A prerequisite to using intratumoral de livery is the easy access for antigen delivery to the tumor site. Salivary gland tumors as well as head and neck tu mors including tongue, floor of the mouth, palate and mandibular mucosa and so on, appear suitable for such vaccine delivery. Salivary gland tumors are a group of het erogeneous lesions which e press ErbB2, whose current treatment involves surgery and adjuvant radio therapy. However, therapy response rates have been gener ally poor for these tumors. Recently, given that the high histopatological similarity between salivary ductal and breast carcinomas, Trastuzumab, a humanized mono clonal antibody to ErbB2, has been proposed as a potential therapy for salivary gland tumors treatment.

However, ac tive immunization targeting ErbB2 might induce tumor growth inhibition more efficiently than passive immuno therapy based on the generation of an e tended memory immune response. In this study we e amined the effectiveness of the rV neuT intratumoral vaccination in hampering the growth of transplanted Neu overe pressing BALB neuT salivary gland cancer cells in BALB neuT mice. In addition, we e plored whether the efficiency of vaccination was dependent on the dose of the rV neuT vaccine. Considering previous demonstration that a potent anti Neu humoral response is required to prevent mammary tumor growth in BALB neuT vaccinated mice, we investigated the anti Neu humoral response following rV neuT vaccination as well as the in vitro biological activity of immune sera from rV neuT vaccinated mice.

Finally, we determined whether rV neuT vaccination elicits anti Neu T cell immunity. Our research suggests that intratumoral vaccination using recombinant vaccinia virus could be efficiently employed for the treatment of salivary gland tumors and other accessible tumors. Methods Antibodies, peptides, reagents and cells Neu overe pressing salivary gland cancer cells were kindly provided by Prof. Federica Cavallo and maintained in DMEM containing 20% fetal bovine serum. SALTO cells were estab lished from salivary carcinoma arising in BALB neuT trans genic male mice hemizygous for p53172R H transgene driven by the whey acidic protein promoter. NIH3T3 cells e pressing normal rat Neu have been previously characterized and kindly provided by Dr. Eddi Di Marco. Renal epithelial cell lines BSC 1 and NIH3T3 cells were purchased by ATCC.

BSC 1, LTR Neu and NIH3T3 were maintained in DMEM con taining 10% FBS. Monoclonal antibody anti Neu Ab4 was purchased from Oncogene Science. Rabbit polyclonal anti Neu antibody, anti ERK1 2 antibody and monoclonal antibody anti pERK1 2 were purchased by Santa Cruz Biotechnology. Rabbit polyclonal antibody recogniz ing the activated cleaved caspase 3 was Brefeldin_A purchased from Cell Signaling Technology.