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資料番号: AA0046920002

資料番号: AA0045981019

第24回大気圏シンポジウム（2010年2月18日-19日. 宇宙航空研究開発機構宇宙科学研究本部（JAXA)(ISAS)）, 相模原市, 神奈川県, 24th Atmospheric Science Symposium (February 18-19, 2010. Institute of Space and Astronautical Science, Japan Aerospace Exploration Agency (JAXA)(ISAS)), Sagamihara, Kanagawa Japan, 資料番号: SA6000073016, レポート番号: Ⅲ-4

第23回大気圏シンポジウム（2009年2月26日-27日. 宇宙航空研究開発機構宇宙科学研究本部（JAXA)(ISAS)）, 相模原市, 神奈川県, 23rd Atmospheric Science Symposium (February 26-27, 2009. Institute of Space and Astronautical Science, Japan Aerospace Exploration Agency (JAXA)(ISAS)), Sagamihara, Kanagawa Japan, 資料番号: SA6000074014, レポート番号: Ⅲ-1

Resume Une technique rapide d'isolement des basophiles humains permet d'obtenir une suspension cellulaire 10 a 20 fois plus riche en basophiles que le sang circulant. A partir de cette suspension, un test de degranulation des basophiles est pratique sur lame a l'aide de microquantites (10 mm 3 ). Apres fixation des cellules par le glutaraldehyde et coloration, le compte comparatif des basophiles, parfaitement identifiables grâce a leur metachromasie, est effectue sur des plages temoins et sur des plages avec antigenes a diverses concentrations. Ceci permet d'etablir l'index de degranulation. Cette technique est applicable aux diagnostics des allergies reaginiques telles les allergies medicamenteuses.

Resume Un test de degranulation des basophiles humains, pratique sur une suspension cellulaire enrichie, a ete applique a 28 patients, dont 14 allergiques a Dermatophagoides pteronyssinus (D.pt) et 14 temoins. Chez les 14 temoins, aucune degranulation significative n'a ete observee avec les diverses dilutions de D.pt utilisees (index toujours inferieur a 20 p. cent). Chez 13 allergiques sur 14, l'index a ete superieur a 40 p. cent (p

Resume Chez vingt sujets, presentant une urticaire generalisee, un compte absolu des eosinophiles (CAE) a ete pratique moins de douze heures apres le debut des signes cliniques et avant tout traitement. Dix-sept malades presentaient une eosinopenie (inferieure a 76/mm 3 ). La valeur moyenne du CAE etait de 25/mm 3 (7–78). Chez trente-neuf sujets temoins, elle etait de 99/mm 3 (76–131). La difference est statistiquement significative (p 3 , p 50 ) n'etait pas different de celui du groupe temoin. Chez deux malades, un facteur serique anticomplementaire assimile a la presence d'immuns complexes circulants a ete detecte. Le compte absolu des eosinophiles s'avere un examen simple et precis qui peut apporter un argument en faveur d'un mecanisme anaphylactique.

Resume Nous avons mis au point une nouvelle methode permettant de quantifier le degre de stimulation des lymphocytes T : ceux-ci, apres transformation, forment des megarosettes avec les globules rouges de mouton. Celles-ci ont une vitesse de sedimentation tres elevee. Separees des rosettes E par sedimentation differentielle, les megarosettes sont hemolysees et la densite optique est mesuree. Les resultats montrent une bonne correlation avec ceux obtenus par le TTL utilisant l'incorporation de thymidine tritiee. Cette technique peut etre appliquee en immunologie clinique pour l'etude in vitro des affections immunitaires et aussi pour tester la sensibilisation d'un individu a un antigene donne.

Resume Une simplification de la methodologie du test de degranulation des basophiles humains est rapportee. Elle permet d'obtenir le resultat de l'examen en moins de deux heures et se prete a une utilisation en routine pour le diagnostic en allergologie et en parasitologie (helminthiase).

Total eosinophil count (TEC) was done in 13 normal subjects, in 8 subjects before and 90 mn after a "French" breakfast and in 29 asthmatic patients. The geometric mean is higher (p less than 0,001) in asthmatic than in normal subjects: 304 (231--391) versus 144 (101--197). The breakfast does not interfer with the result of the TEC. In asthmatic subjects, a significant inverse correlation appears (r = 0,57, p less than 0,01) between the logarithm of TEC and the quotient: formula; see text. This correlation persists whatever may be the subject's age, the duration of the asthma or age at which he first contracted the illness. Asthma appeared after 40 years in 4 of the subjects: their TEC was higher (p less than 0,01) than in subjects who had become asthmatic before the age of forty. The respiratory function was lowered more in these 4 patients than in others who had been ill for the same time (p less than 0,001). The TEC is not related to results of skin testine with usuel inhalant allergens (grass pollen, house dust, cat's or dog's dander) or by result of a provocation test (by inhalation of allergen). Eosinophilia and bronchial obstruction are connected by one (or more) factors that may be inhalant allergens in extrinsec asthma and circulating immune complexes in intrinsec asthma.

In stimulated cultures, a proportion of human T lymphocytes form multilayer rosettes with sheep red blood cells (SRBC) and they can be easily separated from common E rosettes by velocity sedimentation. We have developed a simplified method for harvesting multilayer rosettes and quantifying them. Practical applications of this technique are discussed.

Basophils disappear after challenge with specific antigen. A human basophil degranulation test has been carried out on a slide using an enriched cell suspension. On each slide, the same volume of cell suspension was deposited in wells with antigen diluted in buffer or with buffer only (control). The basophil count was made on an equal number of randomly distributed microscopic fields either by eye (optical) or by image analyser (automatic). In 35 subjects, the correlation coefficient between the numbers of non-degranulated basophils counted by eye and by image-analyser on control wells was found to be r = 0.91, P < 10−8. In 12 patients (6 with hydatidosis and 6 widi schistosomiasis), the percentage of degranulation with three antigen dilutions was measured. The correlation between the results obtained by eye and by image-analyser reached 0.76 (P < 10−7). The authors now use the automated measurement of human basphil degranulation routinely.

A total eosinophil count (TEC) and a white blood cell count (WBCC) were performed simultaneously in 213 patients. Discrepancies were found between TEC and WBCC. WBCC did not allow detection of hypereosinophilia (TEC exceeding 300 eosinophils/microliter) in 45 patients. Patients were classified in three groups : controls (n = 27), benign disease (n = 65), and apparently cured cancer (n = 36). There was no significant difference between total eosinophil counts for these three groups (Wilcoxon non-parametric test). The incidence of hypereosinophilia (TEC exceeding 300) was similar in the three groups (11%, 14% and 6%). Average TEC levels were determined in 20 patients with decompensated hepatic cirrhosis, in 31 patients with active cancer, and in 14 patients with infection. These levels were significantly different from those found in each of the three previous groups. (Wilcoxon test). The incidence of hypereosinophilia (greater than 300/microliter) was significantly higher among patients with cirrhosis (50%) or active cancer (58%). The possible mechanisms of this hypereosinophilia and it's eventual prognostic utility are considered.

Total eosinophil count (TEC) was done in 13 normal subjects, in 8 subjects before and 90 mn after a "French" breakfast and in 29 asthmatic patients. The geometric mean is higher (p less than 0,001) in asthmatic than in normal subjects : 304 (231-391) versus 144 (101-197). The breakfast does not interfer with the result of the TEC. In asthmatic subjects, a significant inverse correlation appears (r = 0,57, p less than 0,01) between the logarithm of TEC forced expiratory volume in one second and the quotient : vital capacity. This correlation persists whatever may be the subject's age, the duration of the asthma or age at which he first contracted the illness. Asthma appeared after 40 years in 4 of the subjects : their TEC was higher (p less than 0,01) than in subjects who had become asthmatic before the age of forty. The respiratory function was lowered more in these 4 patients than in others who had been ill for the same time (p less than 0,001). The TEC is not related to results of skin testing with usual inhalant allergens (grass pollen, house dust, cat's or dog's dander) or by result of a provocation test (by inhalation of allergen). Eosinophilia and bronchial obstruction are connected by one (or more) factors that may be inhalant allergens in extrinsec asthma and circulating immune complexes in intrinsec asthma.