Yanyan Han¹, Jianting Long², Sheng Ye², Ran Tao³, Dongyun Wu³, Jin Li³, Xiangjun Zhou³, Ping Chen³

¹Department of Infectious Diseases, Nanfang Hospital, Southern Medical University;SYZ Cell Therapy Co., ²Dept. of Medicinal Oncology, The First Affiliated Hospital of Sun Yat-sen University, ³SYZ Cell Therapy Co.

Objective:Cervical cancer is the second most common Gynecologic malignant tumor and frequently occurs in patients with human papilloma virus (HPV) infection. Effective treatment for cervical cancer (including surgery and concurrent chemoradiation) can yield cures in 80% of women with early stage disease (stage I-II). However, vascular invasion, incomplete lymphadenectomy are most common factors predicting poor prognosis in early stage cervical cancer. Patients with vascular invasion confirmed by pathological specimens are more likely to develop metastatic disease in the near future post-surgery. Here we present an immunotherapy named MASCT (Multiple Antigen Stimulating Cellular Therapy), to treat a HPV+ metastatic cervical cancer patient with multiple tumor antigen pulsed dendritic cells (DCs) and Tlymphocytes stimulated by these DCs. Method: A patient was pathologically diagnosed as cervical squamous cell carcinoma when she was 41 years old. Her specimens showed vascular invasion and insufficient lymph node dissection even though she was considered as having stage I disease. And she was also tested positive for the HPV infection. Although she had received radical resection of primary tumor, and subsequent adjuvant chemoradiation therapy, she was still detected metastasis on the right Sacroiliac Joint 33 months after surgery. She was then given local irradiation treatments for 1 month, and MASCT treatments every month for the first 6 months and every 2 months afterwards. To prepare the DCs and activated T lymphocytes, the monocytes from patient`s PBMCs were firstly differentiated into immature dendritic cells (iDCs)and then pulsed with multiple synthetic peptide antigens including tumor-associated antigens and HPV specific antigens. The semi-mature DCs were further stimulated by diverse TLR ligands to differentiate into mature DCs (mDCs). Half of the semDCs were subcutaneous injected to the patient (1-5x106cells/injection), and the other half were co-cultured with the maintaining non-adherent T cells for another 7-9 days before infusion back to the patient (5-10x107cells/kgbody weight). Both the mDCs and the resulting activated T cells were tested negative for fungi, mycoplasma and endotoxin contamination. Result: The mDCs expressed high level of HLA-DR, CD80, CD86, CD83, and CCR7 on the surface. And the activated T cells were almost exclusively CD3+ T cells, with a major part beingCD3+CD8+ cells and a minor proportion being CD3+CD4+and CD3+CD56+ cells, while there were insignificant amounts of CD4+CD25+Foxp3+ regulatory T cells. After 3 times MASCT treatments (6 months after metastasis), the patient was evaluated as partial remission by Tc-99m-oxidronatewhole body bone SPECT (single-photon emission computed tomography). After another 5 times MASCT treatments (12 months after metastasis), the patient remained partial remission. Moreover, specific responses against tumor antigens were also detected in the patients` PBMCs by IFNg ELISPOT assay, such as telomerase, p53, CEA,RGS5, and HPV18/58. We have precisely adjusted her antigen peptide pool by saving the antigens which had induced specific responses and deleting the ones had not. After 3 times MASCT treatments with the new antigen pool, the specific responses against tumor antigens and HPV18/58were clearly enhanced. Conclusion: Our study provides MASCT as a safe treatment, which has reduced the metastasis of the cervical cancer patient. Moreover, tumor antigens specific T cell responses could be robustly raised in cervical cancer patients after MASCT treatment, and were even further boosted after adjustment of the tumor antigen pool. On this basis, this precise strategy of immunocellular therapy could be applied for patients with diverse cancers.

Key Words: MASCT, Cervical cancer, Tumor antigens, specific T cell