Baran Joanna, Kuryk Łukasz, Garofalo Mariangela, Pancer Katarzyna, Wieczorek Magdalena, Kazek Michalina, Staniszewska Monika
Centre for Advanced Materials and Technologies, Warsaw University of Technology, Warszawa, Poland.
National Institute of Public Health National Institute of Hygiene (NIH) - National Institute of Research, Warszawa, Poland.
BioTechnologia (Pozn). 2024 Sep 30;105(3):263-272. doi: 10.5114/bta.2024.141805. eCollection 2024.
Screening for effective vaccines requires broad studies on their immunogenicity and . We used a PBMC-based system to assess changes in CD4 T cells, CD8 T cells, and CD19 B cells upon stimulation with different combinations of antigens and adjuvants. We studied the activation mechanism using flow cytometry and two different adenoviral adjuvants characterized by the presence or absence of costimulatory ligands for the ICOS and CD40 receptors. Our studies identified the cellular targets and molecular mechanisms driving ongoing switched-antibody diversification. Class-switched memory B cells were the main precursor cells (95.03% ± 0.38 vs. mock 82.33% ± 0.45, < 0.05) after treatment with the immunogenic formula: adenovirus armed (MIX1) or not (MIX2) with the ICOS and CD40 ligand, the recombinant receptor binding domain (rRBD), and Lentifect™ SARS-CoV-2 spike-pseudotyped lentivirus (GeneCopoeia, USA). Bcell class-switching towards the IgGIgM- positive phenotypes was noted (50-fold increase vs. mock, < 0.05). A significant increase was observed in the CD8T population of the MIX1 (2-fold, < 0.05) and MIX2 (~4.7-fold, < 0.05) treated samples. CD8T increased when PBMCs were treated with MIX2 (9.63% ± 0.90, < 0.05) vs. mock (2.63% ± 1.96). Class-switched memory B cells were the dominant antigen-specific cells in primary reactions. We indicated a correlation between the protection offered by vaccine regimens and their ability to induce high frequencies of multifunctional T cells.
筛选有效的疫苗需要对其免疫原性等进行广泛研究。我们使用基于外周血单个核细胞(PBMC)的系统来评估在用不同抗原和佐剂组合刺激后CD4 T细胞、CD8 T细胞和CD19 B细胞的变化。我们使用流式细胞术以及两种不同的腺病毒佐剂(其特征在于是否存在用于ICOS和CD40受体的共刺激配体)研究了激活机制。我们的研究确定了驱动正在进行的类别转换抗体多样化的细胞靶点和分子机制。在用免疫原配方处理后,类别转换记忆B细胞是主要的前体细胞(95.03%±0.38,而假处理组为82.33%±0.45,P<0.05):用ICOS和CD40配体、重组受体结合结构域(rRBD)以及Lentifect™ SARS-CoV-2刺突假型慢病毒(美国GeneCopoeia公司)武装(MIX1)或未武装(MIX2)的腺病毒。观察到B细胞向IgGIgM阳性表型的类别转换(与假处理组相比增加约50倍,P<0.05)。在MIX1处理的样本(约2倍,P<0.05)和MIX2处理的样本(约4.7倍,P<0.05)中,CD8 T细胞群体显著增加。当PBMC用MIX2处理时,CD8 T细胞增加(9.63%±0.90,P<0.05),而假处理组为(2.63%±1.96)。类别转换记忆B细胞是初次反应中主要的抗原特异性细胞。我们指出了疫苗方案提供的保护与其诱导高频多功能T细胞能力之间的相关性。