Volume 6, Issue 1, Pages (January 2016)

Slides:



Advertisements
Similar presentations
Joseph H. Chewning, Weiwei Zhang, David A. Randolph, C
Advertisements

by Jad I. Belle, David Langlais, Jessica C
Single Targeted Exon Mutation Creates a True Congenic Mouse for Competitive Hematopoietic Stem Cell Transplantation: The C57BL/6-CD45.1STEM Mouse  Francois.
Induction of Immunity to Neuroblastoma Early after Syngeneic Hematopoietic Stem Cell Transplantation Using a Novel Mouse Tumor Vaccine  Weiqing Jing,
Volume 7, Issue 3, Pages (September 2016)
Yumi Matsuzaki, Kentaro Kinjo, Richard C Mulligan, Hideyuki Okano 
Volume 25, Issue 9, Pages (September 2017)
Volume 8, Issue 5, Pages (May 2017)
Francesca Ficara, Mark J. Murphy, Min Lin, Michael L. Cleary 
Volume 6, Issue 4, Pages (April 1997)
by Hyung-Gyoon Kim, Kyoko Kojima, C. Scott Swindle, Claudiu V
Low c-Kit Expression Level Induced by Stem Cell Factor Does Not Compromise Transplantation of Hematopoietic Stem Cells  Chia-Ling Chen, Katerina Faltusova,
Cited2 Is an Essential Regulator of Adult Hematopoietic Stem Cells
by Xue Li, Jared Sipple, Qishen Pang, and Wei Du
Volume 26, Issue 4, Pages (April 2007)
Volume 3, Issue 5, Pages (November 2014)
by Ute Koch, Anne Wilson, Monica Cobas, Rolf Kemler, H
Volume 8, Issue 6, Pages (June 2017)
Volume 15, Issue 6, Pages (June 2014)
Volume 19, Issue 2, Pages (August 2003)
Marking of definitive HSC precursors in E7. 5–E8
Cytotoxic CD8+ T Cells Stimulate Hematopoietic Progenitors by Promoting Cytokine Release from Bone Marrow Mesenchymal Stromal Cells  Christian M. Schürch,
Volume 2, Issue 4, Pages (April 2008)
Volume 4, Issue 2, Pages (February 2003)
Volume 25, Issue 9, Pages (September 2017)
Mark J. Kiel, Melih Acar, Glenn L. Radice, Sean J. Morrison 
Volume 19, Issue 5, Pages (November 2003)
T Cell and B Cell Immunity can be Reconstituted with Mismatched Hematopoietic Stem Cell Transplantation Without Alkylator Therapy in Artemis-Deficient.
Volume 10, Issue 5, Pages (November 2004)
Volume 11, Issue 1, Pages (July 2018)
Volume 22, Issue 4, Pages e4 (April 2018)
Motonari Kondo, Irving L. Weissman, Koichi Akashi  Cell 
Volume 14, Issue 12, Pages (March 2016)
Volume 43, Issue 2, Pages (August 2015)
Volume 16, Issue 5, Pages (May 2002)
Ravindra Majeti, Christopher Y. Park, Irving L. Weissman 
Volume 6, Issue 3, Pages (March 2016)
Volume 2, Issue 4, Pages (April 2014)
Volume 15, Issue 3, Pages (April 2016)
Deletion of the Scl +19 enhancer increases the blood stem cell compartment without affecting the formation of mature blood lineages  Dominik Spensberger,
Volume 1, Issue 3, Pages (September 2007)
Volume 11, Issue 3, Pages (September 2012)
Volume 11, Issue 2, Pages (August 2018)
Volume 16, Issue 12, Pages (September 2016)
Volume 5, Issue 5, Pages (November 2015)
Volume 6, Issue 3, Pages (March 2010)
Volume 7, Issue 6, Pages (December 2016)
Potential Pitfalls of the Mx1-Cre System: Implications for Experimental Modeling of Normal and Malignant Hematopoiesis  Talia Velasco-Hernandez, Petter.
Cytokines and cytotoxic pathways in engraftment resistance to purified allogeneic hematopoietic stem cells  Christian Scheffold, Yolanda C. Scheffold,
Volume 4, Issue 2, Pages (February 2009)
Volume 17, Issue 4, Pages (October 2002)
Volume 15, Issue 4, Pages (October 2001)
Donor antigen-presenting cells regulate T-cell expansion and antitumor activity after allogeneic bone marrow transplantation  Jian-Ming Li, Edmund K.
Volume 1, Issue 6, Pages (December 2007)
Volume 9, Issue 4, Pages (October 2017)
Volume 15, Issue 3, Pages (September 2001)
Volume 5, Issue 5, Pages (November 2015)
Volume 29, Issue 4, Pages (October 2008)
Volume 1, Issue 6, Pages (December 2007)
Volume 2, Issue 1, Pages (January 2008)
Kiran Batta, Magdalena Florkowska, Valerie Kouskoff, Georges Lacaud 
Volume 11, Issue 6, Pages (June 2005)
Volume 33, Issue 5, Pages (November 2010)
SLAM Family Markers Resolve Functionally Distinct Subpopulations of Hematopoietic Stem Cells and Multipotent Progenitors  Hideyuki Oguro, Lei Ding, Sean J.
Volume 8, Issue 6, Pages (June 2017)
Volume 17, Issue 2, Pages (August 2002)
Volume 6, Issue 4, Pages (April 1997)
Volume 23, Issue 4, Pages (October 2005)
B1b Lymphocytes Confer T Cell-Independent Long-Lasting Immunity
Alicia G Arroyo, Joy T Yang, Helen Rayburn, Richard O Hynes  Cell 
Presentation transcript:

Volume 6, Issue 1, Pages 137-149 (January 2016) Fetal Hematopoietic Stem Cell Transplantation Fails to Fully Regenerate the B- Lymphocyte Compartment  Eliver Eid Bou Ghosn, Jeffrey Waters, Megan Phillips, Ryo Yamamoto, Brian R. Long, Yang Yang, Rachel Gerstein, Cheryl A. Stoddart, Hiromitsu Nakauchi, Leonore A. Herzenberg  Stem Cell Reports  Volume 6, Issue 1, Pages 137-149 (January 2016) DOI: 10.1016/j.stemcr.2015.11.011 Copyright © 2016 The Authors Terms and Conditions

Figure 1 Fetal Liver LT-HSCs Provide Stable Long-Term Reconstitution of All Major Blood Subsets (A) Experimental design. ∼E15 livers were processed into single-cell suspensions and stained as described in the Experimental Procedures. RFP+ LT-HSCs were sorted and injected intravenously along with rescue RFP–GFP+ BM cells into lethally irradiated C57BL/6. (B) Gating strategy to purify fetal liver LT-HSCs (Lin–, C-KIThi, SCA-1hi, CD150+, CD48–, CD41–, CD45+) (see also Figure S1). KLS: C-KIThi, Lineage–, SCA-1hi cells. (C) Levels of blood chimerism (% RFP+ donor cells) in mice that received ∼100 sorted fetal liver LT-HSCs (∼E15), ∼1.5 × 106 unsorted fetal liver (∼E15), and ∼1.5 × 106 unsorted adult (>8 weeks old) BM. Recipient mice were bled several times to determine the percentage of donor-derived B cells, T cells, erythrocytes, and myeloid-granulocytes. (D) Gating strategy identifying all major peripheral blood subsets derived from fetal liver LT-HSCs (RFP+). Data are representative of six recipient mice from three independent experiments. Stem Cell Reports 2016 6, 137-149DOI: (10.1016/j.stemcr.2015.11.011) Copyright © 2016 The Authors Terms and Conditions

Figure 2 Fetal Liver LT-HSCs Selectively Fail to Reconstitute B-1a in Peritoneal Cavity (A) B-cell subsets from adult intact (unmanipulated) C57BL/6 were compared with B cells from mice that received RFP+ sorted fetal liver LT-HSCs, unsorted fetal liver, and sorted fetal liver KLS CD150– transplants (∼E15 embryos). Total donor B cells (RFP+, CD19+) were analyzed for the expression of CD5 and CD23 to identify B-1a (CD5+CD23–), B-1b (CD5–CD23–), and B-2 (CD5–CD23+). (B) Percentage of B-cell subsets in PerC of mice that received sorted or unsorted fetal liver transplants compared with adult unmanipulated C57BL/6. ∗p < 0.0001, ∗∗p = 0.001. (C) Absolute numbers of B-1a cells in PerC of mice that received sorted or unsorted fetal liver transplants. ∗p < 0.0001, ∗∗p = 0.0009, ∗∗∗p = 0.03. Error bars represent the mean and SD for B-cell numbers obtained from the fully chimeric recipient mice that showed the highest levels of B-cell chimerism (six mice for LT-HSCs, seven mice for unsorted fetal liver, and four mice for KLS CD150–) from three independent experiments. p Value for means comparison using Student's t test, α = 0.05 (JMP; SAS Institute). (D) Percentage of blood cell subsets (RFP+) detected in recipients ∼7.5 months after LT-HSC transplantation, when recipient PerC cells were harvested. (E) BM from recipient mouse ∼7.5 months after transplantation. RFP– cells are derived from rescue BM co-transplanted with RFP+ LT-HSCs. KLS: C-KIThi, Lineage–, SCA-1hi population enriched for LT-HSCs. Stem Cell Reports 2016 6, 137-149DOI: (10.1016/j.stemcr.2015.11.011) Copyright © 2016 The Authors Terms and Conditions

Figure 3 Fetal Liver LT-HSCs Effectively Reconstitute FO and MZ B Cells, but Fail to Reconstitute B-1a in Spleen (A) Splenic B-cell subsets in adult intact (unmanipulated) C57BL/6 compared with B cells that were reconstituted in mice that received RFP+ sorted fetal liver LT-HSCs, unsorted fetal liver, and sorted fetal liver KLS CD150– transplants (∼E15 embryos). Live, single cells were analyzed for the expression of CD5 and CD19 to identify T and B cells, respectively. Spleen from recipient mice were first gated on RFP+ to include all progeny derived from the donor transplants. CD21 and CD23 expression levels on B cells (CD19+) distinguished MZ (CD21hiCD23–), FO (CD23hiCD21lo), and immature + B-1a (CD21lo/–CD23lo/–). B-1a were identified as CD5+B220lo/−. (B) Percentage of B-cell subsets in spleen of mice that received sorted or unsorted fetal liver transplants compared to adult unmanipulated C57BL/6. ∗p < 0.0001, ∗∗p = 0.001, ∗∗∗p = 0.05. (C) Absolute numbers of B-1a cells in spleen of mice that received sorted or unsorted fetal liver transplants. ∗p < 0.0001, ∗∗p = 0.03, ∗∗∗p = 0.05. Error bars represent the mean and SD for the B-cell numbers obtained from the fully chimeric recipient mice that showed the highest levels of B-cell chimerism (six mice for LT-HSCs, seven mice for unsorted fetal liver, and four mice for KLS CD150–) from three independent experiments. p Value for means comparison using Student's t test, α = 0.05. Stem Cell Reports 2016 6, 137-149DOI: (10.1016/j.stemcr.2015.11.011) Copyright © 2016 The Authors Terms and Conditions

Figure 4 Anti-Phospholipid VH11 Antibody Repertoire Is Absent in Mice that Received LT-HSC Transplantation (A) PerC cells from adult (unmanipulated) C57BL/6, and mice that received RFP+ sorted LT-HSCs and unsorted fetal liver (∼E15) were stained for anti-VH11 in an 18-par Hi-D FACS panel. Cells were first gated on RFP+ to include all progeny derived from donor transplants. Donor-derived B cells (RFP+, CD19+) were analyzed for the expression of surface VH11 and IGM. B cells expressing IGM VH11 repertoire form a diagonal cluster in the FACS plot. Recipient mice were analyzed ∼7.5 months after transplantation. (B) Percentage of VH11+IGM+ B cells in PerC of mice that received sorted (LT-HSCs) or unsorted fetal liver transplants compared to adult unmanipulated C57BL/6. ∗p < 0.0001, ∗∗p = 0.0007. Error bars represent the mean and SD. p Value for means comparison using Student's t test, α = 0.05. Data are representative of the six recipient mice (in each group) that showed the highest levels of B-cell chimerism in three independent experiments. Stem Cell Reports 2016 6, 137-149DOI: (10.1016/j.stemcr.2015.11.011) Copyright © 2016 The Authors Terms and Conditions