Anti-PD-1 synergizes with cyclophosphamide to induce potent anti-tumor vaccine effects through novel mechanisms

Programmed death-1 receptor (PD-1) is expressed on T cells following TCR activation. Binding of this receptor to its cognate ligands, programmed death ligand (PDL)-1 and PDL-2, down-regulates signals by the TCR, promoting T-cell anergy and apoptosis, thus leading to immune suppression. Here, we find that using an anti-PD-1 antibody (CT-011) with Treg-cell depletion by low-dose cyclophosphamide (CPM), combined with a tumor vaccine, induces synergistic antigen-specific immune responses and reveals novel activities of each agent in this combination. This strategy led to complete regression of established tumors in a significant percentage of treated animals, with survival prolongation. We show for the first time that combining CT-011 and CPM significantly increases the number of vaccine-induced tumor-infiltrating CD8(+) T cells, with simultaneous decrease in infiltrating Treg cells. Interestingly, we find that CT-011 prolongs Treg-cell inhibition induced by CPM, leading to a sustainable significant synergistic decrease of splenic and tumor-infiltrated Treg cells. Surprisingly, we find that the anti-tumor effect elicited by the combination of CT-011 and CPM is dependent on both CD8(+) and CD4(+) T-cell responses, although the antigen we used is a class I MHC-restricted peptide. Thus, we describe a novel and effective therapeutic approach by combining multiple strategies to target several tumor-mediated immune inhibitory mechanisms.     

Tetrodotoxin-, dihydropyridine-, and riluzole-resistant persistent inward current: novel sodium channels in rodent spinal neurons

Recently, we reported the tetrodotoxin (TTX)- and dihydropyridine (DHP)-resistant (TDR) inward currents in neonatal mouse spinal neurons. In this study, we further characterized these currents in the presence of 1-5 μM TTX and 20-30 μM DHP (nifedipine, nimodipine, or isradipine). TDR inward currents were recorded by voltage ramp (persistent inward current, TDR-PIC) and step (TDR-I(p)) protocols. TDR-PIC and TDR-I(p) were found in 80.2% of recorded neurons (101/126) crossing laminae I to X from T12 to L6. TDR-PIC activated at -28.6 ± 13 mV with an amplitude of 80.6 ± 75 pA and time constant of 470.6 ± 240 ms (n = 75). TDR-I(p) had an amplitude of 151.2 ± 151 pA and a voltage threshold of -17.0 ± 9 mV (n = 54) with a wide range of kinetics parameters. The half-maximal activation was -21.5 ± 8 mV (-37 to -12 mV, n = 29) with a time constant of 5.2 ± 2 ms (1.2-11.2 ms, n = 19), whereas the half-maximal inactivation was -26.9 ± 9 mV (-39 to -18 mV, n = 14) with a time constant of 1.4 ± 0.4 s (0.5-2.2 s, n = 19). TDR-PIC and TDR-I(p) could be reduced by 60% in zero calcium and completely removed in zero sodium solutions, suggesting that they were mediated by sodium ions. Furthermore, the reversal potential of TDR-I(p) was estimated as 56.6 ± 3 mV (n = 10). TDR-PIC and TDR-I(p) persisted in 1-205 μM TTX, 20-100 μM DHP, 3-30 μM riluzole, 50-300 μM flufenamic acid, and 2-30 mM intracellular BAPTA. They also persisted with T-, N-, P/Q-, and R-type calcium channel blockers. In conclusion, we demonstrated novel TTX-, DHP-, and riluzole-resistant sodium channels in neonatal rodent spinal neurons. The unique pharmacological and electrophysiological properties would allow these channels to play a functional role in spinal motor system.     

Low-dose megavoltage cone-beam CT imaging using thick, segmented scintillators

Megavoltage, cone-beam computed tomography (MV CBCT) employing an electronic portal imaging device (EPID) is a highly promising technique for providing soft-tissue visualization in image-guided radiotherapy. However, current EPIDs based on active matrix flat-panel imagers (AMFPIs), which are regarded as the gold standard for portal imaging and referred to as conventional MV AMFPIs, require high radiation doses to achieve this goal due to poor x-ray detection efficiency (～2% at 6 MV). To overcome this limitation, the incorporation of thick, segmented, crystalline scintillators, as a replacement for the phosphor screens used in these AMFPIs, has been shown to significantly improve the detective quantum efficiency (DQE) performance, leading to improved image quality for projection imaging at low dose. Toward the realization of practical AMFPIs capable of low dose, soft-tissue visualization using MV CBCT imaging, two prototype AMFPIs incorporating segmented scintillators with ～11 mm thick CsI:Tl and Bi(4)Ge(3)O(12) (BGO) crystals were evaluated. Each scintillator consists of 120 × 60 crystalline elements separated by reflective septal walls, with an element-to-element pitch of 1.016 mm. The prototypes were evaluated using a bench-top CBCT system, allowing the acquisition of 180 projection, 360° tomographic scans with a 6 MV radiotherapy photon beam. Reconstructed images of a spatial resolution phantom, as well as of a water-equivalent phantom, embedded with tissue equivalent objects having electron densities (relative to water) varying from ～0.28 to ～1.70, were obtained down to one beam pulse per projection image, corresponding to a scan dose of ～4 cGy--a dose similar to that required for a single portal image obtained from a conventional MV AMFPI. By virtue of their significantly improved DQE, the prototypes provided low contrast visualization, allowing clear delineation of an object with an electron density difference of ～2.76%. Results of contrast, noise and contrast-to-noise ratio are presented as a function of dose and compared to those from a conventional MV AMFPI.     

Self-Efficacy and Self-Care: Missing Ingredients in Health and Healthcare among Adults with Serious Mental Illnesses

To help inform the design of a self-management intervention for improving the physical health of adults with serious mental illnesses, we conducted focus groups about their perceived medical care and physical health needs. Adults with serious mental illnesses participated in four semi-structured focus groups conducted at a transitional living facility, a social club, and a Hispanic outpatient mental health clinic. Questions included their recent experiences of seeking medical care, the effect of having a mental illnesses diagnosis, strategies for active self-care, and perceived barriers to better physical health. In addition to various systemic barriers to better medical care, participants articulated limited knowledge and self-efficacy regarding active self-management of their physical health. Despite their interest in learning more about health promotion, most participants expressed a sense of personal futility and powerlessness in improving their health. These data suggest that any effort to improve the wellbeing of these adults will need to address self-efficacy in the hope of improving self-care for their physical health needs.