No abstract available.
Animals ; Colitis/physiopathology/*therapy ; Cytokines/*secretion ; *Lactobacillus ; Probiotics/*therapeutic use

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

In order to explore a new way to study allogeneic reactive T lymphocytes, detection of cytokine-secreting T lymphocytes after allogeneic peripheral blood mononuclear cells (PBMNCs) stimulation and investigation of its clinical significance were performed. A novel cytokine secretion assay (CKSA) was first applied to detect T lymphocytes secreting cytokine including IFN-gamma, IL-4 and IL-10 at single cell level in human mixed lymphocytes reaction. IFN-gamma-secreting T cells from PBMNCs were then evaluated in 2 patients with acute graft versus host disease (aGVHD) after allogeneic bone marrow transplantation. The results showed that compared with IL-4 and IL-10 (which were 0.12 +/- 0.03% and 0.10 +/- 0.03% respectively), a sizable proportion of IFN-gamma-secreting T lymphocytes could be detected (1.12 +/- 0.13)% after allogeneic PBMNCs stimulation. Preliminary results indicated that frequency of IFN-gamma-secreting T lymphocytes correlated with the onset and severity of clinical aGVHD. In conclusion, it is feasible to detect IFN-gamma secreting T lymphocytes after allogeneic PBMNCs stimulation and to apply the CKSA technique for clinical identification of aGVHD.
Acute Disease ; Cytokines ; secretion ; Graft vs Host Disease ; etiology ; Humans ; Interferon-gamma ; secretion ; Interleukin-10 ; secretion ; Interleukin-4 ; secretion ; T-Lymphocytes ; immunology ; Transplantation, Homologous ; immunology

OBJECTIVETo detect the expression of cytokines by acute promyelocytic leukemia (APL) cells before and after exposure to arsenic trioxide.

METHODSDiagnoses were performed according to the FAB cytological classification criteria and cytogenetic criteria. Bone marrow or blood samples from APL patients were collected in heparinized tubes, then primary APL cells were separated by traditional Ficoll-Hypaque density centrifugation and purified after adherence to plastic surfaces. IL-1(beta), IL-6, IL-8, TNF alpha and G-CSF levels in the leukemia cell culture supernatants were detected by ELISA. At the same time, nitro blue tetrazolium (NBT) reduction test was used to detect the differentiation of APL cells.

RESULTSAfter 96 hours exposure to arsenic trioxide, 10 - 6 mol/L in vitro or 10 mg/d in vivo, APL cells showed a significant increase of IL-1(beta) (P < 0.05) and G-CSF (P < 0.05) production, and a significant decrease of IL-6 (P < 0.05) and IL-8 (P < 0.05). However, there was no obvious variation of TNF alpha when compared with APL cells without exposure to arsenic trioxide. On the other hand, the proliferation ratio of APL cells in vitro was statistically correlated to the IL-1(beta) secretion ratio or G-CSF secretion ratio. The cell number ratio in patients with detectable IL-1(beta) or G-CSF was higher than that without detectable IL-1(beta) or G-CSF.

CONCLUSIONIL-1(beta) and G-CSF secretion may play an important role in the proliferation of APL cells after exposure to arsenic trioxide.

Arsenicals ; pharmacology ; Cells, Cultured ; Cytokines ; secretion ; Granulocyte Colony-Stimulating Factor ; secretion ; Humans ; Interleukin-1 ; secretion ; Interleukin-6 ; secretion ; Interleukin-8 ; secretion ; Leukemia, Promyelocytic, Acute ; metabolism ; Oxides ; pharmacology ; Tumor Necrosis Factor-alpha ; secretion

OBJECTIVETo study the effects of Aloe coarse polysaccharide on the levels of growth factors (EGF, TGF-alpha, TGF-beta1) and interleukins (IL-1beta, IL-6, IL-8) and tumor necrosis factor (TNF) in cultured keratinocytes.

METHODThe cultured keratinocytes were treated with Aloe coarse polysaccharide at concentrations of 75, 150, 300, 600, 1 200 mg x L(-1) land the equal volume of media as control group. The levels of EGF, TGF-alpha, TGF-beta1, IL-1beta, IL-6, IL-8 and TNF in the supernatants of cultured keratinocytes were assayed by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA).

RESULTCompared with the control group, the levels of EGF, TGF-alpha, IL-1beta, IL-6 and IL-8 were significantly increased by treatment with Aloe coarse polysaccharide (P < 0.05, P < 0.01) and in a dose dependent manner, and the levels of TGF-beta1 and TNF were also increased but no statistical significance.

CONCLUSIONAloe coarse polysaccharide may promote keratinocytes to secrete EGF, TGF-alpha, IL-1beta, IL-6 and IL-8.

Aloe ; chemistry ; Cells, Cultured ; Cytokines ; secretion ; Dose-Response Relationship, Drug ; Epidermal Growth Factor ; secretion ; Humans ; Interleukin-1beta ; secretion ; Interleukin-6 ; secretion ; Interleukin-8 ; secretion ; Keratinocytes ; secretion ; Plants, Medicinal ; chemistry ; Polysaccharides ; administration & dosage ; isolation & purification ; pharmacology ; Transforming Growth Factor alpha ; secretion ; Transforming Growth Factor beta1 ; secretion ; Tumor Necrosis Factors ; secretion

ISG15 is a well-known intracellular ubiquitin-like molecule involved in ISGylation. However, a recent study has revived the notion first put forward two decades ago that ISG15 is also a secreted molecule. Human neutrophils, monocytes and lymphocytes can release ISG15, even though this protein has no detectable signal peptide sequence. ISG15 has also been found in the secretory granules of granulocytes. The mechanism underlying ISG15 secretion is unknown. Secreted ISG15 acts on at least T and natural killer (NK) lymphocytes, in which it induces interferon (IFN)-gamma production. However, the mechanism by which ISG15 stimulates these cells also remains unclear. ISG15 and IFN-gamma seem to define an innate circuit that operates preferentially, but not exclusively, between granulocytes and NK cells. Inherited ISG15 deficiency is associated with severe mycobacterial disease in both mice and humans. This infectious phenotype probably results from the lack of secreted ISG15, because patients and mice with other inborn errors of IFN-gamma immunity also display mycobacterial diseases. In addition to raising mechanistic issues, the studies described here pave the way for clinical studies of various aspects, ranging from the use of recombinant ISG15 in patients with infectious diseases to the use of ISG15-blocking agents in patients with inflammatory diseases.
Amino Acid Sequence ; Animals ; Cytokines/chemistry/*secretion ; Humans ; Interferon-gamma/secretion ; Metabolism, Inborn Errors/metabolism ; Models, Biological ; Molecular Sequence Data

OBJECTIVETo investigate the influence of pleiotrophin (PTN) on the growth of rat hepatocytes.

METHODSPrimary rat hepatocytes were isolated from male Sprague-Dawley rats and divided into three groups: group A (negative control), cultivated in normal culture medium; group B (positive control), cultivated with culture medium supplemented with supernatant from the embryonic fibroblast 3T3 cell line; group C (experimental), cultivated with culture medium supplemented with human recombinant (hr) PTN (100 ng/ml). The hepatocytes' growth rate and level of secreted albumin (ALB) were evaluated by microscopy and biochemical assay, respectively. Significance of between-group differences were assessed by one-way ANOVA, and pairwise comparisons were performed by the least significant difference test.

RESULTSThe growth rates of hepatocytes in groups A, B and C were 2.800+/-0.084%, 4.300+/-0.132% and 3.800+/-0.053%, respectively. The growth rate of group B was significantly higher than the other two groups (F = 333.735, P less than 0.05). For all groups, the highest levels of secreted ALB were detected between the second and sixth day of culture, with g/L concentrations at day 2, 4 and 6 of: group A, 0.550+/-0.010, 0.900+/-0.030 and 0.300+/-0.040; group B, 0.900+/-0.030, 1.300+/-0.020 and 1.400+/-0.030; group C, 0.900+/-0.010, 1.160+/-0.010 and 0.700+/-0.050. The secreted ALB of group B was significantly higher than that of the other two groups (F = 651.355, 338.831 and 863.205, P less than 0.05 ).

CONCLUSIONPTN can benefit in vitro culturing of rat hepatocytes by stimulating growth and enhancing their ability to secrete albumin.

Albumins ; secretion ; Animals ; Carrier Proteins ; pharmacology ; Cells, Cultured ; Cytokines ; pharmacology ; Hepatocytes ; cytology ; drug effects ; secretion ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo examine whether the enhanced expression of CD40L cDNA on murine ovarian cancer (OVHM) cells could induce the secretion of Th1 cytokines from dendritic cells (DC).

METHODSOVHM cells were transfected with the full-length mouse CD40L cDNA by lipofectamine 2000 and then G418 resistant cells as positive cells were selected. They were examined for their expression of CD40L with flow cytometry. Bone marrow cells were firstly depleted of erythrocytes, macrophages, T and B cells with PE-conjugated magnetic beads, and then cultured in 10% FCS RPMI 1640 medium supplemented with recombinant mouse GM-CSF and IL-4 for 10 days. PKH67-labeled tumor cells were cultured with DC, and then the stained cells were analyzed for the expression of MHC-I, MHC-II, CD80, CD86, CCR7 in DC with flow cytometry. The expression of p40, p19, p35, p28, EBI3 subunits, IL-18, IFN-gamma, Mig gene in cocultured DC-tumor cells were detected by RT-PCR.

RESULTSThe CD40L cDNA was successfully transfected into OVHM cells. Bone marrow-derived DCs, when cultured with CD40L/OVHM, formed clusters with the tumors and showed an upregulated expression of MHC- I, MHC-II, CD80, CD86, CCR7. Expression of the IL-12, IL-23, IL-27, IL-18, interferon-gamma (IFN-gamma) and Mig (monokine induced by IFN-gamma) genes was induced in the DCs that were cultured with CD40L/OVHM but not with OVHM cells.

CONCLUSIONThese data directly showed that the expression of CD40L on ovarian cancer cells facilitates the interaction between DCs and tumors, enhances the maturation of DCs, induces secretion of Th1 cytokines, especially for IL-12, IL-23 and IL-27, which maybe one of the possible antitumor mechanism for CD40L-transfected ovarian cancer cell line.

Animals ; CD40 Ligand ; genetics ; metabolism ; Cell Line, Tumor ; Cells, Cultured ; Coculture Techniques ; Cytokines ; secretion ; DNA, Complementary ; genetics ; Dendritic Cells ; cytology ; metabolism ; Female ; Interleukin-12 ; secretion ; Interleukin-23 ; secretion ; Interleukins ; secretion ; Mice ; Ovarian Neoplasms ; metabolism ; pathology ; Th1 Cells ; secretion ; Transfection

It has become increasingly clear that cytokines play an important role in modulating neuroendocrine regulation, especially in the secretion of corticotropin (ACTH) in the pituitary. Oncostatin M (OSM), a cytokine of IL-6 family has been reported to increase ACTH secretion and pro-opiomelanocortin (POMC) transcription in murine corticotroph pituitary tumor cells (AtT20 cells). The present study was undertaken to determine the effects of OSM on hormonal release in primary culture of rat pituitary cells. Growth hormone or prolactin release was not affected by OSM. OSM (1 nM) stimulated ACTH release (35.1% increase versus control, p>0.001) in dispersed pituitary cells of rat to a lesser extent than in AtT20 cells. Corticotropin releasing hormone (CRH) (10 nM) also induced a 2.3-fold increase of ACTH secretion (p>0.001), but co-treatment of OSM and CRH did not exhibit any synergistic effect on ACTH secretion. We conclude OSM has a stimulatory effect on ACTH secretion in normal rat pituitary cell cultures, and OSM acts mainly on corticotroph, supporting the potential role of OSM to modulate immune-endocrine regulation in the pituitary.
Animal ; Cells, Cultured ; Corticotropin/secretion* ; Cytokines/pharmacology ; Cytokines/metabolism* ; Inflammation Mediators/pharmacology ; Inflammation Mediators/metabolism* ; Male ; Peptides/pharmacology ; Peptides/metabolism* ; Pituitary Gland/metabolism* ; Pituitary Gland/drug effects ; Pituitary Gland/cytology ; Prolactin/secretion* ; Rats ; Rats, Inbred WF ; Somatotropin/secretion*

It has become increasingly clear that cytokines play an important role in modulating neuroendocrine regulation, especially in the secretion of corticotropin (ACTH) in the pituitary. Oncostatin M (OSM), a cytokine of IL-6 family has been reported to increase ACTH secretion and pro-opiomelanocortin (POMC) transcription in murine corticotroph pituitary tumor cells (AtT20 cells). The present study was undertaken to determine the effects of OSM on hormonal release in primary culture of rat pituitary cells. Growth hormone or prolactin release was not affected by OSM. OSM (1 nM) stimulated ACTH release (35.1% increase versus control, p>0.001) in dispersed pituitary cells of rat to a lesser extent than in AtT20 cells. Corticotropin releasing hormone (CRH) (10 nM) also induced a 2.3-fold increase of ACTH secretion (p>0.001), but co-treatment of OSM and CRH did not exhibit any synergistic effect on ACTH secretion. We conclude OSM has a stimulatory effect on ACTH secretion in normal rat pituitary cell cultures, and OSM acts mainly on corticotroph, supporting the potential role of OSM to modulate immune-endocrine regulation in the pituitary.
Animal ; Cells, Cultured ; Corticotropin/secretion* ; Cytokines/pharmacology ; Cytokines/metabolism* ; Inflammation Mediators/pharmacology ; Inflammation Mediators/metabolism* ; Male ; Peptides/pharmacology ; Peptides/metabolism* ; Pituitary Gland/metabolism* ; Pituitary Gland/drug effects ; Pituitary Gland/cytology ; Prolactin/secretion* ; Rats ; Rats, Inbred WF ; Somatotropin/secretion*