Bone marrow mesenchymal stem cells (BM-MSC) have the characteristics of self-renewal and multipotency. They secrete a variety of cytokines and provided and ideal microenvironment for the division, proliferation and differentiation of hematopoietic stem cells (HSC) through the interaction with other stromal cells. Previous studies indicated that MSC could enhance engrafment and alleviate GVHD in allo-HSC and MSC co-transplantation. This effect on transplantation immunity may associate with escaping MHC compatible HSC from antigen recognition and suppressing activation and proliferation of nonspecific lymphocytes. In this paper, the characteristics of BM-MSC influencing transplantation immunity and its mechanism were reviewed.
Bone Marrow Cells ; cytology ; immunology ; Graft vs Host Disease ; immunology ; prevention & control ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; cytology ; immunology ; Humans ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; immunology ; Transplantation Immunology

OBJECTIVETo study whether human placenta contains hematopoietic stem/progenitor cells (HSPCs), and analyze phenotypes of lymphocyte subpopulations in the placenta.

METHODSNucleated cells from fresh human placenta were analyzed for phenotypes of HSPCs and lymphocyte subpopulations by flow cytometry (FCM). And CD(34)(+) cells were sorted from human placenta nucleated cells by FCM or MiniMACS.

RESULTS(1) CD(34)(+) cells, CD(34)(+)/CD(38)(+) cells, and CD(34)(+)/CD(38)(-) cells from a human placenta were 8.8, 4.6 and 11.9 times higher than those from umbilical cord blood (UCB), respectively. (2) The yields and purity of CD(34)(+) cells isolated from human placenta by FCM sorting system were (63.05 +/- 10.14)% and (86.39 +/- 11.27)%, respectively. (3) Lymphocytes, T cells (CD(3)(+)/CD(2)(+)), B cells (CD(19)(+)), Th cells (CD(3)(+), CD(4)(+)), and Th/Ts ratio in the placenta tissue were apparently lower than those in the UCB, while the CD(8)(+)/CD(28)(-) T suppressor cells were higher in the placenta than in the UCB.

CONCLUSIONSHuman placenta is rich in HSPCs, and has important hematopoietic function in ontogeny. It is probable that human placenta would be graft resource for HSPCs transplantation. CD(8)(+)/CD(28)(-) T suppressor cells might play an important role in feto-maternal immunologic tolerance.

Cells, Cultured ; Female ; Hematopoietic Stem Cells ; cytology ; immunology ; Humans ; Lymphocyte Count ; Lymphocyte Subsets ; cytology ; immunology ; Male ; Placenta ; cytology ; immunology ; Pregnancy

This study was purposed to investigate the accelerating effect of newborn mouse cord blood transplantation combined with hematopoietic progenitor cells of bone marrow (BM) on the early hematopoietic reconstitution after transplantation, and the long-term chimerism of cord blood-derived cells, so as to develop a combined transplantation method for accelerating the early hematopoietic reconstitution. The lin(-)sca-1(-)c-kit(+) (c-kit(+)) cells and lin(-)sca-1(+) (sca-1(+)) cells in the bone marrow of BDF1 mice were isolated by MACS method. Biological characteristics in vitro of isolated fractions were observed and compared by semisolid colony culture combined with Giemsa staining. After transplantation of cord blood (CB) alone, or together with graded numbers of either c-kit(+) or sca-1(+) cells isolated from BDF1 mice (CD45.1) bone marrow into lethally irradiated CD45.2 congenic BDF1 mice, numbers of WBC and platelet were measured within 22 days of post-transplantation. The proportion of chimerism on granulocyte, T and B cell was dynamically measured by flow cytometry within 60 weeks of post-transplantation. The results showed that the number of colony from BM c-kit(+) cells cultured in semi-solid agar medium was significantly smaller than that from BM sca-1(+) population, which showed low proliferative potential in vitro and morphological characteristics of medium- or large-sized blast-like cells. The co transplantation of CB and BM c-kit(+) cells or sca-1(+) cells at the dosages of 1 × 10(4) or 2.5 × 10(4) or 5 × 10(4) to recipient mice leads to the quantity of WBC and platelets increased to 1 × 10(9)/L and 1 × 10(12)/L at day 12, whereas the injection of CB alone resulted at day 17. When mice were transplanted with CB together with BM c-kit(+)cells, and the CB-donor type cells in the peripheral blood increased progressively, while congenic donor BM-derived stem cells decreased gradually. After cotransplantation with CB and BM c-kit(+) cells for 60 weeks, a frequency of complete chimerism in CB-derived cells was continually maintained in granulates (96.68 ± 2.68)% and B lymphocytes (92.55 ± 3.04)%, while T lymphocytes (67.96 ± 7.91)% were dominantly derived from CB. On the other hand, congenic bone marrow or residual-derived cells were the dominant population, and the ratio of CB-derived cells in the peripheral blood was less than 10% (6.19 ± 7.62)% after cotransplantation with CB and sca-1(+)cells for 60 weeks. It is concluded that the cotransplantation of CB and BM congenic c-kit(+) cells is able to accelerate early hematopoietic reconstitution of recipient mice due to congenic marrow cells. Complete or main chimerism of cord blood is formed in long-term multilineage reconstitution of granulocytes, B cells and T lymphocytes.
Animals ; Bone Marrow Transplantation ; immunology ; Fetal Blood ; cytology ; transplantation ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; cytology ; immunology ; Mice ; Mice, Inbred C57BL ; Proto-Oncogene Proteins c-kit ; immunology

Much of the success of hematopoietic stem cell transplantation (HSCT) has been due to the ability to overcome posttransplant complications, by performing HLA genotypically matched or even mismatched sibling donor HSCT or unrelated donor HSCT. Based on the promising results with using vigorously T-cell-depleted high-dose peripheral blood stem cell transplants by the Perugia University group, several methods to eradicate refractory leukemic cells have been employed worldwide. However, there are limitations of the existing data regarding haploidentical HSCT, that is, the small numbers of patients and the heterogeneous patient populations in most of the published series. Also, researchers have not exactly demonstrated the effect of natural killer (NK) cell alloreactivity in various settings of allogeneic HSCT. In reality, haploidentical HSCT is possible without T-cell depletion. However, it isn't clear whether reduced-intensity HSCT from a haploidentical related donor or a mismatched unrelated donor is feasible. Anyhow, successfully overcoming the major histocompatibility barriers with using related or unrelated donors means that virtually all patients would have an immediately available donor for desperately needed HSCT. The full potential of haploidentical HSCT may be ultimately achieved through a better understanding of the transplant immunology, including the Korean specificity of killer cell immunoglobulin-like receptor polymorphism. Further study and better support from Korean government insurance coverage that would offer HSCT to more patients in need of transplant and cultivating an optimal NK alloreaction without detrimental complications is urgently required.
Allergy and Immunology ; Hematopoietic Stem Cell Transplantation* ; Hematopoietic Stem Cells* ; Histocompatibility ; Humans ; Insurance Coverage ; Receptors, KIR ; Sensitivity and Specificity ; Siblings ; Stem Cells ; T-Lymphocytes ; Tissue Donors ; Unrelated Donors

In recent years, the interaction between HLA class I alleles and receptors of NK cells have been paid more attention in HLA-identical sibling hematopoietic stem cell transplantation (HLA-identical sibling HSCT). NK cells mediate alloreactions when the allogeneic targets do not express HLA class I alleles that block inhibitory receptors of NK cells. Alloreactive donor-derived NK cells reduce GVHD by eliminating host-type antigen-presenting cells, but favor engraftment by killing host lymphocyte cells and reduce the risk of relapse by eradicating leukemia cells. Moreover, activating receptors also influence the outcome of HLA-identical sibling HSCT. In this review, the advance of the impact of both inhibitory and activating KIR (killer cell immunoglobulin-like receptor, KIR) on the outcome of HLA-identical sibling HSCT is briefly summarized.
HLA Antigens ; immunology ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Killer Cells, Natural ; immunology ; Prognosis ; Receptors, KIR ; immunology ; Siblings ; Transplantation, Homologous

ZCH-2B8a (IgG2a) is a novel monoclonal antibody (McAb) generated in laboratory of Children Hospital of Medical College, Zhejiang University recently using human myeloblastic leukemia cell line KG1a as immunogen. This antibody has been submitted to the 8th International Workshop and Conference on Human Leukocyte Differentiation Antigens (HLDA8) and the results showed that the antibody recognized an unknown molecule on the surface of some blood cells. The aim of this study was to investigate the reactivity of this antibody on normal blood cells and malignant cell lines and to explore its possible application in clinical practice. The multi-parameter flow cytometry was used to analyze the expression pattern of 2B8a antigen in triplicate on normal blood components including T cells, B cells, natural killers (NK), neutrophils, monocytes, dendritic cells (DC), red blood cells (RBC), platelets (Plt), hematopoietic stem/progenitor cells derived from either bone marrow or G-CSF mobilized peripheral blood CD34(+) cells and malignant cell lines including 14 hematopoietic, 5 neuroblastoma, 1 colon cancer and 1 amniotic epithelium cell lines. The amount of positive cells > or = 20% was considered as positivity. The results showed that 2B8a antibody reacted to 3/3 specimens of blood B cells with a positive rate of 26.29% and 2/3 specimens of monocytes with an average positive rate of 59.84%. 2B8a was weakly reactive to neutrophils (23.72%) and negative for T cells, NK, DC, RBC and Plt. The antibody reacted to all 3 marrow CD34(+) cells with an average positive rate of 39.33% while it was negative for G-CSF-mobilized CD34(+) peripheral blood stem/progenitor cells (PBSC, 1.25%). Cell line analysis showed that the antibody notably reacted to three out of 4 cell lines (Raji, SMS-SB, Nalm-6 and Nall-1) with the positive rates of 98.78%, 98.61%, 94.93% respectively and weakly to one of them with 5.68% in B lineage cell lines and monoblastic cell line (U937, 67.78%) while it was only weakly positive or negative for other myeloid leukemia cell lines including Meg01 (33.40%), HL-60 (29.70%), K562 (28.19%), KG1a (16.23%) and HEL92.1.7 (8.02%). Among 4 T lineage leukemia, 5 neuroblastoma and 1 colon cancer cell lines tested, only Molt-3 was found weakly positive (31.40%) for 2B8a, while the remaining 3 T cell lines (Molt4, JM and CCRF-CEM), 5 neuroblastoma cell lines (LA-N1, KCNR, BE, SK-N-SH, SK-N-AS) and the colon cancer cell line (HR8348) tested were negative. An amniotic epithelium cell line (FL) was showed positive for the antibody (45.03%). It is concluded that 2B8a antibody primarily reacts to B lineage and monocytic lineage cells which may bear the diagnostic and therapeutic applications among different types of hematopoietic malignancies.
Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibody Specificity ; Antigen-Antibody Reactions ; Antigens, Neoplasm ; analysis ; immunology ; Antigens, Surface ; immunology ; B-Lymphocytes ; cytology ; immunology ; HLA Antigens ; immunology ; Hematopoietic Stem Cells ; cytology ; immunology ; Hematopoietic System ; cytology ; Humans ; Leukemia, Myeloid, Acute ; immunology ; pathology ; Tumor Cells, Cultured

Natural killer cells are important innate immune effector cells with broad applications in the treatment of malignancies due to its cytotoxicity against cancer cells without prior immune sensitization. Insight progress in the field of NK cell activating, inhibitory receptors and signals modulating NK cell activity has revolutionized our understanding of the selective killing of cancer cells by NK cells. Recently, much progress has been achieved in the field of NK cell purification and ex vivo expansion. NK cells were used both either directly or in combination with other treatment modalities in the transplantation and non transplantation settings. To strengthen the efficiency of NK cell for therapeutic aim, the biology of NK cells and crucial issues required to development more successful NK-cell-based therapies are discussed in this article.
Hematopoietic Stem Cell Transplantation ; Humans ; Immunotherapy, Adoptive ; Killer Cells, Natural ; immunology

OBJECTIVETo cultivate hematopoietic stem/progenitor cells (CD34(+)CD38(-)) isolated from umbilical cord blood (UCB) long for the observation of cell growth and expansion in vitro, surface marker expression, and chromosomal complements.

METHODSBy flow cytometry CD34-FITC and CD38-PE labeled CD34(+) and CD38(-) stem/progenitor cells were isolated from UCB. The cells were cultivated in vitro for 6 months in a stem cell culture system with addition of six kinds of cell growth factors (IL-3, IL-6, GM-CSF, Epo, SCF, IGF-1). One month after cultivation, cultured cells were investigated for surface marker expression by flow cytometry and karyotype by G banding method.

RESULTSAfter 7-12 days cultivation, the CD34(+)CD38(-) stem/progenitor cells began proliferation. The proliferation rate and the peak proliferation duration were greater in 1 cell/well cultivation conditions than in 10 cells/well. The cells remained CD34(+)CD38(-) and their karyotypic characteristics remained unchanged.

CONCLUSIONCD34(+)CD38(-) stem/progenitor cells from UCB may provide a larger than original amount of stem/progenitor cells for transplantation after long-term cultivation in vitro.

ADP-ribosyl Cyclase 1 ; immunology ; Adult ; Antigens, CD34 ; immunology ; Cell Proliferation ; Cells, Cultured ; Female ; Fetal Blood ; cytology ; immunology ; Hematopoietic Stem Cells ; cytology ; immunology ; Humans ; Immunophenotyping ; Karyotyping

Bone marrow hematopoietic microenvironment occupies the medullary cavities of bones throughout the skeleton and provides support for hematopoiesis and immune cells development. Bone-resorbing osteoclasts in bone marrow environment are specialized cells derived from the hematopoietic stem cells and play a pivotal role in process termed as bone remodeling that involves break down and build-up of bone. It is only recently that studies have provided a novel basis for understanding potential role of osteoclasts in homeostasis, stress-induced mobilization of hematopoietic progenitors and osteoimmunology. Further exploration on the interaction of osteoclasts with others in bone marrow hematopoietic microenvironment may contribute to future clinical treatments for hematopoietic and bone-related immunologic disorders including cancer. In this review the origin and identification of osteoclasts and regulation of mobilizing hematopoietic stem cells, as well as osteoclasts and osteoimmunity were mainly concerned.
Bone Marrow Cells ; physiology ; Bone Remodeling ; immunology ; Hematopoiesis ; physiology ; Hematopoietic Stem Cells ; cytology ; immunology ; Homeostasis ; immunology ; Humans ; Osteoclasts ; cytology ; immunology ; physiology

The purpose of this review is to provide an overview of the effect of (lymph)angiogenic cytokines on hematopoietic cells involved in acute myeloid leukemia (AML). Like angiogenesis, lymphangiogenesis occurs in pathophysiological conditions but not in healthy adults. AML is closely associated with the vasculature system, and the interplay between lymphangiogenic cytokines maintains leukemic blast survival in the bone marrow (BM). Once AML is induced, proangiogenic cytokines function as angiogenic or lymphangiogenic factors and affect hematopoietic cells, including BM-derived immune cells. Simultaneously, the representative cytokines, VEGFs and their receptors are expressed on AML blasts in vascular and osteoblast niches in both the BM and the peripheral circulation. After exposure to (lymph)angiogenic cytokines in leukemogenesis and infiltration, immune cell phenotypes and functions are affected. These dynamic behaviors in the BM reflect the clinical features of AML. In this review, we note the importance of lymphangiogenic factors and their receptors in hematopoietic cells in AML. Understanding the functional characterization of (lymph)angiogenic factors in the BM niche in AML will also be helpful in interrupting the engraftment of leukemic stem cells and for enhancing immune cell function by modulating the tumor microenvironment.
Animals ; Cytokines/*immunology ; Hematopoietic Stem Cells/immunology/*pathology ; Humans ; Immunity, Cellular ; Leukemia, Myeloid, Acute/immunology/*physiopathology ; *Lymphangiogenesis ; Lymphatic Vessels/immunology/*physiopathology ; Vascular Endothelial Growth Factor A/immunology