Cloning and expression of a cDNA encoding a non-classical MHC class I antigen (HLA-E) in eosinophils from hypereosinophilic patients.

International audienceA cDNA library, constructed from purified blood eosinophils, was screened with the B cell CD23 cDNA probe. A clone designated EO15 has been isolated and found to encode a non-classical HLA class I gene transcript. EO15 was compared with HLA-E and found to be 99.9% similar at the nucleotide level and to extend further in the 3' untranslated region. The presence of an additional polyadenylation signal in the EO15 3' end suggests that EO15 clone represents a copy of the 3.3-kb mRNA species detected in Northern blot analyses. HLA-E transcripts of 1.9 and 3.3 kb have been described in a variety of cell types. The two EO15 mRNA species, similar in size to the previously defined HLA-E mRNA, were present at high levels in blood leukocyte populations and at variable levels in different cell lines. The EO15 transcripts were found at abundant levels in hypodense and normodense eosinophils from hypereosinophilic patients. In situ hybridization confirmed the expression of EO15 mRNA in eosinophils. Neutrophils and lymphocytes from normal donors of from patients with hypereosinophilia also strongly expressed EO15 mRNA. Among the cell lines studied, the highest levels of EO15 transcripts were detected in B and monocytic cell lines, whereas intermediate and lower levels were found in eosinophilic, NK-like, megakaryocytic, and T cell lines, respectively. Similar to its effect on classical HLA class I transcripts, IFN-gamma increased the levels of EO15 mRNA in eosinophils and neutrophils from hypereosinophilic patients. These results suggest that purified blood eosinophils as well as neutrophils express EO15/HLA-E mRNA; however, further experiments are needed to investigate the localization and the function of EO15 protein products

Induction of CD23, CD25 and CD4 expression on an eosinophilic cell line (EoL-3) by interleukin-3 (IL-3), granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-5 (IL-5).

International audienceThe effects of IL-3, GM-CSF and IL-5 on the expression of CD23 (Fc epsilon RII), CD25 (IL-2R/p55) and CD4 on an eosinophilic cell line (EoL-3) were investigated by flow cytometry. A separate incubation with IL-3, GM-CSF or IL-5 alone, did not induce the expression of CD23, CD25, or CD4. However, a sequential incubation with IL-3 for 6 days, then with IL-3 and GM-CSF for the following 6 days, induced a significant expression of CD23 and CD25. After a further incubation for 6 days with IL-3, GM-CSF and IL-5, CD4 was then expressed, while CD23 and CD25 expression still increased. The kinetics of expression of CR3/CD11b were parallel to that of CD23, but the expression of the transferrin receptor (CD71) remained negative. Northern blot analysis revealed the presence of mRNA encoding CD23, CD25 and CD4 in EoL-3 stimulated by IL-3, GM-CSF and IL-5. Culture with GM-CSF induced the binding of radiolabeled IL-5 to EoL-3 cells, with an increased affinity after incubation with IL-3, GM-CSF and IL-5. These data indicate that IL-3, GM-CSF and IL-5, might be involved in the expression of functional markers on eosinophil membrane

Mesurer le niveau de résistance quantitative des variétés de tournesol face au mildiou dans les processus de sélection et d’évaluation variétale : un enjeu fort pour la gestion durable du risque

La résistance quantitative du tournesol (Helianthus annuus) au mildiou (Plasmopara halstedii), mise en évidence récemment, est difficile à évaluer. Un réseau de onze expérimentations associant la GEVESSNES, deux laboratoires de l’INRA et Terres Inovia a été construit afin de proposer une méthodologie permettant de mieux caractériser et quantifier cette résistance. En plus de lignées témoins, 17 lignées recombinantes obtenues par l’INRA et portant entre 0 et 3 QTL (quantitative trait loci) impliqués dans la résistance quantitative du tournesol au mildiou ont été testées face à 5 pathotypes de mildiou : 710, 704, 714, 304 et 334. Bien que des différences aient été observées entre laboratoires, les résultats ont permis de poser les bases d’un futur protocole utilisable en routine pour valoriser ce type de résistance dès la création variétale jusqu’à l’inscription puis la post-inscription des variétés.A quantitative resistance of sunflower (Helianthus annuus) to downy mildew (Plasmopara halstedii) has been identified a few years ago. An experimental network involving several labs (GEVES-SNES, INRA and Terres Inovia) over several years aimed at describing and quantifying such a quantitative response. It was hoped to propose a methodology useable to improve sustainable resistance. In addition to check varieties and breeding lines, 17 recombinant inbred lines carrying 0 to 3 quantitative trait loci alleles for quantitative resistance were tested with five downy mildew pathotypes (710, 704, 714, 304, 334). While the experimental results revealed some discrepancies between experiments, this study provided a first experimental protocol to evaluate quantitative resistance both for breeding and for variety registration and developmen