In vitro cultivation of Brugia malayi, a parasitic nematode that causes human lymphatic filariasis.

The in vitro cultivation of the filarial nematode Brugia malayi from the infective stage to the fourth and the young adult stage is described. Different culture conditions including cell-free systems and coculture with different human lymphatic cell lines were compared. Cocultivation with each of the cell lines used enhanced the survival of the parasites.

Analysis of IgG subclass and IgE response in allergic disease caused by Aspergillus fumigatus by immunoblotting techniques.

Analysis of sera from patients suffering from allergic asthma, extrinsic allergic alveolitis (EAA) due to Aspergillus fumigatus or allergic bronchopulmonary aspergillosis (ABPA) by means of IEF immunoprint and Western blot showed characteristic IgG subclasses and IgE responses. Asthmatic patients revealed specific IgE antibodies. IgG subclass reactivities in asthmatics and healthy blood donors were weak and uncharacteristic.

Major allergen Phl p Va (timothy grass) bears at least two different IgE-reactive epitopes.

It is established that most grass pollen allergens consist of several isoforms of which the function is mainly still unknown. A number of these allergens belonging to group V have been cloned, sequenced, and expressed. Antigenic sites and IgE-reactive epitopes of the major allergen Phl p Va, are unknown.

[The Th1/Th2 concept--its importance for regulation of IgE].

T helper type 2 (TH2) cells are a major source of interleukin- (IL-)4 which plays a critical role in the induction of the IgE synthesis. Their counterplayers, the T helper type 1 (TH1) cells produce interferon- (IFN-)gamma which inhibits the IL-4-induced IgE synthesis. Thus, the TH2/TH1 ratio plays an important role in the regulation of the IgE synthesis.

Epitope analysis of isoforms of the major allergen Phl p V by fingerprinting and microsequencing.

The major allergen of timothy grass pollen (Phleum pratense), designated as Phl p V, consists of isoallergenic components of 38 and 32 kDa with pl values of 5.2-7.5 and 4.

Characterization of grass group I allergens in timothy grass pollen.

Using Phl p V-depleted timothy grass pollen extract (Phleum pratense) as immunogen, we obtained a monoclonal antibody, QG 4, which recognized proteins of 33, 35, and 37 kd as determined by Western blotting. The antibody cross-reacted with pollen proteins of other grass species in the molecular weight range of 30 to 37 kd. By means of two-dimensional polyacrylamide gel electrophoresis blot of timothy grass pollen extract, we demonstrated at least seven protein spots: two of 37 kd with isoelectric points of 6.

Comparison of four grass pollen species concerning their allergens of grass group V by 2D immunoblotting and microsequencing.

The identification and characterization of allergenic components is a vital step towards improving diagnosis and therapy. Members of the grass family (Poaceae) reveal a high cross-reactivity among each other caused by the close phylogenetical relationship. In order to investigate the variability between allergenic components, we studied the allergen grass group V, one of the major allergens.

Monocyte-mediated restoration of density-dependent growth control and induction of cell death in tumor cell populations.

It is shown in a serum-free system with human cells that during monocyte/tumor cell interactions signals are generated which restore the principles of density-dependent inhibition of growth in tumor cell populations. Tumor cells that fail to comply with these principles are induced to die. This cell death is an active process in target cells involving protein biosynthesis and tumor cell-derived signals.

[Food allergy: identifying and characterizing peanut allergens with patient sera and monoclonal antibodies].

The purpose of this study is to improve the diagnosis of peanut allergy. In order to standardize test substances for in vivo and in vitro diagnostic, the type I allergy-associated single components of peanuts have been identified and characterized with the aid of patients' sera and monoclonal antibodies. For allergen detection IEF-immunoprint-, SDS-PAGE-immunoblot- and 2-D electrophoresis-techniques have been used.

Successful technique for the selective production of monoclonal antibodies against a major allergenic component in timothy pollen extract.

Monoclonal antibodies (MAbs) were selectively raised against a major allergenic component of 38 kD in timothy grass pollen (Phleum pratense). We used a special prefractionating technique to isolate the 38 kD allergen, because immunizations with crude pollen extract had resulted in a wide variety of antibodies against other determinants, and cross-reactions between different-sized proteins occurred. Pollen extract was separated by Western blotting.

[Circadian rhythm in cytokines].

The well-known human circadian rhythm (CR) in body temperature may be due to CR in endogenous pyrogens such as interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF). We, therefore, investigated the daily variation in spontaneous and LPS-induced monokine release by peripheral blood cells (whole blood assay) in 10 healthy volunteers. Spontaneous release of monokines (IL-1, IL-6, TNF) showed a simultaneous significant peak at noon, whereas LPS-induced release of IL-1 and IL-6 was strictly correlated with the CR in body temperature with its significant maximum at 8.

[Characterization of allergens of timothy grass (Phleum pratense)].

By means of 2-D electrophoresis and subsequent immunoblotting allergens of timothy grass pollen are separated as single components. Via N-terminal sequencing, it is possible to classify these components as isoallergens of defined groups according to the allergen nomenclature. We are able to identify the allergens as Phl p I, Phl p IV, Phl p V and Phl p VI.

Inhibition of endogenous TNF formation by pentoxifylline.

During the last decade cytokines were recognized as focal components in acute and chronic inflammatory processes. The growing knowledge about these agents stimulated efforts to pharmacologically control their synthesis and action in clinical situations. Various rational approaches to these issues including selective antibodies or receptor antagonists are at present under clinical investigation.

Sensitivity to aspirin: a new serological diagnostic method.

Certain adverse reactions to aspirin (ASA), nonsteroidal anti-inflammatory drugs (NSAIDs) and pyrazolone derivatives resemble IgE-mediated hypersensitivity. However, convincing evidence of antigen-antibody interactions or inhibition of the cyclooxygenase pathway of arachidonic acid metabolism, stimulating the production of leukotrienes (LTs) and decreasing the production of prostaglandins (PGs), has not been presented. In this study, two types of specific IgE antibodies have been found in six serum samples from eight ASA-sensitive patients with salicyloyl and O-methylsalicyloyl disks using the Radio Allergo Sorbent Test (RAST), whereas no positive result could be found with acetylsalicyloyl disks.

Detection of IgE antibodies specific for 1-phenyl-2,3-dimethyl-3-pyrazoline-5-one by RAST: a serological diagnostic method for sensitivity to pyrazoline drugs.

Certain adverse reactions to pyrazoline drugs resemble IgE-mediated hypersensitivity. However, convincing evidence of antigen-antibody interactions is not fully demonstrated. In this study, IgE antibodies specific for 1-phenyl-2,3-dimethyl-3-pyrazoline-5-one have been found in 17 out of 19 serum samples from individuals sensitive to pyrazoline drugs with 4-aminoantipyrine discs by Radio Allergo Sorbent Test (RAST).

Epitope analysis of the allergen ovalbumin (Gal d II) with monoclonal antibodies and patients' IgE.

Ovalbumin (OVA) is a major allergen (Gal d II) of hen egg white and is often the cause of hypersensitivity reactions to food. Further knowledge of the antigenic and allergenic epitopes of allergens will provide better treatment of this disease. To analyse these epitopes we produced a panel of monoclonal antibodies (mAbs) against native OVA.

Cytokines involved in monocyte mediated tumor cell death and growth inhibition in serum-free medium.

In a serum-free culture system, the release of TNF, lI-1, lI-6, IFN-alpha, and IFN-beta during interaction of elutriated human monocytes (MO) with human tumor cells (TC) was studied by ELISA-technique. Contributions of these cytokines to inhibition of TC-growth and to induction of TC-death by supernatants (SU) gained from such MO/TC-interaction cultures were investigated using affinity chromatography for removal of individual cytokines. Although the TC used are relatively insensitive to recombinant human TNF, withdrawal of TNF causes 50% to 75% reduction of SU-induced TC-death rates, suggesting that susceptibility to TNF is raised during MO/TC-interaction by the other cytokines.

Examination of microheterogeneity in grass pollen allergens.

The separation of timothy pollen extract by two-dimensional immunoblotting revealed microheterogeneity of the major allergens PhI p I and PhI p V. There was not only a diversity in size, 38 and 32 kDa for PhI p V and 37, 35, and 33 kDa for PhI p I, but also a separation into proteins of identical sizes but different pIs. Since former studies on the protein structure by amino acid analysis and N-terminal microsequencing did not reveal any differences, we examined other possibilities that might cause microheterogeneity.

[Pentoxifylline--an inhibitor of the synthesis of tumor necrosis factor alpha].

The growing knowledge on the pathophysiological role of cytokines in acute and chronic inflammatory processes stimulated efforts to control their synthesis and action pharmacologically in clinical situations. Recently, in our institute evidence was raised that pentoxifylline (POF) is able to suppress the synthesis of tumor necrosis factor-alpha (TNF) in cell cultures, in vivo, and to protect experimental animals against endotoxin shock. Studies in human experimental endotoxemia showed that pentoxifylline decreased circulating TNF without affecting endogenous formation of interleukins.

The presence of specific IgE to salicyloyl and O-methylsalicyloyl in aspirin-sensitive patients.

Certain adverse reactions to aspirin (ASA), nonsteroidal anti-inflammatory drugs (NSAIDs) and pyrazoline derivatives resemble IgE-mediated hypersensitivity. However, convincing evidence of antigen-antibody interactions or of the inhibition of the cyclooxygenase pathway of arachidonic acid metabolism leading to an increase in the generation of leukotrienes (LTs) and a decrease in the generation of prostaglandins (PGs) was not fully demonstrated. In this study, two types of specific IgE antibodies have been found in 27 serum samples from 28 ASA-sensitive patients with salicyloyl-discs and O-methylsalicyloyl-discs by Radio Allergo Sorbent Tests (RAST).