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A monoclonal antibody reacting with human basophils.

Bsp-1 is an IgM murine monoclonal antibody raised against the human erythroblastic leukemia cell line (HEL) that reacts with basophils but not neutrophils or eosinophils. Western blotting techniques showed that Bsp-1 reacts with a 45-kilodalton surface antigen on HEL cells. The distribution of Bsp-1 antigen on leukemic cells is confined to a basophilic leukemia cell line, KU812, chronic myeloid leukemia with basophilia, and some cases of acute undifferentiated leukemia. Bsp-1 might therefore be a useful reagent for the study of basophil function and differentiation.

Antibodies, Monoclonal↗

Modulation of human basophil histamine release by hemopoietic growth factors.

To determine what kinds of CSF modulate human basophil function, recombinant or purified hemopoietic growth factors were tested for the effect on histamine release from basophils. Both granulocyte (G)-macrophage (M) CSF and IL-3 markedly enhanced histamine release upon stimulation with anti-IgE in a dose-dependent manner (maximal enhancement 25.5% by GM-CSF and 30.8% by IL-3 as expressed as percent increase against total cellular histamine content), whereas G-CSF, M-CSF, and IL-4 had no effect. Enhancing action of these factors was still observed in the highly enriched basophil population, suggesting that this action was not via contaminating cells. Enhancement of histamine release by both factors was not necessarily IgE mediated, because they also amplified histamine release upon stimulation with FMLP and ionophore A23187. The enhancement by both factors was temperature dependent, and took place rapidly and reached plateau levels in 15 min. GM-CSF and IL-3 achieved the similar plateau level of augmentation and no additive effects were observed between them. This finding suggests that they enhance histamine release by sharing the same pathway in the release reaction.

Adjuvants, Immunologic↗

[The basophilic histamine liberation test as a diagnostic method in Hymenoptera venom allergy].

In 181 patients with systemic reactions to hymenoptera stings as a result of immediate-type allergy to bee or wasp venom confirmed by history, skin testing and demonstration of hymenoptera venom-specific IgE antibodies on radioallergosorbent testing (RAST), in vitro histamine release from peripheral basophilic granulocytes following stimulation with bee or wasp venom was evaluated. Suspensions of washed peripheral leukocytes at a density of 2 x 10(6)/ml were incubated with four different concentrations of hymenoptera venoms. Histamine was measured by spectrofluorometry, and histamine release was calculated as a percentage of the total histamine content. Tests performed with peripheral leukocytes obtained from 18 non-allergic individuals served as controls. In both bee venom allergy and wasp venom allergy the corresponding allergen induced concentration-dependent histamine release. Upper normal limits of histamine release were defined for the venom concentrations used. When these were used as reference values the basophil histamine release test exhibited a specificity of 94% and a sensitivity of 82% in the diagnosis of bee venom allergy, and a specificity of 83% and a sensitivity of 68% in the diagnosis of wasp venom allergy. Since there was no relevant significant correlation between the results of the basophil histamine release test on one hand and the severity of sting reactions and the prick test and RAST results on the other, it seems that the histamine release test determined additional parameters of sensitization. Thus, the method is a valuable adjunct to the in vitro methods available for the diagnosis of hymenoptera venom allergy.

Adolescent↗

Complement- and IgE-mediated release of histamine from basophils in vitro. V. Differential effects of drugs modulating arachidonic acid metabolism.

Basophil degranulation probably plays a significant role in the pathogenesis of different hypersensitivity reactions. These cells can be stimulated to secrete lysosomal histamine in vitro in response to various secretagogues. We compared the effects of drugs, modulating arachidonic acid (AA) metabolism, on histamine release (HR) from human basophils stimulated by anti-IgE antibody or C5a anaphylatoxin. Leukocytes from normal donors were preincubated with drug for 15 min at 22 degrees C, followed by the addition of C5a or anti-IgE for 30 min at 37 degrees C. Bromophenacyl bromide, an inhibitor of AA formation by phospholipase, blocked the effects of C5a and anti-IgE (greater than or equal to 3.3 X 10(-6) M, p less than 0.05 and p less than 0.01, respectively). 3-Amino-1-(3-trifluoromethylphenyl)-2-pyrazoline hydrochloride (BW755C, greater than or equal to 3.3 X 10(-5) M) and 5, 8, 11, 14-eicosatetraynoic acid (greater than or equal to 3.3 X 10(-4) M), known inhibitors of both cyclooxygenase (COX) and lipoxygenase (LPX) pathways of AA metabolism, blocked both C5a- and anti-IgE-induced HR (p less than 0.01). Nordihydroguaiaretic acid, an inhibitor of LPX, decreased HR induced by anti-IgE (greater than or equal to 3.3 X 10(-6) M, p less than 0.01) and allergens, but reduced C5a-initiated HR only at a higher concentration (greater than or equal to 7 X 10(-5) M, p less than 0.01). Indomethacin (INDO), an inhibitor of COX, significantly reduced HR caused by C5a (greater than or equal to 3.3 X 10(-8) M, p less than 0.01) and its degradation product C5adesArg, but had no effect or caused slight enhancement of HR initiated by anti-IgE. We confirmed that INDO augments allergen-induced HR. Our findings suggest that there are basic differences in the regulation of C5a- and IgE-mediated basophil degranulation.

4,5-Dihydro-1-(3-(trifluoromethyl)phenyl)-1H-pyraz↗

Granulopoiesis in tadpoles of Rana esculenta. Ultrastructural observations on the morphology and development of heterophil and basophil granules.

The morphology and development of heterophil and basophil granules from the trunk kidneys of Rana esculenta tadpoles were studied at the electron microscopic level. Cells of the heterophils series contain granules displaying either spheroid profiles with homogeneous content (Type A), or elongate profiles with a crystalloid interior (Type B). Type A granules apparently originate directly from Golgi-derived vesicles, which, gaining slightly in size and density, transform into the mature granules. Type B granules could be traced back to vacuolate structures showing an irregular content. Their development could be traced through increasingly elongated forms with the appearance of a characteristic crystalloid core. Fully developed basophil granules are considerably larger in size than heterophil granules and contain heterogeneous interna showing a central-cortical organisation: a finely stippled medullary zone of varying density is surrounded by a sickle-shaped and lamellate cortex (Type L), or a moderately dense and uniformly stippled medulla is enclosed by two diametrically opposed, cap-shaped, filamentous regions (Type F). The heterophil and basophil granules are compared to those in other vertebrates and possible phylogenetic aspects are discussed.

Animals↗

Basophils (Bsp-1+) derive from the leukemic clone in human myeloid leukemias involving the chromosome breakpoint 9q34.

The monoclonal antibody (MoAb) Bsp-1 was used to purify basophilic cells from leukemic blood of five patients with Philadelphia chromosome (Ph') positive chronic myeloid leukemia (CML) and two patients with acute myeloid leukemia (AML) characterized by the chromosomal translocation t(6;9)(p23;q34). When cultured, Bsp-1 positive cells from all CML and AML patients showed the same clonal karyotype changes observed in diagnostic buffy coat preparations, indicating that the basophilic cells were of leukemic origin. In contrast, T lymphocytes from four of five CML patients cultured in the presence of interleukin-2 (IL-2) showed a normal karyotype and were therefore not derived from the leukemic clone. Bsp-1 staining correlated with toluidine blue-positive basophils in chronic phase CML and with toluidine blue-negative blast cells expressing an immature myeloid phenotype in blast crisis CML and AML. Chromosome in situ hybridization showed that the ABL oncogene was translocated from chromosome 9 to chromosome 22 in the CML patients but remained on chromosome 9 in the AML patients. These results indicate that the breakpoint at 9q34 in CML is 5' of ABL, whereas the breakpoint at 9q34 in AML is 3' of ABL. Field inversion gel electrophoresis showed that the 9q34 breakpoint was not within 200 kb 3' of ABL in one of the AML patients, nor was there any rearrangement of the PIM oncogene locus at 6p21.

Antibodies, Monoclonal↗

The in vivo effects of recombinant human interleukin-3: demonstration of basophil differentiation factor, histamine-producing activity, and priming of GM-CSF-responsive progenitors in nonhuman primates.

Recently human interleukin-3 (IL-3) produced by molecular cloning was characterized as a growth factor for basophils and eosinophils in human bone marrow cultures. Since we found a similar activity of the human factor on simian bone marrow cells, we investigated the in vivo effects of recombinant human (rh) IL-3 in healthy rhesus monkeys (n = 10). rh IL-3 was administered subcutaneously (SC) to monkeys at different doses (11, 33, and 100 micrograms/kg/d) for 14 days followed by subsequent rh GM-CSF administration (5.5 micrograms/kg/d SC) for another two weeks. During the second week of rh IL-3 administration monkeys responded with a twofold to threefold increase of WBCs caused by a dose-dependent elevation of basophils (up to 40% of WBCs) and eosinophils. rh IL-3 also induced a dose-dependent increase of histamine (up to 700-fold above normal values) in monkey blood cells. Administration of rh GM-CSF to rh IL-3 pretreated monkeys resulted in a twofold enhanced increase in WBCs (due mainly to eosinophils and neutrophils) compared with animals treated with rh GM-CSF alone. Simultaneous administration of both cytokines (100 micrograms/kg rh IL-3 + 5.5 micrograms/kg rh GM-CSF SC) to two separate monkeys for 14 days induced a WBC elevation similar to that observed in monkeys treated with rh GM-CSF alone. In conclusion, our results indicate that rh IL-3 is a differentiation factor for blood basophils and primes the hematopoietic system for subsequent rh GM-CSF actions.

Animals↗

The role of mast cell and basophil activation in human allergic reactions.

Recently developed techniques of purification of human basophils and mast cells from different anatomical sites have made possible the investigation of the biochemical steps leading to mediator release and the pharmacological control of cells participating in allergic reactions. These techniques have also outlined differences between mast cells and basophils with regard to mediator release, activity of secretagogues and pharmacological modulation. The similarities and analogies between in vivo observations and in vitro findings with human mast cells and basophils suggest that the in vitro findings may represent important models for obtaining insight into the mechanisms and to develop therapeutic agents for the treatment of allergic diseases.

Animals↗

[Histophysiology of tissue basophils of the cerebral dura mater after laser irradiation].

By means of the histochemical and morphometric methods the reaction of the tissue basophils of the brain dura mater has been studied to the one-time and varying-duration (0.5 sec.-3 hr.) irradiation by a helium-neon laser of the wave length 632.8 nm, power density 0.76 mvt/sm2. It has been found, that the laser irradiation had a biostimulating effect upon the tissue basophils; the first peak of activity is in the case of a 3-second continuous irradiation; the second--from 15 min. to 1 hr. In symmetrical parts of the right (irradiated) and left (nonirradiated) regions of the dura very similar changes of the functional activity of the tissue basophils activity were seen.

Animals↗

The cutaneous basophil response to phytohemagglutinin in chickens.

A cellular infiltrate rich in basophils was observed in chickens in skin sites injected locally with the mitogen PHA. These infiltrating metachromatically granulated cells exhibited features common to mast cells of some species (monolobed nucleus), but also resembled basophils of other species in that they circulated in the blood, and they responded to the PHA mitogen by infiltrating skin test sites. Previous literature suggests that the chicken has a single metachromatic granulated cell and our data support this notion. PHA, a known T cell mitogen, may act by causing the stimulated T lymphocyte to produce a lymphokine that is responsible for the attraction of basophils to the test site.

Animals↗

[Basophil leukemia, development of a chronic myeloid leukemia. Apropos of a case and review of the literature].

Basophilic leukemia is a rare disease accounting for less than 2% of all hematopoietic malignancies and little is known about the best therapeutical approach. We report a case of basophilic leukemia occurring as an accelerated phase of chronic myelogenic leukemia in a 50 year old man with a past history of uranium and thorium exposure. He was successfully treated with intensive chemotherapy and total body irradiation followed by autologous peripheral stem cell transplantation. Such an approach has never been reported in this disease. However, the benefit on survival could not be evaluated because of a fatal opportunistic infection. The cases of basophilic leukemia reported in the literature are discussed.

Basophils↗

A newly described activity of guinea pig IgG1 antibodies: transfer of cutaneous basophil reactions.

Hapten-specific delayed time course skin reactions containing predominant accumulations of basophils and eosinophils were elicited in newborn guinea pigs after i.v. transfer of small amounts of oxazolone immune serum. The immune serum was fractionated by column chromatography procedures, and the fractions were examined for their ability in transferring this form of cutaneous basophil hypersensitivity (CBH). Only the 7S IgG-containing peak from Sephadex G-200 columns, and only the IgG1-containing fractions from DEAE columns, transferred CBH. An affinity column of bound oxazolone removed the activity from immune serum, and it could be recovered from the column by eluting with soluble oxazolone. About 35 microgram of purified IgG1 anti-oxazolone antibody could systemically transfer CBH reactivity. An immunoadsorbant column of anti-IgG1 removed this activity, but a column of anti-IgG2 did not. None of the procedures were able to separate activity in transferring CBH from passive cutaneous anaphylactic (PCA) activity classically associated with guinea pig IgG1 antibody. IgG1 from 8-day immune and 31-day hyperimmune donors were both effective. The average association constant of 8-day antibody was 8 X 10(-4) M-1. Transfer of cutaneous basophil reactions can be mediated by low affinity serum 7S IgG1 antibody.

Animals↗

Cell membrane glycoproteins of human mast cells: a biochemical comparison with basophils.

The relationship of mast cells (MCs) to other blood leucocytes, and to basophils in particular, is unclear. The relative distribution and abundance of cell surface glycoproteins of normal and neoplastic blood cells has been established as providing a "fingerprint" allowing assignment to a particular hemopoietic cell lineage. We have examined the major labeled glycoproteins of mast cells, basophils, HL-60 cells (granulocytic), and U937 cells (monocytic), after cell surface tritiation by the periodate-borohydride technique. Mast cells have a characteristic glycoprotein profile, different from that of basophils and other leukocytes; a major feature is the lack of the gp105-115 molecular weight band common to all other white blood cells. The data suggest that the tissue mast cell is more distantly related to other hemopoietic cells than has been previously recognized.

Basophils↗

Differences between basophils and mast cells: failure to detect Charcot-Leyden crystal protein (lysophospholipase) and eosinophil granule major basic protein in human mast cells.

Human eosinophils contain several distinctive proteins including eosinophil granule MBP and the membrane-associated CLC protein (lysophospholipase). Human basophils also contain these proteins, indicating biochemical similarities between eosinophils and basophils. To determine whether MBP or CLC protein is present in connective tissue mast cells, we studied human lung and cutaneous mast cells by immunofluorescence by utilizing specific antibodies to CLC and MBP. Cytocentrifuge slides of enriched lung mast cells and mast cells in sections of formalin-fixed, paraffin-embedded cutaneous tissue from urticaria pigmentosa lesions were stained for CLC and MBP. Neither pulmonary nor cutaneous mast cells stained for CLC protein or MBP. In contrast, lung and cutaneous eosinophils in the same preparations showed bright staining for both proteins. The failure to find CLC protein and MBP in mast cells provides additional evidence of dissimilarity between mast cells and basophils, and an immunochemical means to distinguish between them.

Basophils↗

Quantitative aspects of the T cell proliferation response to antigen P1 from D. pteronyssinus: suppression by added histamine and limited effects of basophil depletion.

We have recently shown that peripheral blood mononuclear cells from patients sensitive to the house dust mite, D. pteronyssinus, will proliferate in vitro in response to the purified major allergen, antigen P1. Such cell populations, separated on Ficoll gradients, were shown to contain basophils, and had an average histamine content of 12 ng/10(6) cells. Incubation with antigen P1 resulted in the release of histamine, and histamine is known to activate T suppressor cells. In the present experiments we observed up to 80% inhibition of proliferation (mean 50-60%) with histamine added at 3.3 X 10(-7)-3.3 X 10(-5) M. Cultures of T cells supplemented with irradiated non-T cells, that had been depleted of cells bearing surface membrane IgG, IgM and IgE, contained on average 63% less histamine than unseparated cultures. However, no consistent difference in the proliferative response to antigen P1 was observed. Depletion of histamine by pre-incubation of the cells with antigen P1 at 10(-3)-10(-4) micrograms/ml followed by washing of the cells before culture also produced no significant change in the proliferative response. Passage of cell population over nylon wool resulted in depletion of basophils, as well as other cell types, and generally led to a decrease in proliferation. We conclude that release of mediators from basophils in cell cultures does not markedly affect the magnitude of the proliferative response to antigen P1. The varied responses seen with cells from different individuals are likely to reflect differences in the numbers of circulating allergen sensitized T cells.

Allergens↗

Ultrastructural heterogeneity of the basophilic cells in the allergic nasal mucosa.

Basophilic cells in scrapings and pieces of nasal mucosa removed from the inferior turbinates of adult patients with house dust nasal allergy plus pieces of the oral mucosa from normal persons were examined electron microscopically. Three distinct types of basophilic cells, ie, blood basophil leukocytes (predominantly in the subepithelium), mucosal mast cells (predominantly in the epithelium), and connective tissue mast cells (predominantly in the deeper lamina propria) were identified in the nose.

Basophils↗

Slow reacting substance (SRS) from ionophore A23187-stimulated human leukemic basophils. I. Evidence for a precursor role of arachidonic acid and initial purification.

SRS was generated from human leukemic basophils upon stimulation with ionophore A23187. Radiolabel from [14C]-AA was incorporated into SRS with continued comigration of radioactivity and bioactivity through several chromatographic systems including DEAE-cellulose, silicic acid, and RP-HPLC. Human basophilic leukemia SRS displayed physiochemical properties similar to those of rat basophilic leukemia cell SRS.

Adult↗

Some quantitative aspects of the passive sensitization of human basophils.

We review the theory for the binding of IgE to Fc receptors on basophil surfaces. We then use the theory to analyze binding experiments of Malveaux et al. and T. Ishizaka on the passive sensitization of basophils from a patient with chronic myelocytic leukemia and 75% basophilia. From their data we determine that the forward rate constant for the binding of human IgE to receptors on these human basophils is (3.0 +/- 1.0) x 10(4) M-1 sec-1.

Basophils↗