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具有不同特应性家族史的新生儿的体外IgE合成

In vitro IgE synthesis in neonates with different family history of atopy.

作者信息

Miceli Sopo S, Pesaresi M A, Minchilli G, Maraglino V, Guerrini B, Rossodivita A, Stabile A

机构信息

Department of Pediatrics, Catholic University of Rome.

出版信息

Allergol Immunopathol (Madr). 1997 Mar-Apr;25(2):73-9.

PMID:9150836
Abstract

In this study we want to correlate family history of atopy and in vitro synthesis of IgE, IL4 and IFN gamma in 5 neonates with biparental (group A), 5 with uniparental (group B) and 5 with absent family history of atopy (group C). An aliquot of neonatal blood mononuclear cells (NBMC) was incubated in presence of PHA in combination with the phorbol ester acetate (TPA). The supernatants of cultures were harvested after 48-72 hours of incubation and stored at -20 degrees C until testing for lymphokine production by ELISA kits. Only one neonate of group B showed detectable in vitro synthesis of IL4 (45 pg/ml) after PHA + TPA stimulation. All the others failed to produce detectable levels either of IL4 or IFN gamma. Another aliquot of NBMC was cultured in the presence of saturating concentrations of rhIL4 for 48 h. After this pre-incubation step, the non-adherent cells were cultured in the presence of: 1) rhIL4; 2) rhIL4 + anti-IL4 antibody (Ab); 3) rhIL4 + anti-IFN gamma Ab; 4) rhIFN gamma, 5) rhIFN gamma + anti-IFN gamma Ab + rhIL4; 6) PWM + rhIL4; 7) unstimulated culture. The supernatants of these cultures were tested for their IgE content. In general, spontaneous IgE production by NBMC was very low, rhIL4 did not induce a significant increase of IgE synthesis. The other modalities of stimulations did not produce significative changes. We did not observe significative differences among the three groups of neonates. On the basis of our results, we conclude that NBMC aren't able to produce significative amounts of IgE in vitro either spontaneously or after IL4 stimulation. This test and the evaluation of IL4 and IFN gamma in vitro production aren't useful markers of atopy predisposition.

摘要

在本研究中,我们想要关联5名双亲均有特应性家族史的新生儿(A组)、5名单亲有特应性家族史的新生儿(B组)和5名无特应性家族史的新生儿(C组)的特应性家族史与IgE、IL4和IFNγ的体外合成情况。将一份新生儿血单核细胞(NBMC)在PHA与佛波酯乙酸盐(TPA)联合存在的情况下进行孵育。孵育48 - 72小时后收集培养物的上清液,并储存在 -20℃直至用ELISA试剂盒检测细胞因子产生情况。B组仅1名新生儿在PHA + TPA刺激后显示可检测到的IL4体外合成(45 pg/ml)。所有其他新生儿均未能产生可检测水平的IL4或IFNγ。另一份NBMC在饱和浓度的rhIL4存在下培养48小时。在此预孵育步骤之后,非贴壁细胞在以下条件下培养:1)rhIL4;2)rhIL4 + 抗IL4抗体(Ab);3)rhIL4 + 抗IFNγ Ab;4)rhIFNγ;5)rhIFNγ + 抗IFNγ Ab + rhIL4;6)PWM + rhIL4;7)未刺激培养物。检测这些培养物上清液的IgE含量。一般来说,NBMC的自发IgE产生非常低,rhIL4未诱导IgE合成的显著增加。其他刺激方式未产生显著变化。我们未观察到三组新生儿之间存在显著差异。根据我们的结果,我们得出结论,NBMC在体外无论是自发还是在IL4刺激后都不能产生大量的IgE。该检测以及IL4和IFNγ体外产生的评估不是特应性易感性的有用标志物。

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