Seroepidemiology of Strongyloides infection in the Southeast Asian refugee population in Canada.

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.

Distribution of antibodies against Coxsackie B viruses, arboviruses and Toxoplasma gondii among patients with endomyocardial fibrosis (EMF) compared with normal subjects from EMF endemic and non-endemic zones of Nigeria.

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.

Does reversed-type anaphylaxis in healthy subjects mimic a real allergic reaction?

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.

Identification of a novel and highly potent eosinophil chemotactic lipid in human eosinophils treated with arachidonic acid.

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.

The neutrophil and chronic allergic inflammation. Immunochemical localization of neutrophil elastase.

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.

Change in expression of Fc gamma RIII (CD16) on neutrophils from human immunodeficiency virus-infected individuals.

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.

Fc gamma and CD11/CD18 receptor expression on normal density and low density human eosinophils.

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.

Evidence of eosinophil granule major basic protein in human placenta.

A protein immunochemically related to the eosinophil granule major basic protein (gMBP) is found in increased concentration in the plasma of pregnant women and has been localized to placental trophoblasts by immunofluorescence. Pregnancy MBP (pMBP) is indistinguishable from gMBP in its reactivity with polyclonal antisera and a panel of 14 mouse mAbs. We report the purification of pMBP from human placenta by: (a) affinity chromatography over mAb immobilized on Sepharose, (b) gel filtration in 6 M guanidine.HCl buffer, and (c) reversed-phase HPLC. Purified pMBP and gMBP are biochemically indistinguishable in that both: (a) bind to DNA, (b) polymerize and bind to carrier proteins via disulfide linkages, (c) have a molecular weight of 14,000, (d) have isoelectric points greater than 10.6, (e) comigrate in two-dimensional gels, (f) coelute during reversed-phase HPLC on C18 columns, (g) have identical peptide maps after three different digestions, and (h) have partial amino acid sequence identity. This physicochemical identity has important implications as to the role of pMBP in human placentation.

Heterogeneity of human eosinophil glucocorticoid receptor expression in hypereosinophilic patients: absence of detectable receptor correlates with resistance to corticotherapy.

Assessment of steroid receptor content in human neoplastic lymphoid cells or mammary tumour cells has been previously used to predict steroid sensitivity in various types of cancers. In the present study, we have evaluated the relationship between glucocorticoid receptor content and the glucocorticoid sensitivity of human eosinophils, since hypereosinophilic patients do not always respond favourably to glucocorticoid, particularly in the hypereosinophilic syndrome (HES). Blood or alveolar eosinophils obtained from seven patients (four with HES without leukaemic markers; two with parasitic diseases; and one with eosinophilic pneumonia) displayed the same specific glucocorticoid receptor content as normal eosinophils (7.58 +/- 1.31 x 10(3) versus 7.76 +/- 0.74 x 10(3) sites/cell). In contrast, glucocorticoid-binding sites were undetectable in purified eosinophils collected from seven HES patients with (n = 3) or without (n = 4) leukaemic markers, whilst their mononuclear cells and/or neutrophils bound glucocorticoid. In one HES patient, kinetic studies showed that blood eosinophils initially positive in glucocorticoid binding assays became negative with the subsequent appearance of leukaemic markers. The absence of specific glucocorticoid binding sites was correlated with the absence of glucocorticoid receptor proteins by the use of a specific anti-glucocorticoid receptor monoclonal antibody. Eosinophil sensitivity to glucocorticoid was investigated by the evaluation of glucocorticoid inhibition of eosinophil chemotaxis and by the clinical outcome of in vivo glucocorticoid therapy. Our data provide evidence of the heterogeneity of eosinophil glucocorticoid receptor expression. In addition, the presence of glucocorticoid receptors is a prerequisite for glucocorticoid activity, in vitro and in vivo, on cells of the eosinophil lineage.

The detection and interpretation of urinary eosinophils.

We determined the clinical diagnosis for 183 patients who had urine samples examined for the presence of eosinophils. Urine samples were examined with both Hansel's stain and Wright's stain. A total of 11% of these patients had eosinophils detected in the urine. A variety of clinical conditions were associated with eosinophiluria. Urinary tract infection and acute interstitial nephritis were most common, each accounting for 25% of the total patients with eosinophiluria. Eosinophiluria proved to be a good predictor of acute interstitial nephritis. Hansel's stain was superior to Wright's stain in detecting eosinophils in urine. In particular, Hansel's stain increased the sensitivity of eosinophiluria for acute interstitial nephritis (63% vs 25%) as well as its positive predictive value (50% vs 25%).

Human eosinophil-derived neurotoxin and eosinophil protein X are likely the same protein.

The human eosinophil granule contains a series of cationic proteins. Two of these, eosinophil-derived neurotoxin (EDN) and eosinophil protein X (EPX), are reported to have similar m.w. and both possess neurotoxic and helminthotoxic activities. Therefore, the properties of these molecules were analyzed to determine whether they differ. EDN was purified from eosinophils of patients with the hypereosinophilic syndrome and EPX from the buffy coat cells of normal individuals. By SDS-PAGE, both proteins showed a major band at 18.7 kDa and a minor band at 21.4 kDa. By two-dimensional non-equilibrium gel electrophoresis the proteins migrated identically. With radiolabeled proteins in reverse phase HPLC, both proteins eluted at the same concentration of acetonitrile and showed identical tryptic maps. Both proteins possessed comparable ribonuclease activity and both were comparably neurotoxic in the rabbit. By immunodiffusion the two proteins showed a reaction of identity; by RIA, with both polyclonal and monoclonal antibodies, the proteins had very similar inhibitory activities. These results indicate that EDN and EPX have virtually identical properties and are probably the same protein.

New series of lipoxins isolated from human eosinophils.

Granulocytes from human eosinophilic donors were incubated with arachidonic acid or 15-hydroxyeicosatetraenoic acid (15-HETE) and stimulated with the ionophore A23187. The eicosanoids were extracted with reversed-phase cartridges and subjected to RP-HPLC analysis. When extracts from eosinophil-enriched populations were analysed and compared with extracts from human neutrophils, three additional peaks were detected which coeluted with 15-hydroxy-delta 13-trans-15H derivatives of leukotriene C4, D4 and E4 in different HPLC systems. The recorded absorbance spectra of the eluted compounds and the standards were identical and showed a maximum at 307 nm which is characteristic for a conjugated tetraene system with a bathochromic shift by the sulfur moiety in alpha-position to the tetraene system. The compound which coeluted with the 15-hydroxy-LTC4 standard was treated with gamma-glutamyltransferase and converted to the corresponding leukotriene D4 derivative. The results indicate that interaction between the 5- and 15-lipoxygenase pathways leads to the formation of a new series of arachidonic acid metabolites in human eosinophils. Since the biosynthetic route is similar to that of lipoxin A4 and lipoxin B4, we suggest the trivial names lipoxin C4, D4 and E4.

Bronchoalveolar lavage in the cat: cytological findings.

Cells recovered by bronchoalveolar lavage from the lungs of specific pathogen-free (SPF) cats from birth to maturity and from adult conventional cats were enumerated and identified. The predominant cell type recovered was the pulmonary alveolar macrophage from all ages of both SPF and conventional cats. Other cell types included eosinophils, neutrophils and lymphocytes. Lavage of conventional cats yielded significantly more eosinophils and neutrophils than were recovered from SPF cats.

[Experimental and clinical study of conjunctivo-scleral tolerance of PTFE (Goretex) sutures compared to polyglactine (Vicryl)]. / Etude expérimentale et clinique de la tolérance conjonctivo-sclérale des sutures en PTFE (Goretex) comparée au polyglactine (Vicryl).

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.

Are blood eosinophil number and activity important for the development of the late asthmatic reaction after allergen challenge?

A role for eosinophils in chronic asthma has recently gained substantial support. We hypothesized that in addition to eosinophil chemoattractants released in the lung, the availability of a sufficient number of blood eosinophils to be recruited and their increased activity are prerequisites for the development of asthma. To test this hypothesis the blood eosinophil number and serum levels of ECP (eosinophil cationic protein were measured before allergen challenge of 13 allergic asthmatics. Each individual was pretreated with either placebo or four inhalant anti-asthmatic drugs at random and blind in order to modulate the late asthmatic reaction (LAR). Irrespective of premedication the blood eosinophil number and serum-ECP levels were significantly higher before challenge when the challenge resulted in LAR. Only corticosteroids abrogated the development of LAR and also significantly reduced eosinophil activity as measured by serum-ECP. Thus the inherent activity of a sufficient number of eosinophils may be important determinants for the expression of allergic asthma.