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Biomedical subjects

H M Jansen

Publications and source records attributed to H M Jansen.

At least 127 records · Page 7Linked to original sources
PubMed1992

Cloning of T lymphocytes from bronchoalveolar lavage fluid.

We have prepared T-cell clones from bronchoalveolar lavage fluid (BALF) from four healthy, nonsmoking persons and from four patients with allergic asthma. T cells were cloned by direct limiting dilution and with the use of a fluorescent activated cell sorter with an automated cell deposition unit. T-cell clones from the blood (PB) were prepared as well. The cloning efficiencies of T cells from BALF ranged from 3 to 40% and were lower than those obtained from PB T cells (18 to 72%). The cloning conditions generated CD4+ as well as CD8+ clones. The very late antigen-4, VLA-4, was more frequently expressed on CD4+ T-cell clones from BALF than from the blood (P < 0.05). CD8+ clones from BALF were more frequently VLA-1+ than those from blood (P < < 0.01). Mitogen- and monoclonal antibody-driven proliferation of CD4+ clones showed that BALF clones were well responsive to proliferation stimuli similar to those from the blood. Analysis of interleukin-4 production by 10 BALF and 10 PB clones showed large variations between individual CD4+ clones (BALF: range, < 100 to 700 pg/ml; PB: range, < 100 to 1,100 pg/ml), indicating the generation of different types of clones, which was also clear from analysis of interferon-gamma production. The analysis of properties of BALF T-cell clones and their regulation will improve insight into immunologic reactions in the lungs.

Adult↗
PubMed1992

Surfactant protein A in bronchoalveolar lavage fluid.

We measured surfactant protein A and phosphatidylcholine in the bronchoalveolar lavage fluid from healthy volunteers and several groups of patients with lung diseases to obtain information on surfactant in the lung. We developed three types of enzyme-linked immunosorbent assays that used combinations of polyclonal antiserum and monoclonal antibodies. Phosphatidylcholine was assessed by enzymatic measurement. The median amounts of surfactant protein A in bronchoalveolar lavage fluid determined by the enzyme-linked immunosorbent assay with monoclonal antibodies were as follows: control subjects (n = 10), 2.82 mg/L (range, 0.92 to 5.17 mg/L); patients with asthma (n = 13), 1.89 mg/L (range, 0.45 to 2.95 mg/L); and patients with pulmonary sarcoidosis (n = 20), 2.98 mg/L (range, 0.68 to 7.02 mg/L). The median phosphatidylcholine concentrations in the bronchoalveolar lavage fluid were as follows: control subjects (n = 10), 20 mumol/L (range, 3 to 37 mumol/L); patients with asthma (n = 12), 24 mumol/L (range, 3 to 55 mumol/L); and patients with pulmonary sarcoidosis (n = 20), 26 mumol/L (range, 4 to 76 mumol/L). As a group, the patients with asthma had less surfactant protein A in bronchoalveolar lavage fluid than did the control subjects (Mann-Whitney U test, p less than 0.05). The surfactant protein A levels measured by the enzyme-linked immunosorbent assay with polyclonal antiserum and by a competitive enzyme-linked immunosorbent assay were also lower in bronchoalveolar lavage fluid from patients with asthma than in that from control subjects. The phosphatidylcholine concentrations in all groups were similar.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1992

Long-term multicentre trial in chronic nonspecific lung disease: methodology and baseline assessment in adult patients. Dutch CNSLD Study Group.

Airways obstruction and airways hyperresponsiveness are two dominant features in patients with chronic nonspecific lung disease (asthma and chronic obstructive pulmonary disease (COPD)). We set up a study to determine whether long-term (3 yrs) therapeutic intervention directed at airways obstruction and hyperresponsiveness is superior to one directed at airways obstruction alone. Patients were selected on functional criteria (age, baseline forced expiratory volume in one second (FEV1), and airways hyperresponsiveness) and, furthermore, extensively characterized by history, smoking habits, allergy, reversibility of airways obstruction and quality of life. The methodology and practical problems of setting up this large multicentre study are outlined, together with an analysis of baseline data. Standardization of methods and techniques and recruitment of patients required much effort, recruitment taking about twice as long as expected. A 3 month feasibility study allowed us to eliminate minor problems in the protocol. Over a 16 month period, 274 adult patients (18-60 yrs) from the out-patient clinics of six university centres entered the study; 99 met the diagnostic criteria for asthma, 51 for COPD, 88 for asthmatic bronchitis, and 36 could not be classified. Their mean (SD) FEV1% pred was 65.1 (15.2)%. Their geometric mean provoking concentration of histamine producing a 20% fall in FEV1 (PC20 histamine) was 0.28 mg.ml-1. In a multiple regression analysis, more severe airways hyperresponsiveness was associated with lower prechallenge FEV1% pred (p less than 0.0001), higher pack-years of smoking (p = 0.0099), blood eosinophil count (p = 0.0004), skin test reactivity (p = 0.0047) and with female sex (p = 0.0302). We conclude that setting up long-term multicentre trials in chronic nonspecific lung disease (CNSLD) is feasible and that these may offer valuable information on treatment and outcome of the disease.

Adult↗
PubMed1991

Human atopen-specific types 1 and 2 T helper cell clones.

Eight representative T lymphocyte clones (TLC) randomly selected from previously described panels of CD4+ housedust mite Dermatophagoides pteronyssinus (Dp)-specific TLC from atopic and nonatopic donors were studied in more detail in a comparative investigation. The TLC from the atopic donors closely resembled murine type 2 Th (Th2) cells by secreting substantial IL-4, IL-5, IL-6, TNF-alpha, and granulocyte-macrophage (GM)-CSF, minimal IFN-gamma, and relatively little IL-2. In contrast, the nonatopic's TLC resembled murine type 1 Th (TH1) cells by secreting substantial IFN-gamma, IL-2, TNF-alpha, and GM-CSF, no IL-4, and little IL-5. A difference with murine Th1 cells was their additional secretion of IL-6. These cytokine profiles were consistent upon stimulation via different activation pathways including stimulation with specific Dp Ag, mitogenic lectins, and antibodies to CD2, CD3, or CD28. The observed differences in IL-2 secretion, however, were most evident upon stimulation with anti-CD28. If TLC cells were cultured with highly purified B cells and stimulated with anti-CD3 in the absence of exogenous IL-4, IgE synthesis was induced only in cultures with the atopics' Th2 clones, which could be completely abrogated by anti-IL-4. The mere presence of exogenous rIL-4, however, did not result in IgE synthesis, nor did unstimulated TLC cells alone. But if unstimulated TLC cells (that proved not to secrete detectable amounts of cytokines) were added together with rIL-4, again IgE synthesis was induced only in cultures with the atopics' Th2 clones, suggesting the involvement of an additional, as yet unidentified accessory helper function of the atopics' Th2 clones for IgE induction. Unstimulated Th2 clones showed a significantly higher expression of CD28 than the Th1 clones, but three days after stimulation, CD28 expression was elevated to comparable levels on both subsets. When added to B cells at this time point, together with rIL-4 and anti-IFN-gamma, still only the atopics' Th2 clones supported IgE synthesis, arguing against a role for CD28 in this accessory helper function. Whereas the atopics' Th2 clones were excellent helper cells for IgE induction, a unique property of the nonatopic's Th1 clones was their cytolytic activity toward autologous APC which could be induced by specific Dp Ag and by anti-CD3. The present data provide clear evidence for the existence of Th1 and Th2 cells in man.

Allergens↗
PubMed1991

Influx of urea during bronchoalveolar lavage depends on the permeability of the respiratory membrane.

In 6 healthy controls and 23 patients with pulmonary diseases the influx of urea during bronchoalveolar lavage was measured by comparing the concentrations of albumin and urea in the sequential samples recovered. It varied between -28 and 151 mumol/l. In the bronchoalveolar lavage fluid and serum we measured alpha-2-macroglobulin (A2M) and ceruloplasmin (CP). The bronchoalveolar lavage fluid to serum ratios were calculated (QCP and QA2M). QA2M/QCP was taken as a measure of the respiratory membrane permeability; it varied between 0.05 and 0.53. Influx of urea during lavage was higher according as the QA2M/QCP ratio was higher. We conclude that concentrations of substances in the epithelial lining fluid calculated with the urea correction method have to be corrected for the influx of urea.

Albumins↗
PubMed1991

[Development of a serological test for tuberculosis].

Antigens secreted by M. tuberculosis in the culture medium and antigens obtained from sonicated M. tuberculosis were characterized, at the laboratory of Tropical Hygiene of the Royal Tropical Institute and the division of Pulmonary Medicine at the Academic Medical Centre, in order to explore which antigens could be valuable in the development of a serological test for tuberculosis. Using murine monoclonal antibodies in immunoblot, a pattern of protein bands distinct from that of the corresponding M. tuberculosis sonicate was found in the culture medium. The major protein bands of the culture medium, of 24 and 12 kD, and the major protein band of 16 kD in sonicate were purified. These antigens were tested in ELISA with sera from 20 patients with tuberculosis, diagnosed by a positive culture of M. tuberculosis, and from 21 control subjects. The ELISA results obtained with these 3 antigens were combined and the mean value obtained in the control group plus 2 times the standard deviation was chosen as the cut-off level. Sixteen of the 20 patients with tuberculosis had antibodies against 1 of the purified antigens, while none of the control subjects had. By combining the results, obtained with these 3 antigens, 17 of the 20 patients with tuberculosis were positive in this serological test and none of the control subjects. The 24, 12 and 16 kD antigens may be valuable for the development of a serological test for tuberculosis.

Antigens, Bacterial↗
PubMed1991

Lactoferrin and secretory IgA in the bronchoalveolar lavage fluid from patients with a stable asthma.

We have measured lactoferrin and secretory IgA (sIgA) in the unconcentrated bronchoalveolar lavage fluid (BALF) from nonsmoking healthy volunteers (n = 10) and nonsmoking patients with stable asthma (n = 14). The median concentrations and the ranges of lactoferrin were controls, 0.13 mg/L (0.01-0.43 mg/L); asthma, 0.41 mg/L (0.07-7.51 mg/L). For sIgA the results were controls, 0.48 mg/L (0.12-1.47 mg/L); asthma, 1.29 mg/L (0.65-14.6 mg/L). The concentrations in the epithelial lining fluid (ELF) were calculated on the basis of urea in BALF and serum. SIgA and lactoferrin levels in the BALF and ELF from the patients with asthma were higher than in controls (Mann-Whitney U-test, p less than 0.03). Our results indicate that in patients with stable asthma the airway epithelial cells are activated, resulting in an enhanced secretion of lactoferrin and enhanced secretory transport of sIgA into the airway lumen.

Adolescent↗
PubMed1991

Functional subsets of allergen-reactive human CD4+ T cells.

After a period of resistance, the concept of human helper T (TH)-cell subsets is gaining currency. This is the result of analyses from a number of laboratories on the cytokine profiles of T-cell clones prepared from chronically-infected and hypersensitive individuals. Here, Martien Kapsenberg and colleagues summarize these studies and speculate on the significance of skewed TH1 and TH2 responses.

Allergens↗
PubMed1991

IgG subclasses in bronchoalveolar lavage fluid from patients with asthma.

We have measured Immunoglobulin G (IgG) subclasses in serum and bronchoalveolar lavage fluid (BALF) from 12 non-smoking patients with stable asthma and 9 non-smoking healthy volunteers to obtain information on their possible role in local immunological reactions. The quotients (concentration of IgG subclass in BALF)/(concentration of IgG subclass in serum) were calculated. In controls QIgG3 were lower than QIgG1, QIgG2 and QIgG4. The IgG subclasses in BALF and epithelial lining fluid (ELF) from patients with asthma were significantly higher than in controls, mainly due to increased leakage from the blood. Again QIgG3 were lower than Q of other subclasses. In the analysis of local production of IgG, albumin or ceruloplasmin was used as reference protein. Several patients showed a local production or a preferential accumulation of one or more IgG subclasses. We conclude that in healthy persons the IgG subclasses in ELF originate from the systemic circulation by passive permeation. In patients with asthma, the permeability of the respiratory membrane may be increased resulting in increased concentrations of subclasses in lung-lining fluid. In some patients with asthma, an additional local production of IgG subclasses occurs.

Adolescent↗
PubMed1991

IgM in the airways of asthma patients.

In order to obtain information on the role of IgM in the pathogenesis of asthma, we assayed IgM in the broncho-alveolar lavage fluid (BALF) from 14 non-smoking patients with asthma and 9 non-smoking healthy persons, using an ELISA. The concentrations of IgM in the epithelial lining fluid (ELF) were calculated on the basis of urea in serum and BALF. The concentrations of IgM in ELF after correction for serum IgM, cQIgM, from the patients as a group were higher than those from controls (Mann-Whitney U test, p less than 0.05). The IgM in BALF and serum, analysed by sucrose density gradient centrifugation was pentameric. The transudation of IgM from blood into ELF was compared with that of alpha 2 macroglobulin (A2M), a protein with a similar molecular mass. In 6 patients the cQIgM/cQA2M value was above the upper value of the range for controls suggesting abnormal local production of IgM. The presence of the secretory component (SC) linked to IgM was determined by ELISA. The percentage of SC-IgM in BALF was increased in 5 out of 7 patients tested. The increased local IgM concentrations and its increased local production may suggest a role for local IgM-mediated reactions in the inflammatory events associated with asthma.

Adolescent↗
PubMed1991

Respiratory membrane permeability and bronchial hyperreactivity in patients with stable asthma. Effects of therapy with inhaled steroids.

In patients with stable asthma, we assayed plasma proteins in the bronchoalveolar lavage fluid to obtain information on plasma exudation into the airways. Fourteen nonsmoking patients with asthma who were in a stable period of their disease and eight nonsmoking healthy volunteers were studied. The ratios of the concentrations of albumin, ceruloplasmin (CP), and alpha-2-macroglobulin (A2M) between blood and epithelial lining fluid were calculated (cQalb, cQCP, and cQA2M). The cQalb was increased in the patients (Mann-Whitney U test, p less than 0.05). In 10 patients the bronchial hyperreactivity was assessed with histamine provocation tests. Significant relationships between the cQalb, cQCP, and cQA2M on the one hand and PC15 on the other hand were found (Spearman's rank correlation: r = -0.62, p less than 0.05; r = -0.61, p less than 0.05; r = -0.79, p less than 0.01, respectively). Fourteen patients were treated with two inhalations of 200 micrograms glucocorticosteroids per day in a 3-month prospective study. Three of them were excluded from further study because of an intercurrent exacerbation of asthmatic symptoms during therapy. In the 11 patients with stable asthma, the cQalb and cQA2M decreased after treatment with inhaled steroids (Wilcoxon's matched pairs signed rank test, p less than 0.03). Our results show that in patients with stable asthma, there is an increased plasma exudation into the airways, most likely caused by an increased respiratory membrane permeability. The plasma exudation correlated with the bronchial hyperreactivity to histamine, and it decreased after corticosteroid therapy.

Administration, Inhalation↗
PubMed1991

Aberrant T cell regulation of IgE production in atopy.

Atopy is associated with elevated serum levels of allergen-specific immunoglobulin E (IgE). IgE production is preferentially induced by the T cell-derived lymphokine interleukin-4 (IL-4) and is suppressed by interferon and prostaglandins, of which interferon-gamma (IFN-gamma) is produced by T cells. We review our studies on the question of whether atopy is associated with CD4+ allergen-specific T cells with an unbalanced lymphokine production. We prepared panels of housedust mite (Dermatophagoides pteronyssinus)-specific T lymphocyte clones (TLC) from atopic and non-atopic individuals and found that TLC from housedust mite-allergic patients produced IL-4 but not IFN-gamma, whereas such TLC from a non-atopic individual produced IFN-gamma and only in some cases small amounts of IL-4. Candida albicans- and tetanus toxoid-specific TLC, established from one of the atopic donors, also produced IFN-gamma without IL-4, suggesting that the atopic state is characterized by a defective accumulation of IL-4-producing CD4+ T lymphocytes into the allergen-specific T cell repertoire.

Allergens↗
PubMed1991

Permeability or local production of immunoglobulins and other inflammatory proteins in asthma.

Immunoglobulins in the epithelial lining fluid of the respiratory tract either originate from the systemic circulation or they are locally produced by plasma-cells in the mucosa. For polymeric immunoglobulins (dimer IgA and IgM) specific receptors on the surface of epithelial cells allow facilitated transport of these immunoglobulins into the lining fluid. Local inflammatory reactions in the lung will damage the respiratory membrane, which results in an increased leakage of proteins into the lining fluid. This increased leakage complicates the differentiation between local production of immunoglobulins and passive permeation. In asthma patients inflammation of the respiratory epithelium is found. This inflammation may be the result of non-immunologically triggered processes as well as of immunological reactions. In the experimental part of this paper we analyse possible local production of immunoglobulins and other proteins within the lungs of patients with asthma. This is performed with the use of steady-state protein concentrations in serum and bronchoalveolar lavage fluid. Information on the permeability of the respiratory membrane can be obtained by the measurement of proteins that are not produced within the lungs. Information on local production can be obtained by comparing the epithelial lining fluid to serum concentration ratios of proteins with identical molecular masses.

Adolescent↗
PubMed1990

Evidence for compartmentalization of functional subsets of CD2+ T lymphocytes in atopic patients.

Lymphokine secretion profiles were studied of human allergen-specific CD4+ T lymphocyte clones (TLC). To this aim, panels of house dust mite Dermatophagoides pteronyssinus (Dp)-specific TLC were generated from two atopic Dp-allergic patients, suffering from severe atopic dermatitis (AD1) and allergic asthma (AD2), respectively, and from a non-atopic individual (NAD). From AD1 additional TLC were cloned specific for tetanus toxoid or Candida albicans, both Ag that were not relevant for the atopic state of this patient. Secretion of IL-2, IL-4, and IFN-gamma was determined after specific stimulation of these TLC, using autologous monocytes as APC. With respect to the production of IL-4 and IFN-gamma, clearly distinct profiles were observed. All Dp-specific TLC from both atopic donors produced IL-4 but not IFN-gamma, whereas the Dp-specific TLC from NAD, as well as the tetanus toxoid- and C. albicans-specific TLC from AD1, all produced IFN-gamma but not or small quantities of IL-4. Most TLC from all panels produced IL-2. These lymphokine profiles were consistent for at least 3 days and were neither dependent on the dose of allergen nor on the atopic or nonatopic state of the donor of APC. The functional consequence of these restricted lymphokine profiles was stressed by the observation that, whereas Dp-specific TLC from AD1 and AD2 supported in vitro IgE production, this support could be abrogated by a Dp-specific TLC from NAD. The present results suggest that CD4+ T lymphocytes that produce IL-4, but not IFN-gamma, occur in high frequencies in the allergen-specific T cell repertoires of atopic donors, which may have important implications for the pathomechanism of atopic disease.

Allergens↗
PubMed1990

Comparison of diversity and function of house dust mite-specific T lymphocyte clones from atopic and non-atopic donors.

Panels of CD4+CD8- T lymphocyte clones (TLC), specific for house dust mite Dermatophagoides pteronyssinus (Dp) proteins, were generated from the peripheral blood of an atopic Dp-allergic donor (AD), suffering from severe atopic dermatitis, and a histocompatible non-atopic donor (NAD). We studied the diversity of TLC within these two panels in search for the possible occurrence of dominant clone types with properties that might be characteristic for the atopic or non-atopic state. TLC with specificities for at least four different Dp proteins were found within the panel from AD "L" and for at least three different Dp proteins within the panel from NAD "K". In addition, both panels showed a considerable but comparable restriction diversity within HLA-DR. Despite the diversity within the panels, all Dp-specific TLC from AD were found to produce IL 4, after HLA class II-restricted Dp-specific stimulation, whereas the TLC from NAD produced no or only minimal amounts of this lymphokine. Only supernatants from stimulated AD TLC could induce IgE secretion by B cells from NAD. Conclusively, these observations do not give evidence for the occurrence of an abnormal Dp-specific T cell repertoire in AD, but rather suggest aberrant secretion of the IgE-inducing lymphokine IL 4 by CD4+ Dp-specific T cells from AD.

Allergens↗
PubMed1990

The measuring of "respiratory-membrane permeability" and local production of immunoglobulins and antibodies by means of an analysis of sputum.

When measuring the exudation of serum proteins and the local production of immunoglobulins and antibodies within the lung by means of an analysis of sputum, the permeability properties of the respiratory membrane should be taken into account. In this paper, we describe the "loss of size selectivity" that usually accompanies an increased permeability on the part of the respiratory membrane. This phenomenon enables us to measure respiratory membrane permeability independently of the sputum water content. Consequences with regard to discrimination between leakage from the circulation and/or local production of immunoglobulins and antibodies are discussed. Sequential studies which take these factors into account may provide insights into the extent of local inflammatory reactions in individual patients.

Antibody Formation↗
PubMed1990

Polycyclic aromatic hydrocarbon-DNA adducts in lung tissue from lung cancer patients.

In an attempt to probe for polycyclic aromatic hydrocarbon (PAH)-DNA adducts in human subjects resulting from smoking (or other chronic environmental exposure), lung tissue and lung tumours were obtained from patients hospitalized for lung cancer. DNA was isolated from the tissue samples and examined both in an ELISA using a polyclonal antibody against (+/-)trans-7,8-dihydroxy-anti-9,10-epoxy-7,8,9,10- tetrahydrobenzo[a]pyrene (BPDE)-DNA as well as by the nuclease P1-mediated modification of the 32P-post-labelling technique. The ELISA results showed BPDE-DNA antigenicity in lung DNA from 6 out of 21 patients, and adduct levels ranged from 2 to 134 adducts per 10(8) nucleotides. For all 21 patients, the autoradiographs of chromatograms of 32P-postlabelled digests of DNA from non-tumorous lung tissue showed a strong diagonal radioactive zone (DRZ). This DRZ was generally absent in tumorous tissue. DNA samples that were positive in the ELISA contained a dominant spot within the DRZ that co-chromatographed with the major BPDE-DNA adduct (BPDE-dG). The quantities of the BPDE-dG spots ranged from 2.1 to 42 adducts in 10(9) nucleotides. These values were lower than the levels found in the ELISA but correlated well with the ELISA results (Kendall W = 0.97; P = 0.00). The levels of the DRZ adducts ranged from 1.9 to 34 adducts in 10(8) nucleotides. Correlations between smoking and DNA adduct levels were poor because of the small number of current smokers (n = 13). However, smokers of filter cigarettes had significantly lower DNA adduct levels compared with smokers of cigarettes without a filter (P = 0.02 by Fischer's exact test).

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗