Animals ; Caffeine/adverse effects* ; Central Nervous System Stimulants/adverse effects* ; Dose-Response Relationship, Drug ; Female ; Fetal Growth Retardation/chemically induced* ; Immune System Diseases/chemically induced* ; Male ; Organ Size/drug effects* ; Pregnancy ; Pregnancy Complications/immunology* ; Rats ; Spleen/growth & development*

C57BL/6N is the most widely used inbred mouse strain applied in a wide variety of research areas including cancer, cardiovascular biology, developmental biology, diabetes and obesity, genetics, immunology, neurobiology, and sensorineural research. To compare the fertilization rates of C57BL/6NKorl mice with two commercial C57BL/6N stocks, differences in reproductive organ structures, sperm and egg numbers, fertilization rates, and embryo development rates among C57BL/6NKorl (Korea FDA source), C57BL/6NA (USA source), and C57BL/6NB (Japan source) mice were determined. Among the stocks, no significant differences were detected in organ weight and histological structure of male and female reproductive organs, although body weight was higher in C57BL/6NKorl mice than that in the other groups. The concentration and morphology of sperm and eggs in C57BL/6NKorl mice were similar to those of C57BL/6NA and C57BL/6NB mice. Furthermore, the three stocks had similar in vitro fertilization and embryo development rates, although these rates tended to be higher in C57BL/6NB mice. Pup body weight was higher in C57BL/6NKorl and C57BL/6NB mice than that in C57BL/6NA mice. The results of the present study suggest that C57BL/6NKorl, C57BL/6NA, and C57BL/6NB mice obtained from three different sources have similar fertilization and embryo development rates, although there were slight differences in the magnitude of their responses rates.
Allergy and Immunology ; Animals ; Biology ; Body Weight ; Developmental Biology ; Eggs ; Embryonic Development ; Female ; Fertilization in Vitro ; Fertilization* ; Genetics ; Humans ; Male ; Mice* ; Mice, Inbred Strains ; Neurobiology ; Obesity ; Organ Size ; Ovum ; Pregnancy ; Spermatozoa

This study aims to elucidate the mechanisms by which dexmedetomidine alleviates pulmonary edema in rats with acute lung injury induced by lipopolysaccharide (LPS). Male Wistar rats were randomly divided into five groups: normal saline control (NS) group, receiving intravenous 0.9% normal saline (5 mL/kg); LPS group, receiving intravenous LPS (10 mg/kg); small-dose dexmedetomidine (S) group, treated with a small dose of dexmedetomidine (0.5 μg · kg(-1) · h(-1)); medium-dose dexmedetomidine (M) group, treated with a medium dose of dexmedetomidine (2.5 μg · kg(-1) · h(-1)); high-dose dexmedetomidine (H) group, treated with a high dose of dexmedetomidine (5 μg · kg(-1) · h(-1)). The rats were sacrificed 6 h after intravenous injection of LPS or NS, and the lungs were removed for evaluating histological characteristics and determining the lung wet/dry weight ratio (W/D). The levels of tumor necrosis factor-alpha (TNF-α) and interleukin-1β (IL-1β) in the lung tissues were assessed by enzyme- linked immunosorbent assay (ELISA). The mRNA and protein expression levels of aquaporin-1 (AQP1) and aquaporin-5 (AQP5) were detected by RT-PCR, immunohistochemistry, and Western blotting. The lung tissues from the LPS groups were significantly damaged, which were less pronounced in the H group but not in the small-dose dexmedetomidine group or medium-dose dexmedetomidine group. The W/D and the concentrations of TNF-α and IL-1β in the pulmonary tissues were increased in the LPS group as compared with those in NS group, which were reduced in the H group but not in S group or M group (P<0.01). The expression of AQP1 and AQP5 was lower in the LPS group than in the NS group, and significantly increased in the H group but not in the S group or M group (P<0.01). Our findings suggest that dexmedetomidine may alleviate pulmonary edema by increasing the expression of AQP-1 and AQP-5.
Acute Lung Injury ; chemically induced ; drug therapy ; genetics ; pathology ; Adrenergic alpha-2 Receptor Agonists ; pharmacology ; Animals ; Aquaporin 1 ; agonists ; genetics ; immunology ; Aquaporin 5 ; agonists ; genetics ; immunology ; Dexmedetomidine ; pharmacology ; Dose-Response Relationship, Drug ; Drug Administration Schedule ; Gene Expression Regulation ; Injections, Intravenous ; Interleukin-1beta ; antagonists & inhibitors ; genetics ; immunology ; Lipopolysaccharides ; Lung ; drug effects ; immunology ; pathology ; Male ; Organ Size ; drug effects ; Pulmonary Edema ; chemically induced ; drug therapy ; genetics ; pathology ; Rats ; Rats, Wistar ; Signal Transduction ; Transcription, Genetic ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors ; genetics ; immunology

OBJECTIVETo investigate the effect and mechanism of B16F10-ESAT-6-gpi/IL-21 tumor cell vaccine on pulmonary metastasis in mouse model of melanoma.

METHODSTwelve 8-week old female C57BL/6 mice were used in this study. The mice were injected with wild-type B16F10 cells through tail vein after immunization with B16F10-ESAT-6-gpi/IL-21 tumor cell vaccine, and the pulmonary metastasis was observed. The CD4(+) and CD8(+) T cells were isolated by magnetic activated cell sorting, and then used for the detection of CFSE/7-AAD cytotoxicity by flow cytometry. Serum from the mice immunized with tumor-cell vaccine was used to detect IFN-γ expression by ELISA. The expression of TGF-β2, ZEB1, E-cadherin, and N-cadherin of tumor tissues was detected by RT-PCR and immunofluorescence, respectively.

RESULTSThe mice vaccinated with B16F10-ESAT-6-gpi/IL-21 had significantly fewer nodules in the lung and lower lung weight [(285.8 ± 19.01) mg vs. (406.3 ± 27.12) mg], with lower levels of TGF-β2, ZEB1 and N-cadherin proteins but higher level of E-cadherin protein within the tumor tissue, as compared with the control mice. Meanwhile, the immunized mice had significantly increased CD8(+) T cell killing activity [(42.62 ± 3.465)% vs. (22.29 ± 1.804)%] and IFN-γ expression level [(55.200 ± 7.173) pg/ml vs. (6.435 ± 1.339) pg/ml] over the control mice.

CONCLUSIONSThe B16F10-ESAT-6-gpi/IL-21 vaccine can inhibit the metastasis of melanoma in the lung in vaccinated melanoma-bearing mice. This inhibitory effect is associated with CD8(+) T cell immune response and a higher level of IFN-γ, which may influence on the mesenchymal-epithelial transition of tumor cells.

Animals ; CD8-Positive T-Lymphocytes ; immunology ; Cadherins ; metabolism ; Cancer Vaccines ; immunology ; Cell Line, Tumor ; Epithelial-Mesenchymal Transition ; Female ; Homeodomain Proteins ; metabolism ; Humans ; Interferon-gamma ; metabolism ; Interleukins ; immunology ; Lung ; pathology ; Lung Neoplasms ; metabolism ; secondary ; Melanoma ; metabolism ; pathology ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Organ Size ; Transcription Factors ; metabolism ; Transforming Growth Factor beta2 ; metabolism ; Zinc Finger E-box-Binding Homeobox 1

OBJECTIVETo evaluate the immunotoxicological effects of genetically modified wheat with TaDREB4 gene in female BALB/c mice.

METHODSFemale mice weighing 18-22 g were divided into five groups (10 mice/group), which were set as negative control group, common wheat group, parental wheat group, genetically modified wheat group and cyclophosphamide positive control group, respectively. Mice in negative control group and positive control group were fed with AIN93G diet, mice in common wheat group, non-genetically modified parental wheat group and genetically modified wheat group were fed with feedstuffs added corresponding wheat (the proportion is 76%) for 30 days, then body weight, absolute and relative weight of spleen and thymus, white blood cell count, histological examination of immune organ, peripheral blood lymphocytes phenotyping, serum cytokine, serum immunoglobulin, antibody plaque-forming cell, serum half hemolysis value, mitogen-induced splenocyte proliferation, delayed-type hypersensitivity reaction and phagocytic activities of phagocytes were detected.

RESULTSNo immunotoxicological effects related to the consumption of the genetically modified wheat were observed in BALB/c mice when compared with parental wheat group, common wheat group and negative control group.

CONCLUSIONFrom the immunotoxicological point of view, results from this study demonstrate that genetically modified wheat with TaDREB4 gene is as safe as the parental wheat.

Animals ; Antibody-Producing Cells ; immunology ; Body Weight ; Cytokines ; blood ; Female ; Genes, Plant ; Hemolysis ; Hypersensitivity, Delayed ; Immune System ; drug effects ; Immunoglobulins ; blood ; Mice ; Mice, Inbred BALB C ; Organ Size ; Phagocytosis ; Plants, Genetically Modified ; toxicity ; Spleen ; immunology ; Thymus Gland ; immunology ; Triticum ; genetics

OBJECTIVETo study the effect of polysaccharides in processed Sibiraea on the immunologic function of immunosuppression mice.

METHODThe immunosuppressed mice were induced by cyclophosphamide. After the treatment, the organ weight index and the delayed type hypersensitivity of the mice were investigated. The humoral immune function was determined by serum hemolysin assay. Non-specific immune function was determined by carbon clearance method. Cellular immune function was determined by spleen lymphocyte proliferation test. Two hundred kunming mice were randomly divided into five groups: normal controls, model group, low-dose group (110 mg x kg(-1)), middle-dose group (220 mg x kg(-1)), high-dose group (440 mg x kg(-1)). Drugs were given to the mice by oral gavage every day.

RESULTThe immunosuppressed mice treated with Sibiraea polysibcharide at intragastrica dose of 110-440 mg x kg(-1) have increased weight of the immune organs, increased content of DTH and content in serum hemolysin lgG and lgM. Mean while the rate of carbon clearance was enhanced and the proliferation of spleen lymphocyte was increased.

CONCLUSIONPolysaccharides in processed Sibiraea can increase the weight of the immune organs. At the same time, non-specific immune, DTH, humoral immune and cellular immune function were enhanced significantly.

Animals ; Disease Models, Animal ; Female ; Humans ; Immunity ; drug effects ; Immunocompromised Host ; drug effects ; Male ; Mice ; Organ Size ; drug effects ; Polysaccharides ; administration & dosage ; Random Allocation ; Rosaceae ; chemistry ; Spleen ; drug effects ; immunology

The aim of this study was to examine the efficacy of in ovo prime-boost vaccination against infectious bursal disease virus (IBDV) using a DNA vaccine to prime in ovo followed by a killed-vaccine boost post hatching. In addition, the adjuvant effects of plasmid-encoded chicken interleukin-2 and chicken interferon-gamma were tested in conjunction with the vaccine. A plasmid DNA vaccine (pcDNA-VP243) encoding the VP2, VP4, and VP3 proteins of the very virulent IBDV (vvIBDV) SH/92 strain was injected into the amniotic sac alone or in combination with a plasmid encoding chicken IL-2 (ChIL-2) or chicken IFN-gamma (ChIFN-gamma) at embryonation day 18, followed by an intramuscular injection of a commercial killed IBD vaccine at 1 week of age. The chickens were orally challenged with the vvIBDV SH/92 strain at 3 weeks of age and observed for 10 days. In ovo DNA immunization followed by a killed-vaccine boost provided significantly better immunity than the other options. No mortality was observed in this group after a challenge with the vvIBDV. The prime-boost strategy was moderately effective against bursal damage, which was measured by the bursa weight/body weight ratio, the presence of IBDV RNA, and the bursal lesion score. In ovo DNA vaccination with no boost did not provide sufficient immunity, and the addition of ChIL-2 or ChIFN-gamma did not enhance protective immunity. In the ConA-induced lymphocyte proliferation assay of peripheral blood lymphocyte collected 10 days post-challenge, there was greater proliferation responses in the DNA vaccine plus boost and DNA vaccine with ChIL-2 plus boost groups compared to the other groups. These findings suggest that priming with DNA vaccine and boosting with killed vaccine is an effective strategy for protecting chickens against vvIBDV.
Adjuvants, Immunologic/pharmacology ; Animals ; Antibodies, Viral/blood ; Birnaviridae Infections/immunology/prevention & control/*veterinary/virology ; Body Weight/immunology ; Bursa of Fabricius/immunology ; Chick Embryo ; *Chickens ; Histocytochemistry/veterinary ; Immunization/*veterinary ; Infectious bursal disease virus/genetics/*immunology ; Interferon-gamma/pharmacology ; Interleukin-2/pharmacology ; Organ Size/immunology ; Poultry Diseases/immunology/*prevention & control/virology ; RNA, Viral/chemistry/genetics ; Random Allocation ; Reverse Transcriptase Polymerase Chain Reaction/veterinary ; Specific Pathogen-Free Organisms ; Vaccines, DNA/*administration & dosage/immunology ; Vaccines, Inactivated/administration & dosage/immunology ; Viral Vaccines/*administration & dosage/immunology

BACKGROUNDErythropoietin elicits protective effects in lung tissue injury induced by ischaemic reperfusion and hyperoxia. We investigated the protective roles of erythropoietin in pulmonary inflammation and lung injury during acute endotoxaemia.

METHODSA total of 32 male Sprague-Dawley rats were randomly assigned to four groups: saline group, erythropoietin + saline group, saline + lipopolysaccharide group and erythropoietin + lipopolysaccharide group. Rats were treated with erythropoietin (3000 U/kg, i.p.) or saline, 30 minutes prior to lipopolysaccharide administration (6 mg/kg, i.v.). Four hours after lipopolysaccharide injection, samples of pulmonary tissue were collected. Optical microscopy was performed to examine pathological changes in lungs. Wet/dry (W/D) ratios, myeloperoxidase activity, malondialdehyde concentrations and tumour necrosis factor-alpha (TNF-alpha) as well as interleukin 1 beta (IL-1beta) levels in lungs were measured. The pulmonary expression of nuclear factor kappaB (NF-kappaB) p65 was evaluated by Western blotting. Differences between the different groups were analysed by one-way analysis of variance (ANOVA).

RESULTSThe lung tissues from the saline + lipopolysaccharide group were significantly damaged, which were less pronounced in the erythropoietin + lipopolysaccharide group. The W/D ratio increased significantly in the saline + lipopolysaccharide group (5.75 +/- 0.22) as compared with the saline group (3.85 +/- 0.20) (P < 0.01), which was significantly reduced in the erythropoietin + lipopolysaccharide group (4.50 +/- 0.35) (P < 0.01). Myeloperoxidase activity and malondialdehyde levels increased significantly in the saline + lipopolysaccharide group compared with the saline group, which was reduced in the erythropoietin + lipopolysaccharide group. The TNF-alpha level of pulmonary tissue increased significantly in the saline + lipopolysaccharide group ((9.80 +/- 0.82) pg/mg protein) compared with the saline group ((4.20 +/- 0.42) pg/mg protein, P < 0.01). However, the increase of TNF-alpha level of pulmonary tissue was significantly reduced in the erythropoietin + lipopolysaccharide group ((6.50 +/- 0.66) pg/mg protein, P < 0.01). Similarly, pulmonary IL-1beta levels were elevated markedly in the saline + lipopolysaccharide group in contrast to the saline group, whereas the elevation was much less in the erythropoietin + lipopolysaccharide group. The nuclear localization of p65 increased markedly in the saline + lipopolysaccharide group and this enhancement of nuclear p65 expression was much less in the erythropoietin + lipopolysaccharide group.

CONCLUSIONErythropoietin attenuates pulmonary inflammation and suppresses TNF-alpha and IL-1beta overproduction during acute endotoxaemia, which is partially mediated by inhibition of NF-kappaB.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Blotting, Western ; Endotoxemia ; immunology ; metabolism ; pathology ; Erythropoietin ; pharmacology ; Interleukin-1beta ; metabolism ; Lung ; drug effects ; immunology ; metabolism ; pathology ; Lung Injury ; chemically induced ; immunology ; Male ; Malondialdehyde ; metabolism ; NF-kappa B ; metabolism ; Organ Size ; Peroxidase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha

OBJECTIVETo observe the effects of marine protein peptide (MPP) on immunomodulating in mice and its possible mechanism.

METHODSFemale ICR mice (6-8 weeks old) were administered the MPP for 4 weeks with the dose of 0.22 g/kgBW, 0.45 g/kgBW and 1.35 g/kgBW. Spleen and thymus were weighted and cell-mediated immune functions, humoral immune functions, phagocytic functions of mononuclear phagocyte, NK cell activity assays, the T cell subpopulation of the spleen tissue by the flow cytometer and the concentrations of cytokines in serum by cytometric bead array were examined.

RESULTSThe capacity of lymphocyte proliferation induced by Con A (0.33 +/- 0.21), DTH response (0.36 +/- 0.11) mm in MPP 0.22 g/kgBW group were significantly increased in comparison with these values in control group (0.15 +/- 0.10) and (0.21 +/- 0.10)mm, respectively, P < 0.05. IgM-PFC number of MPP 0.22 g/kgBW group (1.64 +/- 0.06), 0.45 g/kgBW group (1.59 +/- 0.05) and 1.35 g/kgBW group (1.56 +/- 0.10) were higher than those in control group (1.38 +/- 0.10), P < 0.01; and the level of serum HC50 of MPP 0.22 g/kgBW group (141.00 +/- 23.00) and 0.45 g/kgBW group (130.40 +/- 33.20) were greater than the control (100.30 +/- 19.40) , P < 0.01. The activity of NK cells in MPP 0.22 g/kgBW group (1.672 +/- 0.142) was significantly elevated in comparison with this value in control group (1.392 +/- 0.182), P < 0.05. The percentage of CD4 T helper (Th) cell in spleen of MPP 0.22 g/kgBW group (32.84 +/- 3.776)% and 0.45 g/kgBW group (32.42 +/- 3.507) % was higher than those in control group (25.06 +/- 0.354) %, P < 0.05. The concentrations of IL-2 in serum of MPP 0.22 g/kgBW group 181.06 pg/ml, 0.45 g/kgBW group 94.84 pg/ml and 1.35 g/kgBW group 102.61 pg/ml were higher than those in control group 0.50 pg/ml, P < 0.05; and the level of IL-5 of MPP 0.22 g/kgBW group (38.31) pg/ml was greater than the control 0.50 pg/ml, P < 0.05. Nevertheless, no obvious effects on weight increasing, the ratio of immune organ and body weight and phagocytosis capacity were observed in our study.

CONCLUSIONMPP could improve the immune functions in mice, and might be by the mechanism of enhancing the function of Th cells stimulating the secretion of Th1 and Th2 type cell cytokines.

Animals ; CD4-CD8 Ratio ; Female ; Killer Cells, Natural ; metabolism ; Macrophages ; immunology ; Marine Biology ; Mice ; Organ Size ; Peptides ; pharmacology ; Phagocytosis ; Spleen ; immunology ; Th1 Cells ; Th2 Cells

The studies were conducted on Balb/c mice exposed to restraint stress twice for 12 h at 24 h intervals. Prior to restraint stress the mice were treated with sodium diethyldithiocarbamate (DTC) i.p. at a dose of 20 mg/kg five times at 48 h intervals. DTC was used per se or with zinc ions interaction, by adding zinc sulfate to drinking water at a dose of 72 microgram/mouse daily. The results obtained in the study show that restraint stress causes involution of lymphatic organs, decreased the percentage of immature (CD4+CD8+) and, mature (CD4+) thymocytes and CD4+, CD8+and CD19 + splenocytes and proliferative response of thymocytes stimulated in vitro with concanavalin A (Con A) and phytohemagglutinin (PHA). The restraint stress decreased also interleukin-1 (IL-1) production by murine intraperitoneal macrophages stimulated in vitro with lipopolysaccharide (LPS) from E. coli. Pretreatment with DTC counteracted restraint stress-induced immunosuppression, which is expressed as partial normalisation of the total number of thymocytes, splenocytes and IL-1 production, accelerated regeneration of thymus and spleen, shorter suppressive action of restraint stress on the percentage of CD4+CD8+thymocytes and in total normalisation of the CD4+thymocytes and splenocytes. DTC administered prior to restraint stress augmented the proliferative response of thymocytes to two mitogens. The immunocorrecting action of DTC is enhanced by zinc supplementation, expressed in the increased percentage of CD4+thymocytes and splenocytes, CD19 + splenocytes, proliferative activity of thymocytes stimulated with PHA and IL-1 production. The obtained results show that DTC administration can be supplemented with zinc in order to restore the immune system impaired by stress.
Adjuvants, Immunologic/*pharmacology ; Animals ; Ditiocarb/*pharmacology ; Female ; Immunity, Cellular/*drug effects ; Interleukin-1/biosynthesis ; Macrophages, Peritoneal/immunology ; Male ; Mice ; Mice, Inbred BALB C ; Mitogens/biosynthesis ; Organ Size/drug effects ; Restraint, Physical ; Spleen/cytology/drug effects ; Stress/etiology/*immunology ; T-Lymphocyte Subsets/drug effects ; Thymus Gland/cytology/drug effects ; Zinc Sulfate/*pharmacology