Related Experiment Video
Updated: May 2, 2026
Use of Interferon-γ Enzyme-linked Immunospot Assay to Characterize Novel T-cell Epitopes of Human Papillomavirus
Published on: March 8, 2012
HPV16E7 silencing enhances susceptibility of CaSki cells to natural killer cells
Huimin Guo1, Ruili Hu1, Xinlei Guan1
1Department of Obstetrics and Gynecology, The First Affiliated Hospital of Xinxiang Medical University, Weihui, Henan 453100, P.R. China.
Abstract:
The aim of the present study was to investigate the cytotoxicity of natural killer (NK) cells to CaSki cells following knockdown of the E7 protein of the human papillomavirus type 16 (HPV16E7). Recombinant adenovirus-short hairpin-E7 protein of the human panillomavirus type 16 (Ad‑sh‑HPV16E7) was constructed and used to infect CaSki cells. The expression of HPV16E7 in CaSki cells was assessed using western blot analysis. The expression of cell surface molecule major histocompatibility complex‑I (MHC‑I) in CaSki cells infected with Ad‑sh‑HPV16E7 was examined using flow cytometry. The cytotoxicity of NK cells isolated and expanded from healthy volunteers on Ad‑sh‑HPV16E7‑infected CaSki cells was assessed using the lactate dehydrogenase (LDH) release assay. Ad‑sh‑HPV16E7 was successfully constructed and able to inhibit HPV16E7 the expression in CaSki cells. The expression of major histocompa-tibility complex I (MHC‑I), a surface molecule, in CaSki cells was increased after infection with Ad‑sh‑HPV16E7. Compared with the controls, the cytotoxicity of NK cells on CaSki cells, which were infected with Ad‑sh‑HPV16E7, was decreased (p<0.05). In conclusion, HPV16E7 suppresses the expression of MHC‑I on CaSki cells to evade cytotoxic T‑cell (CTL) response. However, it was possible to enhance the cytotoxicity of expanded NK cells to cervical cancer cells or HPV16‑infected cells in vitro, indicating that NK cells may be used for immunotherapy of cervical cancer.
Insights
Human papillomavirus type 16 E7 protein (HPV16E7) suppresses major histocompatibility complex-I (MHC-I) on CaSki cells. Knockdown of HPV16E7 enhanced natural killer (NK) cell cytotoxicity against cervical cancer cells, suggesting potential for immunotherapy.
Area of Science:
- Immunology
- Virology
- Oncology
Background:
- Human papillomavirus type 16 (HPV16) is a major cause of cervical cancer.
- The HPV16 E7 oncoprotein plays a crucial role in viral oncogenesis.
- Understanding immune evasion mechanisms is vital for developing effective cancer therapies.
Purpose of the Study:
- To investigate the effect of HPV16E7 knockdown on natural killer (NK) cell cytotoxicity against CaSki cervical cancer cells.
- To determine if targeting HPV16E7 can enhance NK cell-mediated killing of cancer cells.
Main Methods:
- Constructed recombinant adenovirus carrying short hairpin RNA against HPV16E7 (Ad-sh-HPV16E7).
- Infected CaSki cells with Ad-sh-HPV16E7 and assessed HPV16E7 expression via Western blot.
- Analyzed major histocompatibility complex-I (MHC-I) expression on infected cells using flow cytometry.
- Measured NK cell cytotoxicity using the lactate dehydrogenase (LDH) release assay.
Main Results:
- Successful construction and delivery of Ad-sh-HPV16E7, leading to HPV16E7 knockdown in CaSki cells.
- Increased expression of MHC-I on CaSki cells following Ad-sh-HPV16E7 infection.
- Significantly decreased NK cell cytotoxicity against Ad-sh-HPV16E7-infected CaSki cells compared to controls (p<0.05).
Conclusions:
- HPV16E7 suppresses MHC-I expression on CaSki cells, facilitating immune evasion.
- Despite reduced NK cell cytotoxicity in this specific in vitro model, the study highlights the potential of targeting HPV16-infected cells.
- Expanded NK cells show promise for the immunotherapy of cervical cancer.
More Related Videos
Related Concept Videos
01:27Cytotoxic T Cells-mediated Immune Response
Immunological surveillance is the ability of immune cells to monitor and eliminate infected cells with intracellular pathogens, neoplastically transformed cells, and cells with non-self antigens. Cytotoxic T cells and NK...
01:28Cells of the Innate Immune Response
Phagocytes
Phagocytes police the peripheral tissues by removing cellular debris and responding to the invasion of foreign substances or pathogens. Many phagocytes attack and remove microorganisms even before lymphocytes detect them. The human body has two general...
01:29Immune Response Against Viral Pathogens
NK Cells
NK cells are a crucial part of our innate immune system, acting as the first line of defense against viral infections. These cells can recognize and kill infected cells without prior exposure to the virus, effectively slowing down the spread of infection. Additionally, NK cells produce proinflammatory...