Melanie Hayden

Enhanced presentation of MHC class Ia, Ib and class II-restricted peptides encapsulated in biodegradable nanoparticles: a promising strategy for tumor immunotherapy.

BackgroundMany peptide-based cancer vaccines have been tested in clinical trials with a limited success, mostly due to difficulties associated with peptide stability and delivery, resulting in inefficient antigen presentation. Therefore, the development of suitable and efficient vaccine carrier systems remains a major challenge.MethodsTo address this issue, we have engineered polylactic-co-glycolic acid (PLGA) nanoparticles incorporating: (i) two MHC class I-restricted clinically-relevant peptides, (ii) a MHC class II-binding peptide, and (iii) a non-classical MHC class I-binding peptide. We formulated the nanoparticles utilizing a double emulsion-solvent evaporation technique and characterized their surface morphology, size, zeta potential and peptide content. We also loaded human and murine dendritic cells (DC) with the peptide-containing nanoparticles and determined their ability to present the encapsulated peptide antigens and to induce tumor-specific cytotoxic T lymphocytes (CTL) in vitro.ResultsWe confirmed that the nanoparticles are not toxic to either mouse or human dendritic cells, and do not have any effect on the DC maturation. We also demonstrated a significantly enhanced presentation of the encapsulated peptides upon internalization of the nanoparticles by DC, and confirmed that the improved peptide presentation is actually associated with more efficient generation of peptide-specific CTL and T helper cell responses.ConclusionEncapsulating antigens in PLGA nanoparticles offers unique advantages such as higher efficiency of antigen loading, prolonged presentation of the antigens, prevention of peptide degradation, specific targeting of antigens to antigen presenting cells, improved shelf life of the antigens, and easy scale up for pharmaceutical production. Therefore, these findings are highly significant to the development of synthetic vaccines, and the induction of CTL for adoptive immunotherapy.

Pediatric concussions in sports; a simple and rapid assessment tool for concussive injury in children and adults

BackgroundWe established a routine protocol for concussion evaluation in athletes for nonmedical personnel. The evaluation, management guidelines, and return-to-play recommendations were summarized with a memorable mnemonic on a convenient handheld card.Materials and methodsThe ability to remember the return-to-play mnemonic and effectively apply it to corresponding guidelines was evaluated in 194 sports personnel without medical training. The participants were given three clinical scenarios, each including age, pertinent history, sporting event, description of an injury, symptoms, signs, and a brief neurological exam. Subsequently, the subject’s ability to recall the return-to-play mnemonic and the Standard Assessment of Concussion, to describe the corresponding guidelines, and to advise return-to-play was evaluated.ConclusionOur “return-to-play” mnemonic was found to be simple and memorable, allowing for a high recall percentage and accurate evaluation of concussion cases. High intersubject agreement suggests that this method has both standardization and generalization potential.

Analyzing T cell responses

This book represents the first comprehensive description and evaluation of the most important assays utilized to monitor immune responses against tumor associated antigens. Each chapter, prepared by leading investigators in tumor immunology and immunotherapy, details a specific immune topic or analytical method. Specifically tailored to the needs of clinical researchers, basic science investigators, and biotechnology innovators, this book provides information that is both sufficiently detailed and eloquently succinct. All discussions are substantiated with a comprehensive literature review. The categorization and organization of these sections is such that one can either pursue the entire book or quickly find necessary information on a specific assay or concept of interest. An in-depth description of the methods utilized in the identification of tumor associated antigens is complemented by extensive lists of HLA class I and class II restricted antigens. This book also addresses the essential topic of tumor cells’ ability to evade immune recognition and destruction. The potential mechanisms regarding the disparity between the patients’ immunological and clinical response following immunization are reviewed. Importantly, potential therapeutic strategies to overcome the immune escape mechanisms have been discussed. Having described an appropriate amount of background information in order to establish a fundamental understanding of the antigen-specific immune responses, the book then moves to specifically address the particular assays utilized by the tumor immunologists in the following chapters: Cytotoxicity Assays focuses on evaluating the cellular immune responses to tumors and the monitoring of killer lymphocytes in cancer patients. In addition, the killing mechanisms utilized by the effector lymphocytes and the different types of CTL-inducing cancer vaccines are discussed. Monitoring T Cell Proliferation highlights the importance of proliferation as a significant indicator of lymphocyte activation. Specifically, important regulators and principles of T cell proliferation are reviewed. A variety of monitoring techniques and methods for data analysis are also discussed. ELISPOT Assay concentrates on the principle, performance, result interpretation and limitations of this important and widely used method for monitoring the immune status of patients receiving cancer vaccines. A very helpful section on the assay’s problems and suggested solutions is also included. Modified ELISPOT describes some modifications of this assay specifically developed for T cell monitoring in cancer vaccine trials. These modifications, Granzyme B ELISPOT and Autologous Tumor IFN-γ ELISPOT, allow better assessment of low frequency tumor-specific CTL and their functions in cancer vaccine trials. Intracellular Cytokine Staining focuses on the technique of this sensitive assay for analysis of T cell responses. In addition, monitoring of spontaneous tumor immunity and vaccination studies in patients with cancer are also discussed. Cytometric Cytokine Secretion Assay provides a detailed description of this innovative method for the analysis and enrichment of viable cells according to the secreted cytokines. The cytokine secretion assay has proven to be especially useful for the detection and isolation of viable antigen-specific T cells after a short restimulation with specific antigen in vitro to induce secretion of cytokines. This chapter provides comprehensive background information and very useful practical considerations on the detection and isolation of cytokine-secreting, antigenspecific T cells. Cancer Metastasis Rev (2006) 25:501–502 DOI 10.1007/s10555-006-9015-1

Cerebellopontine angle cyst compressing the vagus nerve: case report.

OBJECTIVEThe cerebellopontine angle (CPA) is a rare location for an arachnoid cyst. We describe a patient with a CPA arachnoid cyst who presented with hoarseness (unilateral vocal cord paralysis) and dysphagia secondary to isolated compression of the vagus nerve. This rare presentation of a CPA arachnoid cyst has not been reported previously. CLINICAL PRESENTATIONThe patient described is a 50-year-old man who experienced a precipitous onset of hoarseness and dsyphagia. An otolaryngological evaluation revealed right-sided vocal cord paralysis. Brain magnetic resonance images displayed a cystic mass at the right CPA and anterior displacement of the vagus nerve. INTERVENTIONThe patient underwent retrosigmoidal craniectomy with cyst fenestration, which was well tolerated. Intraoperatively, Cranial Nerve X was found splayed over the cyst and was consequently decompressed. CONCLUSIONPostoperatively, the patients dysphagia completely resolved. However, the results of a laryngeal electromyocardiogram revealed minimal evidence of recovery in the affected vocal fold, and the patient continued to suffer from dysphonia. Although CPA arachnoid cysts are rare, they should be considered when a patient presents with an isolated cranial nerve palsy. Treatment options include cyst fenestration and cranial nerve decompression.

214 Fractal Structure in Volumetric Contrast Enhancement of Malignant Gliomas Correlates With Oxidative Metabolic Pathway Gene Expression.

INTRODUCTION Fractal structure is found throughout many processes in nature, and often arises from sets of simple rules. We examined the contrast enhancement pattern in glioblastoma brain tumor MRIs for evidence of fractal structure, which might then be compared with expression of specific gene sets obtained from surgical specimens of each tumor. METHODS Volumetric T1 postcontrast imaging was obtained in 39 patients prior to surgical resection of pathology-confirmed glioblastoma lesions. For each tumor, we calculated the fractal dimension (Minkowski Bouligand dimension) using a box-counting (cubic scaling) approach. RNA expression microarray data from resected tissue were explored by gene set enrichment analysis (GSEA). RESULTS We found robust evidence for fractal structure in the contrast enhancement pattern, with an average fractal dimension of 2.17 ± 0.10, with a visually apparent split at 2.10. GSEA analysis showed a definitive association between this split in fractal dimension and 6 gene sets (of 4080), all 6 of which are linked to mitochondrial respiration/ATP production pathways. CONCLUSION There is fractal structure in the volumetric enhancement pattern of glioblastoma tumors, with dimension approximately 2.15. Variation in this fractal dimension, and therefore the complexity of contrast enhancement it reflects, is specifically associated with genetic correlates of a shift to glycolytic metabolism in tumor cells. Drugs that shift glioblastoma to oxidative metabolism have recently been identified as independent therapeutic agents as well as sensitizing agents for irradiation. Therefore, a radiogenomic marker of glucose metabolism, such as this fractal structure in enhancement, might help to guide individualized therapy.

Book review: analyzing T cell responses

tion and evaluation of the most important assays utilized to monitor immune responses against tumor associated antigens. Each chapter, prepared by leading investigators in tumor immunology and immunotherapy, details a specific immune topic or analytical method. Specifically tailored to the needs of clinical researchers, basic scientist investigators, and biotechnology innovators, this book provides information that is both sufficiently detailed and eloquently succinct. All discussions are substantiated with a comprehensive literature review. The categorization and organization of these sections is such that one can either pursue the entire book or quickly find necessary information on a specific assay or concept of interest. An in depth description of the methods utilized in the identification of tumor associated antigens is complemented by extensive lists of HLA class I, and class II restricted antigens. This book also addresses the essential topic of tumor cells’ ability to evade immune recognition and destruction. The potential mechanisms regarding the disparity between the patients’ immunological and clinical response following immunization are reviewed. Importantly, potential therapeutic strategies to overcome the immune escape mechanisms have been discussed. Having described an appropriate amount of background information in order to establish a fundamental understanding of the antigen-specific immune responses, the book then moves to specifically address the particular assays utilized by the tumor immunologists in the following chapters: CYTOTOXICITY ASSAYS focuses on evaluating the cellular immune responses to tumors and the monitoring of killer lymphocytes in cancer patients. In addition, the killing mechanisms utilized by the effector lymphocytes and the different types of CTLinducing cancer vaccines are discussed. MONITORING T CELL PROLIFERATION highlights the importance of proliferation as an important indicator of lymphocyte activation. Specifically, important regulators and principles of T cell proliferation are reviewed. A variety of monitoring techniques and methods for data analysis are also discussed. ELISPOT ASSAY concentrates on the principle, performance, result interpretation and limitations of this important and widely used method for monitoring the immune status of patients receiving cancer vaccines. A very helpful section on the assay’s problems and suggested solutions is also included. MODIFIED ELISPOT describes some modifications of this assay specifically developed for T cell monitoring in cancer vaccine trials. These modifications, Granzyme B ELISPOT and Autologous Tumor IFN-c ELISPOT, allow better assessment of low frequency tumor-specific CTL and their functions in cancer vaccine trials. INTRACELLULAR CYTOKINE STAINING focuses on the technique of this sensitive assay for analysis of T cell responses. In addition, monitoring of spontaneous tumor immunity and vaccination studies in patients with cancer are also discussed. CYTOMETRIC CYTOKINE SECRETION ASSAY provides a detailed description of this innovative M. Hayden S. Schroter B. R. Minev (&) Moores UCSD Cancer Center , Bldg. MCCT, Room 4313 3855 Health Sciences Drive, #0820, La Jolla, CA 92093-0820, USA e-mail: bminev@ucsd.edu Clin Exp Metastasis (2007) 24:67–68 DOI 10.1007/s10585-006-9039-5

Analyzing T cell responses: how to analyze cellular immune responses against tumor associated antigens

This book represents the first comprehensive description and evaluation of the most important assays utilized to monitor immune responses against tumor associated antigens. Each chapter, prepared by leading investigators in tumor immunology and immunotherapy, details a specific immune topic or analytical method. Specifically tailored to the needs of clinical researchers, basic scientist investigators, and biotechnology innovators, this book provides information that is both sufficiently detailed and eloquently succinct. All discussions are substantiated with a comprehensive literature review. The categorization and organization of these sections is such that one can either pursue the entire book or quickly find necessary information on a specific assay or concept of interest. An in depth description of the methods utilized in the identification of tumor associated antigens is complemented by extensive lists of HLA class I, and class II restricted antigens. This book also addresses the essential topic of tumor cells’ ability to evade immune recognition and destruction. The potential mechanisms regarding the disparity between the patients’ immunological and clinical response following immunization are reviewed. Importantly, potential therapeutic strategies to overcome the immune escape mechanisms have been discussed. Having described an appropriate amount of background information in order to establish a fundamental understanding of the antigen-specific immune responses, the book then moves to specifically address the particular assays utilized by the tumor immunologists in the following chapters: CYTOTOXICITY ASSAYS focuses on evaluating the cellular immune responses to tumors and the monitoring

Dirk Nagorsen and F. M. Marincola (eds): Analyzing T Cell Responses: How to analyze cellular immune responses against tumor associated antigens

tion, and evaluation of the most important assays utilized to monitor immune responses against tumor associated antigens. Each chapter, prepared by leading investigators in tumor immunology and immunotherapy, details a specific immune topic or analytical method. Specifically tailored to the needs of clinical researchers, basic science investigators, and biotechnology innovators, this book provides information that is both sufficiently detailed and eloquently succinct. All discussions are substantiated with a comprehensive literature review. The categorization and organization of these sections is such that one can either pursue the entire book or quickly find necessary information on a specific assay or concept of interest. An in depth description of the methods utilized in the identification of tumor associated antigens is complemented by extensive lists of HLA class I, and class II restricted antigens. This book also addresses the essential topic of tumor cells’ ability to evade immune recognition and destruction. The potential mechanisms regarding the disparity between the patients’ immunological and clinical response following immunization are reviewed. Importantly, potential therapeutic strategies to overcome the immune escape mechanisms have been discussed. Having described, an appropriate amount of background information in order to establish a fundamental understanding of the antigen-specific immune responses, the book then moves to specifically addressing the particular assays utilized by the tumor immunologists in the following chapters: CYTOTOXICITY ASSAYS focuses on evaluating the cellular immune responses to tumors and the monitoring of killer lymphocytes in cancer patients. In addition, the killing mechanisms utilized by the effector lymphocytes and the different types of CTLinducing cancer vaccines are discussed. MONITORING T CELL PROLIFERATION highlights the importance of proliferation as a significant indicator of lymphocyte activation. Specifically, important regulators and principles of T cell proliferation are reviewed. A variety of monitoring techniques and methods for data analysis are also discussed. ELISPOT ASSAY concentrates on the principle, performance, result interpretation and limitations of this important and widely used method for monitoring the immune status of patients receiving cancer vaccines. A very helpful section on the assay’s problems and suggested solutions is also included. MODIFIED ELISPOT describes some modifications of this assay specifically developed for T cell monitoring in cancer vaccine trials. These modifications, Granzyme B ELISPOT and Autologous Tumor IFN-c ELISPOT, allow better assessment of low frequency tumor-specific CTL and their functions in cancer vaccine trials. INTRACELLULAR CYTOKINE STAINING focuses on the technique of this sensitive assay for analysis of T cell responses. In addition, monitoring of spontaneous tumor immunity and vaccination studies in patients with cancer are also discussed. CYTOMETRIC CYTOKINE SECRETION ASSAY provides a detailed description of this innovative M. Hayden Æ S. Schroter Æ B. Minev (&) Moores UCSD Cancer Center, Bldg. MCCT, Room 4313, 3855 Health Sciences Drive, #0820, La Jolla 92093-0820 CA, USA e-mail: bminev@ucsd.edu Angiogenesis (2006) 9:153–154 DOI 10.1007/s10456-006-9041-1