A - Human Necessities – 61 – K
Patent
A - Human Necessities
61
K
A61K 39/00 (2006.01) C07K 14/705 (2006.01) C07K 16/28 (2006.01) C12Q 1/68 (2006.01) G01N 33/574 (2006.01)
Patent
CA 2321222
The invention concerns a method for selecting tumours expressing HLA-G, sensitive to an anticancer treatment, which inhibits or prevents the HLA-G activity of said tumours and the uses thereof. Said method enable to establish either the HLA-G transcription profile of a solid tumour or the HLA-G expression profile of a solid tumour. The method for establishing the HLA-G transcription profile consists in: (i) drawing a tumoral sample; (ii) extracting the mRNA; (iii) reverse transcription (RT) of said RNA; (iv) successive or simultaneous amplification of the cDNA's obtained in (iii) in the presence of primers specific to each HLA-G isoform and analysing the resulting amplification products, by electrophoresis and/or specific hybridisation and (v) establishing said sample HLA-G transcription profile. The method for establishing the HLA-G expression profile consists in: (i) drawing a tumoral sample; (ii) optionally marking said sample cells; (iii) carrying out a lysis of the cells; (iv) contacting said cells which have been subjected to lysis with different antibodies directed against the class I HLA-G antigens, to form, optionally HLA-G isoform/antibodies complexes; and (v) establishing said sample HLA-G expression profile by detecting the complexes formed in step (iv).
Méthode de sélection de tumeurs solides exprimant HLA-G, sensibles à un traitement anticancéreux, qui inhibe ou prévient l'activité HLA-G desdites tumeurs solides et ses applications. Ladite méthode permet d'établir soit le profil de transcription HLA-G d'une tumeur solide, soit le profil d'expression HLA-G d'une tumeur solide. La méthode d'établissement du profil de transcription HLA-G comprend: (i) le prélèvement d'un échantillon tumoral; (ii) l'extraction de l'ARNm; (iii) la transcription inverse (RT) dudit ARN; (iv) les amplifications successives ou concomitantes des ADNc obtenus en (iii), en présence d'amorces spécifiques de chaque isoforme d'HLA-G et l'analyse des produits d'amplification obtenus, par électrophorèse et/ou hybridation spécifique et (v) l'établissement du profil de transcription HLA-G dudit échantillon. La méthode d'établissement du profil d'expression HLA-G comprend: le prélèvement d'un échantillon tumoral, (ii) éventuellement le marquage des cellules dudit échantillon, (iii) la lyse des cellules, (iv) la mise en contact des cellules lysées avec différents anticorps dirigés contre les antigènes HLA de classe I, pour former, éventuellement des complexes isoforme d'HLA-G/anticorps et (v) l'établissement du profil d'expression HLA-G dudit échantillon, par détection des complexes formés à l'étape (iv).
Carosella Edgardo Delfino
Dausset Jean
Moreau Philippe
Paul Pascale
Rouas-Freiss Nathalie
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