Antisense anti IGF-I cellular therapy of malignant tumours: immune response in cancer patients.

The treatment of cancer by antisense anti-IGF-I cellular therapy inducing immune response has evoked interest among many promising strategies. Here, we reported some results obtained from patients with cancer, mainly glioblastoma treated by this strategy, which was also extended to patients with colon carcinoma, ovary cystadenocarcinoma and prostate adenocarcinoma. It was shown that, in the phase I of clinical trial, patients vaccinated with their own tumour cells treated by antisense IGF-I presented a slight increase of temperature. Their peripheral blood lymphocytes showed a shift in the percentage of CD8 effector cells as judged by expression of cell surface markers CD8+ CD28+. Particularly, in two treated patients with glioblastoma, the survival time was 19 and 24 months respectively in comparison to the range of 12 to 15 months observed in the case of classical treatment such as surgery, radiation or chemotherapy. These results, although preliminary, gave indication that the reported strategy could deserve consideration owing to its safety. Furthermore, the increase in the percentage of peripheral blood monomorphonucleated cells (PBMNCs) with effector phenotype, i.e., CD8+ CD28+ in vaccinated patients might explain their prolonged survival time.

Real-time PCR quantification of plasma DNA in non-small cell lung cancer patients and healthy controls.

INTRODUCTION: Free-circulating DNA is present in minute amounts in plasma of healthy individuals, whereas increased levels are found in a number of malignant pathologies including non-small cell lung cancer (NSCLC). The objective of this research was the evaluation of the plasma DNA quantification capacity to distinguish between healthy subjects and non-small cell lung cancer (NSCLC) patients. MATERIAL AND METHODS: Plasma samples were collected prospectively from 16 healthy volunteers and 30 untreated NSCLC patients (I-IIIA). Subsequently, free-circulating DNA extraction and quantitative real-time PCR analysis were performed. RESULTS: The values of plasma DNA concentration ranged from 0.9 up to 7.0 ng/ml in healthy individuals and from 1.5 up to 50 ng/ml in NSCLC patients before treatment. Cancer group showed several-fold higher mean free-circulating DNA concentration than that present in healthy subjects (mean 12.00 vs. 2.65 ng/ml; P<0.001). A greater variability of plasma DNA concentrations was observed in NSCLC patients than in controls (SD 14.50 vs. 2.02, respectively). The area under the ROC curve was 0.87 (95% CI, 0.744 to 0.954, P<0.001). CONCLUSION: Non-small cell lung cancer is associated with elevated levels of cell-free DNA in plasma with respect to healthy controls. Real-time PCR method proved its utility in effective free-circulating DNA detection and quantification.

Evaluation of angiogenesis, p-53 tissue protein expression and serum VEGF in patients with endometrial cancer.

Endometrial carcinoma occurs mostly in post-menopausal women. Classical methods of prognostication, as FIGO stage and histopathologic grade, could be improved by applying additional techniques, utilizing molecular biology and immunochemistry. p-53 tumor suppressor gene, the most commonly mutated gene in human cancers has been shown to play an important role in the biology of gynecologic carcinomas. Angiogenesis, a process of formation of new vessels, being connected to tumors progression and metastatic potential was shown to be linked with tumor suppressor genes expression. The aim of the study was to evaluate relationships between intensity of tumor angiogenesis, serum levels of Vascular Endothelial Growth Factor (VEGF) and tissue p-53 protein expression in endometrial adenocarcinoma. Angiogenic Point's Density (APD) was calculated in hot spots areas using the morphometric appliance. For detection of p53 protein in tumor samples, LSAB + Kit Alkaline Phosphatase (DAKO) was used. VEGF levels were assessed in patient's blood sampled before the operation. Overexpression of p53 protein was found in tumor tissue in 35.2% of cases and mean angiogenic points density was greater in p53 positive cases. Serum levels of VEGF were above the cut off level in 54.5% of patients, in those cases angiogenesis was also elevated. In cases of p53 overexpression, VEGF levels tended to be greater as compared with p53 negative cases. In conclusion, our study demonstrated that angiogenesis was more intensive in p53 positive cases, confirming the hypothesis of tumor suppressor-gene regulation of the process of neovascularization. Serum levels of VEGF were borderline-significantly higher in cases of p53 overexpression, they were also correlated to the angiogenesis. Joint assessment of angiogenesis and tumor suppressor genes expression may contribute to reliable evaluation of the biology of endometrial carcinoma.

Presence of MHC-I in rat glioma cells expressing antisense IGF-I-receptor RNA.

The supposed immunogenic character of glioma cells transfected with antisense IGF-I-Receptor (IGF-I-R) expression vector was tested for the presence of MHC-I currently present in cells of IGF-I antisense type. C6 rat glioma cell line was comparatively transfected in vitro with IGF I antisense (pMT-Anti-IGF I) or IGF I Receptor antisense (pMT-Anti-IGF I R) expression vectors. The wild and transfected cells were examined for the presence of IGF-I and MHC-I molecules. Using RT PCR technique, the transfected "antisens" cells showed total inhibition of IGF-I. The both transfected cultures of IGF-I and of IGF-I-R type were positively stained for MHC-I. Moreover "antisense IGF-I-R" cells as compared to "IGF-I antisense" cells showed slightly higher expression of MHC-I. The transfected cells showed also the feature of apoptosis in 60% of cells. The immunogenicity of IGF-I-R antisense glioma cells is related to MHC-I presence; therefore both approaches of antisense IGF-I and of antisense IGF-I-R could be use in paralel for cellular therapy of glioblastoma.

IGF-I triple helix gene therapy of rat and human gliomas.

PURPOSE: IGF-I anti-gene technology was applied in treatment of rat and human gliomas using IGF-I triple helix approach. MATERIAL AND METHODS: CNS-1 rat glioma cell and primary human glioblastoma cell lines established from surgically removed glioblastomas multiforme were transfected in vitro with IGF-I antisense (pMT-Anti-IGF-I) or IGF-I triple helix (pMT-AG-TH) expression vectors. The transfected cells were examined for immunogenicity (immunocytochemistry and flow cytometry analysis) and apoptosis phenomena (electron microscopy). 3 x 10(6) transfected cells were inoculated subcutaneously either into transgenic Lewis rats or in patients with glioblastoma. The peripheral blood lymphocytes (PBL) derived from "vaccinated" patients were immunophenotyped for the set of CD antigens (CD4, CD8 etc). RESULTS: Using immunocytochemistry and Northern blot techniques, the transfected "antisense" and "triple-helix" cells showed total inhibition of IGF. Transfected cultures were positively stained either for both MHC-I and B7 antigens--60% of cloned lines, or for MHC-I only--40% of cloned lines. Moreover "triple helix" cells as compared to "antisense" cells showed slightly higher expression of MHC-I or B7. Transfected cells also showed the feature of apoptosis in 60%-70% of cells. In in vivo experiments with rats bearing tumors, the injection of "triple helix" cells expressing both MHC-I and B7 interrupted tumor growth in 80% of cases. In contrast, transfected cells expressing only MHC-I stopped development in 30% of tumors. In five patients with surgically resected glioblastoma who were inoculated with "triple helix" cells, PBL showed an increased percentage of CD4 + CD25+ and CD8 + CD11b-cells, following two vaccinations. CONCLUSIONS: The anti-tumor effectiveness of IGF-I anti-gene technology may be related to both MHC-I and B7 expression in cells used for therapy. The IGF-I antigene therapy of human glioblastoma multiforme increases immune response of treated patients.

Growth hormone treatment in pituitary insufficiency: selected cases of children with craniopharyngioma and medulloblastoma.

PURPOSE: The work concerns the substitution treatment with growth hormone (GH) in hypopituitary children, including cases that occurred in the course of tumor disease, craniopharyngioma (CP) and medulloblastoma (MB). MATERIAL AND METHODS: The studied population concerned 117 children who presented either somatotropic or polyhormonal pituitary insufficiency (the average age was 12.6 years for girls and 13.6 years for boys). The diagnosis of somatotropic pituitary insufficiency (SPI) was based on insulin and clonidin stimulation tests evaluating GH reserve of hypophysis. The computer tomography (CT) and nuclear magnetic resonance (NMR) examinations were carried out before GH substitution in all children. The tumors (four CP cases and one case of MB) were all found in boys and they were treated with surgery and/or radiotherapy. All studied children, including CP and MB operated patients were treated with human GH (hGH)--Genotropin 16 IU, administered in subcutaneous injections. The daily dose was calculated as 0.5 IU/kg/week. RESULTS: The annual increase of children height before GH therapy was about 3.2 cm. In the first year of GH therapy the difference in children growth between the CP/MB group as compared with the rest of patients was less than 1.0 cm: 9.4 and 10.2 cm/year, resp. During the second year of hormone substitution the growth became slower: average values were 8.2 cm and 7.4 cm/year, resp. In CP and MB patients the height increase calculated as SDS values was significant (2.7 and 1.0 resp.). Control NMR examination performed in CP/MB patients treated with surgery with subsequent hGH therapy did not demonstrate any recurrence of tumor. CONCLUSIONS: After two years of hGH therapy the final height of hypopituitary children, including CP patients, nearly reached the values observed in healthy children. GH therapy did not induce a recurrence of neoplasm in CP and MB patients.

[Cytoimmunologic changes in material obtained from bronchoalveolar lavage (BAL) in asymptomatic individuals chronically exposed to silica dust]. / Zmiany cytoimmunologiczne w materiale z plukania oskrzelowo-pecherzykowego (bronchoalveolar lavage, BAL) u osób bez objawów klinicznych przewlekle eksponowanych na pyly krzemionki.

The aim of this study was to determine and to evaluate silica induced lung cell reactivity--if any--in bronchoalveolar space, before clinical changes develop. Bronchoalveolar lavage (BAL) was carried out in 15 nonsmoking individuals with chronic professional silica exposure, free of lung signs and symptoms. Controls were healthy nonsmokers. Routine BAL cytology (HE, MGG) was completed by mast cell staining (toluidine blue). BAL lymphocyte subsets were phenotyped by direct two- and three-color immunofluorescence (applied DAKO A/S monoclonal antibodies: anti-CD3, CD4, CD8, CD11b, CD14, CD15, CD16 + 56, CD19, CD25, CD45, HLA-DR). Parallel staining was performed in peripheral blood. In individuals with chronic silica exposure we found: significant increase in alveolar macrophage (362 +/- 45 vs 160 +/- 33 x 10(3) cells/ml, p < 0.05), lymphocyte (61 +/- 9 vs 24 +/- 5 x 10(3) cells/ml, p < 0.05) and BAL total cell (415 +/- 76 vs 187 +/- 34 x 10(3) cells/ml, p < 0.05) numbers; significant increase in mast cell (0.4 +/- 0.1 vs 0.2 +/- 0.1, p < 0.05), NK cell (7.0 +/- 1.8 vs 3.6 +/- 1.0, p < 0.05) and Th early activated lymphocyte percent (CD4 + CD25+ calculated as percentage of CD4+ cells: 15.1 +/- 1.5 vs 7.8 +/- 1.6, p < 0.01). All results were presented as median +/- SEM. Bronchoalveolar space of people with chronic silica exposure usually shows pathological reaction (especially macrophagic alveolitis), although they are free of manifested pulmonary disease. Th early activated lymphocytes, NK cells and mast cells seem to play important role in the early interstitial lung tissue reaction to silica.

[Changes in lymphocyte subsets in bronchoalveolar lavage fluid in patients with systemic sclerosis]. / Zmiany w subpopulacjach limfocytów w plynie z plukania oskrzelowo-pecherzykowego u chorych na twardzine ukladowa.

The aim of this study was to estimate if alterations of lymphocyte subsets obtained by broncholaveolar lavage (BAL) were related to clinical data observed in nonsmoking patients with systemic sclerosis (SSc). Clinical examination included chest X-rays, spirometry and arterial blood gasometry. Patients were divided into group A (pulmonary changes present, n = 15) and B (without any changes, n = 7). Healthy subjects constituted the control group (n = 10). BAL lymphocytes were phenotyped using monoclonal antibodies coupling CD4, CD8 (both in coexpression with CD25), CD19 and HLA-DR human antigens and flow cytometer FACStar (Becton-Dickinson). Parallel staining was performed in peripheral blood. BAL lymphocyte typing was completed by BAL routine cytology. In SSc patients we found increased BAL total cell number, percentage of neutrophils, eosinophils and macrophage giant cells, as well as high percent of CD25+ and HLA-DR+ lymphocytes. In the group A neutrophilic alveolitis was observed in nearly half of cases: total lymphocyte number (per 1 ml of BAL fluid) and significantly reduced CD4/CD8 ratio were found. In the group B, as compared with controls, we found significantly elevated lymphocyte total cell number per 1 ml of BAL fluid (including particular subsets: CD3+, CD4+, CD8+). Also significantly high CD4+25+ lymphocyte percent was observed. Summing up, cytological and/or immunological alterations were observed in all examined SSc patients. The intensity of these alterations seems to be related to the clinical data. A decreased value of CD4/CD8 ratio may play a role in the local appearance of pulmonary changes in the course of systemic sclerosis.

[Principles of optimal preparation of material from bronchoalveolar lavage (BAL)for cytoimmunologic examinations in interstitial lung diseases ]. / Podstawy optymalnego przygotowania materialu z plukania oskrzelowo-pecherzykowego (bronchoalveolar lavage, BAL) do badan cytoimmunologicznych w chorobach sródmiazszowych pluc.

In this paper we considered the value of correctly performed bronchoalveolar lavage (BAL) in diagnosis of interstitial lung diseases and in assessment of the activity of pathological process. We indicated the conditions of exact interpretation of the results of BAL cytoimmunological examination, i.e. fine standard handling of BAL material, including its collection, elaboration and choice of lavage site, as well as regarding external (e.g. cigarette smoking) and internal additional factors. We described the influence of BAL fluid recovery and of the method of staining on obtained results. We emphasized routinely performed BAL to be the valuable diagnostic and research tool in pulmonology however, the method may have limited usefulness and unnecessarily burden the patient, if technical guidelines are not observed.

[Phenotype of alveolar lymphocytes--theoretical and practical implications]. / Fenotyp limfocytów pecherzykowych--przeslanki teoretyczne i uwagi praktyczne.

Actual characteristics of alveolar lymphocytes was presented, including author's own experience, i.e. bronchoalveolar lavage (BAL) lymphocyte typing in over 450 patients with interstitial lung diseases (ILD) and bronchial asthma. In healthy individuals as well as in the patients, alveolar lymphocyte population is dominated by T cell subset. Alveolar T lymphocytes present primed memory cell phenotype and usually do not proliferate; in normal conditions apoptosis occurs very seldom. T cytotoxic cell phenotype is characteristic for CD8 subset. Percent of B and NK cells found in BAL is very low--B cells are localized in interstitial space around the pulmonary alveoli, while NK cells do not seem to play the important role in the local pulmonary defense. CD4/CD8 ratio evaluation in BAL material is an auxiliary tool in ILD diagnostics; it reflects local imbalance between T helper primed memory cells and primed T cytotoxic cells.

Flow cytometric evaluation of lymphocyte subpopulations in BALF of healthy smokers and nonsmokers.

The purpose of this work was to evaluate the normal lymphocyte phenotype in the bronchoalveolar lavage fluid (BALF). BAL was carried out in 12 untreated healthy nonsmoking volunteers and in 9 cigarette smokers. For the analysis of lymphocyte subsets by two-color flow cytometry, the monoclonal antibodies used were directed anti: CD3, CD4, CD8, CD16, CD19, D25, CD45, CD56 and anti HLA-DR. An increase in the total number of cells in BALF of smoking persons and increased proportion of macrophages was observed. The percentage of CD8+ lymphocytes was 1.7 times higher, whereas the proportions of CD4+ cells, and a CD4+/CD8+ ratio were lower 1.5 and 2.6 times, respectively, in the BALF of cigarette smoking persons when compared with nonsmoking volunteers. The changes did not depend on the age of the person. In conclusion, we suggest that the decreased CD4/CD8 ratio and the elevated CD8 T cell subset may be regarded as a potential risk factor associated with clinically asymptomatic lung cancer. Moreover, in the interpretation of BALF from patients with pulmonary diseases cell proportions of nonsmoking and of smoking persons should be compared with the respective controls.

Pulmonary infiltrates with masto- and lymphocytosis in BALF associated with arechine.

Pulmonary infiltrates developed in a 40-year-old man while receiving arechine. The hyperplasia of mast cells, hyperlymphocytosis, a predominance of cytotoxic T-cells of the CD8 type with the very low CD4/CD8 ratio in BAL were observed. The chest roentgenogram findings resolved within a month after discontinuing arechine therapy. A hypersensitivity reaction with the onset of lung fibrosis due to arechine seemed to be the likely cause.

[Use of flow cytometry in evaluation of cellular changes in interstitial lung diseases]. / Zastosowanie cytometrii przeplywowej w ocenie zmian komórkowych w sródmiazszowych chorobach pluc.

Flow cytometry (FC) usefulness for pulmonary reactive cell changes examination in interstitial lung diseases was assessed. Bronchoalveolar lavage (BAL) lymphocyte direct (two-color and three-color) phenotyping was carried out in 63 patients with sarcoidosis (subdivided according to the disease radiological stage and smoking status), 23 patients with systemic sclerosis (SSc) and 12 individuals exposed to silica dusts. 33 healthy volunteers were used as controls. Routine BAL cytology and peripheral blood lymphocyte (PBL) typing was performed in all tested subjects. FC results quality control was performed according to BD Simultest IMK Plus criteria (for PBL) modified by the author. The conventional lymphocytic gate (PBL light scatter based) was excluded, as the source of error in L-BAL subset analysis. Two-color L-BAL phenotyping and alveolar lymphocyte CD45FITC/SSC gating was found to be usually sufficient. However, in poor-lymphocyte materials (e.g. in healthy smokers) three-color method, using anti-CD45 PECy5 monoclonal antibody for staining each patient sample, should be considered. No significant changes in lymphocyte subsets were found in FC as compared with alkaline phosphatase immunocytochemical method. The prevalence of CD3+ cells with only a few lymphocytes expressing NK (CD3-CD16 or 56+), T suppressor (CD8+ 11b+) or B cell phenotype--as compared with PBL, was found in BAL of all examined subjects. CD8+ cell subset was dominated by T cytotoxic cell phenotype (CD8+ CD11b-) in all tested groups. L-BAL T cells expressed sensitized memory cell phenotype--CD4+ (CD8+)CD45RO+ in sarcoidosis, SSc and controls (the staining not performed in silica exposed persons). Characteristic BAL cytological and immunological pattern (lymphocytic alveolitis, increased CD4/CD8, high CD3(CD4)+HLA-DR+ percentage) was observed in pulmonary sarcoidosis. SSc patients showed reduced BAL CD4/CD8 ratio, due to T cytotoxic cell predominance (the changes were more distinct in the group with clinical pulmonary changes). Early activation marker (CD25) was expressed on the elevated percentage of BAL T helper cells in SSc. High CD4+CD25+ cell percentage was the most characteristic sign of lymphocyte activation in individuals exposed to silica. Summing up, the reference values for L-BAL subsets were proposed. CD4/CD8 ratio alterations in interstitial lung disorders should be interpreted as the local imbalance between T helper memory cells and sensitized T cytotoxic lymphocytes. FC characterization of alveolar lymphocytes provides insight into the pathogenesis of pulmonary diseases.

[Bronchitis and alveolitis in the course of Chlamydia pneumoniae infection]. / Sródmiazszowe zapalenie pluc oraz zapalenie oskrzelików w przebiegu zakazenia Chlamydia pneumoniae.

A case of a 38-year old male with respiratory failure in the course of infection by Chlamydia pneumoniae has been described. Inflammation of bronchioles and alveoli was diagnosed on the basis of clinical examination and analysis of cellular components of bronchoalveolar lavage fluid (BALF). The diagnosis was confirmed by bacteriological examination of the culture infected by the material collected from the patient. Therapy with doxycycline was instituted which resulted in remission of symptoms. The case described confirms the findings of other authors that infection by Chlamydia pneumoniae may be the cause of serious respiratory distress.

[Diagnosis of interstitial lung disease (ILD) in patients with systemic sclerosis. The significance of broncho-alveolar lavage (BAL) --personal observations]. / Diagnostyka choroby sródmiazszowej pluc (interstitial lung disease, ILD) u chorych na twardzine ukladowa. Znaczenie plukania oskrzelowo-pecherzykowego (BAL)--obserwacje wlasne.

Review of current literature on pathogenesis and diagnostic approach to interstitial lung disease (ILD) in systemic sclerosis (SSc) was presented. The review focused in particular on the bronchoalveolar lavage (BAL). The experimental study was aimed to established whether early performed BAL is corresponding with clinical data, especially within a group with signs and symptoms of overt ILD. BAL was performed in 25 non-smoking subjects (22 women, 3 men) with SSc (according to ARA) with no systemic steroids. Diagnosis of lung involvement (presented in 18 patients) was based on history and physical examination, chest X-ray, lung function tests and arterial blood gas determination. BAL was routinely stained and assessed. Changes in BAL cytological examination were observed in all patients. An increased total cell number as well as increased percentage of neutrophils and eosinophils was noted. A lymphocyte number rise was not statistically significant. A lung involvement in group with ILD was more advanced than in group without ILD and controls, i.e. neutrophilic alveolitis in half cases (9/18 vs. 0/7 in group with no ILD) and oesinophilic alveolitis in 33% cases (6/18 vs. 2/7). Lymphocytic alveolitis was found in one patient with ILD and in two patients without ILD. The value of BAL in a diagnostic approach to the ILD in SSc was emphasized. Sensitivity of BAL in case of early ILD seems to be comparable with sensitivity of lung function tests (e.g. DLCO) and computerized tomography. The answer to the question which of the above mentioned methods in most appropriate to detect ILD risk in SSc remains unknown.

Interferon-gamma in the treatment of atopic dermatitis: influence on T-cell activation.

Atopic dermatitis (AD) is a chronic relapsing skin disease characterized by various immunologic abnormalities. Its pathogenesis remains obscure and its treatment difficult. We have studied the efficacy of systemic recombinant human interferon-gamma (rhIFN-gamma) treatment (0.05 mg/m2 sc on 3 consecutive days, during 4 weeks) in 10 patients with severe AD. Marked clinical improvement was observed starting from the third week of treatment. Erythema, dryness, and lichenification were the most responsive symptoms. Serum immunoglobulin E and IgG4 levels did not change during treatment. Blood eosinophil count decreased only transiently at the end of the first and second series of injections (days 4 and 11; P = 0.02). Patients with AD showed an increase in CD25-positive cells (11.0% vs 4.88%; P = 0.0001) as compared to 10 age-matched healthy controls. Moreover, in parallel with clinical improvement, a distinct decrease in CD25-positive lymphocytes was observed on days 32 and 50 (P = 0.002 and P = 0.006, respectively). We suggest that in AD the beneficial effect of rhIFN-gamma might be related to the inhibition of excessive T-cell activation, perhaps of the subpopulations, producing interleukin (IL)-4 and IL-5.

[Human recombinant interferon gamma in the treatment of atopic dermatitis]. / Ludzki rekombinowany interferon gamma w leczeniu atopowego zapalenia skóry.

Atopic dermatitis (AD) is a chronic relapsing skin disease characterized by various immunologic abnormalities. We have studied the efficacy of recombinant human interferon gamma (rhINF-gamma) administered subcutaneously at a dose of 0.05 mg/m2 in ten patients with severe AD. Patients were treated for 4 weeks. They have shown marked clinical improvement starting from the third week of treatment. The efficacy of the drug varied, with erythema, dryness and lichenification being the most responsive symptoms. There was no change in serum immunoglobulin E and IgG4 levels. Whole blood eosinophil count decreased only transiently and was accompanied by a tendency to lower values of serum eosinophil cationic protein. Patient with AD showed an increased expression of a T-cell surface activation marker CD 25 as compared to healthy controls. Moreover, clinical improvement was roughly paralleled by the decrease in this T-cell activation marker. We conclude that rhINF-gamma is a novel efficacious therapeutic approach in severe AD. We suggest that its primary action might be related to the inhibition of T-cell activation.

Reactivity of bronchoalveolar space cells in lung asbestosis.

Preliminary studies to evaluate lymphocyte subsets bronchoalveolar lavage (BAL) and blood were carried out in a group of patients with lung asbestosis. The assessments were made by an indirect immunofluorescent method using monoclonal antibodies as a marker. The results were recorded on a flow cytofluorometer FASC-can. As compared with a control group, the patients with lung asbestosis showed a slight decrease in the total level of T lymphocytes (CD3) in BAL, however with a clearly disturbed proportion between T-helper (CD4) and T-suppressor (CD8) lymphocytes, which led to a decreased CD4/CD8 ratio. The blood level of T lymphocytes and their subsets in these patients approximated that in the controls. Heavy smoking was also found to enhance disorders in their number and proportion. The assessment of B lymphocytes (by using polyclonal antihuman immunoglobulins (sIg/FITC) showed a significantly high level of these lymphocytes in BAL with simultaneously low levels in blood in the same patients with lung asbestosis. We did not find a clear synergic effect of cigarette smoking on the levels of B lymphocytes in the studied group.

[A preliminary assessment of the incidence of atopy at a workplace in Cracow]. / Wstepna ocena czestosci wystepowania atopii w jednym z krakowskich zakladów pracy.

Skin tests, IgE level and eosinophils percentage were determined in 337 employees of the "Hydrokop" Works in Cracow. These investigations aimed at seeking more specific markers of allergy than anamnesis and skin tests. Allergy was reported by 108 individuals (38%). Some features of atopy were confirmed in 7.5%. Skin tests were positive in 36 individuals and were related to atopy in 55%. IgE levels were 10 IU/ml--greater than 1000 IU/ml with the distribution similar to log-normal. Mean IgE concentration was relatively high (202 IU/ml, in the individuals with confirmed congenital allergy. Percentage of eosinophils exceeded 3% in 39 subjects out of which 17 reported the symptoms of allergy. IgE level and percentage of eosinophils are controversial as the markers of atopy due to the contribution of various non-allergic factors while skin tests correlate well with allergic diseases which were relatively frequent in the examined group.