Wang B y, El-Badri N S, Good R A
Department of Pediatrics, University of South Florida, All Children's Hospital, 801 Sixth Street South, St. Petersburg, FL 33701, USA.
Proc Natl Acad Sci U S A. 1997 Dec 23;94(26):14632-6. doi: 10.1073/pnas.94.26.14632.
We report herein the successful long term engraftment of highly purified hematopoietic stem cells (HSCs) without any facilitating cells in fully allogeneic recipient mice across the entire major histocompatibility complex (MHC) transplantation barrier. This finding challenges the assumption that highly purified marrow HSCs alone cannot produce long-lived allogeneic bone marrow chimeras across the MHC barrier. In the present experiments, 1 x 10(5) HSCs from 5-fluorouracil (5-FU)-treated donors, without any facilitating cells, have been found to repopulate lethally irradiated fully allogeneic recipients. Low density, lineage-negative (CD4-, CD8-, B220-, Mac-1-, Gr-1-), CD71-negative, class I highly positive, FACS-sorted cells from 5-FU-treated C57BL/6 (B6) donor mice were transplanted into lethally irradiated BALB/c recipients. (BALB/c --> BALB/c) --> BALB/c T cell-depleted marrow cells used as compromised cells were also transplanted into the recipients to permit experiments to be pursued over a long period of time. Cells of donor origin in all recognized lineages of hematopoietic cells developed in these allogeneic chimeras. One thousand HSCs were sufficient to repopulate hemiallogeneic recipients, but 1 x 10(4) HSCs alone from 5-FU-treated donors failed to repopulate the fully allogeneic recipients. Transplantation of primary marrow stromal cells or bones of the donor strain into recipient, together with 1 x 10(4) HSCs, also failed to reconstitute fully allogeneic recipients. Suppression of resistance of recipients by thymectomy or injections of granulocyte colony-stimulating factor before stem cell transplantation enhanced the engraftment of allogeneic HSCs. Our experiments show that reconstitution of all lymphohematopoietic lineages across the entire MHC transplantation barriers may be achieved by transplanting allogeneic HSCs alone, without any facilitating cells, as long as a sufficient number of HSCs is transplanted.
我们在此报告，在完全异基因受体小鼠中，跨越整个主要组织相容性复合体（MHC）移植屏障，高度纯化的造血干细胞（HSC）在没有任何辅助细胞的情况下成功实现了长期植入。这一发现挑战了仅高度纯化的骨髓HSC无法跨越MHC屏障产生长期存活的异基因骨髓嵌合体的假设。在本实验中，已发现来自经5-氟尿嘧啶（5-FU）处理的供体的1×10⁵个HSC，在没有任何辅助细胞的情况下，能够使接受致死性照射的完全异基因受体实现造血重建。从经5-FU处理的C57BL/6（B6）供体小鼠中通过荧光激活细胞分选术分选得到的低密度、谱系阴性（CD4⁻、CD8⁻、B220⁻、Mac-1⁻、Gr-1⁻）、CD71阴性、I类高度阳性的细胞，被移植到接受致死性照射的BALB/c受体中。（BALB/c→BALB/c）→用作受损细胞的BALB/c T细胞清除的骨髓细胞也被移植到受体中，以便能够长期进行实验。在这些异基因嵌合体中，造血细胞的所有公认谱系中均出现了供体来源的细胞。1000个HSC足以使半异基因受体实现造血重建，但仅来自经5-FU处理的供体的1×10⁴个HSC未能使完全异基因受体实现造血重建。将供体品系的原代骨髓基质细胞或骨骼与1×10⁴个HSC一起移植到受体中，也未能使完全异基因受体实现造血重建。在干细胞移植前通过胸腺切除或注射粒细胞集落刺激因子来抑制受体的抵抗力，可增强异基因HSC的植入。我们的实验表明，只要移植足够数量的异基因HSC，在没有任何辅助细胞的情况下，单独移植异基因HSC就可能跨越整个MHC移植屏障实现所有淋巴造血谱系的重建。
