Thorac Cardiovasc Surg 2011; 59 - eP165
DOI: 10.1055/s-0030-1269393

Human immune system development in NOD/LtSz-scid IL2Rgnull mice after engraftment with purified cord blood derived stem cells

A Andersson 1, S Brösel 1, D Lange 1, C Knosalla 2, LD Shultz 3, D Egger 4, HD Volk 1, 5, I Schmitt-Knosalla 1
  • 1Charité-Universitätsmedizin Berlin, Institute of Medical Immunology, Berlin, Germany
  • 2Deutsches Herzzentrum Berlin, Department of Cardiothoracic and Vascular Surgery, Berlin, Germany
  • 3The Jackson Laboratory, Bar Harbor, Maine, United States
  • 4Vita 34 AG, Leipzig, Germany
  • 5Charité-Universitätsmedizin Berlin, Berlin-Brandenburg Center for Regenerative Therapies, Berlin, Germany

Objectives: The NOD/LtSz-scid IL2Rgnull mice presents an in vivo model for human HSC engraftment, differentiation and analysis of human immune system responses. This study aimed to investigate the time course of HSC engraftment in NOD/LtSz-scid IL2Rgnull mice using purified cord blood derived stem cells.

Methods: Purified human CD34+ cells derived from frozen cord blood were transplanted into non-lethally irradiated newborn mice. Peripheral blood was collected for flow cytometric analysis using two novel eleven colour flow cytometric panel for T-/NK-cells and B-/DC-cells to analyze subtypes and differentiation status of human cells in the peripheral blood and lymphoid organs. Spleen, thymus, lymph nodes and bone marrow were analyzed by flow cytometry and plasma by ELISA measurements of human IgG, IgM, IgA.

Results: At week eight after transplantation human IgM-plasma level became detectable and human lymphocytes became detectable, with low frequency of human T-cells and high frequency of human B-cells in peripheral blood. Between week 16 and 22 huCD45+ cells reached >40%. Over time, the rate of the human T cells increased.. At week 22 after transplantation 40% of thymic cells are human T cells. Immune histology of bone marrow, thymus, lymph nodes and spleen demonstrated presence of huCD45+, huCD4+, huCD8+, huCD11+, huCD20+ and huCD23+ cells. Spleen and lymph nodes showed distinctive human lymphoid structures.

Conclusion: NOD/LtSz-scid IL2Rgnull mice present a robust in vivo model of human HSC engraftment, differentiation and development of human lymphoid structures. This model allows studies of human biological processes in viv.