OBJECTIVE: The aim of this study was to investigate the effect of saliva of patients with chronic periodontitis (CPD) on the differentiation, activation, and secretion of osteoclast-maturing mediators of macrophages. METHODS: A total of 40 saliva samples were collected from healthy donors (n=20) and severe periodontitis patients (n=20). Peripheral blood mononuclear cells (PBMCs) and THP-1 monocyte line cells were challenged with 15% saliva for 5 days. The phenotype, surface marker, and phagocytosis of macrophages were analyzed by flow cytometry and microscopy. Osteoclast-maturing mediators were assayed by using enzyme-linked immunosorbent assay (ELISA) kits. RESULTS: When PBMCs were treated with CPD saliva for 5 days, 61.25%±11.33% of cells were transformed into large granular cells; 86.78%±13.69% of large granular cells were identified as CD14⁺⁺CD16⁺ macrophages. When THP-1 cells were treated with CPD saliva, most cells attached to the bottom of cell culture plates, thereby exhibiting macrophage morphology and releasing additional osteoclast-maturing mediators. Furthermore, the phagocytosis of THP-1 cells considerably increased in the presence of CPD saliva (66.35%±9.67%) compared with medium control (33.33%±7.52%), or healthy saliva (40.71%±3.52%). CONCLUSIONS Saliva from patients with CPD can induce macrophage differentiation, activate phagocytose microorganisms, and secrete osteoclast-maturing mediators.
Cell Differentiation ; Humans ; Leukocytes, Mononuclear ; Macrophages ; Monocytes ; Periodontitis ; immunology ; Saliva

Familial Mediterranean fever (FMF) is an autosomal recessive disease. Although the possibility of multiple immunologic mechanisms have been studied, the actual mechanism is still unresolved. Forty-one patients with FMF (24 males and 17 females with a mean age and disease duration of 17.8 +/- 4.1 and 4.7 +/- 2.3 years, respectively) and 14 healthy controls (10 males and 4 females with a mean age 23.2 +/- 5.1) were involved in the study. A phagotest was studied in both the patients and control groups with a FACScalibur Flow. All patients were in the acute stages of the disease and had not undergone colchicine treatment for 2 months. The percentage blood phagocytic activity of both granulocytes and monocytes were 84.23 +/- 8.76 and 67.28 +/- 10.15 in the patient group and 94.68 +/- 3.24 and 76.23 +/- 5.7 in the control group, respectively. There was no statistically significant difference in the percentage of phagocytic activity of the granulocytes and monocytes between the FMF patients and healthy controls (p > 0.05 and p > 0.05, respectively).
Adolescence ; Adult ; Chemotaxis, Leukocyte ; Familial Mediterranean Fever/immunology* ; Female ; Human ; Male ; Monocytes/immunology ; Neutrophils/immunology ; Phagocytosis*

OBJECTIVETo explore the characteristics of B cell-activating factor (BAFF) secreted by peripheral blood monocyte-derived dendritic cell (MoDC) in chronic idiopathic thrombocytopenic purpura (cITP) and the function of MoDC on B cell proliferation.

METHODSTen cITP patients were studied dynamically before and after treatment. The BAFF levels in serum and the supernatant of LPS stimulated MoDC were tested with ELISA. The BAFF gene expression in LPS stimulated MoDC was tested with RQ-PCR, the B cell proliferation co-cultured with the supernatant of LPS stimulated MoDC for 5 days was tested with flow cytometry for CFSE and (3)H thymidine incorporation.

RESULTSThe BAFF level in serum (serum BAFF) \[(2461 ± 483) ng/L\], and supernatant of LPS stimulated MoDC (supernatant BAFF) \[(1113 ± 113) ng/L\] and BAFF mRNA in LPS stimulated MoDC (BAFF mRNA) (1.70 ± 0.23) before treatment were higher than that after treatment \[(621 ± 53) ng/L, (490 ± 49) ng/L and 0.37 ± 0.12\] and normal group \[(742 ± 77) ng/L, (582 ± 63) ng/L and 0.52 ± 0.08\]. There was a positive correlation among serum BAFF, supernatant BAFF and BAFF mRNA, and a negative correlation among serum BAFF, supernatant BAFF and BAFF mRNA and blood platelet count (BPC) in all ITP patients. The supernatant of LPS-stimulated MoDC from untreated patients enhanced B cell proliferation as compared with the supernatant of LPS-stimulated MoDC from treated patients and normal group.

CONCLUSIONBAFF might contribute to disease development in cITP. MoDC may directly increase B cell proliferation by secreting BAFF without T cell help, playing an important role in the antibody production in cITP.

B-Lymphocytes ; immunology ; Dendritic Cells ; immunology ; Humans ; Interleukin-4 ; Monocytes ; Purpura, Thrombocytopenic, Idiopathic ; immunology

Immune and hemopoiesis are one of basic project of experimental hematology. Immune function is a essential activity of white blood cells. It was puzzled for the diversity and complexity of immune response. Polarized immune response of immune cells was discovered 30 years ago, which facilitates the study on differentiation of lymphocyte. Recently recognition on multifunctional polarized immune response of lymphocyte and monocyte/macrophage would promote to elucidate the regulatory network of immune cells, diversity and complexity of immune response as well as the study on hemopoiesis. In this paper the approach of multifunctional polarized immune response of lymphocyte, monocyte/macrophage and dendritic cells were reviewed, and their role, especially in cytokine storm and tumor pro-inflammation condition were discussed.
Cell Differentiation ; Cytokines ; immunology ; Cytotoxicity, Immunologic ; Dendritic Cells ; cytology ; immunology ; Humans ; Monocytes ; cytology ; immunology

Dendritic cells (DCs) are important immune cells linking innate and adaptive immune responses. DCs encounter various self and non-self antigens present in the environment and induce different types of antigen specific adaptive immune responses. DCs can be classified into lymphoid tissue-resident DCs, migratory DCs, non-lymphoid resident DCs, and monocyte derived DCs (moDCs). Recent work has also established that DCs consist of developmentally and functionally distinct subsets that differentially regulate T lymphocyte function. The development of different DC subsets has been found to be regulated by a network of different cytokines and transcriptional factors. Moreover, the response of DC is tightly regulated to maintain the homeostasis of immune system. MicroRNAs (miRNAs) are an important class of cellular regulators that modulate gene expression and thereby influence cell fate and function. In the immune system, miRNAs act at checkpoints during hematopoietic development and cell subset differentiation, they modulate effector cell function, and are implicated in the maintenance of homeostasis. DCs are also regulated by miRNAs. In the past decade, much progress has been made to understand the role of miRNAs in regulating the development and function of DCs. In this review, we summarize the origin and distribution of different mouse DC subsets in both lymphoid and non-lymphoid tissues. The DC subsets identified in human are also described. Recent progress on the function of miRNAs in the development and activation of DCs and their functional relevance to autoimmune diseases are discussed.
Animals ; Autoimmune Diseases ; immunology ; Cell Differentiation ; immunology ; Dendritic Cells ; cytology ; immunology ; Humans ; MicroRNAs ; immunology ; Monocytes ; cytology ; immunology ; T-Lymphocytes ; cytology ; immunology

IL-23 and IL-12 are structurally similar and critical for the generation of efficient cellular immune responses. Toxoplasma gondii induces a strong cell-mediated immune response. However, little is known about IL-23 secretion profiles in T. gondii-infected immune cells in connection with IL-12. We compared the patterns of IL-23 and IL-12 production by THP-1 human monocytic cells in response to stimulation with live or heat-killed T. gondii tachyzoites, or with equivalent quantities of either T. gondii excretory/secretory proteins (ESP) or soluble tachyzoite antigen (STAg). IL-23 and IL-12 were significantly increased from 6 hr after stimulation with T. gondii antigens, and their secretions were increased with parasite dose-dependent manner. IL-23 concentrations were significantly higher than those of IL-12 at the same multiplicity of infection. IL-23 secretion induced by live parasites was significantly higher than that by heat-killed parasites, ESP, or STAg, whereas IL-12 secretion by live parasite was similar to those of ESP or STAg. However, the lowest levels of both cytokines were at stimulation with heat-killed parasites. These data indicate that IL-23 secretion patterns by stimulation with various kinds of T. gondii antigens at THP-1 monocytic cells are similar to those of IL-12, even though the levels of IL-23 induction were significantly higher than those of IL-12. The detailed kinetics induced by each T. gondii antigen were different from each other.
Antigens, Protozoan/*immunology ; Cell Line ; Humans ; Interleukin-12/*secretion ; Interleukin-23/*secretion ; Monocytes/*immunology/*parasitology ; Time Factors ; Toxoplasma/*immunology

OBJECTIVETo investigate the influence of the LPS of Bacteroides fragilis on the secretion of IL-2 and IL-4 from the peripheral blood mononuclear cells of normal individuals, so as to elucidate the mechanism of the infection by Bacteroides fragilis.

METHODSLPS was obtained from both the strains isolated from patients and from standard NCTC9343. Peripheral blood mononuclear cells (PBMCs) were treated with different concentrations of LPS thus obtained. The supernatants from the cell culture of the PBMCs were harvested at 24 PBHs and were subjected to the determination of the IL-2 and IL-4 contents by ELISA method. RESULTS The IL-2 secretion from the PBMCs of normal volunteers was obviously inhibited by the LPS from Bacteroides fragilis (P < 0.01), and the inhibitory effect was dose-dependent. Nevertheless, the IL-4 secretion from the PBMCs of normal volunteers was significantly stimulated by the LPS from Bacteroides Fragilis (P < 0.05), and it was not concentration dependent. There was no difference between the effects of the LPSs from patients and standard strains (P < 0.05).

CONCLUSIONThe LPS from Bacteroides fragilis was inhibitory to the secretion of IL-2 from PBMCs and was stimulative to that of IL-4 from PBMCs of normal human persons.

Bacteroides fragilis ; metabolism ; Cells, Cultured ; Humans ; Interleukin-2 ; immunology ; secretion ; Interleukin-4 ; immunology ; secretion ; Lipopolysaccharides ; pharmacology ; Monocytes ; drug effects ; immunology

The aim of this study was to investigate whether exosomes derived from K562 cells and human monocyte-derived dendritic cells (DCs) transfected with total RNA of K562 cells are capable of inducing antigen-specific cytotoxic T lymphocytes (CTL) responses in vitro. DCs were generated from peripheral blood mononuclear cells (PBMNC) of healthy volunteers in the presence of GM-CSF and IL-4, and then were transfected with K562 RNA by using DOTAP lipofection. Exosomes was extracted from the supernatant of DCs and K562 cells. The T cell were activated to be tumor specific CTL after DCs and exosomes were co-cultured with autologous T cells derived from healthy volunteers' PBMNC. The effect of CTL on K562 cells was detected by MTT assay. The results showed that treatment of T cells with exosomes derived from K562 cells or DCs transfected with total RNA of K562 cells could significantly promote their killing ability on K562 cells as compared with untreated T cells (P < 0.05). The killing ability of T cells treated with exosomes on K562 cells was stronger than on HL-60 cells (P < 0.05). It is concluded that the specific CTL immune response to leukemia cells can be induced by exosomes derived from K562 cells.
Dendritic Cells ; cytology ; immunology ; Endosomes ; immunology ; Exocytosis ; immunology ; Humans ; K562 Cells ; Monocytes ; cytology ; RNA, Neoplasm ; genetics ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection

Cutaneous dendritic cells (DCs), Langerhans cells (LCs) and dermal dendritic cells (DDCs), are present in an immature state. The maturation of DCs is crucial for initiating an immune response. Since HLA-DM has an important role for antigen presentation, an increase in HLA-DM expression according to the maturation of blood monocyte-derived dendritic cells (MoDCs), which have similar characteristics with DDCs, is expected. Therefore, the aim of this study was to determine whether or not HLA-DM expression in MoDCs is related to maturation at each culture day (from day 0 to day 13) by flow cytometry. This was compared with the functional changes related to the maturation of MoDCs. MoDCs were generated by culturing human peripheral blood monocytes in the presence of GM-CSF and IL-4 for 7 days, which were followed by subsequent treatment with a cytokine cocktail (GM-CSF, IL-4, IL-1beta, TNF-alpha, IL-6 and PGE2) for the maturation of MoDCs. The intracellular HLA-DM was expressed in the immature MoDC. A sudden 3 to 8 fold increase in the intracellular HLA-DM expression was observed after treatment with a cytokine cocktail. HLA-DM was weakly expressed on the surface of the immature MoDC, but it seemed to be decreased with maturation. This study indicated that the intracellular HLA-DM expression increased, but not on the MoDC surface during maturation. This was despite the fact that HLA-DM expression was noted not only on the surface but also in the intracellular in the MoDC.
Dendritic Cells/*immunology/physiology ; Endocytosis ; Flow Cytometry ; HLA-D Antigens/*analysis ; Human ; Monocytes/*physiology

OBJECTIVETo test the immunity of peritoneal monocytes against Plasmodium yoelii infected red blood cells (target cells).

METHODSSaponinized Plasmodium yoelii infected red blood cells (SPRBC, Ghost erythrocyte) were used to immunize mice i.p twice. Three weeks later, the infected red blood cells were injected i.p.; 90 min later, the total peritoneal cells were isolated and washed for scanning electromicroscopy to observe the effects of the peritoneal monocyte to the target cell.

RESULTSThe peritoneal cells of the immunized mice were activated after 90 min of the challenge of target cells. The size of the cell was not even and the pili on the cell surface turned to be long and densed. Cell interconnections were found among the cells. In some peritoneal monocytes, their cell plasma were scattered (omlette-like) or with the shape as "cellular bomb". The scattered or the sheeted pili and spredding cell plasma could adhere to the target cells which were perforated densely and damaged.

CONCLUSIONThe protective adaptive immunity exists in the peritoneal monocytes of immunized mice.

Animals ; Antibodies, Protozoan ; immunology ; Erythrocyte Membrane ; parasitology ; Female ; Malaria Vaccines ; immunology ; Mice ; Mice, Inbred BALB C ; Monocytes ; immunology ; ultrastructure ; Peritoneum ; cytology ; Plasmodium yoelii ; immunology ; ultrastructure