<b>OBJECTIVEb>To analyze the characteristics of lymphocyte phenotypes in hepatitis B virus (HBV) transgenic mice and the effect of exogenous interferon-α on virological profiles and lymphocytes phenotypes of the mice.

<b>METHODSb>HBV transgenic mice and wild-type (WT) mice were examined for serum levels of HBsAg, HBcAb, IL-21, and IL-6 using ELISA. The frequencies of CD4(+)T and CD19(+)B cells separated from the liver, spleen, and peripheral blood were detected by flow cytometry. Nine HBV transgenic mice were injected subcutaneously with recombinant mouse interferon alpha (rmIFN-α) and another 9 transgenic mice were injected with PBS, and their HBsAg, HBV DNA, IL-6, and IL-21 levels and frequencies of peripheral blood CD4(+)T and CD19(+)B cells were detected.

<b>RESULTSb>HBV transgenic mice showed a high level of HBsAg with a detectable level of HBcAb and significantly increased serum levels of IL-21 and IL-6 as compared with WT mice (P<0.05). The transgenic mice had a significantly lower frequency of CD4(+) T cells in the peripheral blood, liver and spleen (P<0.05) but a significantly higher frequency of CD19(+) B cells in the liver (P<0.05). An inverse correlation between intrahepatic CD4(+) T cell frequency and serum HBsAg level while a positive correlation between intrahepatic CD19(+) B cell frequency and HBcAb level were found in HBV transgenic mice. Administration of rmIFN-α significantly increased the frequencies of CD4(+) T and CD19(+) B cells in the peripheral blood and the serum level of IL-6 in HBV transgenic mice (P<0.05).

<b>CONCLUSIONb>HBV transgenic mice have lymphocyte subset dysregulation and exogenous interferon-α can modulate the immune function of the mice by regulating the frequencies of lymphocyte subsets.

Animals ; Antiviral Agents ; pharmacology ; B-Lymphocytes ; drug effects ; DNA, Viral ; blood ; Hepatitis B ; drug therapy ; immunology ; Hepatitis B Antibodies ; blood ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; Interferon-alpha ; pharmacology ; Interleukin-6 ; blood ; Interleukins ; blood ; Liver ; immunology ; Lymphocyte Subsets ; cytology ; drug effects ; Mice ; Mice, Transgenic ; Phenotype ; T-Lymphocytes ; drug effects

We evaluated the effectiveness of rhamnogalacturonan II (RG-II)-stimulated bone marrow-derived dendritic cells (BMDCs) vaccination on the induction of antitumor immunity in a mouse lymphoma model using EG7-lymphoma cells expressing ovalbumin (OVA). BMDCs treated with RG-II had an activated phenotype. RG-II induced interleukin (IL)-12, IL-1beta, tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) production during dendritic cell (DC) maturation. BMDCs stimulated with RG-II facilitate the proliferation of CD8+ T cells. Using BMDCs from the mice deficient in Toll-like receptors (TLRs), we revealed that RG-II activity is dependent on TLR4. RG-II showed a preventive effect of immunization with OVA-pulsed BMDCs against EG7 lymphoma. These results suggested that RG-II expedites the DC-based immune response through the TLR4 signaling pathway.
Acute-Phase Proteins/metabolism ; Adaptor Proteins, Vesicular Transport/metabolism ; Animals ; Antigens, CD14/metabolism ; Bone Marrow Cells/cytology/drug effects ; CD8-Positive T-Lymphocytes/*immunology ; Carrier Proteins/metabolism ; Cell Differentiation/drug effects ; Cell Nucleus/drug effects/metabolism ; Cell Proliferation/drug effects ; Cytokines/biosynthesis ; Dendritic Cells/cytology/drug effects/enzymology/*immunology ; Enzyme Activation/drug effects ; Lymphocyte Activation/*drug effects ; Membrane Glycoproteins/metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Mitogen-Activated Protein Kinases/metabolism ; Myeloid Differentiation Factor 88/metabolism ; NF-kappa B/metabolism ; Neoplasms/immunology/*pathology ; Pectins/*pharmacology ; Phenotype ; Protein Transport/drug effects ; Receptors, Chemokine/metabolism ; Signal Transduction/drug effects ; T-Lymphocytes, Cytotoxic/cytology/drug effects ; Toll-Like Receptor 4/*agonists/metabolism

Calreticulin (CRT) is a multifunctional molecule in both intracellular and extracellular environment. We have previously found that a recombinant CRT fragment (rCRT/39-272) could modulate T cell-mediated immunity in mice via activation and expansion of CD1d(hi)CD5⁺ B cells as well as induction of CRT-specific regulatory antibodies. Antibody secreting cells (ASCs) are terminally differentiated B cells responsible for producing antibodies to participate in positive immune response as well as immune regulation. In this study, we demonstrate that rCRT/39-272 differentiates murine CD1d(hi)CD5⁺ B cells into ASCs marked by increased expression of plasma cell-associated transcription factors and production of polyreactive antibodies against DNA and CRT in vitro. Intraperitoneal administration of rCRT/39-272 augmented differentiation of CD1d(hi)CD5⁺ B cells into ASCs in naïve mice or mice with experimental autoimmune encephalomyelitis. Thus, we propose that ASC differentiation and subsequent antibody production of CD1d(hi)CD5⁺ B cells are key steps in CRT-mediated immunoregulation on inflammatory T cell responses.
Animals ; Antigens, CD1d ; metabolism ; Autoantibodies ; biosynthesis ; B-Lymphocytes ; cytology ; drug effects ; immunology ; metabolism ; CD5 Antigens ; metabolism ; Calreticulin ; chemistry ; Cell Differentiation ; drug effects ; Encephalomyelitis, Autoimmune, Experimental ; immunology ; Humans ; Mice ; Peptide Fragments ; chemistry ; pharmacology ; Solubility

<b>OBJECTIVEb>To investigate the effects of ursolic acid (UA) on T-cell proliferation and activation, as well as to examine its effect on nuclear factor-κB (NF-κB) signaling pathway in T cells.

<b>METHODSb>T-cells isolated from BALB/c mice were incubated with UA at concentrations ranging from 5-30 μmol/L in the presence of phorbol 12-myristate 13-acetate (PMA) or PMA plus ionomycin. The proliferation of T cells was measured by the MTT assay. The expressions of CD69, CD25, and CD71 on T-cell surface were analyzed using flow cytometry. The level of interleukin-2 (IL-2) in the culture supernatant of activated T cells was quantified by enzyme-linked immunosorbent assay (ELISA). The level of phosphorylated IκB-α (p-IκB-α) in total protein and p65, a subunit of NF-κB, nuclear translocation were measured by Western blot analysis.

<b>RESULTSb>UA in a dose-dependent manner significantly decreased the proliferation and inhibited the surface expressions of CD69, CD25, and CD71 in murine T lymphocytes upon in vitro activation (P<0.01). Significant reduction of IL-2 production was found in activated T cells treated with UA (P<0.01). The PMA-induced increase in p-IκB-α protein was inhibited, and nuclear translocation of p65 from the cytoplasm was blocked by UA.

<b>CONCLUSIONb>UA is a potent inhibitor for T cell activation and proliferation; these effects are associated with the inhibition of NF-κB signaling pathway.

Animals ; Cell Nucleus ; drug effects ; metabolism ; Cell Proliferation ; drug effects ; I-kappa B Proteins ; metabolism ; Interleukin-2 ; secretion ; Ionomycin ; pharmacology ; Lymphocyte Activation ; drug effects ; Mice ; Mice, Inbred BALB C ; NF-KappaB Inhibitor alpha ; NF-kappa B ; metabolism ; Phosphorylation ; drug effects ; Protein Transport ; drug effects ; Signal Transduction ; drug effects ; T-Lymphocytes ; cytology ; drug effects ; immunology ; secretion ; Tetradecanoylphorbol Acetate ; pharmacology ; Triterpenes ; pharmacology

B-1 cells, which constitute a predominant lymphocyte subset in serosal cavities and produce most of natural antibodies, are subdivided into the CD5+ B-1a and CD5- B-1b cell subpopulations, but the differential roles of B-1a and B-1b cells are not well understood. We report that B-1a cells preferentially migrate out of the peritoneal cavity and upregulate the expression of CXCR4 with heightened sensitivity to CXCL12 and CXCL13 upon LPS treatment compared to B-1b and B-2 cells. Whereas B-1a cells were slightly more abundant than B-1b and B-2 cells in the homeostatic condition, the number of B-1a cells preferentially decreased 48 hr after LPS treatment. The decrease in the peritoneal B-1a cell number was accompanied with increased migration of B-1a cells toward CXCL-12 and CXCL-13 in in vitro transmigration assay using peritoneal B cells from LPS treated mice. The expression level of CXCR4, but not of CXCR5, was also more prominently increased in B-1a cells upon LPS stimulation. LPS-stimulated B-1a cells did not accumulate in omental milky spots in contrast to B-2 cells. These results suggest that B-1a cells actively migrate out of the peritoneal cavity through the regulation of the migratory responsiveness to chemokines and actively participate in systemic immune responses.
Adjuvants, Immunologic/pharmacology ; Animals ; B-Lymphocytes/cytology/*drug effects/immunology ; Cell Movement ; Cells, Cultured ; Chemokine CXCL12/metabolism/*pharmacology ; Chemokine CXCL13/metabolism/pharmacology ; Lipopolysaccharides/*pharmacology ; Mice ; Mice, Inbred C57BL ; Peritoneal Cavity/cytology ; Receptors, CXCR4/*metabolism ; Up-Regulation

Adenine ; administration & dosage ; analogs & derivatives ; therapeutic use ; Adult ; Antiviral Agents ; administration & dosage ; therapeutic use ; Biomarkers ; blood ; Critical Illness ; Cytokines ; blood ; DNA, Viral ; blood ; Follow-Up Studies ; Hepatitis B virus ; drug effects ; immunology ; Hepatitis B, Chronic ; blood ; drug therapy ; immunology ; virology ; Humans ; Lymphocyte Count ; Male ; Organophosphonates ; administration & dosage ; therapeutic use ; Recurrence ; T-Lymphocytes ; cytology ; immunology ; Time Factors ; Treatment Outcome

<b>BACKGROUNDb>Cryptotanshinone (CT) was originally isolated from the dried roots of Salvia militorrhiza, an herb that is used extensively in Asian medicine and the extracts of this herb have been used in the treatment of several pathologies, including cardiovascular diseases, hematological abnormalities, hepatitis, and hyperlipidemia, but no studies had been carried on the treatment for rheumatic diseases with it. This study aimed to investigate the effects of cryptotanshinone on immune functions in rats with adjuvant arthritis (AA).

<b>METHODSb>Complete Freund's adjuvant was used to induce AA in rats. Thymus and spleen was aseptically taken from normal rats and the AA rats. Then a thymus lymphoid cell suspension, splenic lymphoid cell suspension and peritoneal macrophage cell suspension were prepared. After adding CT (0.1 microg/ml, 1.0 microg/ml, 10 microg/ml, 100 microg/ml, 1000 microg/ml) into the suspension, T and B lymphocytes proliferation was determined by 3-(4,5-2 dimethylthiazal-2yl)2,5-diphenyltetrazoliumbromide (MTT) assay. And the activities of interleukin-1 (IL-1) and IL-2 were measured by the mouse lymphocytes proliferation assay.

<b>RESULTSb>Thymic T and splenic B lymphocyte proliferation of the AA rat was significantly lower, and could be stored through using CT in vitro. CT (100 microg/ml and 1000 microg/ml) increased T or B lymphocytes proliferation in vitro (P < 0.01). In AA rats, the levels of IL-1 released by abdominal PMPhi significantly increased whereas the level of IL-2 released by T cells decreased in vitro. CT (1000 microg/ml) decreased the production of IL-1 and promoted production of IL-2 in vitro (P < 0.05).

<b>CONCLUSIONSb>CT can ameliorate the abnormal immunological functions in AA rats.

Analysis of Variance ; Animals ; Arthritis, Experimental ; drug therapy ; immunology ; B-Lymphocytes ; drug effects ; immunology ; Interleukin-1 ; metabolism ; Interleukin-2 ; metabolism ; Lymphocyte Activation ; drug effects ; immunology ; Male ; Phenanthrenes ; pharmacology ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Spleen ; cytology ; T-Lymphocytes ; drug effects ; immunology ; Thymus Gland ; cytology

<b>OBJECTIVEb>To study the effect of rapamycin in inducing naïve murine effector T cell (Teff) convert to regulatory T cell (Treg) in vitro.

<b>METHODSb>The forkhead box protein 3 (FoxP3) negative Teff were isolated and purified from the spleen and lymph node of C57 BL/6 murines aged 6-8 weeks, then Teff were cultured in three groups with mature dendritic cells (mDC), B cells, and plate coated Anti-CD3. In addition, the control wells and the test wells were prepared in each group, rapamycin were not added in the control wells but added in the test wells with concentrations of 1, 10, 50, and 100 nmol/L. Percentages of FoxP3 positive Treg were examined by flow cytometry after 4 days in Anti-CD3 group and after 6 days in the other two groups.

<b>RESULTSb>As shown by the flow cytometry, the percentages of FoxP3 positive Treg were as follows in three group: in the mDC group, it was 0.01% in the control well and 0.39%, 0.47%, 0.34%, and 0.26% in test wells; in B cell group, it was 0.01% in the control wells and 5.56%, 5.89%, 7.15%, and 4.72% in the test wells; in Anti-CD3 group, it was 0.93% in the control wells and 1.35%, 1.07%, 1.02%, and 1.19% in test wells. No significant difference was found between the test wells and control wells in the mDC group and Anti-CD3 group; however, the percentages of FoxP3 positive Treg was significantly different between the test wells and control wells in the B cell group (P < 0.01).

<b>CONCLUSIONb>When B cell is acted as the antigen-presenting cell, rapamycin can effectively induce Teff convert to Treg in vitro.

Animals ; B-Lymphocytes ; cytology ; drug effects ; immunology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Dendritic Cells ; cytology ; drug effects ; immunology ; Flow Cytometry ; Forkhead Transcription Factors ; immunology ; Mice ; Mice, Inbred C57BL ; Mice, Inbred DBA ; Precursor Cells, T-Lymphoid ; cytology ; drug effects ; immunology ; Sirolimus ; pharmacology ; T-Lymphocyte Subsets ; cytology ; drug effects ; immunology ; T-Lymphocytes, Regulatory ; cytology ; drug effects ; immunology

<b>OBJECTIVEb>To observe the effect of polysaccharide ATPS-2 from Armillariella tabescens on the immunological liver injury in mice induced by BCG plus LPS.

<b>METHODb>BCG and LPS were adopted to establish BCG plus LPS liver injury model in mice. The content of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and NO, the activity of superoxide dismutase (SOD) and malondiadehyde (MDA) content of liver homogenate in mice were measured by colorimetric method. The content of tumor necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1), on serum were measured by enzyme linked immunosorbent assay (ELISA) , and T- and B-lymphocyte proliferation were measured by MTT. Index of liver, spleen and thymus were calculated after treatment.

<b>RESULTb>Polysaccharide ATPS-2 from A. tabescens (25, 50, 100 mg x kg(-1)) could obviously reduce the high level of ALT, AST, NO and TNF-alpha, IL-1 on serum, inhibit the high level of MDA, increase the low activity of SOD in liver homogenate and enhance T-and B-lymphocyte proliferation, elevate the spleen, thymic index and decrease liver index of the mice to different extent.

<b>CONCLUSIONb>Polysaccharide ATPS-2 from A. tabescens had apparently protective effects in the immunological liver injury mice induced by BCG plus LPS.

Agaricales ; chemistry ; Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; B-Lymphocytes ; cytology ; Cell Proliferation ; drug effects ; Chemical and Drug Induced Liver Injury ; Interleukin-1 ; blood ; Lipopolysaccharides ; Liver ; metabolism ; pathology ; Liver Diseases ; immunology ; metabolism ; pathology ; Male ; Malondialdehyde ; metabolism ; Mice ; Mycobacterium bovis ; Nitric Oxide ; blood ; Polysaccharides ; isolation & purification ; pharmacology ; Protective Agents ; isolation & purification ; pharmacology ; Random Allocation ; Superoxide Dismutase ; metabolism ; T-Lymphocytes ; cytology ; Tumor Necrosis Factor-alpha ; blood

<b>AIMb>To investigate effects of hsBAFF synthesized in Escherichia coli on spleen B lymphocyte immune response and its intracellular free Ca2+ ([Ca2]i]) signaling in mice.

<b>METHODSb>Twenty ICR mice, half males-half females, were chosen and randomly divided into a normal control group (n=10) and a hsBAFF treatment group (n-10). The mice in hsBAFF treatment group were given abdominal cavity injection of hsBAFF solution which was diluted with phosphate buffered saline (PBS) at dosage of 0.1 mg/kg body weight once each day for over eight days. The mice in control group were received abdominal injection of PBS at the same dose and frequency. Spleen B lymphocyte proliferation and its immune response to LPS stimulation in mice were evaluated using an MTT assay, and change of spleen B lymphocyte [Ca2+]i was assayed under a laser scanning confocal microscope.

<b>RESULTSb>B lymphocyte proliferation and its immune response to LPS stimulation were significantly higher in hsBAFF-treated mice than in control mice (P < 0.05). The B lymphocyte [Ca2+]i fluorescence intensity in hsBAFF-treated mice maintained at a relatively high level fluctuation, and its average intensity was significantly higher to that of control mice (P < 0.01), but change rate of the intensity was lower compared to that of control group.

<b>CONCLUSIONb>hsBAFF synthesized in Escherichia coli can enhance immune function in the body by increasing B lymphocyte proliferation and its immune response. hsBAFF-activated B lymphocyte function may be associated with increasing B lymphocytes [Ca2+]i.

Animals ; B-Cell Activating Factor ; immunology ; pharmacology ; B-Lymphocytes ; cytology ; drug effects ; immunology ; Calcium ; metabolism ; Calcium Signaling ; drug effects ; Cell Proliferation ; drug effects ; Female ; Male ; Mice ; Mice, Inbred ICR ; Spleen ; cytology ; drug effects ; immunology