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Immunophenotypic and Functional Characterization of Human Umbilical Cord Blood Mononuclear Cells K Paloczi

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Immunophenotypic and Functional Characterization of Human Umbilical Cord Blood Mononuclear Cells K Paloczi Leukemia (1999) 13, Suppl. 1, 87–89 1999 Stockton Press All rights reserved 0887-6924/99 $12.00 http://www.stockton-press.co.uk/leu Immunophenotypic and functional characterization of human umbilical cord blood mononuclear cells K Paloczi National Institute of Haematology and Immunology, Budapest, Hungary Human umbilical cord blood (CB) represents a unique source GM).7,8 Besides the different culture systems immunofluo- of transplantable hematopoietic progenitor cells. Potential rescence and flow cytometry can be regarded as a reliable advantages of using CB relate to the high number and quality of hematopoietic stem and progenitor cells present in the circu- and rapid method for detection of early stem/progenitor cells. lation at birth and to the relative immune immaturity of the new- The CD34 antigen is of particular interest because it represents born immune cells. Discussed in this review are: (a) Quantity the only cell-surface antigen identified by monoclonal anti- and quality of immature hematopoietic stem and progenitor bodies (MoAb), whose expression within the hematopoietic cells from cord blood; (b) Immune cells in cord blood including system is restricted to primitive progenitor cells of all the number of B- and T-lymphocytes, as well as natural killer 9 cells and characterization of their functional capacities; (c) The lineages. 1,2 need of an international CB transplantation registry and the According to the previous reports, cord blood represents availability of cord blood banks. Although still in its infancy, a rich source of hematopoietic stem/progenitor cells, but human CB progenitor cells hold considerable potential for in CD34+ cell enumeration was not reported for these studies. vitro expansion and to transplant the adult recipients. In However, Broxmeyer and co-workers10 have confirmed that addition, the CB repopulating progenitor cells can serve as tar- gets for gene transfer and long-term treatment of genetic the single primitive CD34-positive human umbilical cord inherited diseases, cancer and some immunodeficiencies. blood hematopoietic progenitors have a very high proliferative and replating potential and may at least in part be associated with the successful hematopoietic repopulating capacity of umbilical cord blood cells. + Introduction Several groups have recently analyzed the CD34 cells present in the different cell sources used for transplantation 10–19 Human placental/umbilical cord blood (CB), as a source of including human CB. These data indicate that using only hematopoietic progenitor cells for bone marrow (BM) recon- one-color CD34 analysis may not provide sufficient infor- + stitution, has been shown to yield successful transplants from mation,12,15 but sensitive detection and enumeration of CD34 siblings1–3 and more recently from unrelated donors.4,5 In cells is required.12,13,15 The widespread use of CD34 comparison with BM, the collection of CB is easier and the expression as a measure of the transplant potential of hemato- transmission of certain infectious agents, particularly cytome- poietic tissue also means that it is important to establish any galovirus, to recipients, is much less common.5 Most antibody- or tissue-related differences in levels of importantly, CB graft may result in decreased incidence of expression.14 As a result of the different levels of detection by acut graft-versus-host disease (GVHD) even in partially mis- different MoAbs, the apparent number of CD34+ cells vary matched cases.3,4 The most important potential disadvantage by approximately six-fold.14 Furthermore, the kinetics of the of CB is the limited nucleated cell content of donation (a mean engraftment seems to be depending on the composition of of 117 mls ± 17 s.d. and a mean of 15.6 × 109/l ± 10.3 CD34 subpopulations, eg the distribution of the early and late nucleated cells).5,6 myeloid and B-lymphoid progenitor cells.15,17 Since the first CB transplantation in 19882 the number of A novel developed in vivo functional assay for primitive transplantations with CB hematopoietic cells is increasing and human hematopoietic cells is based on their ability to repopu- nowadays more than 200 have performed worldwide. There late the BM of severe combined immunodeficient (SCID) and is wide international agreement that a pilot clinical CB trans- non-obese diabetic/SCID (NOD/SCID) mice.18 These results plant study is required to evaluate results in comparison with also confirm that CB is enriched for the primitive hematopo- related and unrelated marrow donor transplants. Within the ietic cells compared with BM or mobilized PB, an observation framework of this program the Eurocord Transplant Group that has important implications for clinical transplantation and was established with the purpose of studying the properties of stem cell expansion strategies. hematopoietic stem cells present in placental blood at birth Engraftment will depend not only on the number of progeni- for clinical use in patients affected predominantly by hematol- tors, but also on the quality of these cells for proliferation and ogical diseases.6 self-renewal. These qualities of stem and progenitor cells appear to be high. It has been demonstrated that committed hematopoietic progenitor cells are at least as numerous in Quantity and quality of immature hematopoietic stem cells cord blood as in normal BM.13,15,16 Cord blood cells can be from cord blood extensively expanded in suspension culture with different combinations of cytokines,20 as well as in NOD/SCID mice17 The use of human cord blood as source of transplantable hem- and other murine model in vivo.21 Thus, immature hematopo- atopoietic stem and progenitor cells began with a multi-insti- ietic stem cells and progenitor cells appear to be of good qual- tutional laboratory study.1,7 A common approach to assess the ity in cord blood.16 quality of stem cell grafts relies on methylcellulose-based col- ony assays for lineage-committed hematopoietic progenitor cells as colony-forming units granulocyte/macrophage (CFU- Abstracts S88 Immune cells in cord blood est that little cytotoxic T cell activity is generated by cord blood cells even after primary, secondary and tertiary stimu- There has been little or no GVHD detected in children receiv- lation by allogeneic cells.10,13,16,31,34 The maximal responses ing HLA-matched or one antigen mismatched sibling cord observed in CB T cells were much less than those of adult T blood,2,3,7,16,22 and results using unrelated CB have been simi- cells, whereas the Con-A-induced proliferative responses of lar.4,5,16 There is evidence that immune cells from CB react these cells showed no significant differences. The decreased differently to those in adults. Monocyte/macrophage and T- responses of CB T cells were due to their lower efficiency in and B-lymphocytes are the major immune response cells. It activating the cellular events in T cell activation and prolifer- has been reported that a low antigen presenting ability found ation phase. Cord blood T cells have significantly less ability in CB monocyte/macrophage is due to their immaturity.16,23 than adult T cells to produce IL-2 and express functional IL- 2R complexes, especially high-affinity IL-2R complexes, and HLA-DR. These functional characteristics may be related to Immunologic characterization of cord blood the immaturity of cord blood T cells. B-lymphocytes Results obtained from our own experiments also indicate a distinct cytokine production profile in CB cells as compared It is known that newborn B cells are capable of producing with PB.13,30 A reduction was observed in the potential of CB amounts of IgM antibody comparable with those of adult B cells to produce the Th-1 cell-derived cytokines, IFN␥ and IL- cells, but their capacity to produce IgG or IgA is remarkably 24 2 upon stimulation with PHA or allogeneic cells. On the con- reduced. In humans, during fetal development the B cell trary, a slight increase was found in the production of IL-6 by populations show distinct phenotypes at different tissue sites. ␥ + + − alloactivated CB cells. The reduction of IFN and IL-2 could Besides conventional B cells (CD20 , CD24 , CD5 ) in fetal be associated with the presence of immature, naive T cells liver and bone marrow a significant fraction of fetal spleno- in CB. cytes are CD5+ B-lymphocytes.25 At birth, most IgM+ human umbilical cord blood B-lymphocytes co-express the CD5 anti- gen, too.26 Gluckman et al published that 68% of the CD19+ Cord blood natural killer (NK) cells: Cord blood NK cells cord blood B-lymphocytes proved to be CD5 antigen posi- − + tive.27 CD5+ B-lymphocytes share many properties of Ly-1 are CD3 CD56 cells expressing CD2, CD7, C11a-c, CD18, CD38, CD45RA and p75 IL-2 receptor.13,26,35 Only small murine B cells including self-replenishment and feedback + + 13,26,35 regulation of development. This unique population of B cells amounts of CD3 CD16 cells exist in CB. Cord blood (CD5+ B cells) is known as B1a lymphocytes preferentially NK cell activity is low or similar to that in adult blood but lymphokine activated killer cell activity is readily induced in generated from fetal/neonatal sources of progenitors. Func- 35,36 tionally, the antibodies produced by B1a cells are charac- CB by cytokines such as IL-2 or IL-12. terized by the predominance of IgM, minimal hypermutation and high frequency of low affinity, polyreactive, self-reactive In summary, umbilical cord blood stem cells are an alternative specificities.28 source of transplantable cells that can be used to reconstitute Our special interest was characterizing the CB B-lympho- hematopoiesis in children with malignant and non-malignant cytes studying differential expression of B cell-associated anti- diseases after treatment with myeloablative doses of chemo- gens on CD5+ B cells (B1a) and CD5− B cells (B1b) using therapy.
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