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As part of a screening and treatment program for intestinal parasite infections offered to newly arrived Southeast Asian refugees in Canada between July 1982 and February 1983, a total of 232 sera were tested for Strongyloides infection using an enzyme-linked immunosorbent assay (immunoglobulin G). These results were compared with coprologic results and eosinophil counts. The seroprevalence was 76.6% (131 of 171) among Kampucheans, 55.6% (15 of 27) among Laotians, and 11.8% (4 of 34) among Vietnamese. A statistically significant relation (p less than 0.001) was found between Strongyloides serology and Strongyloides infection on stool examination (prevalence, 24.7%) among Kampucheans. Eosinophilia (greater than or equal to 10%) was found to be significantly associated with both infection measures. Using coprologic results as the "gold standard," the properties of the serologic test were estimated to be: sensitivity (95%), specificity (29%), positive predictive value (30%), and negative predictive value (95%). These estimates should be regarded as minimal values, as stool examination for Strongyloides infection can be an unreliable diagnostic reference. Further evaluation of the discrepancies observed between coprologic and serologic testing is required to determine the usefulness of these tests in epidemiologic studies.

The sera of eight endomyocardial fibrosis (EMF) subjects, 11 siblings of one of them and 16 normal children matched with the EMF patients for age, sex and socio-economic status from Ogunmakin and Shao/Oloru communities (eight each), situated in EMF-endemic and non-endemic areas of Nigeria respectively, were examined for the presence of antibodies against Coxsackie viruses B1-6, 16 arboviruses and Toxoplasma gondii. Sera from 36 other randomly selected normal children from Ogunmakin and 26 other randomly selected children from Shao/Oloru were also tested for the presence of antibodies against Toxoplasma gondii and the 16 arboviruses. None of the eight EMF subjects nor the 11 siblings of one of them had antibodies against any of the Coxsackie viruses B1-6 in their sera. Two of the 16 matched control subjects, one from each community, had positive antibodies, at equivocal titres against Coxsackie B1 (Ogunmakin) and B4 (Shao/Oloru). There was no significant difference in the distribution of antibody titres to the arboviruses between the EMF patients and matched controls. Normal children from the Shao/Oloru community had higher percentage antibody reactions and higher titres to the arboviruses compared with the children from Ogunmakin. All the eight EMF patients had high antibody titres against Toxoplasma gondii. Seven (87.5%) of the matched controls from Ogunmakin were sero-positive for Toxoplasma gondii compared with three (37.5%) of the matched controls from Shao/Oloru. Of the 36 normal children from Ogunmakin, 32 (88.9%) were sero-positive compared with 11 (42.3%) of the 26 normal children from Shao/Oloru. Four (36.4%) of the 11 siblings of one of the EMF patients had weak sero-positivity. It is therefore concluded that further studies are needed to clarify the role, if any, of Toxoplasma gondii in EMF.

Immediate and late cutaneous reactions were induced in healthy and atopic subjects by anti-IgE challenge and a skin-window technique was used in order to verify if the pattern of cells observed at 24 hr was similar in both groups. The study was first performed under basal conditions, and in a second double-blind cross-over step, it was performed again during treatment with cetirizine 10 mg b.i.d. and terfenadine 60 mg b.i.d. Anti-IgE challenge was followed by a significant eosinophil accumulation in atopic subjects only. Cetirizine significantly inhibited this phenomenon while terfenadine showed a mild non-significant inhibitory effect.

To test whether neutrophils infiltrate and degranulate in areas of chronic respiratory allergic inflammation, we developed an indirect immunofluorescence technique to localize neutrophil elastase in formalin-fixed, paraffin-embedded tissues. The affinity-purified antielastase stained only neutrophils on peripheral blood buffy coat smears, and in lung tissue from patients with pneumonia. We examined tissue specimens from four patients with fatal asthma, 10 patients with chronic sinusitis, and 10 patients with nasal polyposis for the presence of elastase, as well as eosinophil granule major basic protein (MBP). Neutrophil infiltration and extracellular elastase deposition in association with damage to respiratory epithelium were generally sparse in most specimens; the exceptions were one patient with asthma, one patient with chronic sinusitis, and two patients with nasal polyposis. In contrast, eosinophil infiltration and extracellular MBP deposition were generally marked in most specimens; the exceptions were one patient with asthma and one patient with nasal polyps where extracellular MBP deposition did not coincide with damage to respiratory epithelium. The results suggest that the neutrophil does not usually infiltrate tissues showing allergic inflammation; however, on occasion, it may participate in these inflammatory reactions.

Purified human eosinophils generate eosinophil chemotactic lipids (ECL), when incubated with arachidonic acid without any stimulus. Reversed phase HPLC of incubation supernatants revealed major lipid-like eosinophil chemotactic activity eluting in a peak containing 5(S), 15(S)dihydroxy-6,13-trans-8,11-cis-eicosatetraenoic acid (5,15-DiHETE) as well as a 8,15-dihydroxyeicosatetraenoic acid. For further characterization of the ECL, some authentic dihydroxyeicosatetraenoic acids were tested for eosinophil chemotactic activity. Only 5,15-DiHETE as well as 8(S), 15(S)-dihydroxy-5,11-cis-9,13-trans-eicosatetraenoic acid were found to be chemotaxins for human eosinophils, however, with an ED50 near 0.3 microM and 1.5 microM, respectively. The presence of high titer eosinophil chemotactic activity in ECL preparations let us look for a contaminating ECL with higher specific activity. By using a different reversed phase HPLC-system 5,15-DiHETE as well as 8(R), 15(S)-dihydroxy-5,11-cis-9,13-trans-eicosatetraenoic acid could be separated from a highly potent ECL. Final purification of this ECL by the use of straight phase HPLC resulted in a single at 260 nm absorbing peak giving an UV spectrum different from that known for eosinophil chemotactic factors indicating a novel type of eosinophil chemotactic lipid. Eosinophil chemotactic activity of purified ECL has been found to be similar to that seen for platelet-activating factor, the most potent chemotaxin so far known, either in the number of migrating cells or the ED50. Cross-desensitization experiments with ECL, leukotriene B4, and platelet-activating factor revealed the existence of a separate ECL receptor on eosinophils. The production of potent ECL by the responder cells themselves supports the idea that there exists a self-sustaining mechanism of eosinophil accumulation.

Fc gamma RIII on neutrophils is a phosphatidyl inositol glycan (PIG)-anchored protein that can be released from the cells by activation with chemotactic peptides. We have examined the expression of Fc gamma RIII (CD16), CD11b, and Fc gamma RII (CD32) on neutrophils from human immunodeficiency virus (HIV)-I-infected individuals by two-color FACS. In patients with AIDS and AIDS-related complex and in HIV-I positive intravenous drug abusers we observed a substantial population (25%) of neutrophils that were autofluorescent, and did not stain with the anti-Fc gamma RIII mAb 3G8. This population was largely absent (3%) in HIV-I negative control individuals. No changes in the expression of Fc gamma RII, CD11b, or another PIG-anchored protein, decay accelerating factor (CD55) on neutrophils, were found. The presence of the Fc gamma RIII negative neutrophil population may be related to altered functions leading to common bacterial infections in advanced AIDS.

We have studied the expression of Fc gamma receptors and leucocyte integrins (CD11/CD18 family) on human eosinophils using specific monoclonal antibodies (mAb) and flow cytometric analysis. Peripheral blood eosinophils of normal density and low density were compared with neutrophils and monocytes. Several properties of the human eosinophil were established. These were (i) that the eosinophil expressed Fc gamma RII (CDw32) only (unlike monocytes which bear Fc gamma RI and Fc gamma RII and neutrophils which bear Fc gamma RII and Fc gamma RIII); (ii) that the absence of Fc gamma RIII (CD16) on eosinophils served as a basis for distinguishing eosinophil and neutrophil populations by immunofluorescence; (iii) that the leucocyte adhesion glycoproteins, LFA-1 alpha (CD11a), CR3-alpha (CD11b), p150, 95-alpha (CD11c) and the common beta-chain (CD18), were expressed on the eosinophil as well as the neutrophil; (iv) that CD18 expression was significantly reduced on low-density eosinophils from the hypereosinophilic syndrome. Thus, our findings emphasize the unique phenotype of the human eosinophil in terms of Fc gamma receptor expression, the similarity of the eosinophil and neutrophil with regard to the leucocyte integrins and that eosinophils of low density do not differ greatly from those of normal density in terms of receptor expression.

Absolute T and B lymphocytes, assayed by E and EAC rosette techniques, serum IgE levels, total eosinophil counts and delayed hypersensitivity cutaneous reactions were studied in 39 asthmatic subjects (19 extrinsic asthmas and 20 intrinsic asthmas) and in 15 age-matched control subjects. The mean value of T lymphocytes was 1753 +/- 709 mm.3 in the extrinsic group, 1511 +/- +/- 530 mm.3 in the intrinsic, and 1609 +/- 451 mm.3 in the controls. There was no statistical difference between the two groups of patients and the controls (P > 0.05). There was also no statistical difference of B lymphocytes between the two groups of patients and the controls. Serum IgE was elevated in 8 extrinsic asthmatics (42.1%), in one intrinsic (5%), and in none of the control population. The mean value of eosinophils ws significantly higher (P < 0.01) in the extrinsic group as compared to the controls. The same was true of the intrinsic group, but not so significantly (P congruent to 0.02). No correlation was observed between the number of T lymphocytes and the skin tests, serum IgE and total eosinophil counts.

In the uterus of the adult females rats, the luminal epithelial cells and the eosinophil leukocytes are rich in cytoplasmic estrogen receptors. During the estrous cycle, the epithelial estrogen receptor concentration reaches its peak level in proestrus, drops precipitously in estrus, and hits the trough at metestrus. Repopulation of the cytoplasm with estrogen binding sites occurs during diestrus. This pattern of cyclic change is indicative of a rapid turnover of estrogen receptors inthe epithelial cells and its regulation by endogenous estrogens. The concentration of estrogen receptors in the cytoplasm of the eosinophils does not appear to fluctuate during the cycle. But the intrauterine distribution of these leukocytes is clearly cyclic in pattern, ostensibly influenced by estrogens. While progesterone binding activity is consistently demonstrated in tandem with estrogen receptors in the cytoplasm of the epithelial cells, it had not been observed in the eosinophil leukocytes. These findings support the claim that there are two estrogen receptor systems in the rat uterus, one mediating the intracellular events of the genomic response to estrogens, and the other being concerned with non-genomic responses.

The authors report a study of the modifications observed with sutures of PTFE (Goretex) and polyglactin (Vicryl) at the level of the sclera and the conjunctival. Inflammatory cells are fewer with PTFE both in the rabbit eyes (14 cases) from 3 to 110 days and in the human eye in a clinical study of 28 cases.

Complex carbohydrates in eosinophils from human, rabbit, and rat marrow were identified and localized by cytochemical and radioautographic methods. The high iron diamine-thiocarbohydrazide-silver proteinate (HID-TCH-SP), low iron diamine (LID)-TCH-SP, and periodate (PA)-TCH-SP methods were used for the localization of sulfate, sulfate and carboxyl, and vicinal glycol- containing complex carbohydrates, respectively. Golgi vesicles and small precursor granules (0.2-0.4 micrometer in diameter) demonstrated strong HID-TCH-SP staining and labeled intensely after a 10-minute pulse with 35SO4(2). Crystalloid-free or immature specific granules (0.5-0.9 micrometers in diameter) labeled heavily after a 60-minute incubation and 60-minute chase with 35SO4(2). Immature granules were graded according to their HID-TCH-SP staining. Type I granules demonstrated strong rim staining and similar or somewhat less central staining, whereas type 2 granules only demonstrated rim staining, and type 3 granules lacked staining. Fully mature crystalloid-containing granules lacked staining. LID-TCH-SP similarly stained the HID-positive sulfated material in cytoplasmic granules. PA-TCH-SP stained some Golgi vesicles and diffusely stained all precursor granules and type 1 granules. Weaker staining was observed in type 2 granules and staining was very weak or absent in type 3 and crystalloid-containing granules. In early eosinophils, tubulovesicular structures (TVS) were observed rosetting and contacting precursor and type 1 granules. These TVS contained material with strong PA-TCH-SP staining but lacked HID-TCH-SP or LID-TCH-SP-reactive acidic glycoconjugates. Flattened Golgi saccules or early eosinophils stained weakly or not at all with the PA-TCH-SP method. Small granules and TVS in late (bilobed) eosinophils displayed PA-TCH-SP reactivity and lacked HID-TCH-SP staining but differed from TVS in early eosinophils in that they were not associated as rosettes with specific granules. These results indicate that sulfated and vicinal glycol-containing complex carbohydrates are differently disturbed in immature specific granules of eosinophils and presumably become masked to staining as the granule matures.

Leucocyte B12 and B12-binding capacity were measured by Simultrac radioassay in eosinophilic granulocytes, neutrophilic granulocytes and leucocytes obtained from patients with chronic granulocytic and lymphocytic leukaemia. It is shown that (a) eosinophils are a possible source of B12-binding protein similar to neutrophils and (b) granulocytes in myeloproliferative disorders and normal neutrophils have similar B12 and B12-binding capacity indicating that increased B12 and B12-binding capacity in myeloproliferative disorders arise from an increase in myeloid cell turnover.

As the significance of the eosinophilia which is found in some patients with chronic filarial infections is unknown, blood eosinophils were studied in seven patients with tropical (filarial) eosinophilia who had blood eosinophil counts between 5.2 and 56.6 x 10(9)/l. In six of these patients over 20% of the eosinophils were vacuolated, and degranulated eosinophils were found in the three patients with the most severe illness. In four patients serum eosinophil cationic protein levels were raised, and serial studies in two showed that they returned to normal after treatment with diethylcarbamazine. In addition, an increased proportion of eosinophils in these patients were able to bind to complexed IgG suggesting that they were responding to stimuli in the blood, and in one patient the blood eosinophil half-time clearance was prolonged. Labelled cells were cleared into the spleen, liver and bone marrow. These findings support the hypothesis that in patients with tropical (filarial) eosinophilia, blood eosinophils are induced to release their granule constituents into the circulation. It is suggested that these secretion products may interact with microfilariae, and may give rise to some of the clinical features of chronic filarial infections.

The value of three objective tests of the nasal mucosa in 22 patients with perennial non-allergic rhinitis treated with a topical corticosteroid has been investigated. Placebo and three different dosages of a new steroid, budesonide, were administered intranasally using a double-blind, cross-over technique. Nasal airway resistance was not reduced by the steroid treatment as compared with the placebo treatment. Eosinophilia in nasal smears was reduced by budesonide. However, the value of this test was decreased by the fact that eosinophilia was found in less than 50% of the patients at the beginning of the trial. Metacholine-induced nasal secretion was significantly reduced by the budesonide treatment. Symptom scores for nasal obstruction, secretion and sneezing attacks were also significantly reduced by the steroid.

An anion-exchange high-performance liquid chromatography method has been used to quantitate the intracellular purine and pyrimidine nucleotides in extracts of pure lymphocytes, monocytes, neutrophils, eosinophils, erythrocytes, and platelets isolated from the blood of healthy human donors. For accurate and reproducible measurements of the nucleotide profiles in different types of pure leukocytes, the cell suspensions have to be free of platelets and erythrocytes. Incubation of the purified leukocytes for 1 h at 0 degrees C did not alter the nucleotide concentrations but reduced the interdonor variation to 10%. Incubation of purified lymphocytes for 1 h at 37 degrees C caused considerable changes in the relative concentrations of the adenine, guanine, uracil, and cytosine nucleotides. During this incubation the cell viability, the cell number, and the ATP:ADP ratio decreased. Incubation of monocytes and granulocytes for 1 h at 37 degrees C caused considerable loss of cells and/or cell death. For erythrocytes and platelets reproducible nucleotide concentrations were obtained after extraction of freshly isolated cells. During storage of erythrocytes, both at 0 degrees C and at 37 degrees C, a decrease in the ATP:ADP ratio was detected. In all cell types the predominant nucleotides were purine nucleotides, especially adenosine triphosphate. The relative concentrations of the adenine, guanine, uracil, and cytosine nucleotides were very reproducible per cell type and appeared to be characteristic for each cell type. The total nucleotide content was nearly the same for all cell types except erythrocytes, when expressed per microgram of protein. The described methods for purification and storage of blood cells will be useful for comparison of blood cells from healthy donors with those of patients, for example, leukemia patients, in which deviations of the purine and pyrimidine metabolic enzymes have already been described.

Fibrosis complicates a number of chronic inflammatory diseases and occurs in some conditions following chronic hypereosinophilic syndromes. We assessed whether eosinophils might be a source of fibrogenic factors. Extracts of human and guinea pig cell populations enriched for eosinophils contained substances that stimulated tritiated thymidine incorporation by human fibroblasts. Supernatants derived from resting eosinophils and extracts prepared from eosinophil granules also contained fibrogenic factors. Our findings demonstrate a new potential role for eosinophils and suggest a causal relationship between tissue eosinophilia and scar formation in certain parasitic conditions.

A 2 1/2-year-old girl presented with monthly episodes of angioneurotic edema, eruption of pruritic papules, and fever. During acute episodes, white blood cell counts rose as high as 52,100/cu mm with 62% eosinophils, and body weights increased up to 20% of remission weight. Short courses of prednisone acetate caused rapid defervescence, resolution of angioneurotic edema, and lowering of eosinophil counts. In a one-year follow-up no evidence was found for cardiac or other visceral organ involvement. Findings of extensive diagnostic evaluations revealed no evidence for atopy, neoplasm, collagen-vascular disease, or parasitic infestation. Results of immunologic studies were essentially normal with the exception that this patient had a high level of circulating activated helper T cells. Biopsy specimens of the skin lesions revealed dermal infiltration of lymphocytes and eosinophils with deposition of eosinophil major basic protein in the extracellular matrix. Awareness of this clinical entity and its distinction from the hypereosinophilic syndrome is important because of its favorable prognosis and rapid response to corticosteroid therapy.

In this article we present a patient with acute lymphoblastic leukemia (ALL) associated with eosinophilia, in which the eosinophilia preceded a meningeal and bone-marrow relapse of ALL. We analysed the purine and pyrimidine nucleotide content of the eosinophils (92% pure) and compared the nucleotide pattern with that of eosinophils from healthy donors and from patients with eosinophilia not associated with leukemia. The ratios of purine:pyrimidine and of uracil:cytosine nucleotides were decreased compared with those in eosinophils from healthy donors and from patients with eosinophilia with other aetiologies. The total nucleotide concentration was increased, especially the concentration of UDP-sugars and pyrimidine nucleotides. The decrease in these ratios and the increase in concentration of the nucleotides and the UDP-sugars were also detected in leukemic cells of patients with ALL (de Korte et al., Leukemia Res. 10, 389-396 (1986) compared to normal lymphocytes. We suggest a malignant character of the eosinophils in our patient with ALL associated with eosinophilia, in contrast with the non-malignant state suggested previously for these cells.

An immunoelectron microscopic technique using protein A-gold as a specific marker was used for precise intracellular localization of eosinophil granule proteins. Eosinophils from healthy individuals were isolated in metrizamide gradients. Eosinophil cationic protein (ECP) and eosinophil peroxidase (EPO) were clearly located in the matrix of the large crystalloid-containing granules. In addition, ECP was probably present in the small granules of eosinophils. Major basic protein (MBP) was present in the crystalloid structure of specific granules. This method can be applied in studies of eosinophil degranulation to trace the release of biological effector molecules.

Lectin from Griffonia simplicifolia (GSA-I) and soybean (SBA) are reliable markers for human eosinophils. In this study we have shown that fluorochrome labeled GSA-I and SBA can be used for specific labeling of eosinophils in paraffin embedded tissue sections, in peripheral blood smears and in cell suspensions prepared for flow cytometry. These two lectins are useful diagnostic reagents which could be applied for further characterization of cytoplasmic components selectively found in human eosinophils.

In the case of 36 female patients who were anamnestically known to have taken laxatives, semiquantitative histological investigations with laparoscopically obtained liver needle biopsies were effected after the exposition with preparations containing phenolisatine. The time gap until exposition was 12 to 24 h (16 cases), 48 h (8 cases), 72 to 96 h (4 cases) and 7 to 14 days (4 cases). The histological result after the exposition is an acute cholangiolitis of the allergic-hyperergic type with edema and a dense eosinophile infiltration of the portal fields with destruction of the epithelium of preformed bile ducts and portally proliferated ductles. In addition, the parenchyma of the liver shows a pleomorphism of the cells in form and colour with a cellular edema and with disseminated acidophilic necroses and necrobioses of the individual cells as well as with little reactive proliferation of the Kupffer's cell. After a period of 8 days the acute process has more or less subsided. Also, in the majority of cases there are histological signs of an aggressive chronic hepatitis of type IIa, partially in the active stage with piece-meal necroses and partially stabilized or in the process of healing. A transition to the picture of hepatitic cirrhosis is possible. In serious cases the picture of a chronic non-purulent destructive cholangitis can be simulated by the hepatocellular and canalicular damage. Thirty-one bioptic pre-examinations from the same results, whereby the acute cholangiolitical exacerbation can be attributed to an exposition of the patients themselves. The clinical picture of the phenolisatine damage in its entirety is induced by medication and is described as a recurrent chronic cholangiohepatitis. Similarities exist between the liver damages caused by chlorpromazine and arsphenamine. When medication is discontinued, the morphologic substrate recedes leaving behind an inactive fibrosis or cirrhosis. The formal and known causal pathogenetic connections are discussed with regard to this clinically important liver disease. Guidelines are then given for histological diagnosis of this damage caused by medication. 14% of the female patients with a histological picture of aggressive chronic hepatitis and hepatitic cirrhosis are affected by this type of liver damage.

Local cellular and extracellular (acid mucopolysaccharide) reactions in human lung cancers and metastases - prognostic parameters? The role of immunologic reactions in human malignant tumors is still unsettled. In the work presented, 72 primary lung cancers and 17 pulmonary metastases of 8 different organs of origin were studied with histochemical techniques for cellular infiltrates (eosinophils and macrophages) and extracellular acid mucopolysaccharides (AMPS). The results showed a positive correlation between eosinophils and macrophages and a negative correlation between cellular reactions and the deposition of AMPS. The clinical course of radically operable patients with primary lung cancer 2 to 3 1/2 years after operation showed that patients with the reactive pattern "strong local eosinophilia, no AMPS deposition" had the best prognosis with 12 of 14 patients free of tumor. Of the patients with the same tumor stage, but lacking local eosinophilia, more than half died from the tumor or had generalized metastases within the same period. The most frequent pattern in patients with pulmonary metastases was "no local eosinophilia, strong AMPS reaction". These observations suggest that cellular reactions in radically operable primary lung cancer express immunologic responses. These are often absent in metastases.

Skin testing represents a direct method of assessing immune responses in vivo. Twenty-six patients with metastatic cancer of the lung, kidney, or melanoma were treated with adoptive transfers of autologous tumor-infiltrating or blood lymphocytes and continuous infusions of interleukin-2 (IL-2). Prior to therapy, cutaneous anergy to recall antigens was observed in 19 patients (73%), whereas 6 (27%) displayed normal delayed-type hypersensitivity (DTH) responses. When tested again at the end of therapy, DTH responses could not be elicited in any of the patients. Proliferative responses to skin test antigens, lectins, and IL-2 diminished progressively during therapy but returned to baseline values at 1 month. Unexpectedly, 14 of these patients (53%) developed immediate skin test responses to candida antigens and 5 (19%) to mumps antigens. These immediate responses were characterized by local erythema and induration that developed within minutes of injecting antigen. Biopsies displayed marked dermal edema and infiltration by eosinophils. Although serum IgE levels were not increased, immediate reactivity could be transferred by a heat-sensitive serum factor. The implications of this novel response are uncertain, and its development did not correlate directly with the anti-tumor effects of therapy. We conclude that adoptive immunotherapy with IL-2 produces a reduction in cutaneous DTH and diminished responses to mitogens while simultaneously promoting cutaneous allergy. We hypothesize that this may reflect diminished IL-2 production by antigen-specific helper T cells and that other lymphokines may promote these immediate hypersensitivity responses.

The distribution of peroxidase and acid phosphatase activity in the bone marrow of mouse eosinophils was investigated by electron microscopy. Peroxidase activity was found in the perinuclear space, the endoplasmic reticulum, the Golgi complex, non-mature and mature specific crystal-containing granules. In the course of development peroxidase activity disappears from the cisternal system. In mature eosinophils the enzyme is stored in specific granules, but some of these failed to reveal peroxidase activity. A hypothesis is offered concerning the complete condensation into crystal of the enzyme. In cells incubated in peroxidefree media some granular components stained weakly. Their activity probably depended on the presence of endogenous non-organic peroxidase. In developing eosinophils acid phosphatase was found in the Golgi complex and in non-mature specific granules. In the course of development, acid phosphatase disappears from the granules.

In experiments using an immunofluorescent method to localize human eosinophil granule major basic protein (MBP) in cells and tissues, a small number of cells stained for MBP that subsequently could not be identified as eosinophils. Because the Charcot-Leyden crystal protein, another eosinophil protein, was recently identified in basophils, we tested whether MBP might also be a constituent of blood basophils. Highly purified, eosinophil-free basophil suspensions were prepared using the fluorescence-activated cell sorter (FACS) to sort basophil-containing mononuclear cell preparations stained with fluorescein-conjugated sheep IgG anti-human IgE antibody. Using these FACS-purified basophils, we found that: (a) enrichment for surface IgE-positive cells (greater than 95% basophils) by FACS also enriched for cells staining for MBP by immunofluorescence; (b) MBP appeared to be localized in the histamine-, heparin-containing granules; (c) significant quantities of MBP were measurable by radioimmunoassay (RIA) in freeze-thaw detergent extracts of purified basophils; and (d) RIA dose-response curves for extracts of purified eosinophils and basophils had identical slopes. The MBP content of basophils from normal individuals averaged 140 ng/10(6) cells, whereas purified eosinophils from normal donors and patients with the hyper-eosinophilic syndrome averaged 4,979 and 824 ng/10(6) cells, respectively. MBP was also detected by immunofluorescence and RIA in cells obtained from a patient with basophil leukemia, although the MBP content for basophil leukemia cells was lower than that for normal basophils. We conclude that basophils contain a protein that is immunochemically indistinguishable from eosinophil granule MBP.

Five patients with eosinophilic lung diseases and blood hypereosinophilia (PIE syndrome) were investigated clinically and by bronchoalveolar lavage (BAL). Comparative studies on blood and alveolar eosinophils were carried out after purification and selection of eosinophil subpopulations according to their density. A predominant 'hypodense' alveolar eosinophil population was found in BAL fluids of active chronic eosinophilic pneumonia (CEP). In addition, supernatants of alveolar macrophages obtained from CEP are able to enhance spontaneously the generation of eosinophil oxygen metabolites. Such eosinophil stimulation emphasizes a probable tissue cell cooperation. In addition, BAL permitted the study of membrane immunological markers on eosinophilic inflammatory cells endowed with migratory properties. An increase in eosinophils carrying surface IgE was demonstrated in alveolar cells from PIE Syndrome particularly with hypodense eosinophils from CEP patients. Although no specific stimulus is known at the present time, this work underlines the potential implication of IgE-mediated hypersensitivity processes in the pathogenesis of eosinophilic lung diseases.

Local infiltrates of eosinophilic leucocytes and macrophages and the deposition of acid mucopolysaccharides (AMPS) in 72 operable primary lung cancers and 17 isolated pulmonary metastases of known origin were correlated to tumour stage (radically or non-radically operable) and clinical course, by following the patients for 2-3 1/2 years. Half of the primary lung cancers showed strong local eosinophilia which, in combination with either strong macrophage infiltration or absence of AMPS reaction, characterized a very good prognosis in radically operable patients. No eosinophils, together with a strong AMPS reaction, indicated a very poor prognosis, irrespective of tumour stage. 16/17 metastases (7 different histologies) had either no local eosinophilia (13), strong AMPS deposition (12) or both (9). This suggests that malignant clones with great metastatic potential in general are characterized by absence of local eosinophilia and/or a strong AMPS reaction. These observations taken together indicate that local eosinophilia expresses an immune reaction which is, houl metastatic clones. It if does, metastatic success may be due to an escape mechanism based on the elaboration of AMPS.

The central 95 percentile estimates of the normal white cell types (as determined by a standard differential count) were calculated from 777 normal individuals. The results were divided into groups by age and sex and expressed both as percentages and as absolute numbers of cells.A similar survey was made using the prototype of a new automated differential counting method (Hemalog-D(TM)). The two sets of laboratory values showed no statistically significant differences regarding age or sex and were strikingly similar in most cell types with the exception of band and juvenile forms of granulocytes. As expected, the normal range using the manual technique was somewhat broader than that found with the automated method. In addition to being the first comparison of machine counts with visual differentials this study provides additional insight into the age distributions of normal cells.

Incubation of eosinophils with schistosoma of Schistosoma mansoni in vitro induces changes in the eosinophil membrane. Cell surface proteins were blocked with unlabelled iodide and then, after incubation with schistosomula, newly accessible proteins were detected by lactoperoxidase catalysed iodination with radioactive iodide. Normal turnover restores several proteins to the eosinophil surface, but the schistosomula specifically induced the appearance of a protein of mol. wt 18K. This protein has previously been detected when eosinophils interact with antibody coated agar layers. Soluble factors released by schistosomula induced the appearance of enhanced levels of 16 and 18K proteins. It has previously been shown that proteins involved in binding of eosinophils to IgG coated antigen coupled cellulose have mol. wts of 16K and 18K.

Serum IgE levels and nasal/aural secretion eosinophils and basophils cells were studied in 84 patients with ear discharge despite previous radical operation, in 83 patients with postoperatively dry ears, and in 30 normal persons. Serum IgE values suggesting atopy (greater than 100 U/ml, PRIST) were more frequent (p less than 0.05) among patients with draining ears than among persons with healthy ears. Normal IgE concentrations showed reversed frequency distribution. Nasal secretion eosinophils and/or basophilic cells were also found more frequently (p less than 0.05) among patients with discharging ears than in other groups. Analogously, those with both serum IgE of greater than 100 U/ml and 'positive' nasal cytology more often had infected than dry (p less than 0.01) or normal (p less than 0.05) ears. Aural secretions showed eosinophils and/or basophilic cells in 14 (17%) patients; 11 of them had such cells in their nasal secretions too. It would seem that atopy is probably one cause of persistent therapy-resistant otitis media and it must be taken into account when considering the treatment of a chronic ear.

A recent report by Lyttle et al. (Lyttle, C. R., Medlock, R. L., and Sheehan, D. M. (1984) J. Biol. Chem. 259, 2697-2700) suggested that nuclear type II sites in the rat uterus are of eosinophil origin and may represent [3H]estradiol binding to eosinophil peroxidase. To further evaluate this hypothesis we examined the response of nuclear type II sites to estrogen under conditions where eosinophils are not present. Results of our experiments show that physiological levels of estradiol-17 beta (10 nM for 72 h) will stimulate nuclear type II sites in highly purified cultures (21-25 days; 4 passages) of rat uterine stromal and myometrial cells. The magnitude of the response of type II sites to estradiol in these stromal (4-fold) and myometrial (80-fold) cell cultures was essentially identical to that observed in the uterine cell types following in vivo estrogen treatment. Since these highly purified cultures of uterine cells were prepared from the uterus of a 21-day ovariectomized rat which is devoid of eosinophils, we conclude that estradiol stimulation of nuclear type II sites is a direct intracellular response to estrogen which occurs independent of eosinophil accumulation. Furthermore, we have found that type II sites in the rat uterus are not peroxidase. This was demonstrated by experiments which show type II sites are present in the 39,000 X g supernatant fraction of uterine cytosol, whereas peroxidase activity is quantitatively recovered in the crude mitochondrial (39,000 X g) pellet. Likewise, the small amount of peroxidase activity (approximately 10%) in the total homogenate which contaminates our nuclear pellet preparations was extracted (98-100%) with 0.5 M CaCl2. Type II estrogen-binding sites (95-100%) remained associated with the nuclear pellet fraction after peroxidase extraction. Therefore, stimulation of cytosol and nuclear type II sites by estrogen in the rat uterus is a direct intracellular response to the hormone unrelated to eosinophil accumulation and/or peroxidase activity.

The histogenesis of the Reed-Sternberg (R-S) cell in Hodgkin's disease is uncertain. Some have suggested that it is a derivative of the monocyte/macrophage lineage. To explore this possibility, we have searched for the presence of mRNA corresponding to the c-fms proto-oncogene, a marker for cells of the monocyte/macrophage lineage which encodes the colony-stimulating factor-1 receptor. In situ hybridization was performed using a single-stranded c-fms complementary RNA (cRNA) to probe R-S cells, lymphocytes, and eosinophils from touch imprints of a lymph node from a 12-year-old boy with mixed cellularity Hodgkin's disease in relapse. The probe was synthesized from a bacterial plasmid, pSM3, into which a portion of v-fms (a viral-derived oncogene) had been inserted. The plasmid was linearized with a restriction endonuclease, and 35S-labeled cRNA was synthesized from the DNA template using T3 RNA polymerase and the nucleotide analog [35S]UTP. Positive control hybridizations were obtained with the human acute promyelocytic cell line HL-60 induced to monocyte differentiation. R-S cells were clearly negative, supporting a cell of origin other than the monocyte. In situ hybridization is a potentially powerful method for exploring differentiation and assigning cell lineage in R-S cells.

This study is an investigation on whether allergy, and especially food allergy, is more common in patients with secretory otitis media (SOM) than in others. At the same time, the effectiveness of the cytotoxic leucocyte test (CLT) as an aid in the diagnosis of food allergy was also studied. The material comprised 90 patients: 69 suffering from secretory otitis media and 21 without the disease as controls. Patients were studied using allergy history, ear status, tests for nasal and blood eosinophilic cells, total IgE, skin tests for 20 allergens. RAST for milk and wheat, immunoglobulin A, G and M, precipitating antibodies for milk and gluten. Mantoux test, and the CLT for 23 allergens. Allergy was found in 20% of the SOM patients and 10% of the control patients; the difference was not statistically significant. The best methods for verifying allergy were history, skin tests, and tests for nasal eosinophils cells. Laboratory tests could not verify objectively that food allergy was more common among SOM patients than controls, although 16% of the SOM patients compared with none of the controls had a history of food allergy. Of the tests used, the CLT correlated best with a history of allergy, but the results of the different tests were not in accordance wtih each other.

Eosinophilic endomyocardial disease is a complication of the hypereosinophilic syndrome and of several other disorders associated with high blood eosinophil counts. Eosinophil granule proteins may be involved in the development of these lesions. This multi-centre study investigated whether these proteins could be demonstrated within the cardiac tissues of eighteen patients with eosinophilic endomyocardial disease. Serial sections of tissue taken at necropsy or at cardiac biopsy were stained for eosinophil major basic protein by indirect immunofluorescence and for eosinophil cationic protein, eosinophil protein-X, and activated eosinophils by means of alkaline-phosphatase-linked monoclonal antibodies. Activated eosinophils and secreted eosinophil granule proteins were most evident within the necrotic and later stage thrombotic lesions and were found mainly within the areas of acute tissue damage in the endocardium and in the walls of small blood vessels. These findings suggest that eosinophil granule proteins are involved in cardiac injury, producing muscle damage and vascular injury which lead to the development of endomyocardial fibrosis.

Purpose: The concentrations of myeloperoxidase (MPO), a neutrophil granule constituent, and eosinophil cationic protein (ECP), a specific eosinophil granule protein, were measured in jejunal perfusion fluid in an attempt to elucidate the neutrophil and eosinophil involvement of the small bowel in health and disease., Patients and Methods: The control group consisted of 14 males and two females. Ten patients (seven males and three females) with Crohn's disease and seven patients (two males and five females) with celiac disease were also studied; in addition, one patient with relapsing giardiasis, one patient with giardiasis and complete absence of plasma cells in small intestinal lamina propria, and one patient with selective IgA deficiency and no IgA plasma cells in duodeno-jejunal lamina propria were evaluated. Segmental perfusion of the jejunum was performed according to a previously described method. MPO and ECP were measured by radioimmunoassays., Results: In healthy control subjects, the concentrations of both granule proteins were in a narrow range and much higher than would have been anticipated from passive leakage from circulating blood. In patients with celiac disease, the perfusion fluid concentrations of MPO and ECP were on average 3.5 and eight times, respectively, higher than the values seen in the controls. The jejunal segment perfused in patients with Crohn's disease was endoscopically and histologically normal. The perfusion fluid concentrations of MPO and ECP were increased 3.5 and two times, respectively, compared with that in the control subjects. Both patient groups and the control group had similar perfusion fluid concentrations of albumin. Data on MPO and ECP expressed as jejunal secretion rates gave the same differences between patients and controls as just described for the jejunal fluid concentrations. Immunohistochemical studies of jejunal biopsy specimens from another group of patients with celiac disease demonstrated a prominent extracellular deposit of ECP in the lamina propria of the atrophic intestinal mucosa, whereas the release of neutrophil constituents (cathepsin G, MPO) was scarce. In Crohn's disease, an extracellular degranulation of ECP and, to a lesser extent, of cathepsin G was observed in relation to ulcerations only., Conclusion: Data obtained indicate that the local release of neutrophil and eosinophil granule components is enhanced in the jejunal tissue from patients with celiac sprue and Crohn's disease. The prominent extracellular deposit of eosinophil granule constituents with cytotoxic properties at the site of inflammatory intestinal lesions in celiac sprue might reflect a pathophysiologic mechanism.(ABSTRACT TRUNCATED AT 400 WORDS)

A 9-year-old girl had fever, life-threatening interstitial pneumonia, a vesicular skin rash, parotid and submandibular swelling, and marked blood eosinophilia (WBC count 47,000/microL, 39% eosinophils). Results of lung and skin biopsies showed vasculitis with intense eosinophil infiltration. Immunofluorescence analyses of these biopsies, as well as analysis of tissue from the parotid, lip, and trachea, showed striking deposition of eosinophil granule major basic protein associated with areas of tissue damage. Treatment with glucocorticoids and hydroxyurea produced clinical improvement. The association between tissue damage and deposition of the cytotoxic major basic protein suggests that the eosinophil contributed to the pathophysiology of this disease. Recognition of the capacity of the eosinophil to produce tissue damage can be helpful in the selection of therapy.