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4.
Biomaterials ; 32(35): 9263-70, 2011 Dec.
Article in English | MEDLINE | ID: mdl-21908041

ABSTRACT

Cord blood expansion ex vivo can be achieved in liquid suspension through the addition of cytokines at the expense of often undesirable cell differentiation. In order to derive a cytokine-free dynamic culture system, we hypothesised that a three-dimensional (3D) environment in the form of highly porous scaffolds made of poly (D,L-lactide-co-glycolide) (PLGA) or polyurethane (PU) for the biomimetic growth of cord blood mononuclear cells (CBMNCs), would facilitate expansion of hematopoietic cells without exogenous cytokines. Both scaffolds supported cellular expansion ex vivo. Cytokine-free, long-term culture was best in PU coated with collagen type I (54-fold expansion). In contrast, traditional 2D well-plate cultures collapsed within 4 days in the absence of cytokines. CBMNCs cultured in the scaffolds were visualised by scanning electron microscopy and immunophenotypic/immunostaining analysis and the studies validated the presence of a dynamic culture containing erythroid precursors (CD45(-)/CD71(+)/CD235a(+)), hematopoietic stem/progenitor cells (CD38(-)CD34(+), CD117(+)), maturing myeloid cells (CD38(+), MPO(+)), CD4(+) and CD8(+) T-lymphocytes and megakaryocytes (FVIII(+)). Colony forming unit (CFU) assays indicated that BFU-E and CFU-GM increased (p < 0.05) whereas CFU-GEMM were maintained at week 4. In conclusion, this 3D culture system is capable of long-term, cytokine-free expansion of CBMNCs, enabling the study of hematopoiesis and providing a potential platform for drug discovery and therapeutic applications ex vivo.


Subject(s)
Cytokines/pharmacology , Fetal Blood/cytology , Leukocytes, Mononuclear/cytology , Leukocytes, Mononuclear/drug effects , Tissue Scaffolds/chemistry , Cell Lineage/drug effects , Cell Proliferation/drug effects , Cells, Cultured , Clone Cells , Humans , Immunophenotyping , Lactic Acid/pharmacology , Leukocytes, Mononuclear/ultrastructure , Polyglycolic Acid/pharmacology , Polylactic Acid-Polyglycolic Acid Copolymer , Polyurethanes/pharmacology , Time Factors
5.
J Leukoc Biol ; 78(5): 1142-52, 2005 Nov.
Article in English | MEDLINE | ID: mdl-16260587

ABSTRACT

Plasmacytoid dendritic cells (PDC) constitute a distinct subset of DC found in human peripheral lymph nodes (LN), but little is known about their function. Cell suspensions were prepared from tumor draining LN (n=20) and control LN (n=11) of women undergoing surgical resection for primary breast cancer and elective surgery for benign conditions, respectively. Using four-color flow cytometry, human leukocyte antigen-DR+ DC subsets were identified phenotypically. The proportions and numbers of cells innately producing interleukin (IL)-4, IL-10, IL-12, and interferon-gamma (IFN-gamma) were also measured from intracellular accumulation of cytokine after blocking with monensin. All flow cytometry data were collected without compensation and were compensated off-line using the Winlist algorithm (Verity software). This package also provided the subtraction program to calculate percentage positive cells and intensity of staining. PDC (CD11c-, CD123+) expressed more cytokines than did myeloid DC (CD11c+) or CD1a+ putative "migratory" DC (P<0.001). LN PDC from patients with a good prognosis (px; n=11) demonstrated a relative increase in IL-12 and IFN-gamma expression (median IL-10:IL-12 ratio=0.78 and median IL-4:IFN-gamma ratio=0.7), and PDC from LN draining poor px cancer (n=9) showed a relative increase in IL-10 and IL-4 expression (median IL-10:IL-12 ratio=1.31 and median IL-4:IFN-gamma ratio=2.6). The difference in IL-4:IFN-gamma expression between good and poor px cancer groups was significant (P<0.05). Thus, PDC innately producing cytokines were identified in cell suspensions from human LN, and the character of PDC cytokine secretion may differ between two breast cancer prognostic groups. We speculate that a shift towards PDC IL-10 and IL-4 expression could promote tumor tolerance in LN draining poor px breast cancer.


Subject(s)
Breast Neoplasms/immunology , Cytokines/biosynthesis , Dendritic Cells/immunology , Immunity, Innate/immunology , Lymph Nodes/immunology , Adult , Aged , Breast Neoplasms/secondary , Cytokines/analysis , Cytokines/immunology , Dendritic Cells/cytology , Female , Flow Cytometry , Humans , Interferon-gamma/biosynthesis , Interferon-gamma/immunology , Interleukin-10/biosynthesis , Interleukin-10/immunology , Interleukin-12/biosynthesis , Interleukin-12/immunology , Interleukin-4/biosynthesis , Interleukin-4/immunology , Lymph Nodes/cytology , Lymph Nodes/pathology , Lymphatic Metastasis , Middle Aged , Tumor Cells, Cultured , Tumor Escape/immunology
6.
Clin Infect Dis ; 41(8): e80-2, 2005 Oct 15.
Article in English | MEDLINE | ID: mdl-16163623

ABSTRACT

We present a case of Kikuchi's disease causing meningitis associated with fever and treated successfully with corticosteroids alone. This is considered to be rare but provides an instructive example of how the diagnosis may be confused with that of tuberculous meningitis and highlights the importance of histopathological analysis.


Subject(s)
Histiocytic Necrotizing Lymphadenitis/complications , Meningitis/diagnosis , Meningitis/etiology , Adult , Anti-Inflammatory Agents/therapeutic use , Female , Histiocytic Necrotizing Lymphadenitis/drug therapy , Humans , Lymph Nodes/pathology , Prednisolone/therapeutic use
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