Immune humanization of immunodeficient mice using diagnostic bone marrow aspirates from carcinoma patients.
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.
Institute of Immunology, University of Regensburg, Regensburg, Germany.
Melanie
M
Werner-Klein
Institute of Immunology, University of Regensburg, Regensburg, Germany.
Judith
J
Proske
Project Group Personalized Tumor Therapy, Fraunhofer Institute of Toxicology and Experimental Medicine, Regensburg, Germany.
Christian
C
Werno
Experimental Medicine and Therapy Research, University of Regensburg, Regensburg, Germany.
Katharina
K
Schneider
Department of Thoracic Surgery, University Regensburg, Regensburg, Germany.
Hans-Stefan
HS
Hofmann
Department of Gynecology and Obstetrics, Ludwig-Maximilians-Universität München, Munich, Germany.
Brigitte
B
Rack
Department of Gynecology and Obstetrics, University Medical Center Regensburg, Regensburg, Germany.
Stefan
S
Buchholz
Department of Urology, University of Leipzig, Leipzig, Germany.
Roman
R
Ganzer
Department of Urology, Fuerth Hospital, Fuerth, Germany.
Andreas
A
Blana
Clinic of Gynecology and Obstetrics St. Hedwig, University of Regensburg, Regensburg, Germany.
Birgit
B
Seelbach-Göbel
Department of Surgery, Klinikum Rechts der Isar, Technische Universität München, Munich, Germany.
Ulrich
U
Nitsche
Institute of Immunology, University of Regensburg, Regensburg, Germany.
Daniela N
DN
Männel
Project Group Personalized Tumor Therapy, Fraunhofer Institute of Toxicology and Experimental Medicine, Regensburg, Germany; Experimental Medicine and Therapy Research, University of Regensburg, Regensburg, Germany.
Christoph A
CA
Klein
D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D001853:Bone Marrow / Q000276:immunology / Q000473:pathology; D001943:Breast Neoplasms / Q000276:immunology / Q000473:pathology; D004195:Disease Models, Animal; D004938:Esophageal Neoplasms / Q000276:immunology / Q000473:pathology; D005260:Female; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000276:immunology / Q000378:metabolism; D006801:Humans; D008175:Lung Neoplasms / Q000276:immunology / Q000473:pathology; D008297:Male; D008810:Mice, Inbred C57BL; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D011471:Prostatic Neoplasms / Q000276:immunology / Q000473:pathology
D018952:Antigens, CD34