Diagnostic value of intraoperative ultrasonography in assessing thoracic recurrent laryngeal nerve lymph nodes in patients with esophageal cancer.

BACKGROUD: The incidence of recurrent laryngeal nerve (RLN) injury has increased due to RLN lymph node dissection. The aim of this study was to evaluate the ability of intraoperative ultrasonography (IU) to detect RLN nodal metastases in esophageal cancer patients. METHODS: Sixty patients with esophageal cancer underwent IU, computed tomography (CT), and endoscopic ultrasonography (EUS) to assess for RLN nodal metastasis. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were compared. RESULTS: The sensitivities of IU, CT, and EUS in diagnosing right RLN nodal metastases were 71.4, 14.3, and 30.0%, respectively, and a significant difference among these three examinations was observed (χ2 = 10.077, P = .006). The specificities of IU, CT, and EUS for diagnosing right RLN nodal metastasis were 67.4, 97.8, and 95.0%, respectively, and a significant difference was observed (χ2 = 21.725, P < .001). No significant differences in either PPV or NPV were observed when diagnosing right RLN nodal metastases. For diagnosis of left RLN lymph nodal metastases, the sensitivities of IU, CT, and EUS were 91.7, 16.7, and 40.0% respectively. There was a significant difference among these diagnostic sensitivities (χ2 = 14.067, P = .001). The specificities of IU, CT, and EUS for diagnosis of left RLN nodal metastases were 79.2, 100, and 82.5%, respectively and a significant difference was observed (χ2 = 10.819, P = .004). No significant differences were observed in PPV or NPV for these examinations when diagnosing left RLN nodal metastases. CONCLUSION: Intraoperative ultrasonography showed superior sensitivity compared with preoperative CT or EUS in detecting RLN lymph node metastasis in patients with thoracic esophageal cancer.

PLGA-nanoparticle mediated delivery of anti-OX40 monoclonal antibody enhances anti-tumor cytotoxic T cell responses.

OX40 (CD134) is a tumor necrosis factor (TNF) receptor expressed mainly on activated T cells and transmits a potent costimulatory signal once engaged. Agonistic anti-OX40 monoclonal antibody (mAb) enhances tumor immune response leading to therapeutic effects in mouse tumor models. However, when tested in phase I clinical trials it did not show objective clinical activity in cancer patients. In this study, we examined the feasibility of nanoparticle (NP)-mediated delivery of anti-OX40 mAb to efficiently induce cytotoxic T lymphocyte (CTL) responses. The biodegradable poly(DL-lactide-co-glycolide) nanoparticle (PLGA-NP) carrying anti-OX40 mAb, anti-OX40-PLGA-NP, was prepared by double emulsion method and showed an average diameter of 86 nm with a loading efficiency of 25%. We found that anti-OX40-PLGA-NP induced CTL proliferation and tumor antigen-specific cytotoxicity as well as cytokine production more strongly than free anti-OX40 mAb. These results suggest that PLGA-based nanoparticle formulation may provide efficient delivery system of anti-OX40 mAb for cancer immunotherapy.

[Effect of PLGA nanoparticles conjugated with anti-OX40/anti-AFP mAbs on cytotoxicity of CTL cells against hepatocellular carcinoma].

OBJECTIVE: To evaluate the effect of anti-OX40 and anti-AFP antibodies conjugated onto poly(DL-lactide-co-glycolide)-nanoparticles (PLGA-NPs) on the cytotoxic activity of AFP158-166; -specific cytotoxic T lymphocyte (CTL) against hepatocellular carcinoma cells in vitro. METHODS: PLGA-NPs were prepared by oil-in-water single emulsion solvent evaporation method and covalently conjugated with anti-OX40 and anti-AFP monoclonal antibodies. Scanning electron microscopy (SEM) was utilized for the characterization of the surface morphology and estimation of the size of the PLGA-NPs. The mean diameter and zeta potential of the nanoparticles were measured by dynamic light scattering (DLS) performed in a Zetasiser Nano Series ZEN3600. Antibody conjugation efficiency was determined using bicinchoninic acid (BCA) protein assay. Dendritic cells (DCs) were induced from human peripheral blood mononuclear cells (PBMCs) in the presence of GM-CSF and IL-4, and loaded with AFP158-166; peptide to generate AFP-specific CTL (CTL/AFP158-166;). WST-1, ELISA and lactate dehydrogenase (LDH) methods were respectively used to examine the effects of the anti-OX40/anti-AFP-NPs on CTL/AFP158-166; proliferation, IL-2 and IFN-γ production, and cytotoxicity against the tumor cells. RESULTS: The obtained nanoparticles were found to be of regular spherical shape and the smooth surface with an average diameter of (300±42) nm and a negative zeta potential of -(25.12±5.34) mV. Approximately 100 µg antibodies were conjugated to every milligram of the nanoparticles with a conjugation efficiency of about 25% as estimated by BCA protein assay. Proliferation and activation analysis revealed that anti-OX40/anti-AFP mAb-NPs significantly induced CTL proliferation and the secretion of IL-2 and IFN-γ. The cytotoxicity assay showed that anti-OX40/anti-AFP-NPs markedly enhanced CTL/AFP158-166; specific killing on HepG2 cells but had no obvious effect on SMMC-7721 cells. CONCLUSION: Anti-OX40 mAb and anti-AFP mAb conjugated to PLGA-NPs could stimulate CTL/AFP158-166; cell proliferation and cytokine production as well as enhancing their specific killing on AFP-positive hepatocellular carcinoma cells.