Patients and samples
We collected blood samples from 78 women: patients with cervical carcinoma (n=21; median age 44, age range 47.2 +/- 11.5 years), high grade lesions (n= 9; median age 37, age range 42.9 +/- 16.6 years), low grade lesions (n=24; median age 30, age range 30.2 +/- 9.6 years), and healthy donors (n=24; median age 38.5, age range 39.25 +/- 7.2 years). The women with premalignant lesions and cancer had their diagnosis confirmed with colposcopy and histopathological analysis. The cancer group included cervical adenocarcinoma and squamous cell carcinoma in situ (stage 0), and FIGO stages I, II, III and IV. These patients were attended at the Instituto Jalisciense de Cancerología, and the Hospital Civil Nuevo, Guadalajara, Jalisco, México. All study participants gave their written informed consent. This study was performed according to the guidelines of the local institutional ethics committees and in accordance with the 2013 Declaration of Helsinki.
Venous blood was collected from each donor (15 mL). Peripheral Blood Mononuclear Cells (PBMCs) were separated from the blood samples using Lymphoprep™ (STEMCELL Technologies, Vancouver, Canada) density gradient centrifugation. The PBMCs were stored frozen in liquid nitrogen at a temperature below –130°C until the cytometry analysis. The viability of the thawed cells was determined with trypan blue and cells were used when viability was greater than or equal to 90%. Serum from the blood of the same women was isolated and stored at -80 °C.