients. In summary, in contrast to active TB, increased ONOO- concentration may explain the low level of NOx with induction of M. tuberculosis genetic dormancy program via higher Mtb-HSP16 expression in SA.

Pharmacotherapy for sarcoidosis: an example of an off-label procedure.

Sarcoidosis is a granulomatous multiorgan diseases with an unknown etiology, with the predominant lung involvement. Immunosuppressive agents such as corticosteroids, methotrexate, azathioprinum, ciclosporinum A, chlorambucil, cyclophosphamide, hydroxychlorochinum, indomethacin, pentoxyfillinum, thalidomide, leflunomidum, and adalimumab, or infliximab have been used in its treatment. It should be emphasized that the Summary of Products Characteristics (SPC) of these drugs does not specifically recommend their use in the therapy for sarcoidosis. That makes the application of the drugs in sarcoidosis an off-label use, which is not formally accepted by the authorities but is supported by medical bibliography or recommendations given by scientific bodies. Thus the off-label drugs raise legal, but also ethical and medical problems. The dosing regimen and the required duration of therapy for sarcoidosis are missing. In effect the therapy usually follows the recommendations from the American and European Respiratory Societies (ATS/ERS), based on the long-term medical research. The American Food and Drug Administration recognizes the existence of the off-label use. European legislations do not precisely specify the rules for the admissibility of the off-label use. The doctrine of law assumes that the off-label use constitutes a medical experiment. Therefore, the commencement of therapy with such drugs requires patients' informed consent, which must be kept along with other medical records. Insufficient knowledge of the legal regulations may result in civil and professional liability of a physician supervising the therapy of a sarcoidosis patient, especially in case of adverse effects.

Sarcoidosis and tuberculosis: a connection to the human leukocyte antigen system.

Infectious, genetic factors, and autoimmunity have been considered as potential causes of sarcoidosis (SA). Pathological similarities between SA and tuberculosis (TB) suggest M. tuberculosis antigen(s) as causative agent(s). Our published comparative analysis of the human leukocyte antigens (HLA) system in patients with SA or TB in the same ethnic group revealed that some antigens were connected with high risk of developing of SA or TB, but other were comparable in both patient populations. Is it possible that the predominating occurrence of HLA antigens characteristic for TB may cause tuberculosis in patients with SA? To answer this question we evaluated the HLA class I and II alleles frequency by PCR amplification with sequence-specific primers in three women with histopathologically proven pulmonary SA, who developed bacteriologically confirmed TB on a corticosteroids (CS) therapy. Analysis of HLA in every case separately revealed a trend for higher occurrence of both alleles predisposing and protecting from TB than SA, in comparison with healthy individuals in our previously mentioned HLA genotyping study. Overall, the number of alleles predisposing to TB was statistically greater than the number of alleles connected with a high risk of developing SA. Also, the frequency of protecting alleles was statistically higher for TB than for SA. Therefore, SA in these patients developed at first, and the presence of additional environmental factors, e.g., age, CS might decrease an immune response and provoked TB. There is a possibility that the occurrence of HLA antigen more associated with high risk of developing TB than SA causes the development of tuberculosis in our patients with sarcoidosis.

The spontaneous and stimulated nitroblue tetrazolium (NBT) tests in mononuclear cells of patients with tuberculosis.

PURPOSE: The aim of this study was to evaluate the ability of the NBT reduction by non- and BCG-stimulated monocytes isolated from peripheral blood persons with pulmonary tuberculosis (TB) before treatment, after two-month antituberculosis therapy and in an inactive stage of this disease. MATERIAL AND METHODS: The spontaneous and induced NBT tests were done in 24 healthy individuals and 59 patients with pulmonary tuberculosis: 33 before antituberculosis treatment and 26 with inactive stage of TB. Mononuclear cells were isolated from peripheral blood by the Bøyum method and identified by histochemical assay. The abilities of non- and BCG-stimulated monocytes of NBT reduction were estimated by the method according to Park with Szczylik modification. RESULTS: In an active state of TB and after 2 months treatment, the non- and BCG-stimulated monocytes capacity to reduce NBT was found to be significantly increased in comparison to controls. The NBT test parameters in the absence of cell stimulation and after administration of BCG were comparable in active TB and after two-month treatment. In an inactive TB, the ability of NBT reduction by non- and BCG-stimulated monocytes was comparable to the controls. The stimulation of mononuclear cells accompanied by the significantly higher capacity of monocytes to reduce NBT in controls and in TB patients with post-tuberculous changes in the lungs. CONCLUSIONS: These results of the spontaneous and induced NBT tests adequately reflect the status of the host's specific reactivity during tuberculosis and can be a simple, inexpensive and useful method for monitoring antituberculosis treatment.

Analysis of DQB1 allele frequencies in pulmonary tuberculosis: preliminary report.

BACKGROUND: The human leucocyte antigen (HLA) system plays an important role in the modulation of the immune response. An association between HLA and pulmonary tuberculosis (TB) has been examined in several populations but the results have been inconsistent. The aim of this study was to evaluate the correlation of DQB1 alleles with TB patients and healthy controls in the same ethnic group in Poland. METHOD: The DQB1 alleles of 38 patients with TB and 58 healthy university staff volunteers were determined by a PCR-SSP low resolution method. RESULTS: The DQB1*05 allele occurred more frequently (p adjusted for multiple comparison=0.002, OR=2.84, 95% CI 1.57 to 5.15) and the DQB1*02 allele occurred less frequently (p=0.01, OR=0.39, 95% CI 0.21 to 0.71) in patients with TB than in controls. The occurrence of DQB1*03,*04,*06 alleles was similar in the two populations. CONCLUSIONS: The occurrence of specific DQB1 alleles may be linked to susceptibility/resistance to tuberculosis.

[Immunogenic Mycobacterium tuberculosis heat shock protein in tuberculosis]. / Immunogenne bialka szoku termicznego Mycobacterium tuberculosis w gruzlicy.

The heat shock protein (hsp) are produced by prokaryotic and eukaryotic cells in the response to a increase in temperature and a variety of insults. Hsp have been defined by their apparent molecular weight as family: Hsp100, Hsp90, Hsp70, Hsp60, Hsp40 and SHsp (15-25 kD). Analysis of the immune response of M. tuberculosis infected individuals allowed for identification of antigens of tubercle bacilli. Most of them belong to evolutionarily highly conserved hsp, which cross-reactive with the hsp of E. Coli. It is conceivable that in a tuberculous granuloma both pathogenes and host cells are stressed and produce hsp. The heat shock proteins with molecular weight of 71 kD, 65 kD, 10-16 kD are immunogens and a functional role as molecular chaperones. The literature on the immunology of hsp in infectious diseases is complex and confusing and it is still not clear whether host immune responses to these proteins are protective, or damaging, as they may lead to the induction of autoimmunity. The study is engaged in a role of others mycobacterial hsp in host immuno-reactivity and their compliance with treatment of infectious diseases, neoplastic genic therapy or/and with more powerful antimycobacterial vaccine.

The morphological conditions of the permanent pacemaker lead extraction.

Pacemaker lead extraction is the treatment of choice in infectious complications regarding implantation procedure. The purpose of this study was to estimate the safety of the extraction in relation to the morphological changes of the pacing electrode. Research was carried out on materials consisting of 60 human hearts from 45 to 95 years of age (average 63 +/- 15 yrs), with VVI or DDD pacing (pacing duration 84 +/- 26 months) fixed in a formalin solution. Classical macroscopic anatomical methods were applied. In 44 hearts (73.3%) from the investigated group the posterior tricuspid leaflet was thickened only, and in 24 of these hearts the process regarded not only posterior leaflet but also the septal one and especially commissure between them. In 52 hearts (86.6%) inflammatory reaction spread also to the neighbouring part of the electrode. The length of the neointima-inflammatory tissue ranged from 4 to 8 mm (average 5 +/- 2 mm). On the tip of the electrode in the right ventricle cavity in 56 hearts (93.3%) we observed that endocardial leads were surrounded by fibrous thickening, and partially covered by endocardial tissue. We concluded that from the anatomical point of view the extraction of the pacing electrode seems to be questionable, especially in long-term permanent pacing. The experimental traction shows that only recently implanted electrodes were removed without any complications and in others with fraction of the tip, myocardial tissue avulsion or such removal was not successful at all.

HLA-DR antigens in patients with pulmonary tuberculosis in northern Poland. Preliminary report.

The aim of the present study was the analysis of the association between particular class II HLA antigens and the incidence of tuberculosis in northern Poland. HLA-DR antigens in a group of 26 patients with pulmonary tuberculosis (PTB) and 58 healthy volunteers were determined. Histocompatibility typing was performed by the PCR-SSP method using primers from the Dynal company. For statistical analysis, the chi2 test was used with Yates' correction. The probability values were weighted for the number of antigens tested (pc). The relative risk (RR) was calculated by Woolf's method. We found that HLA-DR16(2) antigen expression was significantly higher in patients with tuberculosis than in the tested group of healthy controls (p < 0.001, pc < < 0.01); the highest relative risk (RR = 12.4) of tuberculosis incidence was connected with DR16(2) antigen, the prevalence of HLA-DR13(6) antigen was significantly lower in pulmonary tuberculosis (with RR = 0.09) than the control (p < 0.001, pc < 0.01). The results obtained suggest that the presence of HLA-DR16(2) antigen can extend the risk of developing tuberculosis whereas HLA-DR13(6) antigen occurrence was significantly more rare in pulmonary tuberculosis than in healthy individuals and that the relative risk (RR = 0.09) can be connected to their relation with the genes of insusceptibility to tuberculosis.

HLA-A, B, C antigens in pulmonary sarcoidosis in Polish population.

The aim of this study was to analyze association between HLA class I antigens and sarcoidosis in Poland. HLA-A, B, C antigens in a group of 100 patients suffering from sarcoidosis and in a group of 100 healthy blood donors were determined. Histocompatibility typing was performed by the NIH method using commercially available sera. For statistical analysis chi2 test was used after Yates' correction. The relative risk was calculated by Woolf's method. We found that HLA-B8 and -Cw7 prevalence was significantly higher in patients with sarcoidosis than in healthy controls. HLA-B35, -B40, -Cw2 and -Cw4 antigen expression was significantly lower in pulmonary sarcoidosis than in the tested group of healthy individuals. The highest relative risk of sarcoidosis was connected with HLA-B8 and -Cw7. The results obtained suggest that, in the population suffering from pulmonary sarcoidosis in nothern Poland, as compared with the control group of healthy persons, antigens HLA-B8 and -Cw7 are significantly more frequent. It can be assumed that, the presence of these antigens may be connected with a greater risk of pulmonary sarcoidosis. In the group of patients, as compared with the control population, the occurrence of antigens HLA-B35, -B40, -Cw2 and -Cw4 is significantly more rare.