Impact on antitumor response using a new adjuvant preparation as a component of a human papillomavirus type 16 therapeutic vaccine candidate.

Cervical cancer is a global public health problem and human papillomavirus (HPV) 16 accounts for approximately 50% of cases worldwide. Although there are several types of HPV therapeutic vaccines in clinical research, there are currently not approved for use in humans. We developed the fusion protein LALF32-51-E7 (hereafter denominated CIGB550-E7) defined by a cell-penetrating peptide linked to an E7 mutein for the treatment of HPV16-associated tumors. We have demonstrated previously the benefit on antitumor response induced by the immunization with CIGB550-E7 admixed with very small size proteoliposomes (VSSP) adjuvant compared with the adjuvant-free immunization. In this study, we obtained a similar antitumor response in mice immunized with CIGB550-E7 admixed with the new adjuvant sVSSP that does not contain any animal-derived product. Also, the immunization with the above mentioned vaccine preparation induced a cell-mediated immune response. Our results are encouraging for the future clinical trials with the vaccine candidate CIGB550-E7+sVSSP.

LALF32-51-E7 therapeutic vaccine induces antitumor immunity against human papillomavirus type 16 E7-expressing murine tumor metastases in the lungs.

One important goal of cancer immunotherapy is to prevent and treat tumor metastasis. We have previously reported the significant antitumor effect induced by the immunization with our human papillomavirus therapeutic protein-based vaccine (LALF32-51-E7) without adjuvant and admixed with clinically relevant adjuvants in the subcutaneous TC-1 tumor challenge model. In the present study, we evaluated the efficacy of the above mentioned vaccine formulations in controlling the hematogenous spread of TC-1 tumor cells using a more tumourigenic clone named TC-1* and other intravenous injection site less stressful than the tail vein. We generated a lung metastasis model by injecting TC-1* cells into the retro-orbital venous sinus and this is the first study describing it. Also, this is the first study that demonstrates the efficacy of the immunization with LALF32-51-E7 without adjuvant and admixed with VSSP or Al(OH)3 in controlling metastatic tumors increasing the survival of the mice. Our TC-1 lung metastasis model can be used to test the efficacy of other immunotherapeutic strategies based on E6/E7 antigens.

Adsorption behavior of plasmid DNA onto perfusion chromatographic matrix.

Anion exchange chromatography is the most popular chromatographic method for plasmid separation. POROS RI 50 is a perfusion chromatographic support which is a reversed phase matrix and is an alternative to conventional ones due to its mass transfer properties. The adsorption and elution of the pIDKE2 plasmid onto reversed phase POROS R1 50 was studied. Langmuir isotherm model was adjusted in order to get the maximum adsorption capacity and the dissociation constant for POROS R1 50-plasmid DNA (pDNA) system. Breakthrough curves were obtained for volumetric flows between 0.69-3.33 mL/min, given dynamic capacity up to 2.3 times higher than those reported for ionic exchange matrix used during the purification process of plasmids with similar size to that of pIDKE2. The efficiency was less than 45% for the flow conditions and initial concentration studied, which means that the support will not be operated under saturation circumstances.