Cytokines ; metabolism ; Humans ; Leukocytes, Mononuclear ; immunology ; secretion ; Ubiquitin ; blood

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

OBJECTIVETo study myeloid-derived suppressor cells (MDSC) levels in peripheral blood of infants with recurrent wheezing, and the role of MDSC in the development of recurrent wheezing.

METHODSThirty-one infants with recurrent wheezing at wheezing attacks were randomly enrolled in the study. Twenty-seven infants with bronchopneumonia and 27 preoperative infants (hernia or renal calculus), without infectious or neoplastic diseases, were selected as controls. The proportion of MDSC in peripheral blood mononuclear cells (PBMC) was measured by flow cytometry.

RESULTSThe proportion of MDSC in PBMC in infants with wheezing was significantly higher than in those with bronchopneumonia and preoperative infants (P<0.05).

CONCLUSIONSMDSC levels increase in infants with recurrent wheezing, suggesting that MDSC may play a crucial role in the development of this disorder.

Child, Preschool ; Female ; Humans ; Infant ; Leukocytes, Mononuclear ; immunology ; Male ; Myeloid Cells ; immunology ; Recurrence ; Respiratory Sounds ; immunology

OBJECTIVETo determine the frequencies and significance of myeloid-derived suppressor cells (MDSCs) and T-helper 17 (Th17) cells in peripheral blood of young children with recurrent wheezing.

METHODSThirty young children with an acute exacerbation of recurrent wheezing were randomly enrolled. Twenty age-matched children with bronchopneumonia (pneumonia group) and 23 age-matched preoperative children with non-infectious or non-neoplastic diseases (hernia or renal calculus) (control group) were selected. The frequencies of MDSCs and Th17 cells in the peripheral blood were measured using flow cytometry and their correlation was determined by the Spearman's correlation coefficient.

RESULTSThe percentage of MDSCs in nucleated cells was significantly higher in the wheezing group than in the pneumonia and control groups (P<0.05), and it was significantly higher in the pneumonia group than in the control group (P<0.05). The percentage of Th17 cells in mononuclear cells was significantly higher in the wheezing group than in the pneumonia and control groups (P<0.05), but it showed no significant difference between the pneumonia and control groups (P>0.05). The frequency of MDSCs was positively correlated with the frequency of Th17 cells in the wheezing group (r=0.645, P<0.01).

CONCLUSIONSMDSCs and Th17 cells may contribute to the pathogenesis of recurrent wheezing in young children.

Child, Preschool ; Female ; Humans ; Infant ; Leukocytes, Mononuclear ; immunology ; Male ; Myeloid Cells ; immunology ; Recurrence ; Respiratory Sounds ; immunology ; Th17 Cells ; immunology

This study sought to shed light on the killing effect of peripheral blood mononuclear cells (PBMCs) irradiated by gamma ray at a dose of 1 Gy on cultured human gastric tumor cell line MKN-28. The radiation dose rate of 17 Gy/min was used. The groups in the experiment were MKN-28 cell control group, PBMCs control group, MKN-28 tumor cells with irradiated or non-irradiated PBMCs co-cultured groups. Radiation dosage was one Gray, acridine orange/ethidium bromide (AO/EB) staining was used for observation of the killing effects of PBMCs on tumor cells in different period. Cells were harvested 240 h later and observed by transmission electron microscopy. The result showed the living period of irradiated PBMCs was shorter than that of non-irradiated PBMCs. In the irradiated and non-irradiated groups,a few PBMCs were still alive after being cultured for 240 h, but the cell volume was larger than that of lymphocytes. These cells were identified as monocytes (95%) or DCs (5%) by transmission electron microscopy. The co-culture of irradiated PBMCs and MKN-28 cells showed that tumor cells were eliminated after 96 h. As compared with the non-irradiated goup, the irradiated PBMCs had more potent ability for killing tumor. The results demonstrate that 1 Gy gamma irridiation can improve the killing effect of PBMCs on the tumor cells, and that 1 Gy gamma irritation can also induce shorter living period of lymphocytes in PBMCs cultured in vitro, but such irritation has little effect on the living period of monocytes and DCs in PBMCs.
Cell Survival ; Coculture Techniques ; Gamma Rays ; Humans ; Leukocytes, Mononuclear ; cytology ; immunology ; radiation effects ; Stomach Neoplasms ; immunology ; pathology ; Tumor Cells, Cultured

OBJECTIVETo explore the effect of astragalus polysaccharide (APS) on the function and maturation of chronic myelogenous leukemia (CML) peripheral blood mononuclear cells (PBMC)-derived plasmacytoid dendritic cells (pDCs).

METHODSCML-derived pDCs were sorted by flow cytometry, and then incubated with APS (at 0, 50, 100 and 200 mg/L). After 24 hours, the concentrations of IFN-α, IL-6, TNF-α were detected with ELISA. Five days later, the cultured cells were collected and analyzed for immotype, morphology and ultramicrostructure.

RESULTSThe level of IFN-α, IL-6, TNF-α was significantly higher in samples from CML remission group than that in untreated pDCs, and newly diagnosed pDC (P < 0.05) or untreated group. APS could promote more pDCs differentiating to dendritic cells (DCs) in CML remission group than in untreated-pDCs in a dose-dependant manner (P < 0.05).

CONCLUSIONAPS can enhance the immune function of pDCs, promote differentiation and maturation of pDCs from CML patients.

Cells, Cultured ; Dendritic Cells ; immunology ; Humans ; Interferon-alpha ; pharmacology ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; immunology ; Leukocytes, Mononuclear ; Polysaccharides ; pharmacology

OBJECTIVETo study the surface antigen of the dendritic cells (DC) and their Toll-like receptor 4 (TLR4) expression in patients with idiopathic thrombocytopenic purpura (ITP), and to explore their role in ITP pathogenesis.

METHODSThe peripheral blood mononuclear cells isolated from complete remission patients (CR), non-complete remission patients (n-CR) and normal controls were stimulated by rhGM-CSF and rhIL-4. The surface antigen of the DC was analyzed by flow cytometry. The level of IL-12p70 in the supernatant was detected by enzyme linked immunosorbent assay. The expression of TLR4 mRNA of DC was detected by real time PCR.

RESULTSIn the 21 CR ITP patients, the expression of both CD80 and CD86 in DC was significantly increased compared with that in normal controls \[(51.60 ± 13.47)% vs (36.03 ± 15.43)%, (61.50 ± 15.93)% vs (40.28 ± 11.49)%, respectively\] (P < 0.01). The expression of CD80 and CD86 in n-CR group was also significantly increased \[(53.29 ± 19.49)% and (62.91 ± 18.43)%, respectively\] (P < 0.01). After HD-DXM treatment, both CD80 and CD86 in CR patients were decreased (P < 0.01). There was no difference between the DXM treatment patients and the normal controls. In n-CR group, there was no difference in CD80 and CD86 expression before and after DXM therapy \[(52.30 ± 20.98% and (49.79 ± 20.28)%, respectively\] (P > 0.05). CD80 was still higher than normal (P < 0.05), while CD86 was not changed. The level of IL-12p70 in CR ITP patients before treatment was significantly higher \[(67.52 ± 14.43) pg/ml\] than that of the controls \[(39.78 ± 10.03) pg/ml\](P < 0.01), and after treatment, was significantly decreased to (43.90 ± 8.49) pg/ml, being no difference from that in control. In n-CR group, IL-12p70 was lower after treatment \[(48.45 ± 9.68) pg/ml\] than that before treatment \[(65.35 ± 12.52) pg/ml\] (P < 0.01), but still higher than that in control (P < 0.05). The TLR4 mRNA level in DCs of CR ITP patients before treatment were significantly higher 0.69 ± 0.17 than that of controls (0.31 ± 0.09) (P < 0.01) and after treatment, was reduced to 0.35 ± 0.11, being no difference from that in control. In n-CR group, TLR4 mRNA was decreased from 0.65 ± 0.09 to 0.52 ± 0.21 after treatment (P < 0.01), but still higher than normal (P < 0.01).

CONCLUSIONDC may play an important role in ITP by their Toll-like receptor and cytokine secretion.

Dendritic Cells ; immunology ; Humans ; Interleukin-12 ; metabolism ; Leukocytes, Mononuclear ; Purpura, Thrombocytopenic, Idiopathic ; immunology ; Toll-Like Receptor 4

OBJECTIVETo prepare tumor antigen peptides from HL-60 cells and to induce specific immune response.

METHODSHL-60 antigen peptides were obtained using techniques including freezing and thawing, heat precipitation and acid precipitation. The stimulating effect of the in vitro Hsp70 binding HL-60 peptides on PBMC and the proliferation of stimulated PBMC were observed by T cell activation test. The cytotoxicity of proliferated PBMC is detected by incubating HL-60 cells or K562 cells with PBMC respectively.

RESULTSThe obtained tumor antigen peptides were a peptides mixture. The mixed peptides could activate PBMC and cause PBMC proliferation in vitro after presented by Hsp70. The proliferated PBMC showed specific cytotoxicity to HL-60 cells but not to K562 cells.

CONCLUSIONThe method for preparing of human leukemia tumor antigen peptides used in this paper is simple and easy; the obtained antigen peptides can induce specific immune response in vitro.

Cell Division ; HL-60 Cells ; HSP70 Heat-Shock Proteins ; immunology ; Humans ; K562 Cells ; Killer Cells, Natural ; immunology ; Leukocytes, Mononuclear ; cytology ; immunology ; Neoplasm Proteins ; immunology ; Peptides ; immunology

To investigate the characteristics of interstitial inflammatory cells and possible involvement of nudelta T cells, 16 renal allograft biopsies showing chronic rejection were stained by immunohistochemical method and correlated with the data of peripheral blood evaluated by flow cytometry. For immunophenotyping, fresh frozen sections were stained with monoclonal antibodies against CD3, CD4, CD8, CD68, CD56, TCRdelta1 and HLA DR. Paraffin embedded tissue was stained with CD45RO, CD20-Cy and CD68. Nine cases of nonspecific tubulointerstitial change and 4 cases of nonallograft tubulointerstitial nephritis were used as a control. Inflammatory infiltration was present in all cases studied. T cells predominated in the interstitium of chronic rejection and were followed by macrophages and B cells. The degree of interstitial infiltration of frozen section was not accordant with that of paraffin sections. Allografts with nonspecific tubulointerstitial changes or tubulointerstitial nephritis of native kidneys showed similar distribution pattern in terms of type and degree. However, the degree of infiltrate did not give any statistical significance among groups. The CD4/CD8 ratios in interstitial infiltrates were less than 1.0 in 6 cases and was not accordant with those of peripheral blood. Proportion of nudelta T cells increased over 10% in 2 cases in tissue and in 3 cases in peripheral blood. In 3 cases of chronic rejection in which both tissue and blood results were available, there was no concordance of CD4/CD8 or nudeltaT/CD3 between them. Tubular expression of HLA DR was, however, present only in 4 cases of chronic rejection. In conclusion, T lymphocytes were predominant regardless of diagnosis or disease activity. T lymphocyte subset did not give any suggestion as to the diagnosis or disease activity in chronic rejection. Furthermore nudelta T cells had only limited value. Lymphocytic subsets in peripheral blood would not be predictors of tissue destruction in chronic rejection.
Flow Cytometry ; Graft Rejection/*immunology ; Human ; Kidney/cytology/*immunology ; Kidney Transplantation/*immunology ; Leukocytes, Mononuclear/*immunology ; Phenotype ; Receptors, Antigen, T-Cell, gamma-delta/immunology ; Support, Non-U.S. Gov't

To investigate the characteristics of interstitial inflammatory cells and possible involvement of nudelta T cells, 16 renal allograft biopsies showing chronic rejection were stained by immunohistochemical method and correlated with the data of peripheral blood evaluated by flow cytometry. For immunophenotyping, fresh frozen sections were stained with monoclonal antibodies against CD3, CD4, CD8, CD68, CD56, TCRdelta1 and HLA DR. Paraffin embedded tissue was stained with CD45RO, CD20-Cy and CD68. Nine cases of nonspecific tubulointerstitial change and 4 cases of nonallograft tubulointerstitial nephritis were used as a control. Inflammatory infiltration was present in all cases studied. T cells predominated in the interstitium of chronic rejection and were followed by macrophages and B cells. The degree of interstitial infiltration of frozen section was not accordant with that of paraffin sections. Allografts with nonspecific tubulointerstitial changes or tubulointerstitial nephritis of native kidneys showed similar distribution pattern in terms of type and degree. However, the degree of infiltrate did not give any statistical significance among groups. The CD4/CD8 ratios in interstitial infiltrates were less than 1.0 in 6 cases and was not accordant with those of peripheral blood. Proportion of nudelta T cells increased over 10% in 2 cases in tissue and in 3 cases in peripheral blood. In 3 cases of chronic rejection in which both tissue and blood results were available, there was no concordance of CD4/CD8 or nudeltaT/CD3 between them. Tubular expression of HLA DR was, however, present only in 4 cases of chronic rejection. In conclusion, T lymphocytes were predominant regardless of diagnosis or disease activity. T lymphocyte subset did not give any suggestion as to the diagnosis or disease activity in chronic rejection. Furthermore nudelta T cells had only limited value. Lymphocytic subsets in peripheral blood would not be predictors of tissue destruction in chronic rejection.
Flow Cytometry ; Graft Rejection/*immunology ; Human ; Kidney/cytology/*immunology ; Kidney Transplantation/*immunology ; Leukocytes, Mononuclear/*immunology ; Phenotype ; Receptors, Antigen, T-Cell, gamma-delta/immunology ; Support, Non-U.S. Gov't