title: F8 gene mutation profile and ITT response in a cohort of Italian haemophilia A patients with inhibitors.
abstract: Anti factor VIII (FVIII) antibodies represent the main complication of replacement therapy in severe cases of haemophilia and most patients with inhibitor have gross gene rearrangements or point mutations that hamper the production of normal circulating FVIII. In this study we have investigated 82 haemophilia A patients with inhibitors. Seventy six were severe, three were moderate and three were mild. We screened the patients for the causative mutations using long range PCR for the recurrent intron 22 inversion (invint22), multiplex PCR for intron 1 inversion (invint1) and conformation sensitive gel electrophoresis followed by DNA sequencing for all other mutation types in the F8 gene. Diverse genetic defects were detected in the severe cases (with a predominance of severe molecular defects): F8 gene inversions, large deletions and non-sense mutations account for 71% of the mutations. Only missense and splicing mutations were identified in the non-severe patients and we confirmed that the presence of inhibitors correlates well with the presence of severe mutations, but a proportion of severe patients develops inhibitors despite the presence of diverse less severe mutations. When we have analysed the subgroup of patients who underwent immunetolerance, we have found that F8 gene large deletions are likely to be a high risk factor also for immunetolerance therapy unresponsiveness, while no clear evidence has been demonstrated for other mutation types.
authors.affiliation: Dipartimento di Laboratorio, Servizio Trasfusionale, Centro Regionale per le Malattie del Sangue e Servizio di assistenza agli Emofilici di Castelfranco Veneto (Treviso), Castelfranco Veneto (TV), Italy.
authors.forename: R
authors.initials: R
authors.lastname: Salviato
authors.forename: D
authors.initials: D
authors.lastname: Belvini
authors.forename: P
authors.initials: P
authors.lastname: Radossi
authors.forename: R
authors.initials: R
authors.lastname: Sartori
authors.forename: F
authors.initials: F
authors.lastname: Pierobon
authors.forename: D
authors.initials: D
authors.lastname: Zanotto
authors.forename: E
authors.initials: E
authors.lastname: Zanon
authors.forename: G
authors.initials: G
authors.lastname: Castaman
authors.forename: G
authors.initials: G
authors.lastname: Gandini
authors.forename: G
authors.initials: G
authors.lastname: Tagariello
mesh_terms: D019774:Blood Coagulation Factor Inhibitors / Q000097:blood / Q000235:genetics; D001780:Blood Coagulation Tests; D015331:Cohort Studies; D004252:DNA Mutational Analysis / Q000379:methods*; D005169:Factor VIII / Q000235:genetics* / Q000276:immunology; D006467:Hemophilia A / Q000235:genetics* / Q000276:immunology / Q000628:therapy; D006801:Humans; D007108:Immune Tolerance / Q000235:genetics*; D007165:Immunosuppression Therapy / Q000379:methods; D007558:Italy / Q000208:ethnology; D008297:Male; D009154:Mutation / Q000235:genetics*; D010375:Pedigree
chemical_list: D019774:Blood Coagulation Factor Inhibitors; C078147:F8 protein, human; D005169:Factor VIII