RESUMO
We describe the 2-year follow-up of an open-label trial (CT-AMT-011-01) of AAV1-LPL(S447X) gene therapy for lipoprotein lipase (LPL) deficiency (LPLD), an orphan disease associated with chylomicronemia, severe hypertriglyceridemia, metabolic complications and potentially life-threatening pancreatitis. The LPL(S447X) gene variant, in an adeno-associated viral vector of serotype 1 (alipogene tiparvovec), was administered to 14 adult LPLD patients with a prior history of pancreatitis. Primary objectives were to assess the long-term safety of alipogene tiparvovec and achieve a ≥40% reduction in fasting median plasma triglyceride (TG) at 3-12 weeks compared with baseline. Cohorts 1 (n=2) and 2 (n=4) received 3 × 10(11) gc kg(-1), and cohort 3 (n=8) received 1 × 10(12) gc kg(-1). Cohorts 2 and 3 also received immunosuppressants from the time of alipogene tiparvovec administration and continued for 12 weeks. Alipogene tiparvovec was well tolerated, without emerging safety concerns for 2 years. Half of the patients demonstrated a ≥40% reduction in fasting TG between 3 and 12 weeks. TG subsequently returned to baseline, although sustained LPL(S447X) expression and long-term changes in TG-rich lipoprotein characteristics were noted independently of the effect on fasting plasma TG.
Assuntos
Terapia Genética , Hiperlipoproteinemia Tipo I/terapia , Lipase Lipoproteica/genética , Adulto , Dependovirus/genética , Tolerância a Medicamentos , Jejum/sangue , Feminino , Humanos , Hiperlipoproteinemia Tipo I/complicações , Imunossupressores/administração & dosagem , Imunossupressores/uso terapêutico , Lipase Lipoproteica/metabolismo , Masculino , Pessoa de Meia-Idade , Pancreatite/complicações , Estudos Prospectivos , Resultado do Tratamento , Triglicerídeos/sangueRESUMO
A monoclonal antibody against thymocytes (WCL9; of IgG1 class) was produced by immunization of mice with isolated membrane molecules of carp thymocytes. Flow cytometric and fluorescence microscopic analysis showed that WCL9 was reactive with 30-50% of thymocytes and not with lymphoid cells from blood, pronephros, spleen and intestine. Cryo-sections of thymus showed a WCL9+ and WCL9- region in 3-month-old fish and only WCL9+ cells in 1-week-old fish. Because the WCL9- region is more medulla-like, the WCL9+ cells can be considered as cortical thymocytes. The majority of WCL9+ thymocytes appeared to have a higher density (1.06-1.07 g/mL) than the WCL9- cells (1.02-1.06 g/mL). Immunogold labelling or comparison of both density fractions did not show clear ultrastructural differences between WCL9+ and WCL9- thymocytes. The WCL9- fraction could be stimulated much better with PHA than the WCL9+ fraction. Removal of adherent cells or adding adherent accessory cells did not influence this result. Immunochemical analysis showed that WCL9 reacted with a protein determinant present on two molecules (M(r): 200 and 155 kDa) under reduced and non-reduced conditions. These results, together with the absence of WCL9+ cells in other lymphoid organs, strongly suggest that WCL9 is a specific marker for early thymocytes.
Assuntos
Anticorpos Monoclonais/imunologia , Carpas/imunologia , Linfócitos T/citologia , Animais , Especificidade de Anticorpos , Biomarcadores , Técnicas de Cultura de Células , Citometria de Fluxo , Imuno-Histoquímica , Linfócitos/imunologia , Linfócitos/metabolismo , Camundongos , Microscopia Eletrônica , Linfócitos T/imunologiaRESUMO
A rapid and convenient method of nasal provocation (aerosol provocation combined with passive anterior rhinomanometry) is described. A comparative study between skin tests, bronchial provocations and nasal provocations highlights the usefulness of this method.
