Immunology 2016 / AAI Annual Meeting

Abstract:  Antibodies have gained importance as powerful therapeutic armamentarium for a wide range of human diseases, including many types of cancer. Cancer immunotherapy offers advantages of high specificity, fewer side effects, and long-term benefits in some patients. However, it also creates challenges because subtle changes that make cancer cells immortal are not sufficient enough to elicit an immune reaction. Although the number of therapeutic options are growing, monoclonal antibodies offer a clear advantage of specificity and shorter life-span, which limits undesirable side effects. However, there is still need to understand target pathways and develop validated antibodies for research use to first reveal appropriately differentiated targets. This presentation will discuss some new research antibodies developed by MilliporeSigma to key tumor specific antigens that may be valuable in identifying novel cancer pathway targets.

Abstract:  The complement system’s multiple plasma proteins assist phagocytic cells and antibodies in clearing pathogens. The complement system plays a key role in diseases with an immune component, such as asthma or sepsis, and in autoimmune diseases, including SLE, IBD, RA, JIA, and MS. Understanding how this system affects complex states demands measurement of multiple proteins simultaneously (“complement profiles”) in patient serum or plasma. Using Luminex xMAP® technology, we developed two multiplex panels to quantitate 15 complement proteins simultaneously in serum or plasma. Significant differences were observed for several complement proteins when comparing SLE, JIA, and sepsis patients to healthy subjects. Join our exhibitor workshop for the presentation of these data and tips on using the new MILLIPLEX® map Human Complement Panels to save time, money, and sample, compared to ELISAs.

Abstract:  Imaging flow cytometry applies the speed, sensitivity, and phenotyping abilities of flow cytometry with the detailed imagery and functional insights of microscopy to address complex inquiries in immunology. In this workshop, you will learn how Amnis® imaging flow cytometers have been used to examine the trafficking of MHC class II (MHCII) in dendritic cells. The abundance of antigenic peptide-MCHII complexes on antigen presenting cells is determined by many factors that include the balance between the expression of newly generated complexes, complex internalization and subsequent re-emergence or degradation. However, the molecular mechanisms that govern these interactions are still poorly understood. Here, we discuss how multispectral imaging in flow can be used to visualize MHCII trafficking and elucidate the molecular mechanisms that regulate MHCII homeostasis in primary mouse and human dendritic cells.

Abstract:   Precise regulation of lymphocyte function is critical to mounting a specific immune response; this complex process requires exquisite control over cell migration and expression of effector capabilities. Understanding these mechanisms is essential for development of therapeutics that enhance endogenous immune function. The ELISpot assay offers quantitative assessment of effector function(s) at the single cell level with superior sensitivity. MilliporeSigma and collaborators have significantly improved the venerable but tricky ELISpot technology with advanced membrane plates, multiparameter fluorescent reporting (fluorospot), and optimized protocols.

Cell migration models suffer from the inability to establish continuous gradients and are not amenable to real-time studies. The microfluidic-based ONIX cell culture system offers real–time environmental control, including media perfusion and gradient formation, and can be paired with a microscope for dynamic live cell imaging. Together, these platforms enable functional profiling of single immune cells with unparalleled control and sensitivity.

Abstract:  Multilaser flow cytometers with the capacity for simultaneous measurement of sufficient targets for immunological analysis have traditionally required extensive operator expertise and are often housed in select or shared labs due to cost. Guava® easyCyte systems offer four enhanced-power modulated lasers, including a unique 532 nm green excitation laser, in a benchtop instrument priced for accessibility by individual labs. The microcapillary fluidics employed by all Guava® systems uniquely enable absolute cell counts with small sample volumes in single tube or 96-well plate format, rendering them ideally suited for precious biological samples from patients or transgenic models. The Guava® InCyte software includes a heat map view of high-throughput plate results, making simultaneous visualization of multiple parameters possible and permitting assessment of multisample activation, synapse formation, and other immunomodulation in a single view. Data will be shown demonstrating how signal from up to twelve fluorophores can be accurately resolved for autoimmunity studies, immunogenic/tolerogenic responses, and intracellular immune activity.