Transplantation of CD34 human cells into mice with severe combined immunodeficiency results in functional T cells 4 weeks after transplantation

• Published in: American journal of obstetrics and gynecology Vol. 181; no. 1; pp. 80 - 86
• Main Authors: Polcaro, Joseph, Divon, Michael Y., Bentolila, Eric, Rashbaum, William K., Lyman, William D.
• Format: Journal Article
• Language: English
• Published: Philadelphia, PA Mosby, Inc 01.07.1999; Elsevier
• Subjects: Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Animals; Antigens, CD34; Biological and medical sciences; Bone marrow, stem cells transplantation. Graft versus host reaction; Hematopoietic Stem Cell Transplantation; Humans; Liver - cytology; Medical sciences; Mice; mice with severe combined immunodeficiency; Severe Combined Immunodeficiency; Stem cells; T cells; T-Lymphocytes - metabolism; Time Factors; Transfusions. Complications. Transfusion reactions. Cell and gene therapy; transplantation; United States; North America; America; T cells; mice with severe combined immunodeficiency; Stem cells; transplantation; Human; Immunopathology; Animal model; Stem cell; Rodentia; Hematopoietic cell; Experimental study; Immune deficiency; Vertebrata; Mammalia; Fetal cell; Mouse; Animal; T-Lymphocyte; Evolution; Graft; Comparative study
• ISSN: 0002-9378; 1097-6868
• DOI: 10.1016/S0002-9378(99)70439-4

Objective: Our purpose was to determine whether transplantation of fetal human CD34 + cells into mice with severe combined immunodeficiency results in functional T cells. Study Design: The cells used in this study were isolated from fetal human liver tissue obtained after elective termination of normal 18- to 24-week pregnancies. Women with medical conditions that could confound the outcome were excluded. Cells were labeled with fluorochrome-conjugated antibodies that recognized CD34 or other cell surface antigens. The cells were then sorted with the use of a fluorescein-activated cell sorter. The human sorted cells were injected intraperitoneally in mice with severe combined immunodeficiency. Four groups of mice were studied: group 1, injected with 10 5 CD34 + cells (n = 17); group 2, injected with 10 5 CD34 – cells (n = 14); group 3, injected with 10 6 unsorted cells (n = 19); and group 4, sham-injected with phosphate-buffered saline solution as controls (n = 14). At 1, 2, and 4 weeks after transplantation, the peripheral blood monocytes of the study mice were analyzed for functional T cells. Aliquots of cells (10 5) were incubated for 48 hours with 0, 5, 10, and 20 μg of phytohemagglutinin. Thereafter the cells were treated with 1 μCi of tritiated thymidine. Subsequently the incorporation of tritiated thymidine was determined by liquid scintillation counting. Results: Cells from mice transplanted with either unsorted cells, sorted CD34 + cells, or CD34 – cells showed a response to phytohemagglutinin that varied with time and with the mitogen concentration. Even though unsorted fetal human liver cells had a maximal response at 2 weeks, this posttransplantation response was not statistically significant. CD34 + cell response to phytohemagglutinin was significant at 4 weeks after transplantation. CD34 – cells also had a peripheral blood cell response at 4 weeks after transplantation; however, this response was not statistically significant. In addition, all mice transplanted with fetal human liver cells had some functional T cells at 4 weeks; however, this response was statistically significant only for CD34 + cells. Conclusion: Transplantation of either sorted CD34 (positive or negative) cells or unsorted fetal human liver cell preparations into mice with severe combined immunodeficiency results in functional T cells. However, only the mice with transplanted CD34 + cells demonstrated a statistically significant response. (Am J Obstet Gynecol 1999;181:80-6.)