OBJECTIVETo explore the feasibility of Single Fe(2)O(3)-PLL labeled mouse spleen-derived endothelial progenitor cells (EPCs) detection by 7.0T MR system.

METHODSMononuclear cells (MNCs) were isolated from mouse spleen by density gradient centrifugation and EPCs were obtained by the different adherence of cells.Immunocytochemistry and fluorescent staining were performed to identify EPCs. The EPCs were labeled with Fe(2)O(3)-PLL and the intracellular iron was identified with prussian blue staining. MTT assay was assessed to evaluate proliferation of Fe(2)O(3)-PLL labeled EPCs. The cells underwent MR imaging with different sequences.

RESULTSCultured in vitro, mouse spleen-derived MNCs resulted in EC-like morphology. These cells expressed EPCs-specific antigens, such as CD31, CD34 and vWF, and had the ability to incorporate ac-LDL and bind UEA-1. Between Fe(2)O(3)-PLL labeled EPCs and unlabeled cells, MTT value of light absorption had no statistical significant difference (day4 t = 2.81, day5 t = -1.87, day6 t = -0.298, day7 t = -0.115, all P > 0.05). The signal void induced by labeled single cell is 20.2 pixels in MSME sequence, and 20.2 pixels in 3D-FLASH sequence (t = 15.2, P < 0.05). Single cell could be detected by 7.0 T MR system.

CONCLUSIONMNCs isolated from mouse spleen can differentiate into endothelial cells in vitro and have the specific property of stem cells. The mouse spleen-derived EPCs can be labeled with Fe(2)O(3)-PLL efficiently. The labeled EPCs can be imaged as dispersed single cells.

Animals ; Cell Differentiation ; Cells, Cultured ; Endothelial Cells ; cytology ; Ferric Compounds ; Magnetic Resonance Imaging ; Mice ; Spleen ; cytology ; Stem Cells ; cytology

Immunogenicity for medical devices of animal origin is the key and difficult point during immune safety evaluation for these devices. This paper firstly investigated the effect of collagen sponge of animal origin on mouse splenic lymphocyte transformation and proliferation, and then analyzed the influence factors on the MTT method and CFSE method. The results showed that collagen sponge extract cannot significantly induce transformation and proliferation of mouse splenic lymphocyte in vitro.
Animals ; Cells, Cultured ; Collagen ; pharmacology ; Lymphocyte Activation ; drug effects ; Mice ; Porifera ; chemistry ; Spleen ; cytology

To attempt a rigorous definition of the structure of the accessory spleen (AS) in the Chinese hamster, we examined twenty-one animals, and found AS in 5 animals (23.8%), which were over 7-month-old. The AS had no connection with the main spleen and was seen as a dark red oval organ (0.7 mm x 1.5 mm), which was embedded in the adipose tissue near the tail of the pancreas. It was demarcated from the adipose tissue and some pancreatic tissue. The organ was encapsulated by thin collagenous connective tissue and smooth muscle fibers, and contained lymphatic nodules, reticular fibers, nodular central arterioles, macrophages and megakaryocytes. Notably the incidence of AS appeared to increase with age in the Chinese hamsters.
Adipose Tissue/anatomy & histology ; Age Factors ; Animals ; Connective Tissue/anatomy & histology ; Cricetinae ; Cricetulus/*anatomy & histology ; Erythrocytes/cytology ; Lymphocytes/cytology ; Muscle, Smooth/anatomy & histology ; Pancreas ; Spleen/*anatomy & histology/cytology

BACKGROUNDIncreasing evidence suggests that, by the production of indoleamine 2,3-dioxygenase (IDO), dendritic cells (DC) may reduce the activity of T lymphocytes and inhibit T lymphocyte proliferation-induced immune tolerance. One promising way is inspired by increasing IDO expression in DC cells for immune tolerance after transplantation. The aim of this work was to examine the effect of interferon-γ (IFN-γ) on the expression of IDO by DC.

METHODSSpleen-derived rat DCs were cultured and induced by cytokines, and the expression of OX62 and surface molecules CD80 and CD86 were measured with flow cytometry. After the DCs were induced by IFN-γ at different concentrations (0, 100, 300, 500 U/ml), the expression levels of IDO mRNA were measured with real-time PCR, and the expression levels of IDO protein in DCs were measured with Western blotting. The allogeneic mixed lymphocyte reaction (MLR) was used to test the effects of DCs incubated with different concentrations of IFN-γ on allogeneic T lymphocyte proliferation.

RESULTSUnder the microscope, the DCs induced by IFN-γ showed a typical dendritic morphology. The expression rate of OX62 was above 80% and the positive expression rates of CD80 and CD86 were both about 80%. The expressions of IDO mRNA and IDO protein increased gradually with the increase of IFN-γ concentration, showing statistical significance in the differences between the groups (P < 0.05). Compared with the control DC, the DC incubated with IFN-γ had a notable decrease in allostimulatory activity (P < 0.05). With the increasing IFN-γ concentration, the T lymphocyte proliferation decreased, and the difference between the groups was also statistically significant (P < 0.05).

CONCLUSIONSThe highly purified spleen derived rat DCs can be successfully acquired through the improved adhesion in-vitro method. IFN-γ can induce increased expression of IDO in spleen-derived rat DCs and reduce the spleen DCs' capacity to stimulate the proliferation of allogeneic T cells.

Animals ; Cell Proliferation ; Cells, Cultured ; Dendritic Cells ; cytology ; Enzyme Induction ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; biosynthesis ; Male ; Rats ; Rats, Wistar ; Spleen ; cytology ; T-Lymphocytes ; cytology

OBJECTIVETo explore the mechanism of electroacupuncture of "Shuanggu Yitong" prescription on postponing aging.

METHODSForty 3-month SD rats, male only, 30 rats were made sub-acute aging model by D-galactose s.c. injection continuously for 42 d, and rest of the rats, 10, were divided into a normal control group. After the modeling, the sub-acute aging model rats were randomly into a Shuanggu Yitong group [electroacupuncture at "Guanyuan" (CV 4) and "Zusanli" (ST 36), hand needle at "Baihui" (GV 20)], an acupuncture control group [electroacupuncture at "Weizhong" (BL 40) and "Shuifen" (CV 9), hand needle at "Yintang" (GV 29)] and an aging model group, ten in each one. The treatment was given once in a day, six of which made a course. The rats in the normal control group and aging model group were not received any treatment. After the treatment for three weeks, the rats were put to death and their spleen index, thymus index and the T lymphocytes subgroups (CD8(+) T/T cell and CD8(+) CD28(-) T/CD8(+) T cell) were tested.

RESULTSThe spleen index (1.74 +/- 0.059) and thymus index (0.64 +/- 0.039) in the aging model group was obviously lower than those in the normal control group (1.93 +/- 0.061), (0.81 +/- 0.053) respectively (both P < 0.05); the CD8(+) CD28(-) T/CD8(+) T cell percentages (26.28 +/- 4.69)% and CD8(+) T/T cell percentages (43.33 +/- 2.84)% in the aging model group were both significantly higher than those (15.08 +/- 5.58)% (P < 0.01), (34.70 +/- 4.24)% (P < 0.01) in the normal control group. Compared with the aging model group, the spleen index (1.91 +/- 0.081) and thymus index (0.79 +/- 0.080) in the Shuanggu Yitong group were significantly higher (both P < 0.05), but obviously decreased with the percentage of CD8(+) CD28(-) T/CD8(+) T cell (18.07 +/- 1.73) (P < 0.01); the percentage of CD8(+) CD28(-) T/CD8(+) T cell (18.07 +/- 1.73)% in the acupuncture control group was also lower than the aging model group (P < 0.05), but more obvious reduce for the Shuanggu Yitong group (P < 0.05).

CONCLUSIONThe treatment of Shuanggu Yitong prescription could regulate the proportions of the T lymphocyte subset, and slow down the immunosenescence of subacute aging model rats induced by D-galactose.

Acupuncture Points ; Aging ; physiology ; Animals ; Electroacupuncture ; Flow Cytometry ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Spleen ; cytology ; T-Lymphocyte Subsets ; cytology ; Thymus Gland ; cytology

OBJECTIVETo investigate the association of dendritic cell distribution in the peripheral blood, spleen and arterial wall with intimal hyperplasia in rats with diabetes mellitus.

METHODSDiabetes mellitus was induced in rats by intraperitoneal injection of streptozotocin and high-fat feeding for 8 weeks. Peripheral blood, arterial wall and the spleen were collected from the rats to prepare cell suspensions, in which the proportions of dendritic cells and T cells were determined by flow cytometry.

RESULTSThe tunica intimal hyperplasia was more obvious in diabetic rats with or without high-fat feeding as compared with that of the control rats (P<0.05), and their dendritic cells decreased significantly in the peripheral blood (P<0.05) but increased in the arterial wall. The percentage of T cells was also increased in the peripheral blood and arterial wall of the diabetic rats.

CONCLUSIONChanges in the distribution of dendritic cells and T cells are closely associated with intimal hyperplasia in diabetic rats, suggesting the involvement of dendritic cells and T cells in the formation of atherosclerosis.

Animals ; Dendritic Cells ; cytology ; Diabetes Mellitus, Experimental ; complications ; pathology ; Hyperplasia ; complications ; pathology ; Rats ; Rats, Sprague-Dawley ; Spleen ; cytology ; Streptozocin ; T-Lymphocytes ; cytology ; Tunica Intima ; pathology

Eimeria tenella, an intracellular protozoan parasite infecting the epithelial cells of the ceca of chickens, causes severe diarrhea and bleeding that can lead its host to death. It is of interest that E. tenella first penetrate into the mucosal intraepithelial lymphocytes (IEL) before they parasitize crypt or villous epithelial cells. This in vitro study was undertaken to know whether the penetration of E. tenella into such a lymphoid cell is a beneficial step for the parasite survival and development. Three sequential experiments were performed. First, the in vitro established bovine kidney cell line, MDBK cells, were evaluated for use as host cells for E. tenella, through morphological observation. Second, the degree of parasite development and multiplication in MDBK cells was quantitatively assayed using radioisotope-labelled uracil (3H-uracil). Third, the E. tenella sporozoites viability was assayed after preincubation of them with chicken spleen cells. E. tenella oocysts obtained from the ceca of the infected chickens were used for the source of the sporozoites. Spleen cells (E) obtained from normal chickens (FP strain) were preincubated with the sporozoites (T) at the E:T ratio of 100:1, 50:1 or 25:1 for 4 or 12 hours, and then the mixture was inoculated into the MDBK cell monolayer. Morphologically the infected MDBK cells revealed active schizogonic cycle of E. tenella in 3-4 days, which was characterized by the appearance of trophozoites, and immature and mature schizonts containing merozoites. The 3H-uracil uptake by E. tenella increased gradually in the MDBK cells, which made a plateau after 48-60 hours, and decreased thereafter. The uptake amount of 3H-uracil depended not only upon the inoculum size of the sporozoites but also on the degree of time delay (preincubation; sporozoites only) from excystation to inoculation into MDBK cells. The 3H-uracil uptake became lower as the preincubation time was prolonged. In comparison, after preincubation of sporozoites with spleen cells for 4 or 12 hours, the 3H-uracil uptake was significantly increased compared with that of control group. From the results, it was inferred that, although the penetration of E. tenella sporozoites into the lymphoid cells such as IEL is not an essential step, it should be at least a beneficial one for the survival and development of sporozoites in the chicken intestine.
Cattle- ; Cell-Line ; Cells,-Cultured ; Chickens- ; English-Abstract ; *Eimeria-growth-and-development ; *Kidney-parasitology ; *Lymphocytes-parasitology ; *Spleen-cytology

The study was aimed to explore the diagnostic significance of hemophagocytosis in the patients with hemophagocytic lymphohistiocytosis (HLH). 61 suspected HLH patients from June 2005 to October 2008 were enrolled in the study. The suspected HLH patients were divided into confirmed group (43 out of 61) and excluded group (18 out of 61) according to HLH-2004 diagnostic criteria. The incidences of hemophagocytosis in bone marrow, spleen or lymph nodes were compared in all groups. The results indicated that the hemophagocytosis in bone marrow, spleen or lymph nodes was found in 33 patients of confirmed group, while the hemophagocytosis was found in 4 patients of excluded group. The sensitivity of hemophagocytosis for the diagnosis of HLH was 76.7%, and its specificity was 77.8%. In conclusion, hemophagocytosis is a helpful marker for the diagnosis of most but not all HLH patients, however, the lack of hemophagocytosis does not mean to exclude the diagnosis of HLH.
Biopsy ; Bone Marrow Cells ; cytology ; pathology ; Diagnosis, Differential ; Humans ; Lymph Nodes ; pathology ; Lymphohistiocytosis, Hemophagocytic ; diagnosis ; Sensitivity and Specificity ; Spleen ; pathology

OBJECTIVETo investigate the influence of CD4+ CD25+ regulatory T cells(Treg cells) on mouse gastric cancer.

METHODSTreg cell in mouse spleen bearing gastric tumor was tested in different time points. Magic cell sorting(MACS) method was used to purify mouse Treg cells and the Treg cells were injected into mouse bearing gastric tumor with different dosage. After 3 weeks, the tumor size and tumor cell apoptosis rate were measured.

RESULTSTreg existed in normal mouse spleen with a rate of (3.86+/-0.07)%. In tumor model this percentage increased gradually and was (4.12+/-0.13)% after 3 weeks, which was significantly higher than that in control. When Treg cell applied in mouse reached 2.0 x 10(5), the tumor size enlarged significantly(P=0.013) and tumor cell apoptosis rate decreased significantly (P=0.012).

CONCLUSIONSTreg cell is associated with gastric cancer progress in mouse tumor model. Treg cell can promote gastric cancer growth and decrease tumor apoptosis. The anti- Treg GITR can improve anti- tumor effects.

Animals ; Apoptosis ; Female ; Flow Cytometry ; Male ; Mice ; Mice, Inbred Strains ; Spleen ; cytology ; Stomach Neoplasms ; immunology ; pathology ; T-Lymphocytes, Regulatory ; immunology

AIMTo investigate intermittent hypoxia effects on splenocyte mitogen-induced proliferation.

METHODSRats were exposured to intermittent hypoxia in a hypobaric chamber 4 h/d for 1 d, 2 d, 5 d and 15 d.

RESULTS5 km (10.8% O2) hypoxia for 1 d significantly inhibited ConA-induced splenocytes proliferation by--74.57% +/- 7.33% (P < 0.05). Hypoxia (5 km) for 2 d, 5 d and 15 d did not markedly affect splenocyte proliferation (97.03 +/- 7.18%, 104.5% +/- 8.38%, 99.55% +/- 3.8% respectively). Hypoxia 2 km (16.0% O2) for 1 d, 2 d, 5 d and 15 d had no influence on splenocytes proliferation (93.19% +/- 11.88%, 96.43% +/- 7.9%, 99.03% +/- 10.97%, 100.54% +/- 9.54% respectively). We also demonstrated that acute hypoxia exposure (5 km) 4 h significantly suppressed DNA contents of rat splenocytes by 76.22% +/- 7.06% (P < 0.05). The suppressed DNA synthesis were returned to control level after the hypoxia for 5 d and 15 d.

CONCLUSIONThese results suggest that the acute hypoxia (5 km, 4 h) induces a transient suppression on splenic lymphocyte proliferation, and the intermittent hypoxia may induce an adaptation response of the splenocytes proliferation.

Animals ; Cell Proliferation ; Hypoxia ; immunology ; Lymphocyte Activation ; Lymphocytes ; cytology ; immunology ; Male ; Rats ; Rats, Sprague-Dawley ; Spleen ; immunology