Gonçalves Ana Katherine, Giraldo Paulo César, Machado Paula Renata Lima, Farias Kleber Juvenal Silva, Costa Ana Paula Ferreira, Freitas Janaina Cristiana de Oliveira Crispim, Eleutério José, Witkin Steven S
1 Department of Gynecology and Obstetrics, Federal University of Rio Grande do Norte , Natal, Brazil .
2 Department of Gynecology and Obstetrics, State University of Campinas , Campinas, Brazil .
Viral Immunol. 2015 Jul-Aug;28(6):339-42. doi: 10.1089/vim.2015.0008. Epub 2015 May 12.
The objective of this work was to evaluate the influence of human papillomavirus (HPV) vaccination on peripheral blood mononuclear cell (PBMC) proliferation and cytokine gene transcription. PBMCs isolated after HPV immunization were incubated with HPV vaccine, phytohemagglutinin, or buffer. Cell proliferation was assessed by MTT reduction assay. RNA was extracted from PBMCs, and the relative concentration of cytokine messenger RNA (mRNA) transcripts (IFN-β, IFN-γ, IL-12, TNF-α, IL-6, IL-17, or IL-10) relative to transcription of the β-actin gene was determined by real-time polymerase chain reaction. PBMC proliferation in response to HPV vaccine and PHA were greater than that observed in unstimulated cells (p<0.001). Cytokine mRNAs were upregulated in stimulated PBMC cultures. The median increase in vaccine-stimulated cultures was: IFN-β=334.4-fold; IL-12=46.33-fold; IFN-γ=12.64-fold; IL-6=9.07-fold; IL-17=7.33-fold; IL-10=6.47-fold; and TNF-α=2.36-fold. The IFN-β expression was significantly higher (p<0.05). Proliferative PBMC responses and multiple cytokine gene expression were detected in women who received the HPV vaccine.
这项工作的目的是评估人乳头瘤病毒（HPV）疫苗接种对外周血单个核细胞（PBMC）增殖和细胞因子基因转录的影响。HPV免疫后分离得到的PBMC与HPV疫苗、植物血凝素或缓冲液一起孵育。通过MTT还原试验评估细胞增殖。从PBMC中提取RNA，通过实时聚合酶链反应测定细胞因子信使核糖核酸（mRNA）转录本（IFN-β、IFN-γ、IL-12、TNF-α、IL-6、IL-17或IL-10）相对于β-肌动蛋白基因转录的相对浓度。PBMC对HPV疫苗和PHA的增殖反应大于未刺激细胞中的增殖反应（p<0.001）。细胞因子mRNA在受刺激的PBMC培养物中上调。疫苗刺激培养物中的中位数增加倍数为：IFN-β=334.4倍；IL-12=46.33倍；IFN-γ=12.64倍；IL-6=9.07倍；IL-17=7.33倍；IL-10=6.47倍；TNF-α=2.36倍。IFN-β表达显著更高（p<0.05）。在接种HPV疫苗的女性中检测到增殖性PBMC反应和多种细胞因子基因表达。
