Citations

Abstract Introduction: Immune suppression is a major component of lymphoma pathogenesis. Recent advances in lymphoma treatment have been immunotherapy. We present the preliminary results of our clinical trial testing 2 vaccine strategies in patients with indolent B-cell non-Hodgkin lymphoma (NHL). The primary objective is to determine the safety and feasibility of the vaccine approaches and secondary objectives are to describe clinical responses and identify corresponding immune changes. Methods: Autologous mature dendritic cells (mDC) were manufactured from leukapheresed cells of NHL pts. For patients with tumor lymph nodes deemed amenable to cryoablation by interventional radiologist (arm A), they received cryoablation of a node and injection of mDC into the cryoablated node followed by another 1 to 7 intratumoral mDC injections. Remaining patients had a tumor excised to generate tumor lysate ex vivo. mDC were pulsed with tumor lysate during DC maturation (arm B; DC-TL). The DC-TL vaccines were injected intradermally for 4 to 8 doses. Patients are monitored for one year after vaccines for adverse events and systemic tumor response. Correlative studies include cellular immune phenotype of peripheral blood and T cell intracellular cytokine productions. Planned accrual is 10 patients per arm (total = 20). Results: To date, 10 patients have accrued to arm A and 5 patients to arm B (Table 1). All pts tolerated vaccine treatments without major adverse events. Of the 10 evaluable patients, there were 1 CR (arm B: 1 / 4; total: 1/6) and 4 PR (arm A: 3/6; arm B: 1 / 4; total: 4/10) for an ORR of 50% for both arms (Figure 1). Time to next treatment ranged from 6.5 to 25 months (Figure 2). Correlative studies suggest that immune changes can be used as prognostic biomarkers to predict response. These include changes in vaccine induced cytokine secretion, and in the number and composition of the immune system. In addition, preliminary analysis of >40 immune phenotypes using flow cytometry and hierarchical clustering suggest there are characteristic immune system responses in patients responding to vaccine. Conclusions: Both cryoablation and intratumoral mDC vaccination are feasible and safe in NHL. Treatment responses may correlate with immune system changes. Biosystems analysis method can be used to develop novel assays as predictive biomarkers of treatment response and used to develop novel protocols that allow treating to response. Table 1. Patient Demographics Arm A (n = 10) Arm B (n = 5) Total (n = 15) Age (median, rage) 64 (55 – 80) 60 (31 – 86) 63 (31 – 86) Gender (M/F) 6/4 2/3 8/7 Histology (n, %) Follicular Marginal zone DLBCL 10 (100%) 0 (0%) 0 (0%) 3 (60%) 1 (20%) 1(20%) 13 (86.6%) 1 (6.7%) 1 (6.7%) Stage (N, %) II III IV 1 (10%) 1 (10%) 8 (80%) 2 (40%) 1 (20%) 2 (40%) 3 (20%) 2 (13.3%) 10 (66.7%) No of prior Tx (mean, range) 2.4 (0-8) 3.2 (0-7) 2.7 (0-8) FLIPI (n, %) 0-1 2 ≥ 3 1 (10%) 2 (20%) 7 (70%) 1 (20%) 2 (40%) 2 (45%) 2 (13.3%) 4 (26.7%) 9 (60%) Positive GELF criteria (n, %) 4 (40%) 1 (20%) 5 (30%) Figure 1. Waterfall plot. Figure 1. Waterfall plot. Figure 2. Time to next treatment. Figure 2. Time to next treatment. Disclosures No relevant conflicts of interest to declare.

Abstract Abstract 970 Despite advances in treatments for patients with non-Hodgkin's lymphoma (NHL), the relapse rates remain high and 40% of diffuse large B-cell NHL (DLBCL) patients die of disease. These patients have variable levels of immunodeficiency that may impact their prognosis. Previously we reported that monocytes in relapsed NHL patients suppress both autologous and allogeneic T cell proliferation and that these monocytes were unable to mature into dendritic cells in vitro. These suppressive functions were associated with increased frequency of CD14+HLA-DRneg monocytes in NHL patients (NHL: 29.4 ± 3.2%, n = 36; healthy donors 7.54 ± 0.97%, n = 27; p < 0.0001). This phenotype is distinct from other myeloid suppressors (Lin−CD33+HLA-DR−) or non-classical monocytes (CD16+), neither of which was different in numbers between NHL and healthy donors. We have now expanded our study cohort and found that the percentage of CD14+HLA-DRneg cells among circulating monocytes was increased in patients with more aggressive disease: DLBCL 37.5 ± 5.11%, n = 16 vs SLL and MALT lymphoma 15.8 ± 4.02%, n = 7; p = 0.01. Two newly diagnosed patients and three patients with localized disease amenable to resection all had frequencies of CD14+HLA-DRneg monocytes closer to that of healthy donors whereas 5 of the 6 patients with low grade NHL that transformed to DLBCL had significantly elevated percentage of these suppressive monocytes (49.1 ± 4.70%). To better understand the mechanism of immune suppression by CD14+HLA-DRneg monocytes, we investigated production of IFN-alpha induced by CpG oligodeoxynucleotide. Monocytes from NHL patients were less effective at IFN-alpha production (0.54 ± 0.22%, n = 4) compared to healthy donors (1.37 ± 0.25%, n = 4; p = 0.04). The corresponding culture supernatants also had a 4-fold reduction in IFN-alpha level by ELISA (p = 0.01). Reduced IFN-alpha production and subsequent decreased stimulatory response may contribute to diminished treatment response to CpG in NHL. Finally, we also found that the frequency of CD14+HLA-DRneg monocytes correlated with increasing plasma levels of arginase I, a known mediator of T cell suppression (n = 16, p = 0.06). Such correlation was not observed for circulating VEGF, IL-6 or IL-10. Taken together we have described a population of immunosuppressive CD14+HLA-DRneg monocytes in NHL patients that is more prevalent in aggressive disease and likely suppresses T cell function through multiple mechanisms including arginine metabolism. This work was supported through the University of Iowa and Mayo Clinic Lymphoma SPORE CA097274 and the Gerstner Family Foundation. Disclosures: Witzig: Novartis: Research Funding.

Purpose The aim of this study was to examine 2 aspects of treatment intensity in treatment for childhood apraxia of speech (CAS): practice amount and practice distribution. Method Using an alternating-treatments single-subject design with multiple baselines, we compared high versus low amount of practice, and massed versus distributed practice, in 6 children with CAS. Conditions were manipulated in the context of integral stimulation treatment. Changes in perceptual accuracy, scored by blinded analysts, were quantified with effect sizes. Results Four children showed an advantage for high amount of practice, 1 showed an opposite effect, and 1 showed no condition difference. For distribution, 4 children showed a clear advantage for massed over distributed practice post treatment; 1 showed an opposite pattern, and 1 showed no clear difference. Follow-up revealed a similar pattern. All children demonstrated treatment effects (larger gains for treated than untreated items). Conclusions High practice amount and massed practice were associated with more robust speech motor learning in most children with CAS, compared to low amount and distributed practice, respectively. Variation in effects across children warrants further research to determine factors that predict optimal treatment conditions. Finally, this study adds to the evidence base supporting the efficacy of integral stimulation treatment for CAS. Supplemental Material https://doi.org/10.23641/asha.9630599

ABSTRACT The last decade has seen a sharp increase in the number of scientific publications describing physiological and pathological functions of extracellular vesicles (EVs), a collective term covering various subtypes of cell‐released, membranous structures, called exosomes, microvesicles, microparticles, ectosomes, oncosomes, apoptotic bodies, and many other names. However, specific issues arise when working with these entities, whose size and amount often make them difficult to obtain as relatively pure preparations, and to characterize properly. The International Society for Extracellular Vesicles (ISEV) proposed Minimal Information for Studies of Extracellular Vesicles (“MISEV”) guidelines for the field in 2014. We now update these “MISEV2014” guidelines based on evolution of the collective knowledge in the last four years. An important point to consider is that ascribing a specific function to EVs in general, or to subtypes of EVs, requires reporting of specific information beyond mere description of function in a crude, potentially contaminated, and heterogeneous preparation. For example, claims that exosomes are endowed with exquisite and specific activities remain difficult to support experimentally, given our still limited knowledge of their specific molecular machineries of biogenesis and release, as compared with other biophysically similar EVs. The MISEV2018 guidelines include tables and outlines of suggested protocols and steps to follow to document specific EV‐associated functional activities. Finally, a checklist is provided with summaries of key points.

Background Platelet polyphosphate is an inorganic procoagulant polymer of orthophosphate units that is stored in dense granules and is released upon platelet activation. Here, we describe an assay to measure polyphosphate on the surface of procoagulant human platelets. Methods and Results Recombinant Escherichia coli ‐expressed exopolyphosphatase deletion mutant PPX_Δ12 labeled with fluorescent Alexa488 dye was used as a polyphosphate probe in flow cytometry. PPX_Δ12‐Alexa488‐signal dose‐dependently increased with long‐chain polyphosphate binding to platelets. In contrast, short‐chain polyphosphate that is found in the supernatant of activated platelets, did not bind to the platelet surface. Both exopolyphosphatase treatment and polyphosphate pre‐incubation abolished PPX_Δ12‐Alexa488 binding to polyphosphate on platelets. Stimulation of platelets with thrombin receptor agonist Trap6, and P2Y 12 receptor activator ADP increased polyphosphate accumulation on platelet surfaces and PPX_Δ12‐Alexa488 signal in a dose‐dependent manner. Conclusion This study indicates that long‐chain polyphosphate binds to platelet plasma membranes and presents a promising diagnostic assay to measure this interaction on human platelets in platelet‐rich plasma. Future investigations will aim to determine if polyphosphate can be used as a novel biomarker of thrombosis. © 2016 International Clinical Cytometry Society

Objectives The aim was for the appointed committee of the Academy of Neurological Communication Disorders and Sciences to conduct a systematic review of published intervention studies of acquired apraxia of speech (AOS), updating the previous committee's review article from 2006. Method A systematic search of 11 databases identified 215 articles, with 26 meeting inclusion criteria of (a) stating intention to measure effects of treatment on AOS and (b) data representing treatment effects for at least 1 individual stated to have AOS. Results All studies involved within-participant experimental designs, with sample sizes of 1 to 44 (median = 1). Confidence in diagnosis was rated high to reasonable in 18 of 26 studies. Most studies (24/26) reported on articulatory–kinematic approaches; 2 applied rhythm/rate control methods. Six studies had sufficient experimental control for Class III rating according to the Clinical Practice Guidelines Process Manual (American Academy of Neurology, 2011), with 15 others satisfying all criteria for Class III except use of independent or objective outcome measurement. Conclusions The most important global clinical conclusion from this review is that the weight of evidence supports a strong effect for both articulatory–kinematic and rate/rhythm approaches to AOS treatment. The quantity of work, experimental rigor, and reporting of diagnostic criteria continue to improve and strengthen confidence in the corpus of research.

Abstract This study examined habitat variables associated with 53 active bald eagle Haliaeetus leucocephalus nest sites in the Winona District of the Upper Mississippi River National Wildlife and Fish Refuge, Minnesota and Wisconsin. The Refuge is the most heavily visited refuge in the United States, where breeding eagle populations have been increasing dramatically. During February–April 2009, nest trees were identified and measured, nest heights were determined, distances to nearest water bodies were assessed, and forest inventories were conducted for the standing timber surrounding the nest trees. Nest densities and spacing were assessed within each navigation pool, and land cover types were examined within 100- and 1,000-m radii around known eagle nest sites and random points within the Refuge. Ninety-three percent of nest sites had supercanopy eastern cottonwoods Populus deltoides and silver maples Acer saccharinum as the nest trees. Potential human disturbances from highways, railroads, and commercial barge and recreational boat traffic were present within 400 m of 90% of known nest sites. Eagle nest sites were located an average of 1.52 km from the next nearest nest, with nest densities ranging from 0.32 to 9.72 nests/100 km2 among the four navigation pools of the Winona District. Land cover types around known nest sites and random points differed significantly at both 100- and 1,000 m scales, with wet forest and open water significantly more abundant and agricultural and developed lands significantly less abundant than around randomly selected points. Successful nests that fledged at least one young were spaced significantly further away from other active nests and were located in areas with lower tree density than were unsuccessful nests. Floodplain-nesting bald eagles tended to select the tallest, dominant trees for nest sites, placing nests near the height of the surrounding canopy. Human presence within the Refuge does not appear to be limiting the expansion of nesting bald eagles in this riverine habitat.