Proteómica clínica: análisis de las células Treg naturales CD4CD25 en autoinmunidad y en salud

  • Marti Ripoll, Maria Merce (Principal Investigator)
  • Xufré Ballesteros, Cristina (Scholar)
  • Costa García, Manuela (Investigator)
  • Pizarro Lozano, Eduarda (Investigator)

Project Details

Description

Natural Tregs CD4CD25\super ++\nosuper (nTregs) are thymic emigrants that participate in the immune homeostasis. A defective phenotype of these subpopulation could be crutial in the development of autoimmunity. The absence of an exclusive marker for nTregs impairs their analysis in PBMC samples from patients. The markers described till now also expressed by effector/memory Tcells and inducible T regulatory cells (Tr1). It is therefore still impossible to isolate these cells as a pure subpopulation without contamination of other T cell types, limiting their use as agents for cellular therapy. Comparative proteomics of n Tregs from thymus, cord blood and peripheal blood would allow to establish the n Treg- proteomic profile from generation to adult life. This proteomic profile in health will be used to determine possible differences in protein expression by these cells patients with type 1 Diabetes and Graves' Disease.
MAIN AIM: The comparative-proteomic analysis of n Tregs obtained from thymus, cord blood and peripheral blood from healthy donors. Definition of a pattern of specific makers and a proteomic profile for the study of n Tregs in patients with endocrine diseases.
METHODOLOGY: Phenotype analysis of n Tregs by Flow Cytometry. Cell-sorting or magnetic-beads purification of CD4\super +\nosuper /CD25\super ++\nosuper and CD4\super +\nosuper /CD25\super -\nosuper cells. Cellular proteome comparation with the high sensivity EttanDIGE system; this system quantifies the expressed proteins and stadistically analyzes the diferences obtained. Sequencing by MALDI-Tof or LQC of the differentially expressed spots digested with tripsin. Confirmation of differences by using alternative techniques: real time PCR (qPCR) and Western Blot.
StatusFinished
Effective start/end date16/10/0730/12/10

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