Phase 1 Trial of Vaccination with Autologous Tumor Cells and Antisense Directed Against the Insulin Growth Factor Type 1 Receptor (IGF-1R AS ODN) in Patients with Recurrent Glioblastoma

Background: Extending a previous Phase I study, we report the results of a second Phase I autologous tumor cell vaccination trial for patients with recurrent glioblastomas (IND 14379-101, NCT01550523). Methods: Following surgery, subjects were treated by 24 hour implantation in the rectus sheath of ten biodiffusion chambers containing irradiated autologous tumor cells and IGF-1R AS ODN with the objective of stimulating tumor immunity. Patients were monitored for safety, clinical and radiographic as well as immune responses. Results: There were no Grade 3 toxicities related to protocol treatment and overall median survival from initial diagnosis was 91.4 weeks. Two protocol survival cohorts with median survivals of 48.2 and 10 weeks were identified and predicted by our pre-treatment assessments of immune function, corroborated by post-vaccination pro-inflammatory cytokine profiles. Longer survival subjects had imaging findings including transient elevations in cerebral blood volume (rCBV) and sustained elevations of apparent diffusion coefficient (ADC) interpreted as transient hyperemia and cell loss. Conclusions: The vaccine paradigm was well-tolerated with a favorable median survival. Our data support this as a novel treatment paradigm that promotes anti-tumor immunity

Cerebrospinal Fluid Cytokine and Chemokine Patterns in Central Nervous System Infections, Hemorrhage and Neoplasms

Cytokines and chemokines are soluble proteins that act as regulators of cellular functions throughout the body. Cytokines and chemokines released in the setting of various CNS disorders appear in the CSF compartment where determination of their levels can provide insight into pathogenic processes such as neuroinflammation. We utilized the Millipore HCYTOMAG 60K assay/kit/system to perform multiplex profiling of 42 different cytokines/chemokines in the CSF of patients with a variety of distinct CNS disease processes, including infection, hemorrhage and neoplasia. CNS infections included viral (Chronic Parechovirus type 3 (HPeV3), Enterovirus (EV) 68, Adenovirus, JC virus, West Nile virus), bacterial (Mycobacterium tuberculosis, Borrelia burgdorferi, Propionibacterium acnes, Staphylococcus epidermidis, Streptococcus sp.), fungal (Cryptococcus neoformans) and single celled parasite (Toxoplasma gondii). CSF specimens negative for infectious organisms in noninflammatory conditions were selected as controls. Additional non-infectious samples tested were obtained from patients with subarachnoid hemorrhage (SAH) and following surgery for glioblastoma. The glioblastoma samples were noteworthy in having negligible elevations in the cytokines/chemokines tested. CSF from patients with SAH was elevated in only MCP-1/CCL2. Distinct patterns of cytokine/chemokine expression were detected for each infectious patient population. Picornavirus infections HPeV3 and EV68 were associated with increased levels of the monocyte chemoattractant protein MCP-1/CCL2 when compared to non-infectious, non-inflammatory samples. In contrast to chronic HPeV3 infection, EV68 encephalitis was associated with increased CSF levels of additional cytokines; CCLX1, IL-4 and IL-7. Adenovirus infection was associated with markedly higher levels of fractalkine in CSF when compared to any of the other non-inflammatory, infectious, hemorrhage or tumor cases. CSF from a Mycobacterium tuberculosis infection demonstrated increased levels of a greater variety of cytokines/chemokines than any of the other groups tested. Patterns of cytokine/chemokine expression in the CNS reveal characteristics of the host innate response that provide insight into the disease process and potential targets for therapeutic intervention

Human Parechovirus and Enterovirus Initiate Divergent Innate Immune Responses in the CNS: Pathogenic and Diagnostic Implications

The picornaviruses human parechovirus (HPeV) and enterovirus (EV) cause a wide range of diseases, including CNS infections, which can be severe and potentially fatal. EV causes most cases of pediatric meningoencephalitis worldwide, and HPeV type 3 (HPeV3) is the most common cause of viral meningitis in young infants. Each year in the United States, there are over 75,000 cases of aseptic meningitis. Despite reassuring short-term outcomes, negative neurodevelopmental sequalae are increasingly associated with HPeV and EV. The pathogenesis and severity of HPeV and EV infections are undoubtedly linked to the innate and adaptive immune responses elicited by these viruses. Until this work, the innate immune response mounted against HPeV was largely unknown. Pattern recognition receptors in the CNS, including a number of Toll-like receptors located in different cells and subcellular compartments, detect invading pathogens and cause the release of cytokines and chemokines almost immediately into the CSF compartment at measurable levels. Essentially, this allows for determination of an amplified, infectious agent-specific pattern. These virus specific patterns of innate immune activation may provide insight into the pathogenesis of the corresponding disease states. Also, since these infections have similar clinical presentations, the immune profiles may be useful for rapid pathogen diagnosis in the clinical setting

High Density of Tumor-Associated Macrophage Staining Correlates with Poor Clinicopathologic Markers in Head and Neck Squamous Cell Carcinoma: A Meta-Analysis

Background: Head and neck squamous cell carcinoma (HNSCC) develops within a complex cellular microenvironment that promotes tumor growth, but also represents many potential therapeutic targets. Macrophage presence within that environment has been implicated in the growth, aggression, and persistence of HNSCC. Current literature reports variable degrees of association between tumor-associated macrophage (TAMs) density and clinicopathologic markers of disease.Inconsistent findings may result from grouping of TAM subtypes, which include both M1 (pro-inflammatory) and M2 (immunosuppressive). Our aim is to define the prognostic significance of the phenotypes of tumor-associated macrophages in HNSCC. Methods: We conducted a meta-analysis of the existing publications investigating the relationship between TAMs (total and M2 subtype) and T stage, nodal involvement, vascular invasion, lymphatic invasion, and tumor differentiation. Forest plots and risk ratios were generated to report overall effect. Results: Higher density of both total and M2 subtype of TAMs in the tumor microenvironment is associated with advanced T stage, increased rates of nodal positivity, presence of vascular invasion, and presence of lymphatic invasion (p \u3c 0.0001). There is no significant association between either total or M2 TAM density and tumor differentiation. Conclusion: Increased density of TAMs, including those of the M2 phenotype, correlates with poor clinicopathologic markers in HNSCC, and therefore poor clinical prognosis. It is unknown whether this relationship is causative or correlative. Additional investigation into the mechanisms behind TAM recruitment and differentiation, and effect of TAM population manipulation on tumor behavior will help define the feasibility of TAM-targeted therapies

Quantification of CSF chemokines and cytokines allows for rapid laboratory detection of CNS infections and further discrimination between viral and non-viral pathogens

Background: Prompt diagnosis of central nervous system (CNS) disease is critical to guide intervention and appropriate therapy. Development of novel laboratory approaches to rapidly classify acute-onset CNS disease is in great demand. Serious microbial pathogens, especially viruses, are quickly expanding beyond their historic geographic range and may not even be considered in the clinician’s differential diagnosis. Unlike bacterial cultures, current viral testing targets a limited number of viruses. Additionally, despite diversity in etiology, signs and symptoms of both infectious and non-infectious CNS disorders can be remarkably similar, which can confuse the clinical picture and delay treatment. Bacterial, viral, fungal and protozoan CNS pathogens are sensed by pattern recognition receptors of the immune system, stimulating immediate release of measurable levels of chemokines and cytokines into the CSF. Our objective is to use pathogen-specific chemokine/ cytokine profiles to classify CNS disease as infectious versus non-infectious and further discriminate between viral and non-viral infections. Methods: Levels (pg/ml) of chemokines and cytokines were determined in the CSF of 45 patients with documented infectious meningitis or meningoencephalitis (mean age 19.2 years) and in the CSF of 25 patients who were negative for CNS infection (mean age 27.4 years). MILLIPLEX MAP Human Cytokine/Chemokine Magnetic Bead Panels (Millipore) were used to measure CSF chemokines and cytokines levels (pg/ml). Innate immune analytes quantified included IP-10 (CXCL10), IFNg, IL-15, MDC (CCL22), MCP-1 (CCL2), Fractalkine, and FLT3L. Samples were analyzed in duplicate by a FlexMAP 3D (Luminex). Standard curves were generated for each cytokine and median fluorescent intensities were transformed into concentrations by 5-point, non-linear regression. For univariate analysis, comparisons between groups were made using the Mann-Whitney test. We utilized receiver operating characteristic (ROC) curve analysis to calculate areas under the ROC curve (AUC) for each analyte to access the utility of chemokine/cytokine levels as discriminating tests. The ROC generated sensitivity and specificity values were then used to determine clinically optimal cutoff values for the informative analytes. Results: Univariate analysis utilizing Mann-Whitney tests demonstrated that median values (pg/ml) of IP-10 (CXCL10), IFNg, IL-15, MDC (CCL22), MDC (CCL22), MCP-1 (CCL2), Fractalkine, and FLT3L were all significantly higher in CSF from patients with infectious brain disorders than in CSF from patients with non-infectious disorders (p-value \u3c 0.05). MDC (CCL22) demonstrated statistical significance, when comparing viral infections versus non-viral infections (with the non-viral infection group having higher analyte levels). IP10 (CXCL10) can reliably distinguish between an infectious versus non-infectious CNS process (AUC 0.9778) with an optimal cut-off value of 2023 pg/ml (sensitivity, specificity; 93.0%, 92.0%). In the infectious group, MDC (CCL22) can reliably differentiate between viral and non-viral CNS infection (AUC 0.9545) with an optimal cut-off value of 194 pg/ml (sensitivity, specificity; 91.67%, 87.88%). Conclusion: CSF levels (pg/ml) of IP-10 (CXCL10) can reliably distinguish infectious versus noninfectious CNS disorders, and in the infectious group, MDC (CCL22) can reliably distinguish between viral and non-viral CNS infections. These results suggest that CSF chemokine/cytokine quantification can serve as a useful laboratory tool for the rapid triage of CNS diseases to help guide prompt therapy and further testing