Traumatic Spondylopelvic Dissociation: A Report of Two Cases of Spondylolisthesis at L5-S1 and Review of Literature.

Study Design Retrospective chart review and review of literature. Objective Few case reports of traumatic L5-S1 displacement have been presented in the literature. Here we present two cases of traumatic spondylolisthesis showing both anterior and posterior displacement, the treatment algorithm, and a review of the literature. Methods The authors conducted a retrospective review of representative patients and a literature review of traumatic spondylolisthesis at the L5-S1 junction. Two representative patients were identified with traumatic spondylolisthesis: one with an anterior dissociation, and the other with a posterior dissociation. Results Radiographic, computed tomography, and magnetic resonance imaging illustrated the bony and soft tissue injury found in each patient, as well as the final stabilization and outcomes. Operative stabilization was necessary, and both patients were treated with open reduction internal fixation. The patient with posterior dissociation had complete recovery without neurologic sequelae. The patient with anterior dissociation had persistent bilateral L5-S1 radiculopathy with intact rectal tone, due to neurologic compression. Conclusions Few cases of traumatic spondylopelvic dissociation that are isolated to the L5-S1 disk space are described in the literature. We examined both an anterior and a posterior dissociation and treated both with L5-S1 posterior spinal fusion. The patient with anterior dissociation had persistent L5-S1 root injury; however, the patient with posterior dissociation had no neurologic deficits. This is the opposite of what is expected based on anatomy. These cases offer insight into the management of anterior and posterior L5-S1 spondylopelvic dissociation

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

Ultrasonographic and Radiographic Findings of Polyethylene Component Displacement with Severe Metallosis and Metal-Induced Synovitis Following Total Knee Arthroplasty

Aseptic loosening and wear of arthroplasty is second to only infection as the most common complication of arthroplasty failure. Degeneration of the polyethylene and metal arthroplasty components can lead to metallosis, which can cause a combination of direct cytotoxic effects and an inflammatory response within the synovial and periarticular tissues. This can result in bone resorption and secondary arthroplasty component loosening as well as a metal containing joint effusion and metal-induced synovitis. Little literature exists as to the ultrasonographic findings of metal-induced synovitis and polyethylene component displacement. As the use of musculoskeletal ultrasound significantly increases, being aware of these findings is important. The following is a case report that demonstrates the ultrasonographic imaging findings of metallosis, metal-induced synovitis and polyethylene component displacement. We will also demonstrate the ultrasound-guided aspiration findings as well as radiographic and gross pathologic correlations.https://scholarlycommons.henryford.com/merf2019caserpt/1081/thumbnail.jp

Ultrasonographic and radiographic findings of polyethylene component displacement with severe metallosis and metal-induced synovitis following total knee arthroplasty

Aseptic loosening and wear is second to only infection as the most common cause of arthroplasty failure. Degeneration of the polyethylene and metal arthroplasty components can lead to metallosis, which can cause a combination of direct cytotoxic effects and an inflammatory response within the synovial and periarticular tissues. This can result in bone resorption and secondary arthroplasty component loosening as well as a metal containing joint effusion and metal-induced synovitis. Little literature exists as to the ultrasonographic findings of metal-induced synovitis and polyethylene component displacement. As the use of musculoskeletal ultrasound significantly increases, being aware of these findings is important. The most important ultrasonographic findings include differentiating a joint effusion from synovitis utilizing dynamic compression, identifying areas of echogenic shadowing related to metal deposition and visualizing displaced arthroplasty components. The following is a case report that demonstrates the ultrasonographic imaging findings of metallosis, metal-induced synovitis and polyethylene component displacement. We will also demonstrate the ultrasound-guided aspiration findings as well as radiographic and gross pathologic correlations

Nanoelectroablation of Murine Tumors Triggers a CD8-Dependent Inhibition of Secondary Tumor Growth.

We have used both a rat orthotopic hepatocellular carcinoma model and a mouse allograft tumor model to study liver tumor ablation with nanosecond pulsed electric fields (nsPEF). We confirm that nsPEF treatment triggers apoptosis in rat liver tumor cells as indicated by the appearance of cleaved caspase 3 and 9 within two hours after treatment. Furthermore we provide evidence that nsPEF treatment leads to the translocation of calreticulin (CRT) to the cell surface which is considered a damage-associated molecular pattern indicative of immunogenic cell death. We provide direct evidence that nanoelectroablation triggers a CD8-dependent inhibition of secondary tumor growth by comparing the growth rate of secondary orthotopic liver tumors in nsPEF-treated rats with that in nsPEF-treated rats depleted of CD8+ cytotoxic T-cells. The growth of these secondary tumors was severely inhibited as compared to tumor growth in CD8-depleated rats, with their average size only 3% of the primary tumor size after the same one-week growth period. In contrast, when we depleted CD8+ T-cells the second tumor grew more robustly, reaching 54% of the size of the first tumor. In addition, we demonstrate with immunohistochemistry that CD8+ T-cells are highly enriched in the secondary tumors exhibiting slow growth. We also showed that vaccinating mice with nsPEF-treated isogenic tumor cells stimulates an immune response that inhibits the growth of secondary tumors in a CD8+-dependent manner. We conclude that nanoelectroablation triggers the production of CD8+ cytotoxic T-cells resulting in the inhibition of secondary tumor growth

Nano-Pulse Stimulation is a physical modality that can trigger immunogenic tumor cell death

Abstract Background We have been developing a non-thermal, drug-free tumor therapy called Nano-Pulse Stimulation (NPS) that delivers ultrashort electric pulses to tumor cells which eliminates the tumor and inhibits secondary tumor growth. We hypothesized that the mechanism for inhibiting secondary tumor growth involves stimulating an adaptive immune response via an immunogenic form of apoptosis, commonly known as immunogenic cell death (ICD). ICD is characterized by the emission of danger-associated molecular patterns (DAMPs) that serve to recruit immune cells to the site of the tumor. Here we present evidence that NPS stimulates both caspase 3/7 activation indicative of apoptosis, as well as the emission of three critical DAMPs: ecto-calreticulin (CRT), ATP and HMGB1. Methods After treating three separate cancer cell lines (MCA205, McA-RH7777, Jurkat E6-1) with NPS, cells were incubated at 37 °C. Cell-culture supernatants were collected after three-hours to measure for activated caspases 3/7 and after 24 h to measure CRT, ATP and HMGB1 levels. We measured the changes in caspase-3 activation with Caspase-Glo® by Promega, ecto-CRT with anti-CRT antibody and flow cytometry, ATP by luciferase light generation and HMGB1 by ELISA. Results The initiation of apoptosis in cultured cells is greatest at 15 kV/cm and requires 50 A/cm2. Reducing this current inhibits cell death. Activated caspase-3 increases 8-fold in Jurkat E6-1 cells and 40% in rat hepatocellular carcinoma and mouse fibrosarcoma cells by 3 h post treatment. This increase is non-linear and peaks at 15–20 J/mL for all field strengths. 10 and 30 kV/cm fields exhibited the lowest response and the 12 and 15 kV/cm fields stimulated the largest amount of caspase activation. We measured the three DAMPs 24 h after treatment. The expression of cell surface CRT increased in an energy-dependent manner in the NPS treated samples. Expression levels reached or exceeded the expression levels in the majority of the anthracycline-treated samples at energies between 25 and 50 J/mL. Similar to the caspase response at 3 h, secreted ATP peaked at 15 J/mL and then rapidly declined at 25 J/mL. HMGB1 release increased as treatment energy increased and reached levels comparable to the anthracycline-treated groups between 10 and 25 J/mL. Conclusion Nano-Pulse Stimulation treatment at specific energies was able to trigger the emission of three key DAMPs at levels comparable to Doxorubicin and Mitoxantrone, two known inducers of immunogenic cell death (ICD). Therefore NPS is a physical modality that can trigger immunogenic cell death in tumor cells

Cellular and genomic disease signature of peripheral blood mononuclear cells in patients with malignant pleural mesothelioma

Background: Recent data on the incidence malignant pleural mesothelioma (MPM) and the continued large-scale use of asbestos throughout the developing world portends an epidemic of asbestos-related disease. MPM is an aggressive and fatal cancer with few treatment options. Recent advances in large scale genomic and high throughput cellular analyses now provide the tools to more easily attain markers of disease status and potential responsiveness to immunotherapeutics. Materials and Methods: Here we present pre-treatment cellular and genomic biomarker data on a cohort of chemotherapy-naïve MPM patients, and demographically matched healthy donors (HD). MPM patients were enrolled in a Phase 1b study utilizing CRS-207, a live, attenuated Listeria monocytogenes strain engineered to express the tumor-associated antigen, mesothelin. Four different multi-color flow cytometry panels were used to provide resolution on major immune cell populations of T cells, γδ T cells, B Cells, dendritic cells, monocytes, and natural killer cells. Together, these panels provided deeper resolution on 39 distinct subpopulations of major immune cell subsets. RNA from these cells was used to perform multiplex gene expression analysis on 770 genes using the Nanostring nCounter PanCancer Pathway Panel. Results: FACS analysis yielded numerous subpopulations with statistically significant differences between MPM patients and healthy controls. Differences in immune populations were analyzed by median and significant findings included populations of CD4+ T cells, CD8+ T cells, B cells, classical monocytes, and monocytic myeloid derived suppressor cells*. Class comparison and hierarchical clustering of gene expression data revealed genomic markers that were significantly expressed in MPM compared to healthy controls. Immune subset deconvolution of gene expression data provided similar findings as FACS analysis and corroborated this disease signature across experimental platforms. Conclusions: Understanding a patient’s biological disease signature can aide in diagnosis, as well as in making informed choices about therapies amidst the complex and broadening immunotherapeutic landscape. Until recently, existing biomarker data in MPM has been limited to a small number of serological markers and limited immune analysis. Here, we present the first comprehensive report of a MPM disease signature from the cellular and genomic perspectives. Correlation of patient baseline disease signatures with treatment outcome may yield biomarkers predictive of treatment efficacy. Predictive signatures are being investigated in the on-going Phase 1b study of CRS-207 and chemotherapy, as well as in the Phase 2 study of CRS-207 with pembrolizumab in MPM patients who failed prior treatment. *Inclusion of additional subjects confirmed the significance of all immune cell subsets except for MDSCs

Comparison of 1st and 2^nd tumor surface areas in controls (A) and CD8⁺-depleted rats (B).

<p>We took blood samples from each rat and counted the number of both CD4⁺ and CD8⁺ cells. The percentage of CD8⁺ cells in this total is plotted for each tumor pair. We plotted the tumor surface area on a log scale to highlight the dramatic increase in the rate of growth of the second tumor when CD8⁺ cells were depleted. In two of the controls, we could not find the second tumor so the mean tumor size was calculated from the four visible second tumors. A Mann-Whitney Rank Sum Test indicates a statistically significant difference between the secondary tumor sizes in these two groups. (P = 0.03).</p

Transillumination images of subdermal MCA205 tumors in isogenic B6 albino mice.

<p>Tumors are outlined with dotted lines to aid in visualization. Two top rows show tumors growing from cells injected into mice vaccinated 3 weeks earlier with nsPEF-treated MCA205 cells. The 3^rd row down shows a tumor growing from cells injected into a mouse vaccinated with mitomycin C-treated cells. The fourth row down shows a tumor growing from cells injected into a mouse vaccinated with saline vehicle. The bottom row shows a tumor growing from cells injected into a mouse vaccinated with nsPEF-treated cells one day after injecting anti-CD8 antibodies IP.</p