RESUMO
Nasal application of native cholera toxin (nCT) as a mucosal adjuvant has potential toxicity for the CNS through binding to GM1 gangliosides in the olfactory nerves. Although mutants of cholera toxin (mCTs) have been developed that show mucosal adjuvant activity without toxicity, it still remains unclear whether these mCTs will induce CNS damage. To help overcome these concerns, in this study we created new double mutant CTs (dmCTs) that have two amino acid substitutions in the ADP-ribosyltransferase active center (E112K) and COOH-terminal KDEL (E112K/KDEV or E112K/KDGL). Confocal microscopic analysis showed that intracellular localization of dmCTs differed from that of mCTs and nCTs in intestinal epithelial T84 cells. Furthermore, both dmCTs exhibited very low toxicity in the Y1 cell assay and mouse ileal loop tests. When mucosal adjuvanticity was examined, both dmCTs induced enhanced OVA-specific immune responses in both mucosal and systemic lymphoid tissues. Interestingly, although both dmCT E112K/KDEV and dmCT E112K/KDGL showed high Th2-type and significant Th1-type cytokine responses by OVA-specific CD4+ T cells, dmCT E112K/KDEV exhibited significantly lower Th1-type cytokine responses than did nCT and dmCT E112K/KDGL. These results show that newly developed dmCTs retain strong biological adjuvant activity without CNS toxicity.
Assuntos
Adjuvantes Imunológicos/genética , Toxina da Cólera/genética , Toxina da Cólera/imunologia , Mutação/genética , Adjuvantes Imunológicos/administração & dosagem , Animais , Anticorpos/imunologia , Linfócitos T CD4-Positivos/citologia , Linfócitos T CD4-Positivos/imunologia , Proliferação de Células , Sistema Nervoso Central/efeitos dos fármacos , Sistema Nervoso Central/metabolismo , Toxina da Cólera/administração & dosagem , Toxina da Cólera/toxicidade , Citocinas/metabolismo , Retículo Endoplasmático/metabolismo , Complexo de Golgi/metabolismo , Imunidade nas Mucosas/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Bulbo Olfatório/metabolismo , Ovalbumina/imunologia , Transporte ProteicoRESUMO
OBJECTIVE: To evaluate the occurrence and prognostic significance of perineural invasion (PNI) in squamous cell carcinomas (SCC) of the oral cavity and oropharynx. STUDY DESIGN: A retrospective study of 101 patients with previously untreated SCC of the oral cavity and oropharynx was undertaken to evaluate the occurrence and prognostic significance of PNI in relation to local recurrence, regional recurrence, distant metastasis and survival. The logistic regression test was used for univariate and multivariate analyses. Actuarial survival curves were determined using the Kaplan-Meier method. RESULTS: PNI was present in 26 (25.7%) of 101 patients and was significantly associated with tumor differentiation, lymph node metastasis, and depth of invasion. Univariate analyses showed PNI was associated with local recurrence (P=.005), regional recurrence (P=.007), and distant metastasis (P=.013). In multivariate analysis, PNI was significantly associated with regional recurrence (P=.033) and distant metastasis (P=.021), but not with local recurrence (P=.109). The 5-year disease-specific survival for patients with and without PNI was 56.6% and 94.6%, respectively (P<.0001). CONCLUSION: PNI is an important predictor for outcome of patients with SCC of the oral cavity and oropharynx.
