Abstract
Chlamydia trachomatis (Ct) is one of the most frequently encountered sexual infection all over the world, yielding tremendous reproductive problems (e.g. infertility and ectopic pregnancy) in the women. This work described the design of a plasmid vaccine that protect mice from Ct infection, and reduce productive tract damage by generating effective antibody and cytotoxic T cell immunity. The vaccine, s was composed of MOMP multi-epitope and HPV16L2 genes carried in pcDNA plasmid (i.e. pcDNA3.1/MOMP/HPV16L). In transfection, the vaccine expressed the chimeric genes (i.e. MOMP and HPV16L2), as demonstrated via western blot, RT-PCR and fluorescence imaging. In vitro, the vaccine transfected COS-7 cells and expressed the proteins corresponding to the genes carried in the vaccine. Through intramuscular immunization in BALB/c mice, the vaccine induced higher levels of anti-Ct IgG titer, anti-HPV16L2 IgG titer in serum and IgA titer in local mucosal secretions, compared to plasmid vaccines that carry only Ct MOMP multi-epitope or HPV16L2 chimeric component only. In mice intravaginally challenged with Ct, the vaccines pcDNA3.1/MOMP/HPV16L2 generated a higher level of genital protection compared to other vaccine formulations. Additionally, histochemical staining indicated that pcDNA3.1/MOMP/HPV16L2 eliminated mouse genital tract tissue pathologies induced by Ct infection. This work demonstrated that pcDNA/MOMP/HPV16L2 vaccine can protect against Ct infection by regulating antibody production, cytotoxic T cell killing functions and reducing pathological damage in mice genital tract. This work can potentially offer us a new vaccine platform against Ct infection.
Highlights
Chlamydia trachomatis (Ct) is one of the primary causes in reproductive tract infections worldwide [1]
Upon digesting pcDNA3.1/major outer membrane proteins (MOMP)/Human papillomavirus 16L2 (HPV16L2) with EcoRI and XhoI, we observed a band at 1456 bp position, which is the size HPV16L2 product, indicating HPV16L2 gene was incorporated into the plasmid (Figure 1B Lane 4)
The Ct MOMP multiepitope (GDNENQSTVKTNSVPNMSLDQSVVELY TWQASLALSYRLNMFTPYIGV) that including 28-amino-acid (138–165) and 19-amino-acid sequences (249–268) was selected [6]. These amino acid sequences contain T helper cell (Th) epitopes, and epitopes related to HLA-A2-restricted cytotoxic T lymphocyte (CTLs) and specific B-cells
Summary
Chlamydia trachomatis (Ct) is one of the primary causes in reproductive tract infections worldwide [1]. Ct causes more than 92 million sexually transmitted infections, yielding long term reproductive problems (e.g. infertility and ectopic pregnancy) in the women patients. Ct is regarded as an immunologically sensitizing infection, and human immunity is unable to resist Ct completely due to the complicated evasion mechanisms of. Ct. Ct usually replicates within the epithelial cells, preventing itself from the immune effectors by using a sanctuary shield [2]. Ct employs multiple other strategies to escape human immune systems, such as avoiding the antibody detection by presenting antigenically diverse surface proteins, inducing the apoptosis of activated effector T cells against the infection, etc. Theses tricks in Ct pathobiology require us to design novel effective Ct vaccines by outdoing the immunological and biological barriers [4]
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