New Evidences about Unusual Behavior of HeLa Cells in Stress Environment Concerning Immortality Status

Abstract

To produce an anticancer vaccine, we harnessed a plant viral expression system utilizing transgenic tomato fruit containing the genes HPV16 E2, E6, and E7. Notably, antibodies from the serum of mice orally vaccinated with HPV16 E2 spurred the formation of biofilms in HeLa cells, resembling dendrimer structures initially colored with sublethal trypan blue (TB) dye, which later faded. These biofilms emerged ex vivo upon the introduction of HPV16 E2, L-amino acid oxidase, D-amino acid oxidase, HPV16 E6, and E7, in conjunction with a CRISPR/CAS cassette. Subsequently, HeLa cells adopted a symplast-like structure devoid of cell demarcation, composed of fused membranes encircling the cytoplasm. In a separate experiment, mice spleens, rich in immune cells and red blood cells, were inoculated with HeLa cells. Following isolation, splenocytes underwent Elispot analysis after exposure to HPV16 E2, L-amino acid oxidase, and D-amino acid oxidase as activators. Significantly elevated levels of interferon, T-cell receptors, CD4/CD8 T lymphocytes, and apoptotic enzymes (granzyme B, perforin, and granulysin) were detected. Furthermore, splenocytes derived from HeLa-treated spleens exhibited the ability to induce regression of mice lung tumors ex vivo. These findings suggest that splenocytes, when exposed to HeLa cells, may undergo a form of training or education, facilitating the development of a microenvironment.