重组E.coli. LLO/OVA明显增强小鼠CD11c细胞活性及抗肿瘤免疫

目的 探讨重组E.coli. LLO/OVA 诱导C57BL/6小鼠机体免疫的途径,观察其免疫后小鼠机体抑制B16-OVA黑色素瘤的效果.方法 磁珠分离E.coli. LLO/OVA及E.coli. OVA免疫后小鼠脾脏CD11c、CD4 和CD8 T细胞并检测CD11c细胞诱导同源CD4 和CD8 T增殖水平和细胞因子分泌程度;流式细胞检测小鼠脾脏内肿瘤抗原OVA257-264 SIINFEKL特异的细胞毒T细胞含量.比较两种E.coli. 免疫后,恶性黑色素瘤B16-OVA在小鼠肺内形成瘤结节的数量.结果 与E.coli. OVA相比, E.coli. LLO/OVA免疫后小鼠脾脏CD11c细胞诱导同源CD4 T细胞增殖作用增强、IL-2分泌增高;同时诱导CD8 T细胞增殖和IFN-γ分泌的作用也明显增强;OVA257-264 SIINFEKL特异的CD8 T细胞含量也明显增高;小鼠肺内形成B16-OVA瘤结节平均数明显减少. 结论 E.coli. LLO/OVA有效地诱导小鼠CD11c细胞活化,增强其对CD4 T细胞增殖和IL-2分泌以及对CD8 T细胞增殖、IFN-γ分泌的作用,诱导了更多的OVA特异的CD8 T细胞,使机体产生了更强的抗肿瘤免疫.     

重组人白细胞介素-1受体拮抗药对豚鼠离体呼吸道平滑肌的影响

目的　观察人重组白细胞介素 1受体拮抗剂 (IL 1ra)对正常和卵白蛋白致敏豚鼠离体肺条、气管平滑肌的影响。方法　应用离体器官装置、张力换能器、MedLab记录系统测定肺条和气管平滑肌的张力。结果　①IL 1ra对正常豚鼠离体肺条和气管平滑肌有直接松弛作用 ,EC50 分别为1 2 9ⅹ 10 -7mol·L-1和 8 0 6× 10 -8mol·L-1;并对卵白蛋白致敏的豚鼠肺条和气管平滑肌也有直接的松弛作用 ,EC50 分别为 2 6 1× 10 -7mol·L-1和 5 88× 10 -7mol·L-1,但致敏豚鼠呼吸道平滑肌对IL 1ra的敏感性要比正常豚鼠低。②IL 1ra(10 -9～ 10 -5mol·L-1)可剂量依赖性地抑制致痉剂组胺对正常豚鼠肺条和气管平滑肌的收缩作用 (P <0 0 1)。③IL 1ra能抑制卵白蛋白攻击引起的肺条和气管平滑肌的收缩 ,IC50 分别为 7 83ⅹ 10 -7mol·L-1和 4 4 8ⅹ 10 -7mol·L-1。结论　IL 1ra对正常、痉挛及致敏状态的呼吸道平滑肌均有松弛作用     

艾叶油的呼吸系统药理研究-Ⅱ,抗过敏作用

目的：本文研究艾叶油的抗过敏作用。方法：采用致敏豚鼠气管ＳｃｈｕｌｔｚＤａｌｅ 反应,组胺或氨甲酰胆碱引起的豚鼠气管收缩,大鼠被动皮肤过敏,５羟色胺引起的大鼠皮肤毛细血管通透性增强,豚鼠肺组织释放过敏性慢反应物质（ＳＲＳＡ） ,ＳＲＳＡ 收缩豚鼠回肠等试验。结果：艾叶油抑制致敏豚鼠气管ＳｃｈｕｌｔｚＤａｌｅ 反应（ＩＣ５０ ：９８ ．６ｍｇ／Ｌ） ;１００ｍｇ／Ｌ 明显降低组胺或氨甲酰胆碱引起的豚鼠气管收缩ｐＤ２ 值;明显抑制大鼠被动皮肤过敏（ＩＤ５０ ：０ ．２２ｇ／ｋｇ） 和５羟色胺引起的大鼠皮肤毛细血管通透性增强反应（ＩＤ５０ ：０ ．５２ｇ／ｋｇ） ;抑制豚鼠肺组织释放ＳＲＳＡ（ＩＣ５０ ：４９ ．７ｍｇ／Ｌ） ;拮抗ＳＲＳＡ 对豚鼠回肠的收缩（ＩＣ５０ ：３４ ．９ｍｇ／Ｌ） 。结论：艾叶油具有抗过敏作用,对呼吸道过敏反应有保护作用,是其治疗支气管哮喘和慢性气管炎作用机制之一     

L-454,560, a potent and selective PDE4 inhibitor with in vivo efficacy in animal models of asthma and cognition

Type 4 phosphodiesterases (PDE4) inhibitors are emerging therapeutics in the treatment of a number of chronic disorders including asthma, chronic obstructive pulmonary disease (COPD) and cognitive disorders. This study delineates the preclinical profile of L-454,560, which is a potent, competitive and preferential inhibitor of PDE4A, 4B, and 4D with IC50 values of 1.6, 0.5 and 1.2 nM, respectively. In contrast to the exclusive binding of cilomilast and the preferential binding of roflumilast to the PDE4 holoenzyme state (Mg2+-bound form), L-454,560 binds to both the apo-(Mg2+-free) and holoenzyme states of PDE4. The intrinsic enzyme potency for PDE4 inhibition by L-454,560 also results in an effective blockade of LPS-induced TNFalpha formation in whole blood (IC50 = 161 nM) and is comparable to the human whole blood potency of roflumilast. The cytokine profile of inhibition of L-454,560 is mainly a Th1 profile with significant inhibition of IFNgamma and no detectable inhibition of IL-13 formation up to 1 microM. L-454,560 was also found to be efficacious in two models of airway hyper-reactivity, the ovalbumin (OVA) sensitized and challenged guinea pig and the ascaris sensitized sheep model. Furthermore, L-454560 was also effective in improving performance in the delayed matching to position (DMTP) version of the Morris watermaze, at a dose removed from that associated with potential emesis. Therefore, L-454,560 is a novel PDE4 inhibitor with an overall in vivo efficacy profile at least comparable to roflumilast and clearly superior to cilomilast.     

川椒方对小鼠变应性结膜炎肥大细胞脱颗粒的影响

目的：观察川椒方对小鼠变应性结膜炎肥大细胞脱颗粒的影响。方法：将68只Balb/c小鼠随机分为6组,即正常组8只,模型组（不给药）、阴性对照组（生理盐水ig）、川椒方水煎剂低、中、高剂量组（5.25,10.5,21.0 g·kg^-1,ig,每天1次,ig体积均为10 mL·kg^-1）,每组各12只。用鸡卵清白蛋白免疫小鼠建立变应性结膜炎模型。ELISA检测小鼠血清中前列腺素D2（PGD2）、组胺（HIS）和肿瘤坏死因子（TNF-α）的表达水平,RT-PCR观察结膜组织中白介素-4（IL-4）和TNF-α的表达。结果：与正常组相比,模型组和阴性对照组结膜肥大细胞聚集,脱颗粒明显（P<0.01）,血清中PGD2,HIS和TNF-α的表达升高（P<0.01）,结膜组织中IL-4和TNF-α的表达升高（P<0.01）。与模型组比较,川椒方各组结膜组织中肥大细胞数量减少,脱颗粒率下降（P<0.01）;血清中PGD2,HIS和TNF-α的表达显著降低（P<0.01）;结膜组织中IL-4和TNF-α的表达明显降低（P<0.01）。结论：川椒方能够治疗变应性结膜炎,其作用机制可能是通过稳定肥大细胞,抑制其脱颗粒。     

Phosphorothioated antisense oligodeoxynucleotide suppressing interleukin-10 is a safe and potent vaccine adjuvant

While vaccination is highly effective for the prevention of many infectious diseases, the number of adjuvants licensed for human use is currently very limited. The aim of this study was to evaluate the safety, efficacy, and to clarify the mechanism of a phosphorothioated interleukin (IL)-10-targeted antisense oligonucleotide (ASO) as an immune adjuvant in intradermal vaccination. The cytotoxicity of IL-10 ASO and its ability to promote T cell proliferation were assessed by Cell Counting Kit-8 (CCK-8) assay. The contents of IL-6, IL-8, TNF-α, IL-1β, and IL-10 in inoculated local tissue and the antigen-specific antibody titers in mouse serum samples were determined by ELISA. The target cells of IL-10 ASO were observed using immunofluorescent staining. The results showed that the specific antibody titer of ovalbumin (OVA), a model antigen, was increased 100-fold upon addition of IL-10 ASO as an adjuvant compared to that of OVA alone. IL-10 ASO showed an immunopotentiation efficacy similar to that of Freund’s incomplete adjuvant, with no detectable cell or tissue toxicity. In vitro and in vivo experiments confirmed that IL-10 ASO enhances immune responses by temporarily suppressing IL-10 expression from local dendritic cells and consequently promoting T cell proliferation. In conclusion, IL-10 ASO significantly enhances immune responses against co-delivered vaccine antigens with high efficacy and low toxicity. It has the potential to be developed into a safe and efficient immune adjuvant.     

Organ-specific autoimmunity

The past year has seen advances in our understanding of the mechanisms that control the expression of organ-specific autoimmunity. Attempts to develop therapeutic strategies for the prevention of autoimmune disease have largely been based on deviation of the T-cell response away from an autoaggressive, pro-inflammatory Th1 response. Manipulation of the cytokine environment at the site of antigen uptake or presentation, the source of endogenous antigen, costimulatory molecules or peptides processed and presented has yielded interesting results.     

The expression of murine eutaneous late phase reaction requires both IgE antibodies and CD4 T eells

Background Exposure of atopic patients to a specific allergen evokes an immediate response which is followed, in many cases, by a late phase reaction (LPR) some hours later. Here we have examined the immunological mechanisms required for the expression of cutaneous LPR in mice. Methods BALB/c mice were immunized by i.p. injection of ovalbumin (OVA) and alum actively or by i.v. injection of anti-OVA IgE monoclonal antibody (mAb) passively. After challenge by intradermal injection of OVA into ears, the changes in ear thickness, the number of eosinophils, and the levels of IL4 and IFN-γ protein at the site of antigen challenge were examined. Results Actively immunized mice developed a biphasic response at the site of OVA injection, while mice passively immunized with IgE anti-OVA mAb displayed a strong early response but no LPR. Cell transfer experiments using BALB/c nu/nu mice revealed that both OVA-specific IgE mAb and OVA-primed CD4 T cells were required to evoke LPR. Moreover, LPR was associated with increased levels of IL-4 production concomitant with reduced IFN-γ production and was abolished by pretreatment with anti-IL-4 neutralizing mAb. Conclusion It is suggested that murine cutaneous LPR against OVA is a type 2 inflammatory response in which both IgH antibodies and CD4 T cells play an obligatory role.