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Comprehensive evaluation of single T cell functions such as cytokine secretion and cytolysis of target cells is greatly needed in adoptive cell therapy (ACT) but has never been fully fulfilled by current approaches. Herein, we develop a hierarchical loading microwell chip (HL-Chip) that aligns multiple cells and functionalized beads in a high-throughput microwell array with single-cell/bead precision based on size differences. We demonstrate the potential of the HL-Chip in evaluating single T cell functions by three applications: high-throughput longitudinal secretory profiling of single T cells, large-scale evaluation of cytolytic activity of single T cells, and integrated T cell-tumor cell interactions. The HL-Chip is a simple and robust technology that constructs arrays of defined cell/object combinations for multiple measurements and material retrieval. • A hierarchical loading microwell chip (HL-Chip) pairs objects with high efficiency • A high-throughput assay enables single-cell cytokine detection and cell retrieval • The assay system enables simultaneous detection of cytotoxicity and cytokine • The high-throughput HL-Chip enables screening of functional T cells Zhou et al. describe the hierarchical loading microwell chip, which enables cell/bead pairing with high efficiency in a high-throughput manner. They conduct coupled measurements of cytotoxicity and cytokine secretion from single lymphocytes and uncover the linkage of lymphocytes' functions together with the cell behavior. [ABSTRACT FROM AUTHOR]

Background and Aims: We investigated PTEN expression in primary pancreatic cancer and pancreatic cancer liver metastasis in order to evaluate the interrelationship between PTEN expression and clinicopathological characteristics of pancreatic cancer patients with and without liver metastasis. Methods: Eighty five primary pancreatic cancer specimens without liver metastasis were analyzed as controls. Eighty seven pancreatic cancer specimens and homologous liver metastasis specimens were investigated immunohistochemically, and the correlation between immunohistochemical findings and clinicopathological factors was investigated. Results: A strong PTEN expression was observed in 52 (61.2%) specimens from patients without liver metastasis. In contrast, only 26 (29.9%) specimens were observed in patients with liver metastasis. A strong PTEN expression was apparently associated with low-grade lymph node metastasis (p <0.05) and TNM stage (p <0.05). PTEN expression in patients without liver metastasis was apparently stronger than that with liver metastasis. In addition, among patients with liver metastasis, the 5-year survival rate was markedly higher in patients with strong PTEN expression compared to those with weak PTEN expression. Conclusions: Our results suggest that a high level of PTEN expression is associated with low-grade liver metastasis and satisfactory patient survival in pancreatic cancer. The diagnostic evaluation of PTEN expression may provide valuable prognostic information to aid treatment strategies for pancreatic cancer patients. [ABSTRACT FROM AUTHOR]