RESUMEN
The cytotoxic function of polyclonal expanded ϒ/δ T cells against pamidronate-treated cervical cancer cells in vitro and in vivo were determined. The ϒ/δ T cells were isolated and purified from PBMCs by using miniMACS and were later treated with 10 μM pamidronate. The expansion of ϒ/δ T cells was 15 times more than the non-stimulated cells. Among the expanded ϒ/δ T cells, 47% were Vϒ9/Vδ2 T cells with a purity of 87%. Analyzing the cytotoxic function of ϒ/δ T cells against 3 cervical cancer cells in vitro by LDH cytotoxicity test revealed that the killing efficacy increased if the cervical cancer cells (HeLa, SiHa and CaSki) were pretreated with pamidronate. The presence of CD107 on ϒ/δ T cells indicated the degranulation of perforin and granzyme pathway is one of the mechanisms used by the ϒ/δ T cells to kill cancer cells. The killing ability of ϒ/δ T cells against cancer cells in vivo was preliminary assessed by using mouse baring HeLa cells. The results demonstrated that ϒ/δ T cells induce apoptosis in tumor cells. Our study supports the usefulness of ϒ/δ T cells in future development of immunotherapy for cervical cancer.
RESUMEN
HPV infection is known to be associated with cervical cancer development. Precancerous lesions named cervical intraepithelial neoplasia (CIN) are divided into 3 grades, i.e., CIN-1, CIN-2, and CIN-3. Here, HPV infection determined by PCR and dot hybridization was observed in these 3 different grades of formalin-fixed paraffin embedded tissues. The HPV infection was demonstrated in 33.3 per cent of CIN-1, 36.8 per cent of CIN-2 and 75 per cent of CIN-3. Using type specific probes for HPV-6, 11, 16, 18 and 33, HPV-16 was the most prevalent type (44.44%) followed by HPV-18 (16.05%) in CIN-3. Only one HPV-18 was identified in CIN-1 while CIN-2 contained one HPV-6 and one HPV-18. Mixed infection was found in CIN-3 (12.35%). All of them had HPV-16. The cervicitis cases with normal histopathology were included as control. Only 2.7 per cent of HPV infection was shown. The relative risk of HPV infection was high in CIN-3 (OR = 107.25, 95% CI = 50.29-228.73). Our data confirm the association between high-risk HPV types and development of CIN.