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PLoS One. 2014 May 15;9(5):e97860. doi: 10.1371/journal.pone.0097860. eCollection 2014.

Immune humanization of immunodeficient mice using diagnostic bone marrow aspirates from carcinoma patients.

Author information

1
Institute of Immunology, University of Regensburg, Regensburg, Germany.
2
Project Group Personalized Tumor Therapy, Fraunhofer Institute of Toxicology and Experimental Medicine, Regensburg, Germany.
3
Experimental Medicine and Therapy Research, University of Regensburg, Regensburg, Germany.
4
Department of Thoracic Surgery, University Regensburg, Regensburg, Germany.
5
Department of Gynecology and Obstetrics, Ludwig-Maximilians-Universität München, Munich, Germany.
6
Department of Gynecology and Obstetrics, University Medical Center Regensburg, Regensburg, Germany.
7
Department of Urology, University of Leipzig, Leipzig, Germany.
8
Department of Urology, Fuerth Hospital, Fuerth, Germany.
9
Clinic of Gynecology and Obstetrics St. Hedwig, University of Regensburg, Regensburg, Germany.
10
Department of Surgery, Klinikum Rechts der Isar, Technische Universität München, Munich, Germany.
11
Project Group Personalized Tumor Therapy, Fraunhofer Institute of Toxicology and Experimental Medicine, Regensburg, Germany; Experimental Medicine and Therapy Research, University of Regensburg, Regensburg, Germany.

Abstract

Tumor xenografts in immunodeficient mice, while routinely used in cancer research, preclude studying interactions of immune and cancer cells or, if humanized by allogeneic immune cells, are of limited use for tumor-immunological questions. Here, we explore a novel way to generate cancer models with an autologous humanized immune system. We demonstrate that hematopoietic stem and progenitor cells (HSPCs) from bone marrow aspirates of non-metastasized carcinoma patients, which are taken at specialized centers for diagnostic purposes, can be used to generate a human immune system in NOD-scid IL2rγ(null) (NSG) and HLA-I expressing NSG mice (NSG-HLA-A2/HHD) comprising both, lymphoid and myeloid cell lineages. Using NSG-HLA-A2/HHD mice, we show that responsive and self-tolerant human T cells develop and human antigen presenting cells can activate human T cells. As critical factors we identified the low potential of bone marrow HSPCs to engraft, generally low HSPC numbers in patient-derived bone marrow samples, cryopreservation and routes of cell administration. We provide here an optimized protocol that uses a minimum number of HSPCs, preselects high-quality bone marrow samples defined by the number of initially isolated leukocytes and intra-femoral or intra-venous injection. In conclusion, the use of diagnostic bone marrow aspirates from non-metastasized carcinoma patients for the immunological humanization of immunodeficient mice is feasible and opens the chance for individualized analyses of anti-tumoral T cell responses.

PMID:
24830425
PMCID:
PMC4022674
DOI:
10.1371/journal.pone.0097860
[Indexed for MEDLINE]
Free PMC Article

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