Systemic human hematopoietic cell engraftment is compromised following direct bone marrow transplantation in NSG mice

2016 ◽  
Vol 44 (9) ◽  
pp. S72
Author(s):  
Kathryn Futrega ◽  
Bill Lott ◽  
Michael Doran
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Transplantation ◽  
Marrow Transplantation ◽  
Hematopoietic Cell ◽  
Nsg Mice ◽  
Cell Engraftment

scholarly journals Lsh/HELLS is required for B lymphocyte development and immunoglobulin class switch recombination

2020 ◽  
Vol 117 (33) ◽  
pp. 20100-20108
Author(s):  
Yafeng He ◽  
Jianke Ren ◽  
Xiaoping Xu ◽  
Kai Ni ◽  
Andrew Schwader ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Transplantation ◽  
B Cell ◽  
Marrow Transplantation ◽  
Class Switch Recombination ◽  
B Cell Development ◽  
Hematopoietic Cell ◽  
Cell Numbers ◽  
Class Switch

Mutation of HELLS (Helicase, Lymphoid-Specific)/Lsh in human DNA causes a severe immunodeficiency syndrome, but the nature of the defect remains unknown. We assessed here the role of Lsh in hematopoiesis using conditional Lsh knockout mice with expression of Mx1 or Vav Cre-recombinase. Bone marrow transplantation studies revealed that Lsh depletion in hematopoietic stem cells severely reduced B cell numbers and impaired B cell development in a hematopoietic cell-autonomous manner. Lsh-deficient mice without bone marrow transplantation exhibited lower Ig levels in vivo compared to controls despite normal peripheral B cell numbers. Purified B lymphocytes proliferated normally but produced less immunoglobulins in response to in vitro stimulation, indicating a reduced capacity to undergo class switch recombination (CSR). Analysis of germline transcripts, examination of double-stranded breaks using biotin-labeling DNA break assay, and End-seq analysis indicated that the initiation of the recombination process was unscathed. In contrast, digestion–circularization PCR analysis and high-throughput sequencing analyses of CSR junctions and a chromosomal break repair assay indicated an impaired ability of the canonical end-joining pathway in Lsh-deficient B cells. Our data suggest a hematopoietic cell-intrinsic role of Lsh in B cell development and in CSR providing a potential target for immunodeficiency therapy.

Specific Non-Responsiveness to Pig in Baboons Receiving BMT from GalT-KO Pigs.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 5265-5265 ◽  
Author(s):  
Ahmed Ghazi ◽  
Adam Griesemer ◽  
Masayoshi Okumi ◽  
Erica Hirsh ◽  
Diana Lo ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Bone Marrow Transplantation ◽  
Nonhuman Primate ◽  
Marrow Transplantation ◽  
Progenitor Cell ◽  
Cytotoxicity Assay ◽  
Whole Body ◽  
Serum Antibodies ◽  
Cell Engraftment

Abstract Background: The induction of stable hematopoietic cell chimerism through bone marrow transplantation (BMT) has been demonstrated to induce donor-specific tolerance in rodent, porcine, nonhuman primate, and human clinical allogenic models, and has also been successful in concordant rodent and nonhuman primate xenogeneic models, as well as in the pig-to-NOD/SCID humanized mouse xenogenic model. However, stable chimerism and tolerance has been difficult to achieve in the discordant pig-to-baboon xenotransplantation model, possibly due in part to the presence in baboons of pre-formed natural xeno-reactive antibodies to a1,3-galactose (Gal) determinants expressed in pigs, but not in Old World primates and humans. The recent availability of miniature swine homozygous for a disruption in the gene encoding a1,3-galactosyltransferase (GalT-KO pigs) has now made it possible to study pig-to-baboon xenografts in the absence of effects of anti-Gal antibodies. We have investigated the GalT-KO pig-to-baboon model further by modifying the conditioning and immunosuppression regimen to facilitate engraftment and tolerance through bone marrow transplantation. Methods: BM was harvested from GalT-KO swine (n=3). Baboons (n=3) were pre-treated with whole body (3 Gy) and thymic (7 Gy) irradiation, Sangstat rabbit anti-thymocyte globulin (ATG), LoCD2b (rat IgG2b anti-primate CD2) and splenectomy, and received FK506 immunosuppressive and supportive therapy for 28 days. The baboons were monitored for the presence of pig cells by flow cytometry, for porcine progenitor cells in the bone marrow by porcine cytochrome b specific PCR of colony-forming units (CFUs), and for cellular reactivity to pig cells by MLR and CML. Antibody formation to LoCD2b and ATG was tested by enzyme-linked immunosorbent assay (ELISA), and antibody reactivity to GalT-KO pig cells was tested by flow cytometry and antibody mediated cytotoxicity assay. Results: A mean of 1.4 × 109 BM cells/kg was infused into each baboon. Although pig cells were undetectable in the peripheral blood of the baboons by flow cytometry, porcine progenitor cell engraftment as well as chimerism in the bone marrow and thymus was detected by PCR in the first baboon on day 28. ELISA results indicated the presence of antibodies to rat (LoCD2b) and rabbit (ATG) immunoglobulin within two weeks; however, no antibodies to pig cells could be detected by flow cytometry or cytotoxicity assay. The second baboon had undetectable serum antibodies to pig cells for 60 days despite the presence of induced antibodies to rat LoCD2b and rabbit ATG. Porcine progenitor cell engraftment was confirmed by PCR of CFUs on day 60 and MLR showed no response to pig although the animal regained alloresponses by this time. The third baboon, in contrast, had detectable induced serum antibodies to pig cells as well as rat and rabbit immunoglobulin by day 14 following BMT. Conclusions: Engraftment has been achieved following GalT-KO pig-to-baboon BMT with evidence of specific humoral and cellular non-responsiveness to pig cells (2/3 baboons), suggesting the possibility that this protocol may facilitate xenograft tolerance.

scholarly journals Host bone marrow-derived IL-12 enhances donor T cell engraftment in a mouse model of bone marrow transplantation

2014 ◽  
Vol 7 (1) ◽  
pp. 16 ◽  
Author(s):  
Katarzyna A Darlak ◽  
Ying Wang ◽  
Jian-Ming Li ◽  
Wayne AC Harris ◽  
Cynthia R Giver ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Transplantation ◽  
T Cell ◽  
Mouse Model ◽  
Marrow Transplantation ◽  
Host Bone ◽  
Cell Engraftment

scholarly journals Donor–origin cell engraftment after intraosseous or intravenous bone marrow transplantation in a rat model

2007 ◽  
Vol 40 (4) ◽  
pp. 373-380 ◽  
Author(s):  
A Klimczak ◽  
S Unal ◽  
A Jankowska ◽  
C Coburn ◽  
M Siemionow
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Transplantation ◽  
Rat Model ◽  
Marrow Transplantation ◽  
Cell Engraftment

scholarly journals Extrathymic T Cell Deletion and Allogeneic Stem Cell Engraftment Induced with Costimulatory Blockade Is Followed by Central T Cell Tolerance

1998 ◽  
Vol 187 (12) ◽  
pp. 2037-2044 ◽  
Author(s):  
Thomas Wekerle ◽  
Mohamed H. Sayegh ◽  
Joshua Hill ◽  
Yong Zhao ◽  
Anil Chandraker ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Stem Cell ◽  
Bone Marrow Transplantation ◽  
T Cell ◽  
Marrow Transplantation ◽  
T Cell Tolerance ◽  
Cell Tolerance ◽  
Costimulatory Blockade ◽  
Cell Engraftment

A reliable, nontoxic method of inducing transplantation tolerance is needed to overcome the problems of chronic organ graft rejection and immunosuppression-related toxicity. Treatment of mice with single injections of an anti-CD40 ligand antibody and CTLA4Ig, a low dose (3 Gy) of whole body irradiation, plus fully major histocompatibility complex–mismatched allogeneic bone marrow transplantation (BMT) reliably induced high levels (>40%) of stable (>8 mo) multilineage donor hematopoiesis. Chimeric mice permanently accepted donor skin grafts (>100 d), and rapidly rejected third party grafts. Progressive deletion of donor-reactive host T cells occurred among peripheral CD4+ lymphocytes, beginning as early as 1 wk after bone marrow transplantation. Early deletion of peripheral donor-reactive host CD4 cells also occurred in thymectomized, similarly treated marrow recipients, demonstrating a role for peripheral clonal deletion of donor-reactive T cells after allogeneic BMT in the presence of costimulatory blockade. Central intrathymic deletion of newly developing T cells ensued after donor stem cell engraftment had occurred. Thus, we have shown that high levels of chimerism and systemic T cell tolerance can be reliably achieved without myeloablation or T cell depletion of the host. Chronic immunosuppression and rejection are avoided with this powerful, nontoxic approach to inducing tolerance.

scholarly journals IGF-1-mediated osteoblastic niche expansion enhances long-term hematopoietic stem cell engraftment after murine bone marrow transplantation

Stem Cells ◽  
2013 ◽  
Vol 31 (10) ◽  
pp. 2193-2204 ◽  
Author(s):  
Anna Caselli ◽  
Timothy S. Olson ◽  
Satoru Otsuru ◽  
Xiaohua Chen ◽  
Ted J. Hofmann ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stem Cell ◽  
Bone Marrow Transplantation ◽  
Hematopoietic Stem Cell ◽  
Marrow Transplantation ◽  
Hematopoietic Stem ◽  
Niche Expansion ◽  
Murine Bone Marrow ◽  
Cell Engraftment
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