RESUMEN
Cell Squeeze is a novel technology that relies on temporarily disrupting the cell membrane to deliver cargo directly into the cytosol. This approach is applicable to a broad range of cell types (peripheral blood mononuclear cells, red blood cells, hematopoietic stem cells, etc.) and cargos (peptides, proteins, small molecules, nucleic acids, and gene-editing complexes) while minimally disrupting normal cell function. By enabling direct cytosolic delivery, one can use this technology to dramatically enhance major histocompatibility complex (MHC) class I presentation of antigens (Ags) for CD8+ T-cell activation-a longstanding challenge for the therapeutic cancer vaccine field that has generally relied on cross-presentation of endocytosed Ags. In addition, by coupling improved MHC class I presentation with coexpression of additional stimulatory factors or systemic immune modulators, one can further enhance the potential impact of an antitumor CD8 response. Pursuing a more direct cellular engineering strategy, which is independent of viral transduction, genetic manipulation, and expansion steps, enables <24 h manufacturing of autologous cell therapies. Through generation of more sophisticated, multifunctional, cell-based vaccines, clinical testing of this technology will elucidate its potential for impact across multiple tumor types.
RESUMEN
Steady-state photosynthetic responses to leaf temperature of 4-year-old Eucalyptus globulus Labill. and E. nitens (Deane and Maiden) Maiden trees were measured between 10 and 35 degrees C at approximately monthly intervals from early spring to midwinter. The photosynthetic temperature optimum of recently expanded leaves in the sun canopy was linearly related to the average temperature of the preceding week during the 9-month measurement period. The optimum temperature for net photosynthesis of E. globulus increased from 17 to 23 degrees C as the mean daily temperature increased from 7 to 16 degrees C. Similarly, the optimum temperature for net photosynthesis of E. nitens increased from 14 to 20 degrees C as the mean daily temperature increased from 7 to 19 degrees C. The temperature for maximum photosynthetic response of E. globulus and E. nitens was similar at each measurement time, but the photosynthetic performance of E. nitens was less sensitive to temperatures above and below this optimum than that of E. globulus. In December, the apical shoots of branches of E. globulus had a net photosynthetic temperature optimum of between 10 and 15 degrees C. The corresponding values for expanding leaves, fully expanded leaves from the current year's growth, and fully expanded leaves from the previous year's growth were 15, 20 and 20-25 degrees C, respectively. In a second experiment, E. globulus clones taken from four mother plants originating from climatically dissimilar locations within Tasmania were acclimated at day/night temperatures of 10/15, 18/23 and 25/30 degrees C in temperature-controlled greenhouses. Another set of clones was acclimated in a shadehouse where temperatures ranged between 10 and 25 degrees C and with a mean daily temperature of approximately 15 degrees C. Plants grown at 25/30 degrees C had significantly lower net photosynthetic rates when measured at 10 and 20 degrees C than plants grown at lower temperatures. Plants grown at 10/15 degrees C had significantly lower net photosynthetic rates when measured at 30 degrees C than plants grown at higher temperatures. Plants grown at the ambient conditions prevailing in midautumn in Hobart had significantly higher net photosynthetic rates at 20 degrees C than plants raised in the greenhouses and were equal best performers at 10 and 30 degrees C. A comparison of the light response curves of the plants showed that the maximum rate of net photosynthesis was affected by the growth temperature, whereas the apparent quantum efficiency remained unchanged. There were no significant differences in the photosynthetic temperature responses of the four genotypes derived from climatically dissimilar locations within Tasmania. A comparison of temperature response models for E. globulus indicated that incomplete acclimation (defined by a slope value of less than 1 for the linear relationship between the temperature optimum for photosynthesis and the growth temperature) generally resulted in a greater daily carbon uptake than complete acclimation (slope value of 1).