[Concentration of selected cytokines associated with monocytes/macrophages in serum of patients with different clinical stages of pulmonary tuberculosis]. / Stezenie wybranych cytokin zwiazanych z ukladem monocytarno/makrofagowym w surowicy chorych na gruzlice pluc o róznym przebiegu klinicznym.

Monocytes/macrophages are the main effector cells as well as the essential elements in granuloma formation in tuberculosis. Serum concentration of some cytokines connected with monocytes/macrophages like IL-12, MCP-1, TNF-alpha i sTNFRI was assessed in patients with advanced bacteriologically confirmed pulmonary tuberculosis (I group) and in those with minimal tb changes, usually without clinical signs of the disease, and sputum smear-negative (II group). We found that patients from group I had significantly higher concentration of IL-12 and sTNFRI and lower level of MCP-1 in comparison to group II. Significant difference between groups of patients with tuberculosis and the control group was found only while assessing serum concentration of TNFRI.

[The effect of nedocromil sodium on spontaneous production of histamine releasing factor (HRF) by mononuclear blood cells in patients with nonatopic bronchial asthma]. / Wplyw nedokromilu sodu na samoistne wytwarzanie, przez komórki jednojadrowe krwi obwodowej, czynnika uwalniajacego histamine z bazofili (HRF) u chorych na nieatopowa astme oskrzelowa.

The aim of the study was to evaluate the effect of 28-days antiinflammatory treatment with nedocromil sodium (Ned.sod.) (Tilade-8 mg/24 hours) on nonspecific bronchial hyperreactivity (PC20H in mg/ml) and spontaneous HRF activity production by mononuclear blood cells in patients with nonatopic bronchial asthma treated with beclomethasone dipropionate. The correlation between PC20H and HRF was also examined. The study was performed in 10 subjects with mild and moderate chronic, stable asthma in a double-blind, placebo-controlled way. Placebo and Ned. sod were given through 4 weeks in a randomized way with 8 weeks wash-out period. It was shown that Ned. sod. did not influence clinical asthma symptom scores, ventilatory parameters and PC20H. No changes in the spontaneous production of HRF activity were observed. The correlation between HRF activity and PC20H assessed 4 times was not significant. The authors conclude that studies with Ned. sod. in nonatopic asthma should be continued, but the dosage of the drug ought to be bigger and the time of treatment should to be longer.

[Chemotactic serum activity for neutrophils and eosinophils and spontaneous production of histamine releasing factor in patients with seasonal bronchial asthma treated with nedocromil sodium]. / Aktywnosc chemotaktyczna surowicy dla neutrofili i eozynofili oraz samoistna produkcja przez komórki jednojadrowe krwi obwodowej czynnika uwalniajacego histamine u chorych na sezonowa astme oskrzelowa leczonych nedokromilem sodu.

Nedocromil sodium (NS- Tilade) is an effective therapeutic agent against asthma and has been shown to exhibit antiinflammatory activity in vitro. The aim of this study was to assess the effect of NS on the serum neutrophil (NCA) and eosinophil (ECA) chemotactic activity and mononuclear cells-derived histamine releasing factor (HRF) activity in 14 patients with seasonal allergic asthma. The chemotactic activities, HRF and bronchial reactivity to histamine (PC20H in mg/ml) were determined before the grass-pollen season and at the end of the 14-day placebo treatment and 21-day NS treatment during the pollen-season. NCA and ECA were assessed using of the Boyden chamber methods. Blood samples for mononuclear cells culture were collected during the trial. The supernatants were assayed for HRF activity with basophils from a single donor. The NCA index was unaltered during the trial. There was a significant (p < 0.05) increase in the ECA after the NS treatment period (ECA index--2.5) compared with out of season and after placebo treatment (ECA index, 1.88 and 1.82). HRF activity increased after placebo compared with out of season (HRF % activity--38.62 before season, 47.61 after placebo p < 0.05) but NS did not revealed the further effect on HRF activity (49.81 p > 0.05). During placebo the significant decrease in PC20H from 1.81 mg/ml to 0.54 mg/ml (p < 0.05) was observed. NS did not have the effect on nonspecific bronchial reactivity--PC20H--0.74 mg/ml (p > 0.05). The correlation between PC20H and spontaneous production of HRF activity was not found. The results indicate that NS has some effect on inflammatory process which takes place in seasonal asthmatic patients during the natural allergic exposure.

[The effect of peat preparation on the release of histamine from basophils of atopic bronchial asthma]. / Wplyw preparatu torfowego tolpy (PTT) na uwalnianie histaminy z bazofilów u chorych na astme oskrzelowa atopowa.

Antigen-induced basophil histamine releasing (BHR) is an in vitro model of the immediate-type allergic reaction. We studied the influence of immunologically active peat preparation (PTT). On BHR induced with specific antigen or Con A. The highest concentration of PTT enhanced, but lower concentrations decreased antigen-induced BHR. Surprisingly, similar effect was observed with placebo, suggestions that immunologically inactive components of both preparations may be responsible for the modulatory activity.

The effect of preseasonal immunotherapy on the production of histamine-releasing factor (HRF) by mononuclear cells from patients with seasonal asthma: results of a double-blind, placebo-controlled, randomized study.

The effect of immunotherapy on the production of histamine-releasing factor (HRF) by mononuclear cells (MNC) from patients with seasonal asthma was investigated in a double-blind, placebo-controlled, randomized study. Twenty-four patients with asthma were randomly divided into a placebo-treated group and a grass pollen-treated group. In vitro production of HRF by MNCs and the provocative concentration of histamine that causes 20% fall in FEV1 were measured before and after immunotherapy, and symptoms were monitored during the pollen season. MNCs from the patients were either cultured alone, spontaneous HRF production (spHRF), or in the presence of grass allergens (grass-stimulated HRF production), and the supernatants were assayed for HRF activity with basophils from a single donor after the study. We found that MNCs from patients with seasonal asthma to grass pollen spontaneously produce substantial amounts of HRF. The group of patients treated with placebo developed typical symptoms in the pollen season and also an increase in HRF production. In contrast, patients treated with grass pollen demonstrated reduced symptoms and no increase in spHRF production. A high degree of correlation between spHRF productions and symptom scores in both treated groups was noted. After 2 years of immunotherapy, allergen-stimulated HRF production decreased significantly, whereas spHRF production decreased significantly only in the patients who were clinically benefitted. The change in the provocative concentration of histamine that causes 20% fall in FEV1 during the pollen season highly correlated with the change in HRF production. The results of this study suggest that HRF might be involved in the pathogenesis of atopic asthma.

Effect of autogenic bacterial antigens on the production of histamine releasing factor by mononuclear cells from intrinsic asthmatic patients.

The production of histamine releasing factor (HRF) by mononuclear cells (MNC) from intrinsic asthmatic patients has previously been reported. In this study, we investigated the effect of preincubation of lymphocytes with autogenic killed bacteria upon the production of HRF. Bacteria were isolated from the sputum, and nasopharyngeal swab obtained from patients and control subjects. MNC from intrinsic asthmatics and healthy controls were preincubated with killed bacteria for 4 h, then washed and cultured for 18 h. HRF activity of the cell-free supernatants was assayed in the histamine release test using basophils from normal subjects. We found that MNC from the patients spontaneously produce significant amounts of HRF. Preincubation of the cells with autogenic bacterial antigens enhanced HRF production in 12 of 25 patients and only in one of 15 control subjects. No specific bacterial strain was identified as having the sole stimulatory property for HRF production; rather, individual susceptibility predisposes to the ability to produce HRF in response to some common bacteria. When MNC from healthy subjects were preincubated with bacterial antigens isolated from the patients, no enhancement in HRF production was observed. We concluded that MNC from some intrinsic asthmatics are specifically sensitized to certain bacterial antigens and release HRF upon contact with these antigens.

The magnitude of the spontaneous production of histamine-releasing factor (HRF) by lymphocytes in vitro correlates with the state of bronchial hyperreactivity in patients with asthma.

In our previous studies we reported that lymphocytes from patients with asthma spontaneously produce histamine-releasing factor (HRF) in vitro. In an effort to examine whether spontaneous HRF production (SpHRF) by lymphocytes from patients with asthma is related to the state of bronchial hyperreactivity (BHR), 20 patients with mild to severe asthma were studied. Peripheral blood lymphocytes were cultured alone in a serum-free medium for 24 hours, and culture supernatant was assayed for HRF activity in two separate histamine-release tests with autologous basophils and normal basophils from known healthy donors. BHR was measured as bronchial reactivity to inhaled histamine and was expressed as a provocation concentration of histamine required to induce a 20% fall in FEV1 (PC20). The result of this study demonstrated that lymphocyte supernatant from all patients with asthma released significant amount of histamine from both autologous and normal basophils. Very high histamine release was usually induced by lymphocyte supernatant from severely ill patients who had PC20 less than 2 mg/ml. Statistical analysis demonstrated that the magnitude of the SpHRF significantly correlated (r = -0.86; p less than 0.001) with PC20. Since mast cell- and basophil-derived mediators have been implicated in the pathogenesis of BHR, high correlation between PC20 and SpHRF by lymphocytes suggests that the latter may contribute to the development of BHR. Further studies are required to disclose the exact relationship between SpHRF and BHR.

Bacterial antigens stimulate the production of histamine releasing factor (HRF) by lymphocytes from intrinsic asthmatic patients.

Lymphocyte from 12 intrinsic asthmatic patients and 10 healthy controls were studied for their capability to produce histamine releasing factor (HRF) in vitro. Spontaneous HRF production was measured by culturing the lymphocytes alone for 20 h. In another set of experiments lymphocytes were first preincubated separately with phytohaemagglutinin, antigens of Haemophilus influenzae, Streptococcus viridans, Staphylococcus sp. and Neisseria catarrhalis for 4 h then carefully washed three times and cultured alone for an additional 16 h. Cell-free supernatant was assayed for histamine releasing activity using basophils from healthy donors. It was observed that lymphocytes from intrinsic asthmatic patients spontaneously produced HRF. The production of this lymphokine was enhanced following preincubation of lymphocytes with phytohaemagglutinin or bacterial antigens. Results of skin test with bacterial antigens did not correlate with the magnitude of the production of HRF by lymphocytes. At gel chromatography over Sephadex G-75 bacterial antigen-stimulated lymphocyte supernatant revealed two peaks of HRF activity in the molecular weight ranges 35,000-50,000 and 3,000-7,000.

A lymphocyte-derived histamine-releasing factor in asthmatic patients. III. Further studies.

Peripheral blood lymphocytes from atopic asthmatic patients cultured overnight (16 hours) produced a novel lymphokine-histamine releasing factor (HRF) in the culture supernatant. Activity of this lymphokine has been tested in the basophil histamine release test. Supernatant from unstimulated lymphocytes released 29% histamine from basophils. A short incubation of lymphocytes with skin test-positive allergens (grass, mite) enhanced the generation of HRF. Skin test-negative allergens had no effect. HRF appeared in the supernatant as early as four hours after the start of the culture and its production level remained high even at 48 hours. Gel-filtration with Sephadex G-75 revealed an apparent molecular weight for HRF in the range of 10,000 to 15,000. HRF is relatively heat stable at 56 degrees C.

"In vitro" basophil degranulation by polymyxin B in asthmatic patients.

Polymyxin B is a potent mast cell degranulator. Inhalation of polymyxin B produces bronchoconstriction in atopic asthmatic patients. The study was undertaken to find the influence of polymyxin B on human basophils. Thirty atopic asthmatic patients and 10 healthy controls participated in the study. Preparation and counting of basophils taken from the whole venous blood was performed with the human basophils degranulation test (method by Benveniste). No significant degranulation was observed in the group of healthy subjects. Degranulation of basophils by polymyxin B was stated in 24 from 30 atopic asthmatic patients with dose-reaction dependency. Our results point out, that there exists "basophil releasability" to polymyxin B in atopic asthmatic patients.

Bronchial and basophil reaction to polymyxin B in asthmatic patients.

The hypothesis that increased reactivity to polymyxin B in asthmatic patients is not limited to the bronchi but also exists in basophils was investigated in 26 atopic asthmatic subjects. Quantification of basophil degranulation induced by polymyxin B was performed using a flow-cytometric method. Significant degranulation was noted in 17 patients, but in none of 10 healthy controls. There was close relationship between basophil "releasability" to polymyxin B and bronchial reactivity to inhaled polymyxin B. These findings support the hypothesis.

Complement in allergen-induced bronchospasm in house-dust RAST negative asthmatic patients.

We studied a group of asthmatic patients with aspirin intolerance who were RAST-negative for house-dust and Dermatophagoides-pteronyssinus despite having positive skin tests and positive inhalation tests to these allergens. Total haemolytic activity (CH50), C3, C4 and C3PA were measured after allergen-induced bronchospasm in these patients and also in a group of RAST-positive asthmatic patients taken as a control group. We did not find any change in complement level after allergen-induced bronchospasm in both groups of patients.