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The functional fluorescent dye efflux assays are used in the study of the multidrug resistance of the malignant cells to the cytotoxic drugs which is caused by the overexpression of P-glycoprotein (P-gp) or another membrane transport system--a multidrug resistance related protein (MRP). P-glycoprotein and a multidrug resistance related protein are involved in the efflux of cytotoxic drugs out of the cell and are responsible for the resistance. Fluorescent dye efflux mediated by these proteins could be evaluated by the flow cytometry. This test seems to be an optimal approach to study the multidrug resistance. With help of the specific inhibitors such as verapamil and cyclosporine A, the functional capacity of these proteins and the possibility to overcome the multiresistant phenotype can be revealed. The cell line K562 with transfected P-glycoprotein gene serves as a model system for the studying of the transport function with the use of the fluorescent substrates and P-glycoprotein inhibitors.

MALT lymphoma is raised from lymphoid cells of mucosa associated lymphoid tissues in a long-term, multistep process of the accumulation of genomic damage very often under the influence of chronic (e.g. Helicobacter pylori) infection. Cytoimmunofluorometric analysis of cell suspensions obtained from mechanically disintegrated bioptic samples of gut mucosa is a very useful approach to detect the presence of MALT lymphoma cells. MALT lymphoma cells are in a great majority of B cell origin. Malignant cells are characterized by the typical characteristic SS parameter, strong expression of CD20 and CD79b molecules, and by the absence of CD5 and CD23 molecules. The ultimate evidence of clonality is immunoglobulin light chains either kappa or lambda expression determination.

We report a case of a 60-year-old man with acute myocardial infarction (AMI) treated using thrombolysis, and complicated by cardiogenic shock (CS). Plasma interleukin (IL) 1 beta, IL 6, IL 8, tumor necrosis factor alpha (TNF alpha), and soluble adhesion molecule (sICAM 1, sE-selectin) levels were measured at 3-h intervals. This observation showed the effect of AMI and CS on the plasma interleukin levels. Remarkable changes were found in the plasma TNF alpha level, which reached two significant peaks. The peak of the first elevation caused by AMI (80.11 pg/ml, vs. normal value 4.35 pg/ml, SD 21.3 pg/ml) was seen 6 h after the onset of the symptoms. After the period of significant decrease, TNF alpha level was increasing until the end of the observation period because of CS (the last TNF alpha level was 204.1 pg/ml). The plasma IL 1 beta level was continually increased during the period of observation (maximal IL 1 beta level 32.1 pg/ml, normal value < 10 pg/ml). The plasma IL 6 level reached the first peak caused by AMI nine hours after the onset of the symptoms (362.85 pg/ml, normal value (10 pg/ml). Because of CS, after the short period of decrease, the plasma IL 6 level was increasing until the end of the observation period (the last IL 6 level was 859.61 pg/ml). The plasma IL 8 level was also elevated throughout the time of observation (max. value 1652 pg/ml, vs. normal value < 30 pg/ml). The soluble adhesion molecule levels (sE-selectin and sICAM 1) were elevated throughout the period of observation without any significant peak.

Resistance to cytotoxic drugs is a serious drawback in the treatment of patient with tumours, both of the haemopoietic and non-haemopoietic origin. The cytotoxic effect of drugs on the malignant cells manifests as the process of apoptosis. In the resistant malignant cells, apoptosis becomes prevented by several mechanisms. The multidrug resistance (MDR) is one of the principal mechanisms when the cancer cells develop resistance to multiple chemically and functionally unrelated cytotoxic compounds. The decrease of the cytotoxic drug concentration at the molecular target site may come from activation of some efflux membrane systems participating in the transport of cytotoxic drugs out of the cell (e.g. Pgp, MDR, and LRP). Another mechanism of resistance is the increased enzymatic detoxification of the drug by glutathion-S-transferase system. Changes in the molecular target of the cytotoxic drug such as topoisomerase molecule can be also responsible for the resistance. At least two additional mechanisms of resistance of tumour cells were identified. Resistance can come from either deregulation of proapoptotic mechanisms in tumour cells or by increased activity of reparation processes which control the damaged molecule of DNA. Several methods that detect the cause of resistance in distinct cell populations have been developed. The great effort is now focused on both the detection of mechanisms of the resistance and on the clinical procedures of overcoming the resistance to cytotoxic drugs.

Resistance to cytotoxic drugs is a serious drawback in the treatment of patients with tumours, both of the haemopoietic and non-haemopoietic origin. The cytotoxic effect of drugs on the malignant cells manifests as the process of apoptosis. In the resistant malignant cells, apoptosis becomes prevented by several mechanisms. The multidrug resistance (MDR) is one of the principal mechanisms when the cancer cells develop resistance to multiple chemically and functionally unrelated cytotoxic compounds. The decrease of the cytotoxic drug concentration at the molecular target site may come from activation of some efflux membrane systems participating in the transport of cytotoxic drugs out of the cell (e.g. Pgp, MDR, and LRP). Another mechanism of resistance is the increased enzymatic detoxification of the drug by glutathion-s-transferase system. Changes in the molecular target of the cytotoxic drug such as topoisomerase molecule can be also responsible for the resistance. At least two additional mechanisms of resistance of tumour cells were identified. Resistance can come from either deregulation of proapoptotic mechanisms in tumour cells or by increased activity of reparation processes which control the damaged molecule of DNA. Several methods that detect the cause of resistance in distinct cell populations have been developed. The great effort is now focused on both the detection of mechanisms of the resistance and on the clinical procedures of overcoming the resistance to cytotoxic drugs.

Resistance to cytotoxic drugs is a serious drawback in the treatment of patients with tumours, both of the haemopoietic and non-haemopoietic origin. The cytotoxic effect of drugs on the malignant cells manifests as the process of apoptosis. In the resistant malignant cells, apoptosis becomes prevented by several mechanisms. The multidrug resistance (MDR) is one of the principal mechanisms when the cancer cells develop resistance to multiple chemically and functionally unrelated cytotoxic compounds. The decrease of the cytotoxic drug concentration at the molecular target site may come from activation of some efflux membrane systems participating in the transport of cytotoxic drugs out of the cell (e.g. Pgp, MDR, and LRP). Another mechanism of resistance is the increased enzymatic detoxification of the drug by glutathion-S-transferase system. Changes in the molecular target of the cytotoxic drug such as topoisomerase molecule can be also responsible for the resistance. At least two additional mechanisms of resistance of tumour cells were identified. Resistance can come from either deregulation of proapoptic mechanisms in tumour cells or by increased activity of reparation processes which control the damaged molecule of DNA. Several methods that detect the cause of resistance in distinct cell populations have been developed. The great effort is now focused on both the detection of mechanisms of the resistance and on the clinical procedures of overcoming the resistance to cytotoxic drugs.

The diversity and heterogeneity of biologic reactions is a condition sine qua non for the adaptation of live organisms to changes of external environment. Reactions of the immune system enable survival even in extreme conditions. Modern medicine often reaches situations never met by the organism in phylogenesis. Thanks to up-to-date therapeutical approaches many patients survive with grave symptoms such as multiple organ failure and extensive forms of injury. New infectious diseases are emerging, e.g. HIV/AIDS. Notwithstanding the broad array of external insults, it becomes obvious that organisms mobilize defence reactions according to a general scheme. Likewise, exhaustion of the immune potential occurs according to an archetypal pattern. In the case of cell-mediated immunity the faultless course of which is of critical importance for organisms in extreme conditions, the loss begins with the absence of reactivity of T-cells to specific antigens to end with the absence of their reactivity to polyclonal mitogens. The article deals with pathogenic mechanisms underlying the onset of immunodeficiencies caused by both infectious and non-infectious stimuli with special regard being focused on cardiac surgery in extracorporeal circuit.

Background: Detection of membrane molecules by immunophenotypisation is a routine counterpart of the laboratory tests which are needed for the complex diagnostic procedures of hematopoetic malignancies at present. The immunochemical detection of intracellular molecules is essential in such cases, where the expression of surface molecules on malignant cells is poor, or where the results of other diagnostic analyses are unequivocal (cases with ectopic expression of other lineage-specific molecules and cases of biphenotypic leukemias)., Methods and Results: We followed the procedure using 4% solution of paraformaldehyd in phosphate buffered saline for fixation of cells. Lysing solution G (Becton-Dickinson) was used for permeabilisation of cells to detect nuclear and cytoplasmatic molecules. The permeabilised cell suspension was than washed with 0.5% solution of Tween 20 in phosphate buffered saline. This simple and rapid procedure is readily available for the routine detection of intracellular molecules. This approach is important for the diagnosis of hematopoetic malignancies.

The aim of this study was to map normal levels of serum soluble isoforms of adhesion molecules in relation to age and sex in the group of school-aged children. sICAM-1, sVCAM-1 and sE-selectin were determined in the group of 158 normal children subdivided into two subgroups; 6-10 years (68 children, median age 8 years) and 11-15 years (90 children, median age 12 years) and in 70 normal adult blood donors (25 females and 45 males, median age 46 years). The levels of sICAM-1 and sE-selectin fell down significantly over the age range 6-15 years, while the level of sVCAM-1 was remained. Age-related normal ranges were established using correlation analysis and were expressed as the 5%-95% percentiles intervals: sICAM-1 206.8-486.8 ng/ml, sE-selectin 36.7-153.2 ng/ml in the group of 6-10 years old children, sICAM-1 184.1-354.0 ng/ml, sE-selectin 29.9-114.1 ng/ml in group of 11-15 years old children. The levels of sVCAM-1 were 359.6-822.0 ng/ml and were constant within the examined age interval from 6 to 15 years. The influence of sex was also assayed and it was not statistically significant in any age category tested. Normal ranges of sICAM-1 (60.2-218.4 ng/ml), sE-selectin (8.3-116.9 ng/ml) and sVCAM-1 (338.0-1148.0 ng/ml) were established for adult population of healthy blood donors using the same methods.

The advent of new therapeutic approaches to multiple myeloma made necessary the introduction of novel methods for detection of minimal residual disease. Among others approaches residual disease can be detected by the immunofluorescence using flow cytometry. We have examined the co-expression of CD19, CD38, CD45, CD54, CD56, and CD138 molecules in cells of peripheral blood and bone marrow aspirates in patients with multiple myeloma by 3-color flow cytometry. For the detection and characterization of multiple myeloma cells, combinations of following antibodies were used: anti-CD19 FITC, anti-CD38 FITC, anti-CD38 PE, anti-CD54 FITC, anti-CD56 PE-Cy5, anti-CD45 PE, anti-CD45 PE-Cy5 (Immunotech) and anti CD138 PE (Serotec). The samples were analyzed using EPICS XL (Coulter) flow cytometer, and the analysis was based on at least 10,000 events. Samples from 17 patients were analyzed. The percentage of multiple myeloma cells ranged between 0.3% and 54.2% in bone marrow aspirates and between 0.0 and 11.8% in periferal blood. The expression of CD138, CD38, CD54 and CD56 molecules was found in 100%, 100%, 85% and 68% of examined cases, respectively. In our opinion, multiple myeloma cells are best characterized by following combinations of antibodies: CD38 FITC/CD138 PE/CD45 PE-Cy5, CD54 FITC/CD138 PE/CD56 PE-Cy5 or CD54 FITC/CD38 PE/CD 56 PE-Cy5. The identification of a malignant clone is the first and the most important step in the characterization of the disease, determination of its prognosis and the detection of residual disease after treatment. Three-color flow cytometry represents a method which can meet these goals.

The plasma levels of interleukin 1 beta (IL 1beta), interleukin 6 (IL 6), interleukin 8 (IL 8), tumor necrosis factor alpha (TNF-alpha), E-selectin, ICAM 1 and C-reactive protein (CRP) have been studied in 24 patients with acute myocardial infarction in the course of 96 h. The plasma IL 1beta and IL 6 levels were continually elevated during the 96 h study period (the peak of plasma IL 1beta level was 22.2 pg/ml, S.D. 8.6, P < 0.001, normal values of IL 1beta are less than 10 pg/ml, the mean peak plasma concentration of IL 6 was 184.9 pg/ml, S.D. 134.7, vs. normal values of 15.57 pg/ml, S.D. 2.4, P < 0.001). The mean plasma IL 8 level was increased for the duration of the study, the mean plasma IL 8 level was 103.0 pg/ml, S.D. 23.4 (normal value was below 30 pg/l, S.D. 8.0) P < 0.001. The plasma TNF-alpha level was elevated throughout the time of observation without any significant peak. The mean plasma TNF-alpha concentration was 46.8 pg/ml, S.D. 2.13, vs. normal value 4.35 pg/ml, S.D. 1.23, P < 0.001. The plasma E-selectin level reached the mean level of 145.1 ng/ml, S.D. 75.4, vs. normal value 29.1-63.4 ng/ml, P < 0.001 at an interval of 15-42 h after the onset of the symptoms. The plasma ICAM 1 level showed only a slight significant increase during the first 36 h. The plasma CRP concentration increased later than IL 6, and reached a peak at 42 h after the onset of the symptoms (69.2 mg/l, S.D. 29.9, vs. 1.2 mg/l, S.D. 4.7, P < 0.0001). We conclude that cytokines and adhesion molecules can play an important role in the mechanisms of tissue injury in the process of ischemia and reperfusion.

Sjögren's syndrome is a systemic autoimmune disease characterized by focal lymphocytic infiltration of the salivary and lacrimal glands. Expression and up-regulation of adhesion molecules and activation of cellular immune system is essential for the migration of inflammatory cells into tissues. Soluble forms of adhesion molecules sICAM-1, sVCAM-1, sE-selectin and neopterin were analyzed in serum of 17 patients with primary Sjögren's syndrome and 11 patients with secondary Sjögren's syndrome together with 26 age-matched healthy blood donors. There were significantly higher serum concentrations (mean +/- 1SD) of sICAM-1 (362.0 +/- 67.9 ng/ml, p < 0.001), sE-selectin (78.7 +/- 28.1 ng/ml, p < 0.001) and neopterin (17.9 +/- 6.4 nmol/l, p < 0.001) in primary Sjögren's syndrome patients in comparison to control group (sICAM-1: 128.3 +/- 46.9 ng/ml, sE-selectin: 46.3 +/- 39.5 ng/ml, and neopterin: 7.6 +/- 2.3 nmol/l). Sera from patients with secondary Sjögren's disease contained significantly higher levels of sICAM-1 (356.0 +/- 62.4 ng/ml, p < 0.001), sE-selectin (65.5 +/- 27.0 ng/ml, p < 0.05), and neopterin (18.8 +/- 9.8 nmol/l, p < 0.001) in comparison with control group. There were no significant differences between patients with primary and secondary Sjögren's syndrome in any parameters tested. No statistically significant differences in serum levels of sVCAM-1 were found either in patients with primary or secondary SS compared to control group.

Background: CagA antigen of Helicobacter pylori is highly immunogenic in humans. There is an increasing evidence that infection with CagA-positive strains is related to the development of peptic ulcer disease, atrophic gastritis, or gastric cancer. The aim of our study was to assess seropositivity to CagA in a group of 95 clinically symptomatic adults who underwent gastroduodenoscopy and to correlate results to their disease characteristics., Methods and Results: Serum immunoglobulin G antibodies to CagA detected by ELISA kit (Helicobacter p120, Viva Diagnostika, Germany) were compared to standard IgG specific antibodies against a pool of H. pylori antigens Synelisa Pin plate, ELIAS, Germany). Immunoglobulin G antibodies to CagA were present in 5/31 (16%) serum samples from H. pylori negative persons and 10/28 (36%) serum samples from H. pylori positive patients without peptic ulcer disease compared with 8/11 (73%) H. pylori positive patients with peptic ulcer disease in the past, 11/13 (85%) H. pylori positive patients with duodenal ulcers or duodenitis and 4/5 (80%) H. pylori positive (1/7, 14% H. pylori negative) serum samples from patients with gastric resection for peptic ulcers in the past. Serum levels of antibodies to CagA in the groups of patients with peptic ulcer disease in the past, with present duodenal ulcers of duodenitis and in H. pylori infected patients with gastric resection were significantly higher then those of H. pylori infected patients without peptic ulcer disease (P < 0.05). On the other hand, there was no significant difference in the presence of the specific antibodies against at pool of H. pylori antigens between these four groups., Conclusions: These data suggest that serologic response to the CagA antigen is more prevalent in H. pylori positive persons with present or past peptic ulceration than among infected persons without peptic ulcer disease. The presence of antibodies to CagA in H. pylori positive persons may be useful for the identification of patients with higher risk or more severe disease.

Many cell populations are thought to be involved in the etiopathogenesis of bronchial asthma. We examined by flow cytometry the relative and absolute number of CD3+, CD4+, CD8+, alpha beta TcR+ or gamma delta TcR+ T cells, CD19+ B cells; and CD56+ natural killer (NK) cells in the peripheral blood of 26 adult patients with difficult-to-control asthma (DCA) and 22 patients with minimally symptomatic asthma (MSA). Statistically higher relative and absolute numbers of NK cells (18.39 +/- 10.67% and 0.38 +/- 0.17 x 10(9)/l) in comparison with healthy controls (11.77 +/- 8.06% and 0.25 +/- 0.19 x 10(9)/l) and significantly decreased relative and absolute numbers of gamma delta T cells (3.02 +/- 2.16% and 0.06 +/- 0.04 x 10(9)/l) in comparison with controls (5.65 +/- 2.90% and 0.13 +/- 0.08 x 10(9)/l) in the DCA patient group were found. After pooling of data from both MSA and DCA patients and dividing the patients according to the presence of allergy, the relative and absolute numbers of gamma delta T cells were found to be diminished in both the allergy (3.77 +/- 2.98 and 0.07 +/- 0.05 x 10(9)/l) and nonallergy (3.06 +/- 1.78% and 0.06 +/- 0.03 x 10(9)/l) groups in comparison with healthy controls. The reason for the low number of gamma delta T cells in the peripheral blood of patients suffering from bronchial asthma is under investigation.

Interactions between endothelial cells, adherence of endothelial cells to the molecules of extracellular matrix, adhesion of leukocytes and platelets to the activated endothelial cell lining are mediated by adhesion molecules belonging into four families: immunoglobulins, integrins, selectins, and cadherins. Adhesion interactions mediate intercellular signaling. Soluble forms of adhesion molecules modulate adhesion interactions. The determination of soluble adhesion molecules in blood is used for indirect evaluation of function or dysfunction of endothelial cells. It is possible to modulate abnormal function of endothelial cells using monoclonal antibodies against adhesion molecules.

Homeostasis is mediated through interactions of adhesion molecules on the surface of platelets which belong into the families of selectins, integrins and immunoglobulins with soluble plasma proteins, surface molecules of activated endothelial cells and molecules of extracellular matrix. The membrane expression of some adhesion molecules (P-selectin) which are the part of granular system of resting platelet identify activated platelet. Adhesion molecules of platelets could be detected by immunochemistry through flow cytometry. By this approach the inherited defects of adhesion molecules on platelets and the activated platelet could be detected. The detection of activated platelets is very useful in many clinical conditions.

Sjögren's syndrome is a chronic inflammatory disease which is believed to be immunologically mediated. It is marked by inflammation and destruction of the salivary glands and lacrimal glands. We have found significantly higher serum levels of sICAM-1 and beta-2-microglobulin in both primary Sjögren's syndrome patients (429.1 +/- 114.33 ng/ml, 4.4 +/- 1.5 ug/ml resp.) and secondary Sjögren's syndrome patients (424 +/- 140.73 ng/ml, 4.4 +/- 2.67 ug/ml resp.) in comparison with healthy controls (189.1 +/- 42.34 ng/ml, 1.3 +/- 0.38 ug/ml resp.). The serum level of beta-2-microglobulin was significantly correlated with the serum level of sICAM-1 in secondary Sjögren's syndrome patients' group.

Pathogenicity of Helicobacter pylori is based partially on the production of toxic substances. On the other hand the gastric mucosal injury may be caused by the Helicobacter pylori-induced host inflammatory response. Considerable antibody response during Helicobacter pylori infection provides a ground for indirect non-invasive serological detection. Appropriate clinical applications of serological tests are based on a good understanding of the antibody response characteristics during Helicobacter pylori infection.

The paper deals with the types of antigens for detection of anti-Helicobacter pylori antibodies. There are informations about the protein composition of Helicobacter pylori and about the specificity and immunogenicity of particular proteins. The authors review and evaluate different types of antigenic preparations used in serological tests and inform about the commercial kits for serological detection of Helicobacter pylori infection.

Background: Sjögren's syndrome (SS) is considered an autoimmune immunopathological disease, which is characterized by disorders of humoral and cell-mediated immunity. The objective of the work was to assess the ratio of T and B lymphocytes, subpopulations of T lymphocytes and NK cells in the peripheral blood of patients suffering from SS., Methods and Results: The investigated group was formed by 22 patients with the diagnosis of primary SS (mean age 55.7 years, range 39-83 years), 17 patients with the diagnosis of secondary SS (mean age 60.5 years, range 45-79 years). The ration of CD3-, CD4-, CD8-, CD19-, CD5-, CD25- HLA DR- cells in full blood was assessed by the monoclonal antibody technique of direct immunofluorescence, using evaluation by means of a flow cytometer. The results were compared by statistical methods with a control group of blood donors using Student's t-test, A relative increase of the ratio of CD3- T lymphocytes was revealed in patients with primary SS (75.3 +/- 17.9 %, p = 0.380) and secondary SS (73.7 +/- 11.8, p = 0.326), as compared with controls (65.3 +/- 10.3%). The absolute number of the subpopulation of suppressor helper inducer CD4+ T lymphocytes was markedly lower in patients with primary SS (0.056 +/- 0.27 - 10(9)/l, p = 0.0154), as compared with controls (0.81 +/- 0.27, 10(9)/l. The relative number of suppressor cytotoxic CD8- T lymphocytes was significantly higher in patients with primary SS (33.7 +/- 15.0%, p = 0.0001 and secondary SS (30.5 +/- 11.0%, p = 0.0002), as compared with controls (17.2 +/- 6.0%). The absolute number of inhibitory cytotoxic T lymphocytes was significantly higher in patients with primary SS (0.47 +/- 0.3. 10(9)/1, p = 0.0084) and patients with secondary SS (0.48 +/- 0.3, 10(9)/l, p = 0.0072). The relative number of CD57+ NK cells was significantly higher in patients with primary SS (21.1 +/- 11.0%, p = 0.0001) and secondary SS (19.3 +/- 11.7%, p = 0.0023). The absolute number of NK cells was significantly higher in patients with primary SS (0.28 +/- 0.20, 10(9)/l, p = 0.0025 and in patients with secondary SS (0.31 +/- 0.23, 10(9)/l, p = 0.01)., Conclusions: The revealed statistically significant increases in the number of T lymphocytes in patients with Sjögren's syndrome is probably caused by a significant increase of the subpopulation of CD8+ suppressor cytotoxic T lymphocytes. The latter probably overlap with the population of NK cells. These immunological abnormalities are even more marked in patients with primary Sjögren's syndrome due to the decrease of the absolute number of CD4+ helper inducer T lymphocytes.

Unlabelled: The main aim of the present study was a search for a characteristic serum marker of inflammatory activity in the airways of asthmatics with difficult-to-control disease. Therefore, serum levels of interleukin-4 (IL-4), serum low-affinity Fc Epsilon Receptor II (sFcER II), Interferon-gamma (INF-gamma) Immunoglobulin-E (IgE), Interleukin-2 (IL-2), serum Interleukin Receptor 2 (sIL-2R) and Intercellular Adhesion Molecule-1 (sICAM-1) were measured in 2 groups of asthmatics: 1-26 patients with difficult-to-control asthma (DTCA), 2-22 asthmatics, minimally symptomatic (MSA)., Results: No significant difference in either measured parameters between the DTCA and MSA group in peripheral blood samples was found., Conclusion: The above mentioned serum markers of T- and B-cell activation as well as the serum ICAM-1 level are not sensitive enough to determine the type, activity and severity of the inflammatory process in the asthmatic airways.

Aims and Background: Proliferative response of lymphocytes to mitogens is known to be decreased in cancer patients. Depressed proliferation of lymphocytes in other disorders has been linked to systemic inflammatory response., Methods: Lymphocyte proliferation in response to phytohemagglutinin (PHA) and concanavalin A, as well as urinary neopterin, plasma interferon-gamma and interleukin-4, were investigated in 41 patients with solid tumors., Results: The proliferative response to PHA was significantly depressed in cancer patients at most of the concentrations examined. Urinary neopterin and plasma interferon-gamma were significantly elevated in cancer patients, whereas interleukin-4 was undetectable in all the subjects. A negative correlation was observed between urine neopterin and lymphocyte proliferative response to mitogens., Conclusions: Lymphocyte proliferative response to PHA is significantly diminished in cancer patients, and this depression appears to be partly linked to systemic inflammatory response. Plasma interferon-gamma was significantly increased in cancer patients, whereas interleukin-4 was undetectable.

The authors made immunophenotyping analyses of blood cells of 50 patients with the diagnosis of B-CLL. Malignant cells contained in all instances HLA DR and CD24 molecules. The superficial molecule CD5 was expressed on cells in 85.7% molecule CD6 on 66.7% cases. By statistical analysis the relationship between selected clinical parameters and expression of surface molecules CD5, CD6 and CD10 was sought. A statistically significantly higher number of leucocytes was found in patients with expression of molecule CD6.

Severe acute response, the synthesis of human acute-phase proteins and the increase of plasma cytokines and adhesion molecules occur in patients in the course of acute myocardial infarction. We examined the plasma tumor necrosis factor alpha (TNF alpha), plasma creatinkinase (CK) and C-reactive protein (C-RP) levels in patients with acute myocardial infarction (AMI) in the course of 96 hours. Venous blood samples were taken at 3-hour intervals during the first 48 hours, and at 6-hour intervals during the next 48 hours. All patients were treated using thrombolytic therapy (streptokinase). Detection of the reperfusion was based on the method of measuring the time to achieve peak serum creatinkinase activity. The study was done on a group of 24 patients. Plasma levels of the parameters were compared between the group of patients with expected reperfusion versus the group of patients in which reperfusion is not suggested. The plasma TNF alpha level was elevated constantly without any significant peak. The mean plasma TNF alpha concentration was 46.8 pg/ml, SD 2.13, vs. normal level 4.35 pg/ml, p < 0.001. The plasma TNF alpha level in the group of patients with reperfused coronary artery showed a significant decrease especially during the 3rd and 4th day (the mean peak plasma TNF alpha concentration was 35.2 pg/ml, SD 15.8, vs. 66.9 pg/ml, SD 38.3 pg/ml, p < 0.005). The plasma C-RP levels were elevated throughout the time of observation in the both groups. The elevation of the plasma C-RP levels was more significant in the group of patients without successful reperfusion (80.6 mg/ml, SD 31.2, the mean plasma C-RP level of the group of the patients with successful reperfusion was 45.7 mg/ml, SD 18.1, p < 0.005). We conclude, that TNF alpha can play a role in the mechanisms of tissue injury. The successful reperfusion of coronary artery leads to significant decrease of plasma TNF alpha and C-RP levels.

Contemporary therapeutic procedures used in the treatment of malignant diseases are based on the no-selective action of cytotoxic or cytostatic substances. The sensitivity of tumour cells to cytostatics can be, however, reduced by the action of several mechanisms among which expression of the membrane glycoprotein P-170 is the most important. The latter actively transfers the cytotoxic substance from the cell and is responsible for multiple drug resistance-MDR). Glycoprotein P-170 can be assessed on leukaemic cells immunochemically by flow cytometry. A very perspective procedure is assessment of the in vitro sensitivity of leukaemic cells to clinically used cytostatics.

Using immunocytochemical phenotyping, the authors assessed the origin of malignant cells in a patient with malignant centroblastic lymphoma. The lymphoma cells expressed the thymocytic surface molecule CD1, pan T-lymphocytic markers CD2, CD5, CD6, CD7 with concurrent absence of complex CD3 and receptors for antigen on T lymphocytes. The proliferating activity of malignant cells could not be controlled by intensive cytostatic therapy.

The level of interleukin-6 in the serum of 17 patients suffering from the primary Sjögren's syndrome and of 16 with secondary Sjögren's syndrome were determined by means of ELISA. The mean value of IL-6 in primary and secondary Sjögren's syndrome was 2.41 pg/ml (1.45-3.37 pg/ml) and 3.01 pg/ml (1.63-4.39 pg/ml). The normal serum level of IL-6 was below 3.0 pg/ml. The serum level of IL-6 was not elevated in Sjögren's syndrome patient, there was no significant difference in the serum level of IL-6 between both groups of the patients.

The total white blood cell count, percentage and total count of lymphocytes, CD3+, CD4+, and CD8+ subpopulations of T-lymphocytes, population of CD20+ B-lymphocytes, the ability of polymorphonuclears to phagocyte yeasts, the ability of polymorphonuclear cells to kill serum-resistant strain of E. coli and O2-dependent killing capacity of polymorphonuclears were evaluated in a group of 10 patients with the sicca syndrome and in a group of 9 patients with secondary Sjögren's syndrome and in a healthy control group. There was found a significant decrease in the relative and absolute counts of CD4+ helper/inducer T cells and in the activity of phygocytic cells in both groups of patients with Sjögren's syndrome as compared with the control group. There was no significant difference in the evaluated parameters between primary and secondary Sjögren's syndrome patients.

The authors present the results of immunophenotyping analysis of cells from 67 patients with the diagnosis of B-CLL from group of 393 adults examined by immunophenotyping because of a haematological malignity. Characteristic for B-CLL is the finding of surface molecules HLA DR, B lymphocytic superficial molecules and CD5. Aberrant membrane equipment of B-CLL cells is suggested by the rare absence of some pan B lymphocytic sign. Marked reduction of T lymphocyte ratios, their sub-populations and NK cells is associated with immunodeficiency manifestations which are common in patients with B-CLL.

The authors describe a case of myeloid leukemia of the myelomonocytic type (M4, FAB 1976) where blastic cells bear the lymphoblastic cALL sign. These cases of the so-called hybrid phenotype of acute leukemia are rare. The importance of the hybrid phenotype for the course and prognosis of the disease is not known.

Crohn's disease is a disease with unknown etiology of which immunological mechanisms participate. As standard treatment is sometimes ineffective, in 15 patients therapeutic plasmapheresis was used. Before plasmapheresis and immediately after it some immunological parameters were assessed. In the serum a significant drop of all immunoglobulins of the basic classes was recorded: (P less than 0.05) and of circulating immunocomplexes (P less than 0.02). Marked clinical improvement was observed in three patients, which as compared with data in the literature, is somewhat less. The cause is in the author's opinion in the selection of patients (plasmapheresis is usually most successful in patients with marked activity of the disease and non-intestinal symptoms) even in case of less active plasma exchange. Despite this plasmapheresis can be recommended in patients with a poor response to common conservative treatment and in some of them we may expect a marked therapeutic effect with a favourable influence on the prognosis of the disease.

The leukocyte migration inhibition test was performed in 39 patients with Helicobacter pylori infection and in 38 patients without such infection. The culture of Helicobacter pylori was used as antigen. A highly significant inhibitory effect on leukocyte migration was found in patients with Helicobacter pylori infection. The results can be taken as proof of a systemic immune response to helicobacters at the cellular level in patients with Helicobacter pylori infection.

The prerequisite of effective infection with the virus of human immunodeficiency (HIV) are the interaction between the CD4 complex of the susceptible cell and outer glycoproteins of the virus. The CD4 complex is found above all on the sub-population of helper inducer T lymphocytes and cells of the monocyto-macrophage line. These interactions are essential for the penetration of the virus into the cell and participate also in the fusion of infected and non-infected cells which is typical for HIV infection. The outer layer glycoproteins of HIV are immunogenic, however, the antibodies which are formed do not neutralize the virus. It is even probable that antibodies can under certain conditions facilitate the penetration of HIV into cells which have on their surface receptors for the Fc fragment of immunoglobulin, i.e. in the first place into cells of the monocyte-macrophage line. These cells play obviously a key role in HIV infection because they are resistant to the cytopathic action of the virus and during their numerous physiological functions serve above all as cells, presenting antigen to T lymphocytes and thus are repeatedly in close contact with other cellular elements of the immune system and may thus help to spread HIV in the organism.

The authors evaluate clinical experience based on a correlation between the clinical condition and the finding of Campylobacter pylori (CP) in a total of 1027 specimens of the gastric mucosa. They confirmed the high incidence of CP in patients with a fresh duodenal ulcer (g 7%) but also in the inactive stage of duodenal ulcer disease (97.8%) and in patients with a pyloric ulcer (94.4%). An analogous incidence of CP in patients with so-called isolated duodenitis (in the course of the disease no ulcer lesion was ever detected), i.e. in 95.5%, indicates the relationship of this condition with peptic ulcer disease. The authors indicate diagnostic possibilities as regards CP infection by cultivation, rapid urease tests and microscopic examination and they emphasize the practical value of urease tests. Based on their own experience the authors confirm the high eradication effect of bismuth preparations, in particular bismuth subsalicylate and colloid bismuth subcitrate.

Prealbumin, orosomucoid (alpha-1-acid glycoprotein), were studied by single radial immunodiffusion in sera of 216 controls and 188 patients with gynecological carcinomas divided according to diagnosis, activity, and progression of the disease. The level of orosomucoid was found higher in 94 patients with active epithelial ovarian carcinoma (mean = 1.35 g/l), in 38 women with squamous cell carcinoma of the uterine cervix (mean = 1.04 g/l), and in 56 women with endometrial carcinoma (mean = 0.91 g/l), than in 61 blood donors (mean = 0.62 g/l) and 155 patients with benign gynecological diseases (mean = 0.71 g/l). Orosomucoid increased with the progression of ovarian carcinoma. Patients with active localized or advanced carcinoma had higher levels of orosomucoid than patients in remission (mean = 0.88 g/l). This can help in diagnosing ovarian carcinoma relapses. Prealbumin and transferrin decreased in all women with active ovarian carcinoma and in advanced cervical and endometrial carcinomas. Transferrin and prealbumin of patients in remission differed from the values in advanced carcinomas of the cervix and endometrium only and cannot be used in relapse determination. Calculation of the O/P index (orosomucoid to prealbumin ratio) brought no advantage for tumor diagnosis in comparison with the investigation of orosomucoid alone.

The effect of the platinum complexes cis-DDP and CBDCA on the functions of human peripheral blood mononuclear cells (PBMC)--viability, proliferating activity after polyclonal stimulation with phytohemagglutinin is followed in an in vitro study. Both the platinum complexes (at concentrations 10(-5) up to 10(-9) mol/l) inhibit significantly the proliferating activity of PBMC without affecting its viability, i.e. integrity and permeability of the cell membrane structures. As to effectivity, cis-DDP has shown itself a more effective inhibitor of cellular proliferation than CBDCA of which 10-100 times higher concentrations are required to achieve the same inhibitory effect in dependence on the temporal relation to the application of phytohemagglutinin. The highest inhibitory effect is noted when cis-DDP is applied either simultaneously with, or 48 h after PHA, i.e. at the stage of maximum proliferating activity. The results bring support to the assumed mechanism of action of the above platinum complexes--intervention into DNA synthesis. Since the test of blastic transformation of PBMC is held to be an image primarily of the function of T lymphocytes, it may be inferred that platinum cytostatics can affect cell-mediated immunity also in patients.