Immunological unresponsiveness established by the elimination or anergy of self-reactive lymphocyte clones is of importance to immunization against tumorassociated antigens. In this study, we have investigated induction of immunity against the human MUC1 carcinomaassociated antigen in MUC1 transgenic mice unresponsive to The human DF3͞MUC1 glycoprotein is overexpressed and aberrantly glycosylated in breast and other carcinomas (1-4). The finding that lymphocytes from certain patients with carcinomas recognize and lyse MUC1-positive tumor cells (5, 6) has suggested that this antigen is a potential target for anticancer vaccines. Whereas MUC1 is expressed on the apical borders of normal epithelium (1-3) and unresponsiveness to self-antigens is an obstacle to the development of antitumor immunity, MUC1 transgenic (MUC1.Tg) mice provide a potential model to assess the induction of anti-MUC1 immune responses. In this context, MUC1.Tg C57BL6 mice express MUC1 in a pattern and at a level similar to that found in humans (7). Significantly, the MUC1.Tg mice are tolerant to stimulation by MUC1 antigen (7).Dendritic cells (DC) are potent antigen-presenting cells (8) that sensitize CD4 ϩ T cells to specific antigens in a major histocompatibility complex-restricted manner (9, 10) and generate antigen-specific cytotoxic T lymphocytes (CTLs) from naive T cells in vitro (11,12). Moreover, DCs are the only antigen-presenting cells known to prime naive CTLs and to induce antigen-specific CTLs in vivo (13). DCs pulsed with tumor antigens or synthetic peptides derived from such antigens have been effective as vaccines in the induction of CTL responses and antitumor activity (14)(15)(16)(17). Other studies have demonstrated that transduction of DC with recombinant viral vectors expressing tumor antigens generates vaccines that induce antigen-specific antitumor immune responses (18)(19)(20). Fusions resulting in heterokaryons of DC and carcinoma cells as vaccines have provided an alternative strategy for inducing immunity against both known and unidentified tumor antigens (21).The present studies demonstrate that MUC1.Tg mice respond to fusions of DC and MUC1-positive MC-38 carcinoma cells with induction of anti-MUC1 immunity. The findings demonstrate that a DC fusion cell vaccine can reverse unresponsiveness to a tumor-associated antigen and induce the rejection of established metastases. MATERIALS AND METHODS MUC1 Transgenic Mice.A C57BL͞6 mouse strain transgenic for human MUC1 was established as described (7). Tail DNA (500 ng) was subjected to PCR amplification by using MUC1 primers (bp 745-765 and bp 1,086-1,065) to confirm the presence of MUC1 sequences. The PCR product was detected by electrophoresis in a 1% agarose gel (7).Cell Culture and Fusion. Murine (C57BL͞6) MC-38 and MB49 carcinoma cells were stably transfected with a MUC1 cDNA (22-24). Cells were maintained in DMEM supplemented with 10% heat-inactivated fetal calf serum, 2 mM

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Peripheral blood-derived inflammation-based markers, including C-reactive protein (CRP), neutrophil-to-lymphocyte ratio (NLR), lymphocyte-to-monocyte ratio (LMR), and platelet-to-lymphocyte ratio (PLR) are indicators of prognosis in various malignant tumors. The present study aimed to identify the inflammation-based parameters that are most suitable for predicting outcomes in patients with breast cancer. Two hundred ninety-six patients who underwent surgery for localized breast cancer were reviewed retrospectively. The association between clinicopathological factors and inflammation-based parameters were investigated. Univariate and multivariate Cox regression analyses were performed to identify independent prognostic indicators associated with disease-free survival (DFS). The NLR level correlated significantly with tumor size (P<0.05). The PLR level correlated with the expression of estrogen receptor and lymph node involvement (P<0.05). Univariate analysis revealed that lower CRP and PLR values as well as tumor size, lymph node involvement, and nuclear grade were significantly associated with superior DFS (CRP: P<0.01; PLR, tumor size, lymph node involvement, and nuclear grade: P<0.05). On multivariate analysis, CRP (hazard ratio [HR]: 2.85, 95% confidence interval [CI]: 1.03–7.88, P<0.05), PLR (HR: 2.61, 95% CI: 1.07–6.36, P<0.05) and nuclear grade (HR: 3.066, 95% CI: 1.26–7.49, P<0.05) were significant prognostic indicators of DFS in patients with breast cancer. Neither LMR nor NLR significantly predicted DFS. Both preoperative CRP and PLR values were independently associated with poor prognosis in patients with breast carcinoma; these were superior to other inflammation-based scores in terms of prognostic ability.