Study on Lymphocyte Activation and Proliferation Induced by Anti-CD3 McAb

T cell activation and proliferation via CD3-TCR complex were investigated by lymphocyte DNA synthesis in vitro.Several interfering factors were also discussed.The result indicated that lymphocyte activation and proliferation are calciumdependent.A rise of cytoplasmic free Ca2+ quickly following activation with CD3 McAb is mainly due to intracellular mobilization of Ca2+,while lymphocyte proliferation needs both intracellular mobilization of Ca2+ as well as influx of extracellular Ca2+, It was confirmed that CTX sensitive G protein plays a role in regulating T cell proliferation by pretreatment with CTX suppressing lymphocyte H-TdR incorporation obviously.PLC and PKC inhibitor neomycin and P.S.S could also decrease T cell proliferation.

Effect of IL-10 gene transmission on the PTg-stimulated splenocytes proliferation and Th1 cytokines production from experimental autoimminue thyroiditis rats

To investigate the effects of gene therapy with IL-10 on PTg-induced proliferation of splenocytes and Thl cytokine production from PTg-stimulated splenocytes. Methods： EAT rats were divided into four groups ：group A （PBS＋PLL） , group B （pORF＋PLL）, group C （pORFmIL10＋PLL）, and group D （pORFmIL10＋ MEM）. The substances mixed with lipofectamine were injected into the thyroid tissues of rats on the 18th dday after immunization. The rats were sacrificed at the 8th week. In vitro proliferative responses to ConA and different concentration of PTg were measured by culturing 4 ×105 splenocytes pulsed with 18.5KBq of [^3H] thymidine for the final 12h and then harvested for liquid scintillation counting. In vitro splenocytes were cultured with PTg （25 mg/L）. Thl cytokine IFN-γ,TNF-α and IL-2 were detected by ELISA. Results： The proliferative response to PTg was suppressed in group C, compared with that of group A and B （P 〈 0.05）. The levels of IFN-γ,TNF-αand IL-2 in the supernatant of PTg-stimulated splenocytes were 3548.25±779.47 pg/ml, 27.66±10.50 pg/ml and 3617.73±609.15 pg/ml, respectively, which were much lower in group C than those in group A and B（P 〈 0.01, P 〈 0.05, P 〈 0.001, respectively）. Conclusion： IL- 10 gene transmission in thyroid tissues could inhibit PTg specific proliferation of splenocytes from EAT rats and the secretion 6f Th1 cytokines from PTg-stimulated splenocytes.

Enhancement of the Immune Response to Rabbit Hemorrhagic Disease Vaccine in Young Rabbits by Advanced Vaccination and Chinese Herbal Adjuvants

Experiments were conducted to determine the effects of advanced vaccination and Chinese herbal adjuvants （CHA）, containing astragalus polysaccharides （APS） and ginsenosides （GS） on the immune response to rabbit hemorrhagic disease （RHD） vaccine in young rabbits. In experiment 1, 5 New Zealand rabbits of each group at 30, 35, 40, or 45 days of age were injected with 2 mL of inactivated RHD vaccine, respectively. The dynamic changes of antibody titers were tested by the hemagglutination inhibition （HI） method. In experiment 2, 30 New Zealand rabbits at 35 days of age were randomly assigned to 5 treatment groups, representing inoculation with 3 mL of non-adjuvant RHD vaccine, CHA-RHD vaccine, CHA-HA vaccine （half dose antigen）, aluminium adjuvant-RHD vaccine, and PBS, respectively. The dynamic changes of peripheral lymphocyte proliferation and serum antibody titers were tested by the MTT method and the HI method. The results showed that the titer of maternal HI antibody in the 35-day-old rabbits was lower than the protective level of 3 log2, while on days 7 to 49 after the vaccination, the antibody titers were higher than 3 log2. The CHA promoted the lymphocyte proliferation and enhanced the serum antibody titer （P〈 0.05）. These findings from the two experiments suggested that advanced vaccination and Chinese herbal adjuvants significantly enhanced the immune response to vaccine against RHD, and effectively protected the young rabbits against RHD challenge.

Effects of methyl-,phenyl-,ethylmercury and mercurychlorid on immune cells of harbor seals(Phoca vitulina)

Mercury （Hg） is present in the marine environment as a natural metal often enhanced through human activities. Depending on its chemical form, Hg can cause a wide range of immunotoxic effects. In this study, the influence of methyl-, ethyl- and phenylmercury as well as mercurychloride on immune functions was evaluated. Two parameters of cellular immunity, proliferation and mRNA cytokine expression of interleukin-2, -4, and transforming growth factor β, were investigated in harbor seal lymphocytes after in vitro exposure to Hg compounds. While all Hg compounds had a suppressive effect on proliferation, differences between juvenile and adult seals were found. Lymphocytes from juveniles showed a higher susceptibility to the toxic effect compared to lymphocytes from adults. Furthermore, the degree of inhibition of proliferation varied among the four Hg compounds. The organic compounds seem to be more immunotoxic than the inorganic compound. Finally, for the cytokine expression of methylmercury-incubated lymphocytes, timedependent changes were observed, but no dose-dependency was found. Marine mammals of the North Sea are burdened with Hg, and lymphocytes of harbor seals may be functionally impaired by this metal. The present in vitro study provides baseline information for future studies on the immunotoxic effects of Hg on cellular immunity of harbor seals.

The Effect of a Superstrate Layer on the Input Impedance of An Annular-Ring Microstrip Antenna

The immunostimulatory property of a synthetic polyphosphoramidate based on stearyl tyrosine (PPST) is described. The immunostimulatory activity of this negatively charged polymer was investigated in vitro through examining the effect on the mitogen induced lymphocytes proliferation by a tetrazolium based colormetric assay and also through studying the primary plaque forming cells (IgM PFC) responses of Balb/c mice after the intraperitoneal (i.p.) injection of the polymer with sheep red blood cells (SRBC). PPST exhibits specific and high immunostimulatory activity in vitro by the stimulation on lipopolysaccharide(LPS) induced murine B cells proliferation, which is probably due to the interaction between the polymer with LPS responding B lymphocytes. This polymer also makes a significant increase of single antibody producing cells. The result shows that the stimulation rate to IgM PFC responses of Balb/c mice achieved 77.9% ( p <0.01) when 160 mg/kg of PPST was injected 4 h before with SRBC.

Effects of purified zearalenone on selected immunological and histopathologic measurements of spleen in post-weanling gilts

The present study was aimed at investigating the adverse effects of dietary zearalenone(ZEA) on the lymphocyte proliferation rate(LPR), interleukin-2(IL-2), mRNA expressions of pro-inflammatory cytokines, and histopathologic changes of spleen in post-weanling gilts. A total of 20 crossbred piglets(Yorkshire × Landrace × Duroc) with an initial BW of 10.36 ± 1.21 kg(21 d of age) were used in the study.Piglets were fed a basal diet with an addition of 0.1.1,2.0, or 3.2 mg/kg purified ZEA for 18 d ad libitum.The results showed that LPR and IL-2 production of spleen decreased linearly(P < 0.05) as dietary ZEA increased. Splenic mRNA expressions of interleukin-1β(IL-1β) and interleukin-6(IL-6) were linearly upregulated(P < 0.05) as dietary ZEA increased. On the contrary, linear down-regulation(P<0.05) of mRNA expression of interferon-γ(IFN-γ) was observed as dietary ZEA increased. Swelling splenocyte in1.1 mg/kg ZEA treatments, atrophy of white pulp and swelling of red pulp in 2.0 and 3.2 mg/kg ZEA treatments were observed. The cytoplasmic edema in 1.1 mg/kg ZEA treatments, significant chromatin deformation in 2.0 mg/kg ZEA treatment and phagocytosis in 3.2 mg/kg ZEA treatment were observed.Results suggested that dietary ZEA at 1.1 to 3.2 mg/kg can induce splenic damages and negatively affect immune function of spleen in post-weanling gilts.

Effects and mechanisms of Cryptotanshinone on rats with adjuvant arthritis

Background Cryptotanshinone （CT） is the major active constituent of Salvia miltiorrhiza Bunge. The present study was carried out to investigate the effects of CT on rats with adjuvant arthritis （AA）. Methods AA was induced by the metatarsal footpad injection with complete Freund＇s adjuvant in male Sprague-Dawley rats. The secondary inflammatory reaction was evaluated by hind paw swelling and the polyarthritis index. Activity of interleukin-1 （IL-1） was detected by the concanavalin A-induced thymocytes proliferation assay. The lymphocytes proliferation and IL-2 production were assayed by 3-（4,5-2dimethylthiazal-2yl）2,5-diphenyltetrazoliumbromide （MTT） and activated mouse splenocytes proliferation, respectively. Results Intragastric administration of CT （50 and 100 mg/kg） significantly decreased secondary inflammatory reactions and increased the spleen and thymus index. There was a marked immunologic and inflammatory response in the AA model, which was accompanied by the decrease of thymocyte proliferation and IL-2 production as well as the increase of IL-1 production. CT apparently enhanced thymocyte proliferation and decreased IL-1 production in AA rats. Conclusion These results indicate that CT may exert its anti-inflammatory and immunoregulatory effects through inhibiting lymphocyte proliferation and production of pro-inflammatory mediators.

Immunotoxic effects of an industrial waste incineration site on groundwater in rainbow trout (Oncorhynchus mykiss)

The discharge of organic waste from the petrochemical industry into the Mercier lagoons caused major groundwater contamination. The objective of this study was to determine the immunotoxic potential of three groundwater wells at increasing distance from the incinerator dumping site （1.17, 2.74 and 5.40 km）. Rainbow Trout were exposed to increasing concentrations of water from three groundwater wells for 14 days. Immunocompetence was characterized by phagocytosis, mitogen-stimulated proliferation of lymphocytes, cell cycle analysis and apoptosis. A significant increase in innate （phagocytosis） and specific immune response （B lymphocyte proliferation） was observed in trout exposed to water collected from the well at 2.74 km. However, phagocytosis activity was suppressed in groups at 1.17 and 5.40 km. The proportion of lymphocytes in S phase was significantly increased in groups at 2.74 and 5.40 kin, while lymphocytes in G0/G1 phase were decreased in all three exposure groups. Additionally, dexamethasone （DEX）-induced apoptosis of lymphocytes was significantly reduced in the group at 2.74 km, which suggests decreased lymphocyte turnover. Furthermore, the ratio of DEX- induced apoptosis/apoptosis was lower in the groups at 2.74 and 5.40 km. In summary, our experiments have shown that exposure to the mixture of organic compounds present in Mercier groundwater modulates phagocytosis and cell proliferation, disrupts the cell cycle and reduces the ratio of DEX- induced apoptosis/apoptosis. It is concluded that groundwater collected in the vicinity of an incinerator containment field could impact immunocompetence in fish.

Cell-wall-deficient bacteria： a major etiological factor for psoriasis？

Background Psoriasis is a common inflammatory skin disease, yet knowledge of the factors that may induce, trigger, or exacerbate psoriasis is not fully delineated. Recent advances have improved our understanding of the link between psoriasis and cell-wall-deficient bacteria （CWDB） infections. In the present study we assessed the prevalence of CWDB infection in patients with psoriasis. Methods The carriage rate of CWDB in the tonsil or pharynx of psoriasis patients, chronic tonsillitis patients and controls were investigated using hypertonic medium. Psoriasis patients with CWDB were randomly assigned to two groups and respectively treated with antibiotics or systemic therapy without antibiotic. Human peripheral blood mononuclear cells （PBMC） from psoriasis patients, chronic tonsillitis patients and control subjects were stimulated with bacteria antigens and extra-cellular levels of interferon-γ（IFN-γ） and interleukin （IL）-10 were measured in the supernatants using the ELISA technique, in vitro. Meanwhile, the proliferation ability of PBMC to respond to bacteria antigens was detected by MTT assay. Results CWDB were isolated from 74.2% of psoriasis patients, 23.5% of chronic tonsillitis patients and only 6.3% of controls. Antibiotic therapy was appropriate for approximately 80% of psoriasis patients with CWDB infection, and in only 8.9% psoriasis patients CWDB infection was detected after antibiotic therapy. Meanwhile, our study showed that CWDB and wide-type bacteria did remarkably enhance the production of IFN-γ, in vitro, and PBMC proliferation. Conclusion CWDB infection may be a virtual triggering factor in psoriasis by regulating T-cell activation.