旋毛虫Ts87重组蛋白诱导的小鼠黏膜免疫保护性研究

本实验探讨旋毛虫Ts87重组蛋白灌胃免疫小鼠诱导的黏膜免疫保护性作用。实验分对照组、佐剂组(霍乱毒素B亚单位组,CTB)和免疫组(CTB Ts87重组蛋白),灌胃免疫,间隔1周共免疫3次。末次免疫后第7天用400条旋毛虫感染期幼虫攻击,比较3组小鼠肠道成虫数、雌虫生殖力和肌幼虫数。且于末次免疫后第7天刮取肠黏液、取血检测特异性sIgA、IgG抗体水平。结果显示免疫组小鼠成虫减虫率、新生蚴减虫率、肌幼虫减虫率分别是81·34%、67·02%、84·49%;小鼠肠黏液sIgA水平及血清IgG水平显著高于对照组和佐剂组。结果表明Ts87重组蛋白黏膜免疫能够诱导小鼠产生抗旋毛虫的保护性免疫。灌胃免疫能显著提高肠黏液特异性sIgA水平,对促进肠道成虫的排出有明显作用。     

Further analysis of ATP-mediated activation of K+ channels in renal epitheloid Madin Darby canine kidney (MDCK) cells

ATP activates K⁺ channels by increasing intracellular calcium activity in Madin Darby canine kidney (MDCK) cells. The present study has been performed to test for the involvement of G-proteins and of protein kinase C in the intracellular transmission of these effects. To this end, the effect of ATP on intracellular calcium and K⁺ channel activity has been studied in cells pretreated with the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA) and/or pertussis toxin. The ATP-induced increase of intracellular calcium is not significantly affected by pretreatment with pertussis toxin, is significantly blunted by pretreatment with TPA and is abolished by pretreatment with both pertussis toxin and the phorbol ester. The ATP activation of K⁺ channels is similarly blunted by pretreatment with TPA, but is not abolished by pretreatment with both the phorbol ester and pertussis toxin. Furthermore, the ATP-induced hyperpolarization is not abolished in cells pretreated with both pertussis toxin and TPA. In those cells, ATP may activate K⁺ channels by calcium-independent mechanisms or lead to localized increases of intracellular calcium sufficient to activate the K⁺ channels but escaping detection with fura-2 fluorescence.     

rCTB和TT为蛋白载体的A群流脑多糖黏膜免疫初步研究

目的 对重组霍乱毒素B亚单位(rCTB)作为多糖蛋白结合疫苗候选载体的可行性进行分析,并对以破伤风类毒素(TT)与rCTB为蛋白载体的黏膜投递型疫茸的免疫效果进行初步探讨.方法 首先通过基因工程手段获得具有五聚体结构的rCTB.再将rCTB五聚体蛋白利用化学方法(ADH方法)与A群脑膜炎球菌多糖(GAMP)耦联,获得多糖蛋白结合物GAMP-rCTB,并将其与TT为蛋白载体的A群流脑多糖蛋白结合物(GAMP-TT)以滴鼻和注射途径免疫BALB/c小鼠,并对其进行免疫学评价.结果 以rCTB和TT为载体的A群流脑多糖蛋白结合物,通过黏膜投递途径均可在血清中产生相对较高的多糖特异性IgG抗体,在肺部盥洗液和小肠黏膜也产生了相应的特异性IgA抗体.结论 rCTB和TT均可作为黏膜投递型多糖结合疫苗的候选蛋白载体.以rCTB为载体的多糖蛋白结合物,黏膜途径可能在免疫功能方面优于注射途径.     

Cell-surface bound pertussis toxin induces polyclonal T cell responses with high levels of interferon-gamma in the absence of interleukin-12

Pertussis toxin (PTx), an exotoxin produced by Bordetella pertussis, has long been used as a mucosal adjuvant. We examined the T cell stimulatory properties of PTx in order to dissect its mechanisms of adjuvanticity. PTx or the B-oligomer of PTx (PTxB) failed to activate purified murine CD4+ or CD8+ T cells, as measured by a lack of proliferation or expression of early T cell activation markers. However, these T cells proliferated extensively in response to the toxin in the presence of syngeneic DC, and proliferation was accompanied by a high level of IFN-gamma production in the absence of IL-12. Interestingly, such responses were independent of signals mediated by MHC-TCR interaction. Both PTx and PTxB were found to bind stably to the surface of DC, and increased the adherence of DC to surrounding cells. These data suggest that polyclonal T cell responses mediated by the toxin are likely to be caused by the toxin bound on the surface of APC, either cross-linking cell surface molecules on T cells, or directly stimulating T cells together with the co-stimulatory molecules expressed on APC. B. pertussis may use this toxin as a mechanism to evade a specific immune response.     

Dual function of Langerhans cells in skin TSLP-promoted TFH differentiation in mouse atopic dermatitis

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Potential-dependent action ofAnemonia sulcata toxins III and IV on sodium channels in crayfish giant axons

Effects of toxins III and IV (ATX III and IV) from the sea anemoneAnemonia sulcata on the Na current of crayfish giant axons were studied. Both toxins slowed the inactivation of Na channels, producing a maintained Na current during a depolarizing voltage pulse. Using the intensity of the toxin-induced maintained current as an index for the fraction of Na channels to which toxin is bound, the toxin association and dissociation kinetics were analyzed. The dissociation rate of ATX III was increased by two orders of magnitudes by depolarizing the membrane from –70 to –40mV. This increase of the dissociation rate caused a marked decrease in the binding rate of ATX III to Na channels in the same potential range. ATX IV exhibited association and dissociation kinetics that had a potential dependency quite similar to that of ATX III in spite of different ionic charge distribution in these two toxins. The results support the view that the potential-dependent kinetics of these toxins are not due to an electrostatic interaction between the ionic charges of toxins and the membrane potential but result from a modulation of the binding energy depending on the gate configuration of the Na channel.     

A fatal case of necrotizing sinusitis due to toxigenic Corynebacterium ulcerans

A 77-year-old farmer developed cough with sputum production, fever, bloody nasal discharge and a mass in his right maxillary sinus leading to necrotic ulceration of the sinus. Corynebacterium ulcerans, carrying the beta-phage for the diphtheria toxin and secreting the toxin, was detected microscopically and by culture from the sinusoidal and ulcer discharge. Despite immediate antimicrobial chemotherapy the patient died of pulmonary failure associated with the production of large amounts of very viscous sputum. Identification of the causative agent, pathophysiological aspects and risk factors of this unusal infection are discussed.     

Tissue cages for study of experimental streptococcal infection in rabbits. II. Humoral and cell-mediated immune response to erythrogenic toxins

Infection of rabbits with erythrogenic toxin producing streptococcal strains caused a marked increase of humoral antibodies, which was detected by immunoprecipitation and ELISA. An antibody response directed towards the erythrogenic toxin type A was demonstrated by fused rocket immunoelectrophoresis. All toxinogenic reference strains produced ET type A under in vivo conditions despite that this toxin was not always demonstrated under in vitro conditions. The infection resulted in an increase of mitogenic response of peripheral lymphocytes to the initial nonspecific mitogenic erythrogenic toxins, whereas the Con A stimulation was depressed starting 14 days after infection and lasting during a period of 90 days. Since a normal antibody response was evoked, it seems likely that the T helper cell function was not affected.     

Tissue cages for study of experimental streptococcal infection in rabbits. I. Production of erythrogenic toxins in vivo

Tissue cages implanted subcutaneously were used to infect rabbits with erythrogenic toxin (ET) producing streptococci. The in-vivo production of ET was followed during the infection by immunoprecipitation analyses of the tissue cage fluid (TCF). ET types A and C were mainly detected during the first week of infection, but, as late as 4 weeks after the inoculation, ET was occasionally found in TCF. The nonspecific mitogenic activity of ET on human lymphocytes was also used as a biological marker to recognize ET in TCF. Mitogenic activity was detected in 90% of samples during the first week. In order to characterize the mitogenic material released by growing streptococci, TCF was electrofocused in polyacrylamide gel. The eluates of sliced gels were checked for mitogenic activity and compared with a purified ET preparation containing ET types A and C. It could be verified that ET type A was produced under in-vivo conditions by strains NY-5 and SF130, while ET type C was produced by strain T18. Differences between production of toxins in vitro and in vivo might be of significance for the understanding of the pathogenetic mechanisms in streptococcal infection.