181 results on '"Muench, MO"'
Search Results
2. Epigenetic remodeling in B-cell acute lymphoblastic leukemia occurs in two tracks and employs embryonic stem cell-like signatures
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Wiemels, Joseph, Wiencke, John, Wrensch, Margaret, Lee, ST, Muench, MO, Fomin, ME, Xiao, J, Zhou, M, De, A, Martín-Subero, JI, Heath, S, Houseman, EA, and Roy, R
- Abstract
© The Author(s) 2015.We investigated DNA methylomes of pediatric Bcell acute lymphoblastic leukemias (B-ALLs) using whole-genome bisulfite sequencing and highdefinition microarrays, along with RNA expression profiles. Epigenetic alteration of B-ALLs occurr
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- 2015
3. A global DNA methylation and gene expression analysis of early human B-cell development reveals a demethylation signature and transcription factor network
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Wiemels, Joseph, Wiencke, John, Lee, ST, Xiao, Y, Muench, MO, Xiao, J, Fomin, ME, Wiencke, JK, Zheng, S, Dou, X, De, A, and Chokkalingam, A
- Abstract
The epigenetic changes during B-cell development relevant to both normal function and hematologic malignancy are incompletely understood. We examined DNA methylation and RNA expression status during early B-cell development by sorting multiple replicates o
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- 2012
4. Identification of a common T/natural killer cell progenitor in human fetal thymus.
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Sánchez, MJ, Muench, MO, Roncarolo, MG, Lanier, LL, and Phillips, JH
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Stem Cell Research - Nonembryonic - Human ,Biotechnology ,Stem Cell Research ,Animals ,Antigens ,CD ,Antigens ,CD34 ,Cell Differentiation ,Cell Separation ,Flow Cytometry ,Hematopoietic Stem Cells ,Humans ,Interleukin-2 ,Killer Cells ,Natural ,Mice ,Phenotype ,T-Lymphocytes ,Thymus Gland ,Medical and Health Sciences ,Immunology - Abstract
The phenotypic similarities between natural killer (NK) and T cells have led to the hypothesis that these distinctive lymphocyte subsets may be developmentally related and thus may share a common progenitor (Lanier, L. L., H. Spits, and J. H. Phillips, 1992. Immunol. Today. 13:392; Rodewald, H.-R., P. Moingeon, J. L. Lurich, C. Dosiou, P. Lopez, and E. L. Reinherz. 1992. Cell. 69:139). In this report, we have investigated the potential of human CD34+ triple negative thymocytes ([TN] CD3-, CD4-, CD8-) to generate both T cells and NK cells in murine fetal thymic organ cultures (mFTOC) and in vitro clonogenic assays. CD34+ TN thymocytes, the majority of which express prominent cytoplasmic CD3 epsilon (cytoCD3 epsilon) protein, can be divided into high (CD34Bright) and low (CD34Dim) surface expressing populations. CD34Bright TN thymocytes were capable of differentiating into T and NK cells when transferred into mFTOC, and demonstrated high NK cell clonogenic capabilities when cultured in interleukin (IL)-2, IL-7, and stem cell factor (SCF). Likewise, CD34Bright TN thymocyte clones after 5 d in culture were capable of generating NK and T cells when transferred into mFTOC but demonstrated clonogenic NK cell differentiation capabilities when maintained in culture with IL-2. CD34Dim TN thymocytes, however, possessed only T cell differentiation capabilities in mFTOC but were not expandable in clonogenic conditions containing IL-2, IL-7, and SCF. No significant differentiation of other cell lineage was detected in either mFTOC or in clonogenic assays from CD34+ TN thymocytes. These results represent the first definitive evidence of a common T/NK cell progenitor in the human fetal thymus and delineate the point in thymocyte differentiation where T and NK cells diverge.
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- 1994
5. Transplantation of a fetus with paternal Thy-1+CD34+cells for chronic granulomatous disease
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Muench, MO, Rae, J, Bárcena, A, Leemhuis, T, Farrell, J, Humeau, L, Maxwell-Wiggins, JR, Capper, J, Mychaliska, GB, Albanese, CT, Martin, T, Tsukamoto, A, Curnutte, JT, and Harrison, MR
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- 2001
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6. Mid-trimester fetal livers are a rich source of CD34+/++ cells for transplantation
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Golfier, F, Bárcena, A, Cruz, J, Harrison, MR, and Muench, MO
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- 1999
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7. Cover Image
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Togarrati, PP, primary, Dinglasan, N, additional, Desai, S, additional, Ryan, WR, additional, and Muench, MO, additional
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- 2018
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8. CD29 is highly expressed on epithelial, myoepithelial, and mesenchymal stromal cells of human salivary glands
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Togarrati, PP, primary, Dinglasan, N, additional, Desai, S, additional, Ryan, WR, additional, and Muench, MO, additional
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- 2018
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9. The FLK2/FLT3 ligand synergizes with interleukin-7 in promoting stromal-cell-independent expansion and differentiation of human fetal pro-B cells in vitro
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Namikawa R, Muench MO, deVries JE, RONCAROLO , MARIA GRAZIA, Namikawa, R, Muench, Mo, Devries, Je, and Roncarolo, MARIA GRAZIA
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The effects of a novel cytokine FLK2/FLT3 ligand (FL) on human fetal bone marrow-derived CD34(+)CD19(+) pro-B cells were analyzed in a stromal-cell-independent, serum-deprived culture system. FL, like interleukin-3 (IL-3), synergized with IL-7 in promoting pro-B cell growth, and differentiation of these cells into CD34(-)CD19(+)clgM(+)slgM(-) pre-B cells, whereas a small proportion of these cells even differentiate into more mature slgM(+) B cells. In contrast, KIT ligand (KL) and granulocyte-macrophage colony-stimulating factor (GM-CSF) were ineffective in promoting IL-7-dependent pro-B cell growth and differentiation. Maximal levels of pro-B cell expansion, generally resulting in 15- to 30-fold increases in cellularity, were obtained in cultures supplemented with optimal doses of FL + IL-7 + IL-3. The addition of mouse bone marrow stromal cells further enhanced the proliferation and differentiation of pro-B cells obtained in the presence of these three cytokines. Under these conditions, cultures could be maintained for more than 4 weeks, and in general 40- to 50-fold increases in cell numbers were observed by 3 weeks of culture, The percentages of clgM(+) and slgM(+) B cells increased 1.5- to 3-fold and 2-fold, respectively, suggesting that stromal cells may provide additional costimulatory signals for human B-cell growth and differentiation that are different from IL-7, IL-3, and FL. Collectively, our results indicate that FL, in contrast to KL, strongly promotes long-term expansion and differentiation of human pro-B cells in the presence of IL-7 or in combination of IL-7 and IL-3, which is a novel property of this hematopoietic growth factor. (C) 1996 by The American Society of Hematology.
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- 1996
10. TRACING THE EXPRESSION OF CD7 AND OTHER ANTIGENS DURING T-CELL AND MYELOID-CELL DIFFERENTIATION IN THE HUMAN FETAL LIVER AND THYMUS
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BARCENA A, MUENCH MO, SPITS H., RONCAROLO , MARIA GRAZIA, Barcena, A, Muench, Mo, Roncarolo, MARIA GRAZIA, and Spits, H.
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During the last decade, the function/s of the cell membrane CD7 antigen have been investigated in human mature T and NK cells, showing the direct involvement of this molecule in multiple effector functions related with activation, proliferation, production of cytokines and modification of adhesion properties. The CD7 glycoprotein is not only expressed by mature lymphoid cells, but also by early hematopoietic progenitors and several types of leukemias, suggesting a role of CD7 during hematopoiesis. However, the function of CD7 in the early stages of hematopoietic development has not yet been elucidated. CD7 has been classically considered the earliest T-cell specific marker. This assumption was based on data indicating the presence of CD45(+)CD7(+)CD3(-)CD4(-)CD8(-) cells in the human embryonic/fetal liver at the gestational age at which the thymic rudiment is colonized by T-cell progenitors. In the present article, we review recent results obtained by several groups concerning the expression of CD7 and various other cell surface antigens by T-, B- and myeloid-cell progenitors generated in the adult bone marrow and fetal liver. In addition, we present an hypothetical model of hematopoiesis in the fetal liver and thymus.
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- 1995
11. Effects of cytokine administration on human hematopoiesis in the SCID-hu mouse
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Muench MO, Namikawa R., RONCAROLO , MARIA GRAZIA, Muench, Mo, Roncarolo, MARIA GRAZIA, and Namikawa, R.
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- 1995
12. PHENOTYPIC AND FUNCTIONAL-ANALYSIS OF T-CELL PRECURSORS IN THE HUMAN FETAL LIVER AND THYMUS - CD7 EXPRESSION IN THE EARLY STAGES OF T-CELL AND MYELOID-CELL DEVELOPMENT
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BARCENA A, MUENCH MO, GALY AHM, CUPP J, PHILLIPS JH, SPITS H., RONCAROLO , MARIA GRAZIA, Barcena, A, Muench, Mo, Galy, Ahm, Cupp, J, Roncarolo, MARIA GRAZIA, Phillips, Jh, and Spits, H.
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- 1993
13. Expression of Fas/CD95 and Bcl-2 by primitive hematopoietic progenitors freshly isolated from human fetal liver
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Barcena, A, primary, Park, SW, additional, Banapour, B, additional, Muench, MO, additional, and Mechetner, E, additional
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- 1996
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14. The FLK2/FLT3 ligand synergizes with interleukin-7 in promoting stromal- cell-independent expansion and differentiation of human fetal pro-B cells in vitro
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Namikawa, R, primary, Muench, MO, additional, de Vries, JE, additional, and Roncarolo, MG, additional
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- 1996
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15. FLK-2/FLT-3 ligand regulates the growth of early myeloid progenitors isolated from human fetal liver
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Muench, MO, primary, Roncarolo, MG, additional, Menon, S, additional, Xu, Y, additional, Kastelein, R, additional, Zurawski, S, additional, Hannum, CH, additional, Culpepper, J, additional, Lee, F, additional, and Namikawa, R, additional
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- 1995
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16. Expression of CD33, CD38, and HLA-DR on CD34+ human fetal liver progenitors with a high proliferative potential
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Muench, MO, primary, Cupp, J, additional, Polakoff, J, additional, and Roncarolo, MG, additional
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- 1994
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17. Phenotypic and functional analysis of T-cell precursors in the human fetal liver and thymus: CD7 expression in the early stages of T- and myeloid-cell development
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Barcena, A, primary, Muench, MO, additional, Galy, AH, additional, Cupp, J, additional, Roncarolo, MG, additional, Phillips, JH, additional, and Spits, H, additional
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- 1993
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18. Bone marrow transplantation with interleukin-1 plus kit-ligand ex vivo expanded bone marrow accelerates hematopoietic reconstitution in mice without the loss of stem cell lineage and proliferative potential
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Muench, MO, primary, Firpo, MT, additional, and Moore, MA, additional
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- 1993
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19. Purification and partial characterization of a human hematopoietic precursor population
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Smith, C, primary, Gasparetto, C, additional, Collins, N, additional, Gillio, A, additional, Muench, MO, additional, O'Reilly, RJ, additional, and Moore, MA, additional
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- 1991
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20. Requirement of Retinoids for the Expression of CD38 on Human Hematopoietic Progenitors in vitro.
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Muench, MO, Barcena, A, Ohkubo, T, and Harrison, MR
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RETINOIDS , *GENE expression , *HEMATOPOIETIC stem cells , *BONE marrow cells , *HEMATOPOIETIC system , *BLOOD cells , *STEM cells , *CAROTENOIDS , *DITERPENES - Abstract
Background Cells expressing high levels of CD34 and little or no CD38 comprise a primitive compartment of progenitors, thought to include hematopoietic stem cells. In this study we sought to determine the feasibility of using CD34 and CD38 as markers of hematopoietic differentiation in vitro, using retinoids to induce the expression of CD38. Methods The effects over time of culture, sera and retinoids on the expression of CD34 and CD38 were determined using a base-medium lacking serum. Two early progenitor populations, isolated by FACS from human fetal liver, were studied: CD38 - CD34 ++ and CD38 + CD34 ++ cells. Additionally, HL-60 cells were adapted to grow in serum-deprived medium to study factors that control CD38 expression. Colony forming cell (CFC) assays and short-term expansion cultures were used to measure the effects of all-trans-retinoic acid (ATRA) oil the growth of fetal progenitors. Results Fetal progenitors and HL-60 cells grown under serum-deprived conditions exhibited almost no CD38 expression. However, CD34 expression was observed on fetal progenitors and declined slowly over time. Addition of FBS or human serum restored CD38 expression to cultured cells, but at levels below those found on progenitors in vivo. Addition of ATRA or 9-cis-retinoic acid (9CRA) to cultures of fetal progenitors or HL-60 cells, resulted in a time- and dose-dependent increase in CD38 expression, ATRA being the more potent of the two retinoids. However, ATRA inhibited colony formation, reduced the expansion of CFC and accelerated the loss of CD34 expression at doses required for the induction of CD38 expression. Discussion ATRA-induced CD38 expression on cells to levels comparable to those found on progenitors in vivo. ATRA also inhibited the growth of early progenitors, which was partly due to ATRA accelerating the differentiation of the progenitors. These findings indicate that CD34 and CD38 expression may be followed as markers of hematopoietic differentiation in vitro, but at the cost of culture conditions that are less than optimal for maintaining early progenitors. [ABSTRACT FROM AUTHOR]
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- 1999
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21. Regulatory Roles of the Ligand for Flk2/Flt3 Tyrosine Kinase Receptor on Human Hematopoiesis
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Reiko Namikawa, Maria Grazia Roncarolo, Marcus O. Muench, Namikawa, R, Muench, Mo, and Roncarolo, MARIA GRAZIA
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medicine.medical_treatment ,Stem cell factor ,Ligands ,Receptor tyrosine kinase ,Proto-Oncogene Proteins ,medicine ,Humans ,Progenitor cell ,Interleukin 3 ,Stem Cell Factor ,biology ,Growth factor ,Membrane Proteins ,Receptor Protein-Tyrosine Kinases ,Cell Differentiation ,Cell Biology ,Hematopoiesis ,Cell biology ,Haematopoiesis ,fms-Like Tyrosine Kinase 3 ,Fms-Like Tyrosine Kinase 3 ,biology.protein ,Cancer research ,Molecular Medicine ,Stem cell ,Developmental Biology - Abstract
The biological activities of the ligand for the Flk2/Flt3 receptor tyrosine kinase (FL) on human hematopoietic cells are reviewed. In in vitro studies, FL shows relatively few effects by itself on the proliferation and differentiation of hematopoietic cells, but exhibits a potent costimulatory activity in enhancing the proliferation of progenitor cells of multiple lineages. FL promotes the growth of clonogenic myeloid progenitor cells in the presence of other cytokines known to be active on myeloid progenitors, including GM-CSF, interleukin 3 (IL-3), kit ligand (KL), M-CSF and G-CSF. In addition, FL synergizes with IL-7 in inducing the proliferation of pro-B cells, whereas FL has little effect on the growth of clonogenic erythroid progenitors. Furthermore, FL induces the in vitro expansion of the high proliferative potential colony-forming cells (HPP-CFC) and stimulates the proliferation of long-term culture-initiating cells (LTC-IC), suggesting an activity on the proliferation of putative stem cells. Thus, FL plays important roles in regulating the proliferation of hematopoietic progenitor cells and, therefore, may have therapeutic applications.
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- 1996
22. Expression of CD33, CD38, and HLA-DR on CD34+ human fetal liver progenitors with a high proliferative potential
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Josephine Polakoff, Maria Grazia Roncarolo, Marcus O. Muench, James Cupp, Muench, Mo, Cupp, J, Polakoff, J, and Roncarolo, MARIA GRAZIA
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education.field_of_study ,Liver cytology ,Immunology ,Population ,CD34 ,Cell Biology ,Hematology ,Biology ,Biochemistry ,Molecular biology ,Haematopoiesis ,Immunophenotyping ,medicine.anatomical_structure ,medicine ,Bone marrow ,Stem cell ,Progenitor cell ,education - Abstract
High proliferative-potential colony-forming cells (HPP-CFC) have been identified in the bone marrow of mice and adult humans, and have been characterized as a compartment of primitive progenitors possibly including stem cells. In this report we describe the human fetal liver (FL) as a source of HPP-CFC. These FL HPP-CFC develop in clonal cultures in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-3 (IL-3) within 3 to 4 weeks. The median frequency of HPP-CFC in FL tissues between 16 and 21 weeks of gestational age was 1 in 3,000 total FL cells. After 4 weeks of growth, FL HPP-CFC grew to a median colony size of 8.3 x 10(4) cells/colony. Using cell-sorting techniques FL HPP-CFC were shown to be predominantly contained in the CD34+ CD33+ CD38- fraction of FL cells. FL HPP-CFC were heterogeneous for HLA-DR expression, and no differences in proliferative capacities were observed between HLA-DR+ and HLA-DR- HPP- CFC. The CD34+ CD33-HLA-DR- CD38- population, previously suggested to contain stem cells, was observed to be very rare in the FL, representing approximately 1 in 1.7 x 10(5) light-density FL cells and containing almost no CFC. Therefore, it is possible that stem cells are contained in the CD33+ fraction of FL cells. Phenotypic characterization of CD34+ CD33+ CD38- lin -LDFL cells showed that these cells are also CD13+, predominantly Thy-1+, CD45RA-, CD45RO-, CD71-, and heterogenoeous for c-kit expression. These data suggest that FL HPP- CFC represent a heterogeneous compartment of primitive myeloid progenitors that may include stem cells.
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- 1994
23. Administration of Flk2/Flt3 ligand induces expansion of human high-proliferative potential colony-forming cells in the SCID-hu mouse
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Laurent Humeau, Reiko Namikawa, S. Menon, Maria Grazia Roncarolo, Marcus O. Muench, Meri T. Firpo, Yuming Xu, Namikawa, R, Muench, Mo, Firpo, Mt, Humeau, L, Xu, Ym, Menon, S, and Roncarolo, MARIA GRAZIA
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Cancer Research ,Sialic Acid Binding Ig-like Lectin 3 ,Transplantation, Heterologous ,CD33 ,Antigens, Differentiation, Myelomonocytic ,Lewis X Antigen ,Mice, SCID ,CD15 ,Biology ,Bone and Bones ,law.invention ,Mice ,SCID-hu Mouse ,Antigens, CD ,law ,In vivo ,Proto-Oncogene Proteins ,Genetics ,Animals ,Humans ,Flk2-flt3 ligand ,Molecular Biology ,Bone Transplantation ,Receptor Protein-Tyrosine Kinases ,Cell Biology ,Hematology ,Flow Cytometry ,Hematopoietic Stem Cells ,Molecular biology ,Recombinant Proteins ,Haematopoiesis ,fms-Like Tyrosine Kinase 3 ,Immunology ,Recombinant DNA ,Myelopoiesis ,Cell Division - Abstract
The effects of Flk2/Flt3 ligand (FL) administration on human hematopoiesis were investigated using SCID-hu mice transplanted with human fetal bone fragments. Treatment with recombinant human FL induced significant increases in the frequencies of the high-proliferative potential colony-forming cells and low-proliferative potential colony-forming cells in steady-state human bone marrow. FL also promoted the expansion of high-proliferative potential colony-forming cells and low-proliferative potential colony-forming cells in the human bone marrow during the recovery phase after irradiation, which was evident in increases in the frequencies as well as in the absolute numbers of colony-forming cells, Furthermore, higher percentages of CD33(+)CD15(-) cells were found in the marrows treated with FL as compared to that of controls, indicating that FL hastened the recovery of at least some aspect of myelopoiesis after irradiation. These results indicate that FL induces the expansion of primitive hematopoietic progenitor cells in vivo and, therefore, may be useful in treating patients to promote an early hematopoietic recovery after cytoablative therapies. (C) 1999 International Society for Experimental Hematology, Published by Elsevier Science Inc.
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- 1999
24. Phenotypic and functional evidence for the expression of CD4 by hematopoietic stem cells isolated from human fetal liver
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M O, Muench, M G, Roncarolo, R, Namikawa, Muench, Mo, Roncarolo, MARIA GRAZIA, and Namikawa, R.
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Transplantation, Heterologous ,Hematopoietic Stem Cell Transplantation ,Antigens, CD34 ,Bone Marrow Cells ,Cell Differentiation ,Mice, SCID ,Thymus Gland ,Fetal Blood ,Hematopoietic Stem Cells ,Immunophenotyping ,Mice ,Liver ,Bone Marrow ,Fetal Tissue Transplantation ,Radiation Chimera ,CD4 Antigens ,Animals ,Humans ,Cell Lineage - Abstract
Expression of the CD4 antigen was observed on human fetal liver, fetal bone marrow (BM), and umbilical cord blood progenitors expressing high levels of CD34. Using clonal and liquid-culture assays, CD4(+) CD34(++) Lin(-) (lineage = CD3, CD8, CD10, CD14, CD15, CD16, CD19, CD20, and glycophorin A) fetal liver progenitors were found to have a greater proliferative potential than CD4(-) CD34(++) Lin(-) progenitors, whereas the CD4(-) fraction was more enriched for erythroid progenitors. Both the CD4(+) and the CD4(-) progenitor subpopulations also gave rise to multilineage engraftment upon transplantation into human fetal bone fragments, supportive of B-lymphoid and myeloid growth, or into human fetal thymic fragments, supportive of T-cell growth, implanted in scid/scid (SCID) mice. However, in SCID-hu mice transplanted with graded doses of donor cells ranging from 2.0 x 10(2) to 2.0 x 10(4) cells, BM reconstitution by the CD4(+) fraction of CD34(++) Lin(-) cells was more frequent than by the CD4(-) fraction when low numbers of cells were injected. These functional data strongly suggest that stem cells reside among CD4(+) CD34(++) Lin(-) fetal liver cells. This hypothesis was further supported by the observations that CD4(+) CD34(++) Lin(-) fetal liver cells were enriched for CDw90(+) (Thy-1), CD117(+) (kit), CD123(+), HLA-DR(+), CD7(-), CD38(-), CD45RA(-), CD71(-), CD115(-) (fms), and rhodamine 123(dull) cells, a phenotypic profile believed to represent fetal stem cells. Furthermore, all CD4(+) CD34(++) Lin(-) fetal liver cells also expressed CD13 and CD33. (C) 1997 by The American Society of Hematology.
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- 1997
25. Colony-forming cells expressing high levels of CD34 are the main targets for granulocyte colony-stimulating factor and macrophage colony-stimulating factor in the human fetal liver
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Marcus Muench, Roncarolo, M. G., Rosnet, O., Birnbaum, D., Namikawa, R., Muench, Mo, Roncarolo, MARIA GRAZIA, Rosnet, O, Birnbaum, D, and Namikawa, R.
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Liver ,Pregnancy ,Macrophage Colony-Stimulating Factor ,Granulocyte Colony-Stimulating Factor ,Humans ,Antigens, CD34 ,Female ,Cell Separation ,Receptors, Cytokine ,Hematopoietic Stem Cells ,Cell Division ,Immunophenotyping - Abstract
The effects of the granulocyte (G) and macrophage (M) colony-stimulating factors (CSFs) on the growth of purified subpopulations of human fetal liver progenitors were investigated. In contradiction to the characterization of these cytokines as CSFs acting late in the course of hematopoiesis, both G-CSF and M-CSF were most potent in promoting the growth of fetal liver colony-forming cells (CFCs) that express high levels of CD34 and CD38 (CD34(++)CD38(+)) and are depleted of cells expressing a panel of lineage markers (Lin(-)). Cultures of these cells in serum-deprived conditions generated a mean of 11.2 and 39.1 low-proliferative potential (LPP)-CFCs per 1.0 x 10(3) CD34(++)CD38(+)Lin(-) cells grown in G-CSF and M-CSF, respectively. Cultures of more mature progenitors, isolated based on a lower level of CD34 expression (CD34(+) Lin(-)), generated few LPP-CFCs and 6.3 and 4.7 clusters per 1.0 x 10(3) CD34(+)Lin(-) cells in response to G-CSFs and M-CSF, respectively. G-CSF was also found to synergistically enhance colony growth by either kit-ligand (KL) or flt-3/flk-2 ligand (FL) in cultures of CD34(++)CD38(+)Lin(-) cells as well as the more primitive compartment of CD34(++)CD38(-)Lin(-) cells. Synergism between G-CSF and KL or FL was also observed in liquid cultures of CD34(++)CD38(-)Lin(-) cells. The effects of G-CSF on CD34(++)CD38(-)Lin(-) cells were further demonstrated by the ability of G-CSF to support the short-term survival of these cells in clonal cultures. In contrast, M-CSF did not affect the growth or survival of CD34(++)CD38(-)Lin(-) cells, a finding that was also supported by the observation that the receptor for M-CSF (CD115 or fms) was only expressed on CD34(++)CD38(+)Lin(-) cells. G-CSF receptor expression and flt-3/flk-2 expression were detected by flow cytometry on both the CD38(-) and CD38(+) subpopulations of CD34(++)Lin(-) cells, but these receptors were not detected on CD34(+) cells. Receptors for KL (CD117) and interleukin-3 (CD123), for which the ligands are active on a broad range of fetal liver progenitors, were detected on cells expressing both high and low levels of CD34. These data help to define the potential roles of cytokines in human fetal hematopoiesis.
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- 1997
26. FLK-2/FLT-3 LIGAND REGULATES THE GROWTH OF EARLY MYELOID PROGENITORS ISOLATED FROM HUMAN FETAL LIVER
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Frank S. Lee, Robert A. Kastelein, Sandra Zurawski, S. Menon, Maria Grazia Roncarolo, Janice Culpepper, Charles H. Hannum, Yuming Xu, Reiko Namikawa, Marcus O. Muench, Muench, Mo, Roncarolo, MARIA GRAZIA, Menon, S, Xu, Ym, Kastelein, R, Zurawski, S, Hannum, Ch, Culpepper, J, Lee, F, and Namikawa, R.
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education.field_of_study ,Liver cytology ,medicine.medical_treatment ,Immunology ,Population ,Stem cell factor ,Cell Biology ,Hematology ,Biology ,CD38 ,Biochemistry ,Molecular biology ,Haematopoiesis ,Cytokine ,Fms-Like Tyrosine Kinase 3 ,medicine ,Progenitor cell ,education - Abstract
The effects of the recently identified FLK-2/FLT-3 ligand (FL) on the growth of purified human fetal liver progenitors were investigated under serum-deprived culture conditions. FL alone was found to stimulate modest proliferation in shortterm cultures of CD34(++) CD38(+) lineage (Lin)(-) light-density fetal liver (LDFL) cells and the more primitive CD34(++) CD38(-)Lin(-) LDFL cells. However, the low levels of growth induced by FL were insufficient for colony formation in clonal cultures. Synergism between FL and either granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3) or KIT ligand (KL) was observed in promoting the growth of high-proliferative potential (HPP) colony-forming cells (CFC) and/or low-proliferative potential (LPP)-CFC in cultures of CD34(++) CD38(+) Lin(-) and CD34(++) CD38(-) Lin(-) LDFL-cells. FL, alone or in combination with other cytokines, was not found to affect the growth of CD34(+) Lin(-) LDFL cells, the most mature subpopulation of fetal liver progenitors investigated. The growth of the most primitive subset of progenitors studied, CD34(++) CD38(-) Lin(-) LDFL cells, required the interactions of at least two cytokines, because only very low levels of growth were observed in response to either FL, GM-CSF, IL-3 or KL alone. However, the results of delayed cytokine-addition experiments suggested that individually these cytokines did promote the survival of this early population of progenitors. Although two-factor combinations of FL, KL, and GM-CSF were observed to promote the growth of early progenitors in a synergistic manner, neither of these factors was found to make fetal liver progenitors more responsive to suboptimal concentrations of a second cytokine. Only myeloid cells were recovered from liquid cultures of CD34(++) CD38(-) Lin(-) LDFL cells grown in the presence of combinations of FL, KL, and GM-CSF. These results indicate that FL is part of a network of growth factors that regulate the growth and survival of early hematopoietic progenitors. (C) 1995 by The American Society of Hematology.
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- 1995
27. Identification of a common T/natural killer cell progenitor in human fetal thymus
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Maria Grazia Roncarolo, Marcus O. Muench, Joseph H. Phillips, Lewis L. Lanier, María José Sánchez, Sanchez, Mj, Muench, Mo, Roncarolo, MARIA GRAZIA, Lanier, Ll, and Phillips, Jh
- Subjects
Cellular differentiation ,T-Lymphocytes ,Immunology ,Antigens, CD34 ,Thymus Gland ,Cell Separation ,Biology ,Medical and Health Sciences ,Natural killer cell ,Interleukin 21 ,Mice ,Antigens, CD ,Stem Cell Research - Nonembryonic - Human ,medicine ,Immunology and Allergy ,Killer Cells ,Animals ,Humans ,Antigens ,Clonogenic assay ,Cell Differentiation ,Articles ,T lymphocyte ,Stem Cell Research ,Hematopoietic Stem Cells ,Flow Cytometry ,Molecular biology ,CD ,Killer Cells, Natural ,Thymocyte ,medicine.anatomical_structure ,Phenotype ,T cell differentiation ,Natural ,Interleukin-2 ,CD34 ,CD8 ,Biotechnology - Abstract
The phenotypic similarities between natural killer (NK) and T cells have led to the hypothesis that these distinctive lymphocyte subsets may be developmentally related and thus may share a common progenitor (Lanier, L. L., H. Spits, and J. H. Phillips. 1992. Immunol. Today, 13:392; Rodewald, H.-R., P. Moingeon, J. L. Lurich, C. Dosiou, P. Lopez, and E. L. Reinherz. 1992. Cell, 69:139). In this report, we have investigated the potential of human CD34(+) triple negative thymocytes ([TN] CD3(-), CD4(-), CD8(-)) to generate both T cells and NK cells in murine fetal thymic organ cultures (mFTOC) and in vitro clonogenic assays. CD34(+) TN thymocytes, the majority of which express prominent cytoplasmic CD3 epsilon (cytoCD3 epsilon) protein, can be divided into high (CD34(Bright)) and low (CD34(Dim)) surface expressing populations. CD34(Bright) TN thymocytes were capable of differentiating into T and NK cells when transferred into mFTOC, and demonstrated high NK cell clonogenic capabilities when cultured in interleukin (IL)-2, IL-7, and stem cell factor (SCF). Likewise, CD34(Bright) TN thymocyte clones after 5 d in culture were capable of generating NK and T cells when transferred into mFTOC but demonstrated clonogenic NK cell differentiation capabilities when maintained in culture with IL-2. CD34(Dim) TN thymocytes, however, possessed only T cell differentiation capabilities in mFTOC but were not expandable in clonogenic conditions containing IL-2, IL-7, and SCF. No significant differentiation of other cell lineage was detected in either mFTOC or in clonogenic assays from CD34(+) TN thymocytes. These results represent the first definitive evidence of a common T/NK cell progenitor in the human fetal thymus and delineate the point in thymocyte differentiation where T and NK cells diverge.
- Published
- 1994
28. PROGRESS IN THE EX-VIVO EXPANSION OF HEMATOPOIETIC PROGENITORS
- Author
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Maria Grazia Roncarolo, Marcus O. Muench, Malcolm A.S. Moore, Alicia Bárcenat, Reiko Namikawa, Muench, Mo, Roncarolo, MARIA GRAZIA, Namikawa, R, Barcena, A, and Moore, Mas
- Subjects
Cancer Research ,Myeloid ,Genetic enhancement ,Bone Marrow Cells ,Biology ,Mice ,Bone Marrow ,medicine ,Animals ,Humans ,Progenitor cell ,Bone Marrow Transplantation ,Hematology ,Hematopoietic Stem Cells ,Stimulation, Chemical ,Transplantation ,Haematopoiesis ,medicine.anatomical_structure ,Oncology ,Immunology ,Cancer research ,Cytokines ,Bone marrow ,Stem cell ,Cell Division ,Ex vivo - Abstract
In this review we describe how studies on the cytokine-stimulated growth of murine bone marrow (BM) progenitors have lead to the observations that large increases in progenitor numbers can be achieved in short-term cytokine-stimulated liquid cultures. Transplantation of these ex vivo expanded murine BM cells was shown to decrease the number of BM cells required to confer radioprotection and to increase the recovery rate of both myeloid and erythroid peripheral blood cells. The ex vivo expansion of murine BM cells does not, however, markedly diminish stem cells capable of long-term hematopoietic reconstitution. Investiga- tions on the expansion of human BM, peripheral blood, umbilical cord blood and fetal hematopoietic progenitors have demonstrated that clinically useful increases in progenitor numbers from these tissues are possible. Thus, ex vivo progenitor expansion may soon be of use in transplantation protocols to accelerate hematopoietic reconstitution and in gene therapy protocols if hematopoietic stem cells can be maintained during ex vivo culture.
- Published
- 1994
29. Lymphoid and myeloid differentiation of fetal liver CD34+lineage- cells in human thymic organ culture
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Dominique Schols, J E de Vries, Juha Punnonen, Maria Grazia Roncarolo, Alicia Bárcena, Marcus O. Muench, H Spits, A H Galy, Other departments, Barcena, A, Galy, Ahm, Punnonen, J, Muench, Mo, Schols, D, Roncarolo, MARIA GRAZIA, Devries, Je, and Spits, H.
- Subjects
Myeloid ,T-Lymphocytes ,Immunology ,CD34 ,Antigens, CD34 ,Thymus Gland ,Biology ,Fetus ,Organ Culture Techniques ,Antigens, CD ,Pregnancy ,medicine ,Immunology and Allergy ,Humans ,Cells, Cultured ,Interleukin 3 ,B-Lymphocytes ,Lineage markers ,Cell Differentiation ,Articles ,Hematopoietic Stem Cells ,Molecular biology ,Haematopoiesis ,Thymocyte ,medicine.anatomical_structure ,Liver ,Female ,Bone marrow ,CD5 - Abstract
In this article, we report that the human fetal thymus contains CD34(bright) cells (
- Published
- 1994
30. In search of T-cell progenitors in the human foetal liver
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Alicia Bárcena, Hergen Spits, Maria Grazia Roncarolo, Marcus O. Muench, Barcena, A, Muench, Mo, Roncarolo, MARIA GRAZIA, Spits, H., and Other departments
- Subjects
Antigens, Differentiation, T-Lymphocyte ,Fetus ,T cell ,Cellular differentiation ,T-Lymphocytes ,Immunology ,Cell Differentiation ,T lymphocyte ,Antigens, CD7 ,Biology ,Foetal liver ,Hematopoietic Stem Cells ,Phenotype ,medicine.anatomical_structure ,Liver ,Antigens, CD ,medicine ,Humans ,Progenitor cell ,Stem cell - Published
- 1994
31. Testosterone supplementation increases red blood cell susceptibility to oxidative stress, decreases membrane deformability, and decreases survival after cold storage and transfusion.
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Tran J, Jackman RP, Muench MO, Hazegh K, Bean SW, Thomas KA, Fang F, Page G, O'Connor K, Roubinian NH, Anawalt BD, and Kanias T
- Subjects
- Humans, Male, Animals, Mice, Middle Aged, Female, Hemolysis drug effects, Adult, Retrospective Studies, Blood Donors, Cell Survival drug effects, Aged, Testosterone blood, Oxidative Stress drug effects, Blood Preservation, Erythrocytes metabolism, Erythrocytes drug effects, Erythrocyte Deformability drug effects, Erythrocyte Transfusion
- Abstract
Background: Blood collection from donors on testosterone therapy (TT) is restricted to red blood cell (RBC) concentrates to avoid patient exposure to supraphysiological testosterone (T). The objective of this study was to identify TT-related changes in RBC characteristics relevant to transfusion effectiveness in patients., Study Design: This was a two-part study with cohorts of patients and blood donors on TT. In part 1, we conducted longitudinal evaluation of RBCs collected before and at three time points after initiation of T. RBC assays included storage and oxidative hemolysis, membrane deformability (elongation index), and oximetry. In part 2, we evaluated the fate of transfused RBCs from TT donors in immunodeficient mice and by retrospective analyses of NIH's vein-to-vein databases., Results: TT increased oxidative hemolysis (1.45-fold change) and decreased RBC membrane deformability. Plasma free testosterone was positively correlated with oxidative hemolysis (r = .552) and negatively correlated with the elongation index (r = -.472). Stored and gamma-irradiated RBCs from TT donors had lower posttransfusion recovery in mice compared to controls (41.6 ± 12 vs. 55.3 ± 20.5%). Recipients of RBCs from male donors taking T had 25% lower hemoglobin increments compared to recipients of RBCs from non-TT male donors, and had increased incidence (OR, 1.80) of requiring additional RBC transfusions within 48 h of the index transfusion event., Conclusions: TT is associated with altered RBC characteristics and transfusion effectiveness. These results suggest that clinical utilization of TT RBCs may be less effective in recipients who benefit from longer RBC survival, such as chronically transfused patients., (© 2024 AABB.)
- Published
- 2024
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32. A Monocytic Barrier to the Humanization of Immunodeficient Mice.
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Du EJ and Muench MO
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- Animals, Humans, Mice, Hematopoietic Stem Cell Transplantation, Transplantation, Heterologous, Receptors, Immunologic metabolism, Receptors, Immunologic genetics, Disease Models, Animal, Monocytes immunology, Mice, SCID, Mice, Inbred NOD
- Abstract
Mice with severe immunodeficiencies have become very important tools for studying foreign cells in an in vivo environment. Xenotransplants can be used to model cells from many species, although most often, mice are humanized through the transplantation of human cells or tissues to meet the needs of medical research. The development of immunodeficient mice is reviewed leading up to the current state-of-the-art strains, such as the NOD- scid -gamma (NSG) mouse. NSG mice are excellent hosts for human hematopoietic stem cell transplants or immune reconstitution through transfusion of human peripheral blood mononuclear cells. However, barriers to full hematopoietic engraftment still remain; notably, the survival of human cells in the circulation is brief, which limits overall hematological and immune reconstitution. Reports have indicated a critical role for monocytic cells - monocytes, macrophages, and dendritic cells - in the clearance of xenogeneic cells from circulation. Various aspects of the NOD genetic background that affect monocytic cell growth, maturation, and function that are favorable to human cell transplantation are discussed. Important receptors, such as SIRPα, that form a part of the innate immune system and enable the recognition and phagocytosis of foreign cells by monocytic cells are reviewed. The development of humanized mouse models has taken decades of work in creating more immunodeficient mice, genetic modification of these mice to express human genes, and refinement of transplant techniques to optimize engraftment. Future advances may focus on the monocytic cells of the host to find ways for further engraftment and survival of xenogeneic cells., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)
- Published
- 2024
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33. Polyinosinic: polycytidylic acid induced inflammation enhances while lipopolysaccharide diminishes alloimmunity to platelet transfusion in mice.
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Tran JQ, Muench MO, Gaillard B, Darst O, Tomayko MM, and Jackman RP
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- Mice, Animals, Poly C, Mice, Inbred BALB C, Histocompatibility Antigens, Inflammation etiology, Poly I-C pharmacology, Platelet Transfusion adverse effects, Lipopolysaccharides pharmacology
- Abstract
Introduction: Alloimmune responses against platelet antigens, which dominantly target the major histocompatibility complex (MHC), can cause adverse reactions to subsequent platelet transfusions, platelet refractoriness, or rejection of future transplants. Platelet transfusion recipients include individuals experiencing severe bacterial or viral infections, and how their underlying health modulates platelet alloimmunity is not well understood., Methods: This study investigated the effect of underlying inflammation on platelet alloimmunization by modelling viral-like inflammation with polyinosinic-polycytidylic acid (poly(I:C)) or gram-negative bacterial infection with lipopolysaccharide (LPS), hypothesizing that underlying inflammation enhances alloimmunization. Mice were pretreated with poly(I:C), LPS, or nothing, then transfused with non-leukoreduced or leukoreduced platelets. Alloantibodies and allogeneic MHC-specific B cell (allo-B cell) responses were evaluated two weeks later. Rare populations of allo-B cells were identified using MHC tetramers., Results: Relative to platelet transfusion alone, prior exposure to poly(I:C) increased the alloantibody response to allogeneic platelet transfusion whereas prior exposure to LPS diminished responses. Prior exposure to poly(I:C) had equivalent, if not moderately diminished, allo-B cell responses relative to platelet transfusion alone and exhibited more robust allo-B cell memory development. Conversely, prior exposure to LPS resulted in diminished allo-B cell frequency, activation, antigen experience, and germinal center formation and altered memory B cell responses., Discussion: In conclusion, not all inflammatory environments enhance bystander responses and prior inflammation mediated by LPS on gram-negative bacteria may in fact curtail platelet alloimmunization., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2023 Tran, Muench, Gaillard, Darst, Tomayko and Jackman.)
- Published
- 2023
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34. Enhanced alloresponse to platelet transfusion due to immune dysregulation following ablative chemotherapy in mice.
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Jackman RP, Darst O, Gaillard B, Tran JQ, Tomayko MM, and Muench MO
- Subjects
- Mice, Humans, Animals, Blood Transfusion, Isoantibodies, Interleukin-4, Platelet Transfusion adverse effects, Blood Platelets
- Abstract
Introduction: Alloimmunization is common following platelet transfusion and can result in negative outcomes for recipients such as refractoriness to subsequent transfusions and rejection of transplants. Healthy people do not receive blood transfusions, and the diseases and therapies that result in a need to transfuse have significant impacts on the immunological environment to which these alloantigens are introduced. Ablative chemotherapies are common among platelet recipients and have potent immunological effects. In this study, we modeled the impact of chemotherapy on the alloresponse to platelet transfusion. As chemotherapies are generally regarded as immunosuppressive, we hypothesized that that they would result in a diminished alloresponse., Methods: Mice were given a combination chemotherapeutic treatment of cytarabine and doxorubicin followed by transfusion of allogeneic platelets, and compared to controls given no treatment, chemotherapy alone, or transfusion alone. Alloantibody responses were measured 2 weeks after transfusion, and cellular responses and growth factors were monitored over time., Results: Contrary to our hypothesis, we found that chemotherapy led to increased alloantibody responses to allogeneic platelet transfusion. This enhanced response was antigen-specific and was associated with increased CD4
+ and CD8+ T cell responses. Chemotherapy led to rapid lymphocyte depletion followed by reconstitution, non-specific activation of transitional B cells with the highest levels of activation in the least mature subsets, and increased serum levels of B cell activating factor (BAFF)., Conclusion: These data suggest that ablative chemotherapy can increase the risk of alloimmunization and, if confirmed clinically, that additional measures to protect these patient populations may be warranted., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2023 Jackman, Darst, Gaillard, Tran, Tomayko and Muench.)- Published
- 2023
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35. Plasma transfusion-transmission of Zika virus in mice and macaques.
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Van Rompay KKA, Coffey LL, Yee JL, Singapuri A, Stuart J, Lanteri MC, Santa Maria F, Lu K, Singh I, Bakkour S, Stone M, Williamson PC, Muench MO, Busch MP, and Simmons G
- Subjects
- Animals, Female, Humans, Mice, Pregnancy, Blood Component Transfusion, Blood Transfusion, Plasma, RNA, Viral, Zika Virus genetics, Zika Virus Infection epidemiology
- Abstract
Background: Zika virus (ZIKV) epidemics with infections in pregnant women are associated with severe neurological disease in newborns. Although an arbovirus, ZIKV is also blood transfusion-transmitted (TT). Greater knowledge of the efficiency of ZIKV TT would aid decisions on testing and pathogen reduction technologies (PRT)., Study Design and Methods: Plasma units from ZIKV RNA-reactive blood donors were used to study infectivity in vitro, in mice, and in macaques. Furthermore, plasma units were subjected to PRT using amotosalen/ultraviolet light A (A/UVA) before transfusion., Results: In vitro infectivity of ZIKV RNA-reactive plasma varied between 100 and 1000 international units (IU) of ZIKV RNA. Immunodeficient mice were more sensitive with as low as 32 IU sufficient to infect 50% of mice. 50-5500 IU of RNA led to TT in macaques using dose escalation of three different RNA-positive, seronegative plasma units. In contrast, RNA-reactive units collected postseroconversion were not infectious in macaques, even at a dose of 9 million IU RNA. After A/UVA PRT, transfusion of plasma containing up to 18 million IU was no longer infectious in vitro and did not result in ZIKV TT in macaques., Conclusion: Significant risks of ZIKV TT are likely confined to a relatively short viremic window before seroconversion, and that sensitive nucleic acid amplification testing likely identifies the majority of infectious plasma. PRT was demonstrated to be effective at preventing ZIKV TT. Considering that there is no approved ZIKV vaccine, these data are relevant to mitigate the risk of TT during the future ZIKV outbreaks., (© 2023 AABB.)
- Published
- 2023
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36. CD203c is expressed by human fetal hepatoblasts and distinguishes subsets of hepatoblastoma.
- Author
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Muench MO, Fomin ME, Gutierrez AG, López-Terrada D, Gilfanova R, Nosworthy C, Beyer AI, Ostolaza G, Kats D, Matlock KL, Cairo S, and Keller C
- Abstract
Background & Aims: Hepatocytic cells found during prenatal development have unique features compared to their adult counterparts, and are believed to be the precursors of pediatric hepatoblastoma. The cell-surface phenotype of hepatoblasts and hepatoblastoma cell lines was evaluated to discover new markers of these cells and gain insight into the development of hepatocytic cells and the phenotypes and origins of hepatoblastoma., Methods: Human midgestation livers and four pediatric hepatoblastoma cell lines were screened using flow cytometry. Expression of over 300 antigens was evaluated on hepatoblasts defined by their expression of CD326 (EpCAM) and CD14. Also analyzed were hematopoietic cells, expressing CD45, and liver sinusoidal-endothelial cells (LSECs), expressing CD14 but lacking CD45 expression. Select antigens were further examined by fluorescence immunomicroscopy of fetal liver sections. Antigen expression was also confirmed on cultured cells by both methods. Gene expression analysis by liver cells, 6 hepatoblastoma cell lines, and hepatoblastoma cells was performed. Immunohistochemistry was used to evaluate CD203c, CD326, and cytokeratin-19 expression on three hepatoblastoma tumors., Results: Antibody screening identified many cell surface markers commonly or divergently expressed by hematopoietic cells, LSECs, and hepatoblasts. Thirteen novel markers expressed on fetal hepatoblasts were identified including ectonucleotide pyrophosphatase/phosphodiesterase family member 3 (ENPP-3/CD203c), which was found to be expressed by hepatoblasts with widespread expression in the parenchyma of the fetal liver. In culture CD203c
+ CD326++ cells resembled hepatocytic cells with coexpression of albumin and cytokeratin-19 confirming a hepatoblast phenotype. CD203c expression declined rapidly in culture whereas the loss of CD326 was not as pronounced. CD203c and CD326 were co-expressed on a subset of hepatoblastoma cell lines and hepatoblastomas with an embryonal pattern., Conclusions: CD203c is expressed on hepatoblasts and may play a role in purinergic signaling in the developing liver. Hepatoblastoma cell lines were found to consist of two broad phenotypes consisting of a cholangiocyte-like phenotype that expressed CD203c and CD326 and a hepatocyte-like phenotype with diminished expression of these markers. CD203c was expressed by some hepatoblastoma tumors and may represent a marker of a less differentiated embryonal component., Competing Interests: MM and AB are employed by Vitalant Research Institute and MF, AG, and RG were employed by Vitalant Research Institute when they contributed to this study. Christopher Nosworthy and Gregory Ostolaza were student interns and not employees of Vitalant Research Institute. KM is employed by Omics Data Automation. SC was employed by XenTech during the time that work was performed, and the manuscript was drafted. CK has sponsored research agreements with Eli Lily, Roche-Genentech and Cardiff Oncology as well as research collaborations with Novartis, and is co-founder of Tio Companies. Artisan Biopharma is a wholly-owned subsidiary of cc-TDI. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. This study received funding from Vitalant Inc. The funder was not involved in the study design, collection, analysis, interpretation of data, the writing of this article or the decision to submit it for publication., (Copyright © 2023 Muench, Fomin, Gutierrez, López-Terrada, Gilfanova, Nosworthy, Beyer, Ostolaza, Kats, Matlock, Cairo and Keller.)- Published
- 2023
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37. Antibody screening data of human midgestation liver cells with a focus on hematopoietic, liver sinusoidal endothelial, and hepatoblast cell-populations.
- Author
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Muench MO and Nosworthy C
- Subjects
- Humans, Liver, Endothelial Cells, Proteomics
- Abstract
Objectives: Cell-surface antigen screening was performed on human fetal liver cells using flow cytometry. The goal was to provide proteomic expression data on a number of human fetal liver cell populations that can inform studies on developmental hepatology and hematology., Data Description: A 21 weeks' gestation liver was depleted of erythrocytes prior to antibody staining. Screening was performed using phycoerythrin-labelled antibodies against 332 antigens. In addition to these antibodies, all samples were stained for CD14, CD45, CD235a, and CD326 (epithelial cell adhesion molecule - EpCAM). Subpopulations of fetal liver cells were identified using the co-stained antigens. Hematopoietic cells were identified by their expression of CD45 and CD235a; non-hematopoietic cells were further subdivided based on CD14 and CD326 expression. CD326
++ CD14low hepatoblasts and CD14++ liver sinusoidal endothelial cells were analyzed for the frequency and intensity of antigen expression. Analyzed flow cytometry data are presented for the expression of the antigens on hematopoietic cells and on non-hematopoietic cells in the context of CD14 and CD326 expression., (© 2022. The Author(s).)- Published
- 2022
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38. Frequent detection but lack of infectivity of SARS-CoV-2 RNA in presymptomatic, infected blood donor plasma.
- Author
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Saá P, Fink RV, Bakkour S, Jin J, Simmons G, Muench MO, Dawar H, Di Germanio C, Hui AJ, Wright DJ, Krysztof DE, Kleinman SH, Cheung A, Nester T, Kessler DA, Townsend RL, Spencer BR, Kamel H, Vannoy JM, Dave H, Busch MP, Stramer SL, Stone M, Jackman RP, and Norris PJ
- Subjects
- Animals, Blood Donors, Humans, Mice, RNA, Viral, Viremia, COVID-19 diagnosis, SARS-CoV-2 genetics
- Abstract
Respiratory viruses such as influenza do not typically cause viremia; however, SARS-CoV-2 has been detected in the blood of COVID-19 patients with mild and severe symptoms. Detection of SARS-CoV-2 in blood raises questions about its role in pathogenesis as well as transfusion safety concerns. Blood donor reports of symptoms or a diagnosis of COVID-19 after donation (post-donation information, PDI) preceded or coincided with increased general population COVID-19 mortality. Plasma samples from 2,250 blood donors who reported possible COVID-19-related PDI were tested for the presence of SARS-CoV-2 RNA. Detection of RNAemia peaked at 9%-15% of PDI donors in late 2020 to early 2021 and fell to approximately 4% after implementation of widespread vaccination in the population. RNAemic donors were 1.2- to 1.4-fold more likely to report cough or shortness of breath and 1.8-fold more likely to report change in taste or smell compared with infected donors without detectable RNAemia. No infectious virus was detected in plasma from RNAemic donors; inoculation of permissive cell lines produced less than 0.7-7 plaque-forming units (PFU)/mL and in susceptible mice less than 100 PFU/mL in RNA-positive plasma based on limits of detection in these models. These findings suggest that blood transfusions are highly unlikely to transmit SARS-CoV-2 infection.
- Published
- 2022
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39. Reduced dimethyl sulfoxide concentrations successfully cryopreserve human hematopoietic stem cells with multi-lineage long-term engraftment ability in mice.
- Author
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Gilfanova R, Auclair KM, Hui A, Norris PJ, and Muench MO
- Subjects
- Animals, Antigens, CD34, Cell Count, Cryopreservation, Hematopoietic Stem Cells, Humans, Mice, Dimethyl Sulfoxide pharmacology, Hematopoietic Stem Cell Transplantation
- Abstract
Background Aims: The cryopreservation of hematopoietic stem cells (HSCs) in dimethyl sulfoxide (DMSO) is used widely, but DMSO toxicity in transplant patients and the effects of DMSO on the normal function of cryopreserved cells are concerns. To address these issues, in vitro and clinical studies have explored using reduced concentrations of DMSO for cryopreservation. However, the effect of reducing DMSO concentration on the efficient cryopreservation of HSCs has not been directly measured., Methods: Cryopreservation of human bone marrow using 10%, 7.5% and 5% DMSO concentrations was examined. Cell counting, flow cytometry and colony assays were used to analyze different cell populations. The recovery of stem cells was enumerated using extreme limiting dilution analysis of long-term multi-lineage engraftment in immunodeficient mice. Four different methods of analyzing human engraftment were compared to ascertain stem cell engraftment: (i) engraftment of CD33
+ myeloid, CD19+ B-lymphoid, CD235a+ erythroid and CD34+ progenitors; (ii) engraftment of the same four populations plus CD41+ CD42b+ platelets; (iii) engraftment of CD34++ CD133+ cells; and (iv) engraftment of CD34++ CD38- cells., Results: Hematopoietic colony-forming, CD34++/+ , CD34++ CD133+ and CD34++ CD38- cells were as well preserved with 5% DMSO as they were with the higher concentrations tested. The estimates of stem cell frequencies made in the xenogeneic transplant model did not show any significant detrimental effect of using lower concentrations of DMSO. Comparison of the different methods of gauging stem cell engraftment in mice led to different estimates of stem cell numbers, but overall, all measures found that reduced concentrations of DMSO supported the cryopreservation of HSCs., Conclusion: Cryopreservation of HSCs in DMSO concentrations as low as 5% is effective., Competing Interests: Declaration of Competing Interest The authors have no commercial, proprietary or financial interest in the products or companies described in this article., (Copyright © 2021 International Society for Cell & Gene Therapy. Published by Elsevier Inc. All rights reserved.)- Published
- 2021
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40. A bioinspired and chemically defined alternative to dimethyl sulfoxide for the cryopreservation of human hematopoietic stem cells.
- Author
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Gilfanova R, Callegari A, Childs A, Yang G, Luarca M, Gutierrez AG, Medina KI, Mai J, Hui A, Kline M, Wei X, Norris PJ, and Muench MO
- Subjects
- Animals, Antigens, CD34 analysis, Cell Survival, Cryoprotective Agents pharmacology, Hematopoietic Stem Cells, Humans, Mice, Cryopreservation methods, Dimethyl Sulfoxide pharmacology
- Abstract
The cryopreservation of hematopoietic cells using dimethyl sulfoxide (DMSO) and serum is a common procedure used in transplantation. However, DMSO has clinical and biological side effects due to its toxicity, and serum introduces variation and safety risks. Inspired by natural antifreeze proteins, a novel class of ice-interactive cryoprotectants was developed. The corresponding DMSO-, protein-, and serum-free cryopreservation media candidates were screened through a series of biological assays using human cell lines, peripheral blood cells, and bone marrow cells. XT-Thrive-A and XT-Thrive-B were identified as lead candidates to rival cryopreservation with 10% DMSO in serum based on post-thaw cell survival and short-term proliferation assays. The effectiveness of the novel cryopreservation media in freezing hematopoietic stem cells from human whole bone marrow was assessed by extreme limiting dilution analysis in immunodeficient mice. Stem cell frequencies were measured 12 weeks after transplant based on bone marrow engraftment of erythroid, myeloid, B-lymphoid, and CD34
+ progenitors measured by flow cytometry. The recovered numbers of cryopreserved stem cells were similar among XT-Thrive A, XT-Thrive B, and DMSO with serum groups. These findings show that cryoprotectants developed through biomimicry of natural antifreeze proteins offers a substitute for DMSO-based media for the cryopreservation of hematopoietic stem cells., (© 2021. The Author(s).)- Published
- 2021
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41. Generation of recombinant hyperimmune globulins from diverse B-cell repertoires.
- Author
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Keating SM, Mizrahi RA, Adams MS, Asensio MA, Benzie E, Carter KP, Chiang Y, Edgar RC, Gautam BK, Gras A, Leong J, Leong R, Lim YW, Manickam VA, Medina-Cucurella AV, Niedecken AR, Saini J, Simons JF, Spindler MJ, Stadtmiller K, Tinsley B, Wagner EK, Wayham N, Tracy L, Lundberg CV, Büscher D, Terencio JV, Roalfe L, Pearce E, Richardson H, Goldblatt D, Ramjag AT, Carrington CVF, Simmons G, Muench MO, Chamow SM, Monroe B, Olson C, Oguin TH, Lynch H, Jeanfreau R, Mosher RA, Walch MJ, Bartley CR, Ross CA, Meyer EH, Adler AS, and Johnson DS
- Subjects
- Animals, Antibodies, Viral immunology, CHO Cells, Cricetulus, Enzyme-Linked Immunosorbent Assay, Globulins immunology, Humans, Immunization, Passive, Mice, Recombinant Proteins biosynthesis, Recombinant Proteins immunology, Zika Virus immunology, COVID-19 Serotherapy, B-Lymphocytes immunology, COVID-19 therapy, Globulins biosynthesis, SARS-CoV-2 immunology
- Abstract
Plasma-derived polyclonal antibody therapeutics, such as intravenous immunoglobulin, have multiple drawbacks, including low potency, impurities, insufficient supply and batch-to-batch variation. Here we describe a microfluidics and molecular genomics strategy for capturing diverse mammalian antibody repertoires to create recombinant multivalent hyperimmune globulins. Our method generates of diverse mixtures of thousands of recombinant antibodies, enriched for specificity and activity against therapeutic targets. Each hyperimmune globulin product comprised thousands to tens of thousands of antibodies derived from convalescent or vaccinated human donors or from immunized mice. Using this approach, we generated hyperimmune globulins with potent neutralizing activity against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in under 3 months, Fc-engineered hyperimmune globulins specific for Zika virus that lacked antibody-dependent enhancement of disease, and hyperimmune globulins specific for lung pathogens present in patients with primary immune deficiency. To address the limitations of rabbit-derived anti-thymocyte globulin, we generated a recombinant human version and demonstrated its efficacy in mice against graft-versus-host disease., (© 2021. The Author(s), under exclusive licence to Springer Nature America, Inc.)
- Published
- 2021
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42. Blood donor obesity is associated with changes in red blood cell metabolism and susceptibility to hemolysis in cold storage and in response to osmotic and oxidative stress.
- Author
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Hazegh K, Fang F, Bravo MD, Tran JQ, Muench MO, Jackman RP, Roubinian N, Bertolone L, DʼAlessandro A, Dumont L, Page GP, and Kanias T
- Subjects
- Adult, Animals, Body Mass Index, Cold Temperature, Erythrocyte Membrane chemistry, Erythrocyte Transfusion, Erythrocytes cytology, Female, Ferritins blood, Hematologic Tests, Hemolysis physiology, Humans, Leukocyte Reduction Procedures, Male, Membrane Lipids blood, Metabolome, Mice, Mice, Inbred NOD, Nitric Oxide blood, Osmotic Pressure, Oxidative Stress, Blood Donors, Blood Preservation methods, Erythrocytes metabolism, Obesity blood
- Abstract
Background: Obesity is a global pandemic characterized by multiple comorbidities, including cardiovascular and metabolic diseases. The aim of this study was to define the associations between blood donor body mass index (BMI) and RBC measurements of metabolic stress and hemolysis., Study Design and Methods: The associations between donor BMI (<25 kg/m
2 , normal weight; 25-29.9 kg/m2 , overweight; and ≥30 kg/m2 , obese) and hemolysis (storage, osmotic, and oxidative; n = 18 donors) or posttransfusion recovery (n = 14 donors) in immunodeficient mice were determined in stored leukocyte-reduced RBC units. Further evaluations were conducted using the National Heart, Lung, and Blood Institute RBC-Omics blood donor databases of hemolysis (n = 13 317) and metabolomics (n = 203)., Results: Evaluations in 18 donors revealed that BMI was significantly (P < 0.05) and positively associated with storage and osmotic hemolysis. A BMI of 30 kg/m2 or greater was also associated with lower posttransfusion recovery in mice 10 minutes after transfusion (P = 0.026). Multivariable linear regression analyses in RBC-Omics revealed that BMI was a significant modifier for all hemolysis measurements, explaining 4.5%, 4.2%, and 0.2% of the variance in osmotic, oxidative, and storage hemolysis, respectively. In this cohort, obesity was positively associated (P < 0.001) with plasma ferritin (inflammation marker). Metabolomic analyses on RBCs from obese donors (44.1 ± 5.1 kg/m2 ) had altered membrane lipid composition, dysregulation of antioxidant pathways (eg, increased oxidized lipids, methionine sulfoxide, and xanthine), and dysregulation of nitric oxide metabolism, as compared to RBCs from nonobese (20.5 ± 1.0 kg/m2 ) donors., Conclusions: Obesity is associated with significant changes in RBC metabolism and increased susceptibility to hemolysis under routine storage of RBC units. The impact on transfusion efficacy warrants further evaluation., (© 2020 AABB.)- Published
- 2021
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43. Pathogen-reduced PRP blocks T-cell activation, induces Treg cells, and promotes TGF-β expression by cDCs and monocytes in mice.
- Author
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Tran JQ, Muench MO, and Jackman RP
- Subjects
- Animals, Immune Tolerance, Mice, Monocytes, Transforming Growth Factor beta, Platelet-Rich Plasma, T-Lymphocytes, Regulatory
- Abstract
Alloimmunization against platelet-rich plasma (PRP) transfusions can lead to complications such as platelet refractoriness or rejection of subsequent transfusions and transplants. In mice, pathogen reduction treatment of PRP with UVB light and riboflavin (UV+R) prevents alloimmunization and appears to induce partial antigen-specific tolerance to subsequent transfusions. Herein, the in vivo responses of antigen-presenting cells and T cells to transfusion with UV+R-treated allogeneic PRP were evaluated to understand the cellular immune responses leading to antigen-specific tolerance. Mice that received UV+R-treated PRP had significantly increased transforming growth factor β (TGF-β) expression by CD11b+ CD4+ CD11cHi conventional dendritic cells (cDCs) and CD11bHi monocytes (P < .05). While robust T-cell responses to transfusions with untreated allogeneic PRP were observed (P < .05), these were blocked by UV+R treatment. Mice given UV+R-treated PRP followed by untreated PRP showed an early significant (P < .01) enrichment in regulatory T (Treg) cells and associated TGF-β production as well as diminished effector T-cell responses. Adoptive transfer of T-cell-enriched splenocytes from mice given UV+R-treated PRP into naive recipients led to a small but significant reduction of CD8+ T-cell responses to subsequent allogeneic transfusion. These data demonstrate that pathogen reduction with UV+R induces a tolerogenic profile by way of CD11b+ CD4+ cDCs, monocytes, and induction of Treg cells, blocking T-cell activation and reducing secondary T-cell responses to untreated platelets in vivo., (© 2020 by The American Society of Hematology.)
- Published
- 2020
- Full Text
- View/download PDF
44. μ-Lat: A mouse model to evaluate human immunodeficiency virus eradication strategies.
- Author
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Sperber HS, Togarrati PP, Raymond KA, Bouzidi MS, Gilfanova R, Gutierrez AG, Muench MO, and Pillai SK
- Subjects
- Animals, Bone Marrow virology, Female, Green Fluorescent Proteins genetics, Green Fluorescent Proteins metabolism, HIV genetics, HIV pathogenicity, HIV Infections pathology, HIV Infections therapy, Humans, Jurkat Cells, Lung virology, Male, Mice, Mice, Inbred NOD, Proviruses genetics, Spleen virology, Transfection methods, Cell Transplantation methods, Disease Models, Animal, HIV physiology, HIV Infections virology, Virus Latency
- Abstract
A critical barrier to the development of a human immunodeficiency virus (HIV) cure is the lack of a scalable animal model that enables robust evaluation of eradication approaches prior to testing in humans. We established a humanized mouse model of latent HIV infection by transplanting "J-Lat" cells, Jurkat cells harboring a latent HIV provirus encoding an enhanced green fluorescent protein (GFP) reporter, into irradiated adult NOD.Cg-Prkdc
scid Il2rgtm1Wjl /SzJ (NSG) mice. J-Lat cells exhibited successful engraftment in several tissues including spleen, bone barrow, peripheral blood, and lung, in line with the diverse natural tissue tropism of HIV. Administration of tumor necrosis factor (TNF)-α, an established HIV latency reversal agent, significantly induced GFP expression in engrafted cells across tissues, reflecting viral reactivation. These data suggest that our murine latency ("μ-Lat") model enables efficient determination of how effectively viral eradication agents, including latency reversal agents, penetrate, and function in diverse anatomical sites harboring HIV in vivo., (© 2020 Federation of American Societies for Experimental Biology.)- Published
- 2020
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45. Immunodeficient mice are better for modeling the transfusion of human blood components than wild-type mice.
- Author
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Blessinger SA, Tran JQ, Jackman RP, Gilfanova R, Rittenhouse J, Gutierrez AG, Heitman JW, Hazegh K, Kanias T, and Muench MO
- Subjects
- Animals, Erythrocyte Transfusion, Heterografts, Humans, Leukocyte Transfusion, Mice, Mice, Inbred NOD, Mice, SCID, Platelet Transfusion, Blood Component Transfusion methods, Models, Animal
- Abstract
Animal models are vital to the study of transfusion and development of new blood products. Post-transfusion recovery of human blood components can be studied in mice, however, there is a need to identify strains that can best tolerate xenogeneic transfusions, as well as to optimize such protocols. Specifically, the importance of using immunodeficient mice, such as NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice, to study human transfusion has been questioned. In this study, strains of wild-type and NSG mice were compared as hosts for human transfusions with outcomes quantified by flow cytometric analyses of CD235a+ erythrocytes, CD45+ leukocytes, and CD41+CD42b+ platelets. Complete blood counts were evaluated as well as serum cytokines by multiplexing methods. Circulating human blood cells were maintained better in NSG than in wild-type mice. Lethargy and hemoglobinuria were observed in the first hours in wild-type mice along with increased pro-inflammatory cytokines/chemokines such as monocyte chemoattractant protein-1, tumor necrosis factor α, keratinocyte-derived chemokine (KC or CXCL1), and interleukin-6, whereas NSG mice were less severely affected. Whole blood transfusion resulted in rapid sequestration and then release of human cells back into the circulation within several hours. This rebound effect diminished when only erythrocytes were transfused. Nonetheless, human erythrocytes were found in excess of mouse erythrocytes in the liver and lungs and had a shorter half-life in circulation. Variables affecting the outcomes of transfused erythrocytes were cell dose and mouse weight; recipient sex did not affect outcomes. The sensitivity and utility of this xenogeneic model were shown by measuring the effects of erythrocyte damage due to exposure to the oxidizer diamide on post-transfusion recovery. Overall, immunodeficient mice are superior models for xenotransfusion as they maintain improved post-transfusion recovery with negligible immune-associated side effects., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2020
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46. A small allelic variant in donor class I MHC is sufficient to induce alloantibodies following transfusion of standard or pathogen-reduced platelets in mice.
- Author
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Jackman RP, Heitman JW, and Muench MO
- Subjects
- Animals, Female, Male, Mice, Mice, Inbred C57BL, Alleles, Histocompatibility Antigens Class I genetics, Immune Tolerance genetics, Isoantibodies immunology, Platelet Transfusion adverse effects
- Abstract
Background and Objectives: Alloimmunization targeting major histocompatibility (MHC) antigens is common following platelet transfusion. Pathogen reduction of platelets can block alloimmunization to MHC in mice and induce partial antigen-specific tolerance to subsequent transfusions. This study utilized small allelic variants to evaluate the relative contributions of class I and class II MHC to the alloresponse against untreated or pathogen-reduced platelets., Materials and Methods: C57BL/6 (B6) K
bm1 and B6 IAbm12 mice with small variants in the class I Kb and class II IAb alleles, respectively, were used as platelet donors for wild-type B6 recipients. Both untreated and pathogen-reduced platelet-rich plasma (PRP) transfusions were evaluated for immunogenicity by measuring antibody responses and ex vivo cytokine production., Results: Both the Kbm1 and IAbm12 alleles induced antibody responses, though the response to Kbm1 was greater. Pathogen reduction blocked the antibody responses to IAbm12 , but not to Kbm1 . Both the Kbm1 and IAbm12 alleles primed ex vivo cytokine responses that were blocked with pathogen reduction, though responses to IAbm12 were broader and larger (Kbm1 responses: IFN-γ, TNFα, and MIP-1β; IAbm12 responses: IFN-γ, TNFα, IL-1β, IL-10, IL-13, and GM-CSF). Pathogen-reduced Kbm1 PRP did not appear to induce any tolerance to subsequent untreated Kbm1 PRP transfusions., Conclusion: Minor allelic variants in both the class I and class II MHC are capable of inducing an alloresponse to transfusion. The Kbm1 PRP induced alloantibodies even with pathogen reduction and did not show signs of inducing the partial tolerance to subsequent transfusions observed with a larger MHC mismatch., (© 2020 International Society of Blood Transfusion.)- Published
- 2020
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- View/download PDF
47. Pathogen reduction with riboflavin and ultraviolet light induces a quasi-apoptotic state in blood leukocytes.
- Author
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Tran JQ, Muench MO, Heitman JW, and Jackman RP
- Subjects
- Animals, Biomarkers metabolism, Female, Flow Cytometry, Humans, Immune Tolerance, Leukocytes immunology, Leukocytes metabolism, Leukocytes pathology, Male, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Photosensitizing Agents administration & dosage, Platelet Transfusion methods, Platelet-Rich Plasma cytology, Platelet-Rich Plasma immunology, Riboflavin administration & dosage, Apoptosis drug effects, Blood Safety methods, Leukocytes drug effects, Photosensitizing Agents pharmacology, Riboflavin pharmacology, Transfusion Reaction prevention & control, Ultraviolet Rays
- Abstract
Background: Alloimmunization to platelet-rich plasma (PRP) transfusions can cause adverse reactions such as platelet refractoriness or transplant rejection. Pathogen reduction treatment with ultraviolet light and riboflavin (UV + R) of allogeneic PRP was shown to reduce allogeneic antibody responses and confer partial antigen-specific immune tolerance to subsequent transfusions in mice. Studies have shown that UV + R was effective at both rapidly killing donor white blood cells (WBCs) and reducing their ability to stimulate an allogeneic response in vitro. However, the manner in which UV + R induces WBC death and its associated role in the immune response to treated PRP is unknown., Methods and Materials: This study evaluates whether UV + R causes WBC apoptosis by examining phosphatidylserine exposure on the plasma membrane, membrane asymmetry, caspase activity, and chromatin condensation by flow cytometry. The immunogenicity of WBCs killed with UV + R versus apoptotic or necrotic pathways was also examined in vivo., Results: WBCs after UV + R exhibited early apoptotic-like characteristics including phosphatidylserine exposure on the outer leaflet of the plasma membrane and loss of membrane asymmetry, but unlike canonical apoptotic cells, caspase activity and chromatin condensation were not apparent. However, in vivo studies demonstrated, unlike untreated or necrotic WBCs, both apoptotic WBCs and UV + R-treated WBCs failed to prime alloantibody responses to subsequent untreated transfusions., Conclusion: Overall, the mechanism of WBC death following UV + R treatment shares some membrane characteristics of early apoptosis but is distinct from classic apoptosis. Despite these differences, UV + R-treated and apoptotic WBCs both offer some protection from alloimmunization., (© 2019 AABB.)
- Published
- 2019
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48. Potential of Membranes Surrounding the Fetus as Immunoprotective Cell-Carriers for Allogeneic Transplantations.
- Author
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Togarrati PP, Dinglasan N, Yee E, Heitman JW, Jackman RP, Geisberg M, Norris PJ, Bárcena A, and Muench MO
- Abstract
Background: Membranes surrounding the fetus play a crucial role in providing a physical and immunological barrier between a semiallogeneic fetus and mother during pregnancy. In this study, we tested whether cotransplantation of fetal membranes (FMs) and allogeneic donor cells would improve the retention and function of allografts in mice., Methods: Intact and enzyme-digested membranes obtained from E18-E19 pregnant mice were subcutaneously cotransplanted with 10F7MN hybridoma cells that are of BALB/cByJ (Balb) origin and secrete anti-human CD235a antibody. Cells were transplanted into C57BL/6J (B6, allogeneic), Balb (syngeneic), and FVB/NJ (third-party) mice. Serum was collected after 1 and 3 weeks of cell transplantation and tested using flow cytometry for the presence of anti-human CD235a antibody. Immunosuppressive functions of membranes were further investigated by analyzing the cytokine profile of supernatants collected from allo-reactive mixed lymphocyte reactions (MLRs) using a multiplex cytokine assay., Results: B6 mice transplanted with 10F7MN cells along with membranes syngeneic to the host had significantly higher levels of CD235a antibody when compared to B6 mice that received cells without membranes, allogenic membranes, or third-party membranes. Syngeneic membranes significantly inhibited T-cell proliferation in the presence of allogeneic stimuli and suppressed the release of Th1-cytokines such as IFNγ, TNFα, and IL-2 in MLRs. Additionally, increases in the levels of Th2-cytokines were found in MLRs containing membrane-derived cells., Conclusions: Our study highlights the potential use of syngeneic FMs to act as potent cell-carriers that could improve graft retention as well as graft-specific immunoprotection during allograft transplantation., Competing Interests: The authors declare no conflicts of interest.
- Published
- 2019
- Full Text
- View/download PDF
49. Allogeneic major histocompatibility complex antigens are necessary and sufficient for partial tolerance induced by transfusion of pathogen reduced platelets in mice.
- Author
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Tran JQ, Muench MO, Heitman JW, and Jackman RP
- Subjects
- Animals, Blood Platelets drug effects, Blood Platelets radiation effects, Isoantibodies immunology, Mice, Mice, Inbred BALB C, Platelet Transfusion adverse effects, Riboflavin pharmacology, Ultraviolet Rays, Blood Platelets immunology, Immune Tolerance, Major Histocompatibility Complex immunology, Platelet Transfusion methods
- Abstract
Background and Objectives: Alloimmunization is common following transfusion with platelet-rich plasma (PRP) and can cause complications such as platelet refractoriness or transplant rejection. It has previously been shown that pathogen reduction of PRP with riboflavin and UV light (UV+R) can protect against alloimmunization in mice and induce partial tolerance to subsequent transfusions., Materials and Methods: Using B6 H2
d congenic mice, this study evaluated the relative contributions of major histocompatibility complex (MHC) antigens and minor antigens to both the alloresponse to PRP transfusion and the partial tolerance induced by UV+R treatment., Results: Both total and MHC-specific alloantibody responses were highest when both MHC and minor antigens were mismatched, with lower alloantibody responses observed with MHC mismatch alone, demonstrating that allogeneic minor antigens can enhance the response to allogeneic MHC. There was a weak, but significant alloantibody response to minor antigens only. UV+R treatment protected against both major and minor antigen alloimmunization. Both allogeneic MHC and minor antigens primed an enhanced cytokine response ex vivo, though this was weaker with minor antigens, and both responses were blocked with UV+R treatment., Conclusion: Allogeneic MHC is both necessary and sufficient to induce the partial tolerance associated with UV+R treatment., (© 2019 International Society of Blood Transfusion.)- Published
- 2019
- Full Text
- View/download PDF
50. Minimal infectious dose and dynamics of Babesia microti parasitemia in a murine model.
- Author
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Bakkour S, Chafets DM, Wen L, Muench MO, Telford SR 3rd, Erwin JL, Levin AE, Self D, Brès V, Linnen JM, Lee TH, and Busch MP
- Subjects
- Animals, Disease Models, Animal, Female, Mice, Mice, Inbred NOD, Real-Time Polymerase Chain Reaction, Babesia microti metabolism, Babesiosis blood, DNA, Protozoan blood, Erythrocytes parasitology, Parasitemia blood, RNA, Protozoan blood
- Abstract
Background: Babesia microti is a parasite that infects red blood cells (RBCs) in mammals. It is transmitted to humans by tick bites, transfusion, organ transplantation, and congenital acquisition. Although the Babesia natural history and seroprevalence in donors have been well described, gaps in knowledge relevant to transfusion remain., Study Design and Methods: Mice were infected with dilutions of parasitized blood to address the minimal infectious dose and the kinetics of parasitemia by quantitative polymerase chain reaction (qPCR) and of antibodies by enzyme immunoassay., Results: In immunocompetent DBA/2 mice infected with 100 parasitized RBCs (pRBCs) and in immunodeficient NSG mice infected with 63 pRBCs, parasitemia was detectable in five of five mice each. Peak parasitemia up to 2 × 10
7 pRBCs/mL at 2 to 3 weeks or 5 × 108 pRBCs/mL at 6 weeks was observed for DBA/2 and NSG mice, respectively. Protracted fluctuating parasitemia was observed for 8 months in DBA/2 mice, whereas NSG mice exhibited a high-plateau parasitemia. Antibody titers continued to increase until 6 to 18 weeks in DBA/2 mice and remained high through 6 months. This study also investigated the analytical performance of Babesia assays that detect parasite DNA or RNA using a blinded panel. A Babesia assay targeting parasite RNA was approximately 10-fold more sensitive compared to qPCR targeting DNA., Conclusion: The mice in this study were highly susceptible to Babesia infection using as few as 1 to 2 log pRBCs and maintained chronic parasitemia. If the infectious dose in human transfusion recipients is comparably low, a highly sensitive assay targeting parasite RNA may safeguard the blood supply, particularly before antibody detection., (© 2018 AABB.)- Published
- 2018
- Full Text
- View/download PDF
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