Programmable protein arrays for immunoprofiling HPV-associated cancers.

Area of Science:

• Immunology
• Virology
• Oncology

Background:

• Over 600,000 cancers globally are linked to human papillomavirus (HPV) annually.
• Understanding HPV immunobiology is crucial for combating HPV-associated cancers like cervical and oropharyngeal cancer (OPC).
• Current methods face challenges due to the diversity of oncogenic HPV types and the need for multiplexed antigen displays to assess antibody responses.

Purpose of the Study:

• To develop and validate a custom human papillomavirus (HPV) protein microarray for multiplexed detection of HPV-specific antibodies.
• To assess the utility of the HPV protein array in identifying serologic responses to multiple HPV types.

Main Methods:

• Generated a custom HPV protein microarray displaying 98 C-terminal GST fusion proteins from 12 HPV types (2 low-risk, 10 high-risk).
• Utilized a human cell lysate expression system for robust protein expression (96/98 antigens).
• Validated antibody specificities and identified target epitopes using monoclonal antibodies.
• Analyzed sera from OPC patients and controls, and a cervical cancer patient, for IgG seroreactivity using the HPV protein array.

Main Results:

• Demonstrated robust and reproducible expression of 96/98 HPV antigens.
• Identified specificities of four monoclonal antibodies.
• Showed significant IgG seroreactivity to HPV16 E1, E2, and E7 in OPC patients compared to controls (p < 0.005).
• Detected HPV52 E7 antibodies in a cervical cancer patient's serum.

Conclusions:

• The developed HPV protein microarray enables robust and reproducible display of multiple HPV antigens.
• The array facilitates the identification of specific antibody responses to various HPV types in patient sera.
• This platform shows potential for rapid serologic profiling in the context of HPV infection and associated cancers.