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[Frequent positivity of the human basophil degranulation test in idiopathic nephrotic syndromes (minimal glomerular changes and segmental and focal glomerulosclerosis)].

Atopic factors play an important role in minimal changes idiopathic nephrotic syndrome ( MCINS ). We therefore studied human basophil degranulation test ( HBDT ) in MCINS and also in patients with segmental and focal glomerulosclerosis ( SFGSNS ) whose relation with MCINS is debated. HBDT was performed with 5 or 6 allergens chosen according to the history of patients and results of cutaneous tests. HBDT was positive in 16 out of 28 MCINS and in 14 among 18 SFGSNS subjects. The difference versus 29 Immune Complex glomerulonephritis and 11 blood donors was significant with the chi 2 test. Because the basophil count was low (less than 10/microliter) in 5 cases, 4 of which had a severe nephrotic syndrome, the HBDT was performed later. The allergen most frequently involved was house dust. This was responsible for a significant degranulation in 10 MCINS (45%) and in 11 SFGSNS (68%) patients; degranulation was obtained more frequently with 2 allergens than with a single one. Obviously such a basophil sensitisation does not prove the responsibility of these allergens.

Adolescent↗

Platelet-activating factor in anaphylaxis and phagocytosis. I. Release from human peripheral polymorphonuclears and monocytes during the stimulation by ionophore A23187 and phagocytosis but not from degranulating basophils.

Platelet-activating factor (PAF) is a mediator of a anaphylaxis found initially in basophils and later in mouse and rat macrophages. The purpose of this paper was to determine the cellular origin of PAF released from human leucocytes and to establish if phagocytosis is a more important stimulus for PAF release than anaphylactic reactions. Phagocytic leucocytes (monocytes and PMNs) released PAF, physicochemically analogous to the PAF obtained by anaphylactic reactions in rabbits when challenged with zymosan, zymosan coated with complement, immune complexes, immunoglobulin aggregates or calcium ionophore A23187. Basophils failed to release PAF by anti-human IgE antibody, although positive degranulation and histamine liberaton were found. Pre-incubation of phagocytosing leucocytes with cytochalasin B or colchicine produced a diminution of PAF release, whereas beta-glucuronidase liberation was increased. The addition of carboxypeptidase B did not significantly modify PAF or beta-glucuronidase release. These data indicate that PAF obtained from preparations of human leucocytes comes from monocytes and polymorphonuclears; human basophils do not liberate measurable quantities of PAF, either by anaphylactic stimulus or by neutrophil cationic proteins; liberation of PAF and lysosomal content follow different mechanisms as they have different kinetics and are modified in an opposite way by drugs acting on the cytoskeleton.

Anaphylaxis↗

[The human basophils degranulation test (HBDT). Leucoconcentration and statistical calculation applied to the degranulation rate (author's transl)].

The technique using hemolysis and vital coloration described by Benveniste permits the human basophils to be studied in immediate type allergies : HBDT. In almost 25% of the cases, the test cannot be carried out because the basophils are less than 20 per mm(3). In 700 but one case, the HBDT could be carried out using a leucoconcentration technique by means of a 100 to 200 g centrifugation of 10 mn duration, followed by tangential extraction, thus obtaining an increased number of basophils. The authors were lent to apply a statistical calculation to a significant degranulation rate due to the number of observable cells by means of an unilateral statistical test suggesting a distribution according to Poisson's law. Thus tests are interpreted more rapidly and are more reliable. On the other hand, comparison with the results obtained with or without leucoconcentration in different types of allergies due to reagins (drugs, hymenoptera, foods) do not show any cellular fragilization. It may be concluded to the reliability of the degranulation observed in presence of an antigen.

Basophils↗

Immune serum transfer of cutaneous basophil-associated resistance to ticks: mediation by 7SIgG1 antibodies.

Guinea pigs acquired resistance to Amblyomma americanum larval ticks after one infestation, resulting in 46% tick rejection when challenged. Intravenous transfer of immune serum from twice-infested hosts to naive animals conferred a significant level of immunity resulting in 18 to 30% tick rejection. The minimum effective dose of serum was 3 ml per recipient, and heating the serum at 56 degrees C for 4 hr had no effect on serum activity. Fractionation of whole immune serum by gel filtration chromatography (Sephadex G-200) and ion -exchange chromatography (DEAE) demonstrated resistance activity to be in the IgG- and IgG1-containing fractions, respectively. Passage of whole immune serum through a heavy chain-specific rabbit anti-guinea pig IgG1 affinity column removed anti-tick activity and decreased the cutaneous basophil response to tick feeding by 70% in recipients. The ability to transfer both resistance to tick feeding and a significant cutaneous basophil response was eluted from the affinity column with 0.2 M Na2CO3, pH 11.5. In addition, immune serum raised against larval Ixodes dammini ticks, but not larval Rhipicephalus sanguineus ticks, was also effective at reducing feeding by larval Amblyomma americanum ticks, indicating that antibodies mediating resistance can be cross-reactive with antigens of different tick species and genera. This study demonstrates that IgG1 antibodies are responsible for the ability of immune serum to transfer cutaneous basophil-associated immune resistance against tick feeding in guinea pigs.

Animals↗

[Diagnosis of allergic diseases by basophil count and in vitro degranulation using manual and automated tests (author's transl)].

The author has designed a human basophil degranulation test based on a one-step method of basophil staining after exposure to a specific allergen. This very simple test is now commercially available as a ready-to-use kit. It explores allergic sensitivity at cellular level, probably through IgE-dependent sensitization of the basophil. It correlates positively with histamine release and the radio-allergo-sorbent test (RAST) and detects sensitivity to common allergens as well as to foods and chemicals, including drugs. In its present form, the test provides a simple, inexpensive and reliable means of diagnosing allergic diseases in vitro but does not lend itself to mass investigations. It has recently been developed further and can now be performed on the automatic Hemalog D Technicon machine.

Allergens↗

Immunological events at the surface of basophil granulocytes and mast cells which induce degranulation.

IgE molecules combine with basophil granulocytes and mast cells through the Fc portion and sensitize these cells for reaginic hypersensitivity reactions. The number of receptors for IgE per human basophil granulocyte is estimated to be 40,000 to 100,000 and those on rat mast cells is about 300,000. The binding of IgE with the receptor is reversible and does not involve covalent bonding. The affinity of IgE molecules for the receptors is high; the equilibrium constant of the reaction is in the order of 10(9)/mole for both human basophils and rat mast cell systems. Such a high affinity will explain why a minute amount of IgE antibody can sensitize the target cells and why sensitization with the antibody is so persistent. The initial step of the reaginic hypersensitivity reactions is probably bridging of cell-bound IgE molecules by antigen, which activates enzymatic sequences. The release of chemical mediators, however, is controlled by pharmacological factors which have most important rôles in transducing immunological signals into intracellular signals and subsequent biochemical processes.

Animals↗

[Interpretation of results on a human basophil degranulation test (author's transl)].

A statistical analysis of a result on a human basophile degranulation test was performed. The shape of the curve representing the number of basophiles, as a function of the logarithm of the allergenic dosage, was parabolic, in the case of degranulation. This curve permits a calculation of the dosage necessary to provoke a degranulation of 10 per cent, in comparison with the mean of the two control test tubes. The authors also determined the significant degranulation threshold to be 5 per cent, as regards the mean number of cells counted for the controls (example: for 50 control cells counted, the threshold was equal to 28 per cent degranulation). This statistical analysis largely facilitates the interpretation of the granulation test by basophiles, by assuring the value of observed degranulations, and by allowing a better quantification of the phenomenon. The latter might permit a comparison of different batches of allergen, that is, the standardisation of allergens. These calculations can be easily programmed, using a currently available calculator.

Basophils↗

Is non-specific cross-reacting antigen a marker on the surface of human basophils?

Leucocytes challenged with anti-NCA serum (non-specific cross-reacting antigen) can release histamine. This release is Ca++, glucose and temperature dependent as usually described for basophils. Only half of the subjects whose basophils can release histamine when challenged with anti-IgE serum are also able to release histamine with anti-NCA serum. There is no cross-reaction between IgE and anti-NCA serum or between NCA and anti-IgE serum. So it seems likely that NCA might be a marker on the surface of some human basophils.

Adult↗

IgE-independent interleukin-4 expression and induction of a late phase of leukotriene C4 formation in human blood basophils.

T-helper cells can differentiate into at least two subtypes secreting distinct profiles of cytokines, Th1 and Th2, regulating immunoprotection and different immunopathologies. Interleukin-4 (IL-4) is both the product and the inducer of Th2 cells, raising the question whether IL-4 can be produced in response to antigen-independent stimuli. Here we show that human basophils produce IL-4 on stimulation with IL-3 and C5a or C5adesarg in similar amounts as induced by IgE-receptor-cross-linking. C5a-induced IL-4 production requires the presence of IL-3, with little effect of the sequence of stimuli addition. No "Th1-cytokines" (interferon-gamma and IL-2) and even no "Th2-cytokines" (IL-3, IL-5, IL-10, and granulocyte-macrophage colony-stimulating factor) are produced by basophils in response to either IgE-dependent or IgE-independent activation. The generation of leukotriene C4 (LTC4) is regulated in a similar manner. However, C5a induces a rapid, transient burst of leukotriene formation only if added after IL-3. Interestingly, upon prolonged culture, a late phase of continuous LTC4 production is observed, which also requires two signals (IL-3 and C5a), but rather depends on their continuous presence than on their sequence of action. These data describe an antigen-independent pathway of very restricted IL-4 expression. Thus, basophils must be considered as central immunoregulatory cells of the innate immune system. Furthermore, the results show that LTC4 can also be generated more continuously for many hours, a phenomenon that may be of particular importance in chornic allergic inflammation, such as asthma.

Basophils↗

Rapid method for basophil count in domestic fowl.

A simple, rapid, and reproducible method for absolute basophil count in adult domestic fowl is described that uses toluidine-blue stain. The absolute basophil count was variable from bird to bird and breed to breed. The importance of a high basophil count in relation to inflammation and hypersensitivity in birds is discussed.

Animals↗

[The effect of loratadine on skin and bronchial reactivity and histamine liberation from basophils in patients with atopic asthma and allergic rhinitis].

The effect of the therapy with Loratadine on skin and bronchial reactivity to histamine and on a specific histamine release from isolated basophils of allergic patients was assessed. The studies were carried out on a 34 patients (23 with pollen rhinitis and 11 with a mild bronchial asthma). The effect loratadine versus placebo on the skin reactivity to histamine was measured in patients suffering from pollen rhinitis. Outside the pollen season in 12 patients from the above group a histamine release from isolated basophils was evaluated by Shore method, using a-IgE and pollen allergen as liberators, before and after 4 days therapy with loratadine. In the group of the 11 asthmatic patients the effect of the single dose of 10 mg Loratadine on spirometric parameters and on a bronchial response to histamine was studied. The Bronchial Provocation Test with histamine was performed by the Ryan's method. The results were expressed as PC20 FEV1. It was shown that loratadine statistically significant limited of the skin reactivity to histamine and specific histamine release from basophils. The one tablet of Loratadine have not changed of spirometric parameters in the patients with mild asthma, but statistically significant reduced their bronchial response to histamine.

Asthma↗

[Spontaneous histamine release by basophils and bronchial hyperreactivity in patients with hay fever].

Basophil spontaneous histamine release test was performed in seventeen patients suffered from seasonal allergic rhinitis before, 2 weeks after immunotherapy and during pollen season. During pollen season at the same day non-specific bronchial hyperreactivity to histamine was estimated. Values of spontaneous basophil histamine release before immunotherapy were not different from control group but significantly increased after immunotherapy and during pollen season. There was no correlation between degree of nonspecific bronchial hyperreactivity and values of spontaneous histamine release from basophils.

Allergens↗

IL-3-induced histamine release from human basophils. Dissociation from cationic dye binding and down-regulation by Na+ and K+.

The effect of recombinant human IL-3 on human basophils from normal subjects was evaluated by the decrease of toluidine blue-positive basophils (TB+) and by histamine release. IL-3 (0.001 to 100 ng) caused the decrease of TB+ without concomitant histamine release, whereas anti-IgE-induced TB+ decrease was accompanied by histamine release. IL-3 enhanced both anti-IgE-induced TB+ decrease and histamine release. IL-3-induced TB+ decrease was dose- and Ca2+-dependent and related to the time of incubation (detectable after an incubation of 5 min and maximal after incubation of 2 h). When extracellular Na+ was replaced isosmotically with choline, N-methyl-D-glucamine, or glucose, histamine release also was observed after direct stimulation with IL-3. The effect was dose dependent, related to the time of incubation (detectable after 30 min and maximal after 60 min), and required extracellular Ca2+. The increase in extracellular Na+ or K+ concentrations was accompanied by a reduction of histamine release, with a more marked effect on IL-3- than on anti-IgE-induced histamine release. In line with these results, the Na+ ionophore gramicidin D, which increases Na+ influx, and the K+ channel blocker 4-aminopyridine, which decreases K+ efflux, dose-dependently inhibited anti-IgE- and IL-3-induced histamine release. The inhibitory effects on Na+ and K+ were slightly additive, suggesting an action via the same pathway, which most probably is membrane potential. These results indicate that: 1) IL-3 can induce TB+ decrease without concomitant histamine release, 2) basophils from normal subjects can release histamine on challenge with IL-3 once the inhibitory effect of Na+ is removed, and 3) Na+ and K+ down-regulate IL-3 as well as anti-IgE-induced histamine release.

4-Aminopyridine↗

[Acute basophilic leukemia: a case report].

A 56-year-old woman was admitted to our hospital in January, 1990 because of fever and petechiae. Leukocyte count of peripheral blood showed 41,000/microliters with 89% immature cells, and bone marrow was normocellular with 96.2% immature cells. They were medium to large in size, positive for peroxidase staining, CD-13 and CD-33. Half of them contained azurophilic granules. They showed metachromasia by toluidine blue, contained basophilic granules in electron microscopic examination and reacted to G-CSF, G-CSF and IL-3. She was diagnosed as acute basophilic leukemia and treated with BHAC-DMP and B triple-V regimen, but remission was not attained. She died of peritonitis due to gastrointestinal tract perforation and pneumonia in March, 1990. This is the fifteenth case of acute basophilic leukemia reported in Japan, and the hematological examinations performed in this patient were demonstrated.

Bone Marrow↗

[Evaluation of diagnostic value of skin test, venom specific antibodies against EgE and basophil histamine release test in hymenoptera allergy].

The study compares results of skin test and assessments of venom specific IgE levels (FAST) in patients with Hymenoptera sting allergic reactions. Results of the two diagnostic methods show considerable correlation. Positive correlation occurred in 90% of the patients (92.4% bee venom sensitive allergic patients and 80% wasp venom sensitive patients). Negative correlation occurred in 2% of the patients. In the remaining 8% of the group the results did not correlate. Basophil histamine release test was performed in 13 patients with allergic reactions after stinging. 77% of the basophil histamine release test results showed positive correlation with the other two tests. No correlation was found between the size skin test, the class of the FAST test, and the result of basophil histamine release test. No relationship was found either, between results of the described test and the severity of sting reaction classified to Mueller scale.

Adolescent↗

[Basophilic cells/eosinophil differentiation activity in conditioned medium of nasal inverted papilloma].

An increased number of histochemically distinct mast cells and eosinophils can be observed in the tumor mass of nasal inverted papilloma. In order to elucidate the mechanism of this accumulation of mast cells and eosinophils in a local site, we examined the activity of conditioned medium (CM) in inducing differentiation and proliferation of basophilic cells and eosinophils from hematopoietic progenitors of peripheral blood mononuclear cells of the patients with allergic diseases. CM was obtained by the culture of nasal inverted papilloma cells (NIPC) and its activity was measured by determining colony proliferation in methylcellulose assay for 14 days. In the 14 day methylcellulose assay, the number of Eo-type colonies in the presence of 1, 5, 10 and 20% CM was significantly higher than the baseline number of Eo-type colonies (0%). Cytochemical analysis confirmed that the number of colonies of cells with metachromatic granules and histamine positive colonies in Eo-type colonies in the presence of 5, 10 and 20% NIPC-CM was significantly higher than that in the control. The number of ECP positive colonies in Eo-type colonies in the presence of 5% and 10% CM was also significantly higher than in the presence of 0.5% CM. Furthermore both histamine and ECP positive colonies were observed in Eo-type colonies. These results are consistent with the hypothesis that basophilic cells and eosinophils accumulate at nasal inverted papilloma sites at least in part by the recruitment of progenitors from the circulation and subsequent differentiation in situ in response to cytokines. Eosinophils and basophilic cells also commonly originate from committed granulocyte progenitors.

Adolescent↗

Expression of mRNA for the GATA-binding proteins in human eosinophils and basophils: potential role in gene transcription.

The expression of the hematopoietic transcription factors GATA-1, GATA-2, and GATA-3 was studied in eosinophils and basophils. Eosinophils express mRNA for GATA-1, GATA-2, and GATA-3. Basophils express GATA-2 and GATA-3. Treatment of HL-60 eosinophilic sublines with either interleukin-5 or butyric acid increased the expression of GATA-1 mRNA concomitant with the expression of eosinophil-specific genes, whereas levels of GATA-2 mRNA remained relatively constant. The presence of mRNA for these proteins in eosinophils and basophils suggests that gene transcription in these lineages may be regulated by GATA-binding proteins.

Base Sequence↗

Vesicular transport of Charcot-Leyden crystal protein in tumor-promoting phorbol diester-stimulated human basophils.

Secretion from human basophils (HB) that are stimulated with a phorbol ester takes place with slower kinetics than typically regulated secretion, which is stimulated by the IgE-mediated mechanism associated with classical granule exocytosis. Phorbol ester stimulation of HB induces emptying of granule contents (with retention of granule containers) and increases the number of cytoplasmic vesicles, an anatomic process similar to one termed piecemeal degranulation (PMD), which is a secretory process originally described in HB that migrate from the blood in vivo into contact allergy skin lesions. Charcot-Leyden crystal (CLC) protein is a basophil granule-associated protein that is readily imaged by using a postembedding immunogold procedure. This method was used to localize this granule protein in phorbol ester-stimulated, isolated human peripheral blood basophil cytoplasmic vesicles in samples collected within a time frame for which histamine was secreted. The results of this study showed that the proportion of cytoplasmic vesicles that were gold-labeled in stimulated HB, which indicated the presence of CLC protein, increased significantly over unstimulated cells at 2, 5, and 10 minutes after stimulation. Additionally, CLC protein-labeled vesicles in the cells that were stimulated for 10 minutes significantly exceeded the number in stimulated cells at 0, 30, and 45 minutes after exposure to phorbol ester. Thus, transport vesicles carrying a granule-associated protein (CLC protein) were increased in phorbol ester-stimulated HB in the time frame for histamine secretion and the anatomic development of PMD. These findings support vesicular transport as a major mechanism for effecting PMD, which is morphologically the most frequent activation anatomy displayed by HB in human disease in vivo.

Basophils↗