Chauvat Anne, Benhamouda Nadine, Gey Alain, Lemoine Francois M, Paulie Staffan, Carrat Fabrice, Gougeon Marie-Lise, Rozenberg Flore, Krivine Anne, Cherai Mustapha, Lehmann Paul, Quintin-Colonna Françoise, Launay Odile, Tartour Eric
INSERM U970 ; Université Paris Descartes Sorbonne Paris-Cité; Paris, France; Hôpital Européen Georges Pompidou; Service d'Immunologie Biologique; Paris, France; CTL-Europe GmbH; Bonn, Germany.
INSERM U970 ; Université Paris Descartes Sorbonne Paris-Cité; Paris, France; Hôpital Européen Georges Pompidou; Service d'Immunologie Biologique; Paris, France.
Hum Vaccin Immunother. 2014;10(1):104-13. doi: 10.4161/hv.26593. Epub 2013 Oct 1.
The type of T cell polarization and simultaneous production of multiple cytokines have been correlated with vaccine efficacy. ELISpot is a T cell detection technique optimized for the measurement of a secreted cytokine at the single cell level. The FluoroSpot assay differs from ELISpot by the use of multiple fluorescent-labeled anticytokine detection antibodies, allowing optimal measurement of multiple cytokines. In the present study, we show that an IFNγ/IL-10 FluoroSpot assay is more sensitive than flow cytometry to detect Tr1 regulatory T cells, an immunosuppressive T cell population characterized by the production of IL-10 and IFNγ. As many tolerogenic vaccines are designed to induce these Tr1 cells, this FluoroSpot test could represent a standard method for the detection of these cells in the future. The use of an IFNγ/IL-2 FluoroSpot assay during influenza vaccine monitoring showed that the influenza-specific IL-2-producing T-cell response was the dominant response both before and after vaccine administration. This study therefore questions the rationale of using the single-color IFNγ ELISpot as the standard technique to monitor vaccine-specific T-cell response. Using this same test, a trend was also observed between baseline levels of IFNγ T cell response and T cell vaccine response. In addition, a lower IFNγ+IL-2+ T-cell response after vaccine was observed in the group of patients treated with TNFα inhibitors (P=0.08). This study therefore supports the use of the FluoroSpot assay due to its robustness, versatility and the complementary information that it provides compared with ELISpot or flow cytometry to monitor vaccine-specific T-cell responses.
T细胞极化类型和多种细胞因子的同时产生与疫苗效力相关。ELISpot是一种优化用于在单细胞水平测量分泌细胞因子的T细胞检测技术。FluoroSpot检测法与ELISpot的不同之处在于使用了多种荧光标记的抗细胞因子检测抗体，从而能够对多种细胞因子进行最佳测量。在本研究中，我们表明，IFNγ/IL-10 FluoroSpot检测法在检测Tr1调节性T细胞方面比流式细胞术更敏感，Tr1调节性T细胞是一种以产生IL-10和IFNγ为特征的免疫抑制性T细胞群体。由于许多耐受性疫苗旨在诱导这些Tr1细胞，这种FluoroSpot检测法未来可能成为检测这些细胞的标准方法。在流感疫苗监测期间使用IFNγ/IL-2 FluoroSpot检测法表明，流感特异性产生IL-2的T细胞反应在疫苗接种前后都是主要反应。因此，本研究对将单色IFNγ ELISpot作为监测疫苗特异性T细胞反应的标准技术的合理性提出了质疑。使用相同的检测方法，还观察到IFNγ T细胞反应的基线水平与T细胞疫苗反应之间存在一种趋势。此外，在接受TNFα抑制剂治疗的患者组中，观察到疫苗接种后IFNγ+IL-2+ T细胞反应较低（P=0.08）。因此，本研究支持使用FluoroSpot检测法，因为它具有稳健性、多功能性，并且与ELISpot或流式细胞术相比，它在监测疫苗特异性T细胞反应时能提供补充信息。
