Technological platforms

The Center has invested a significant amount of economic resources to amass cutting edge technology and scientific expertise.

Imaging at the atomic level

Atomic visualization of the three-dimensional structures of proteins is essential for a complete understanding of the molecular basis underlying immune cell functions and development of infectious diseases. For this purpose, the Center has all required expertise in structural biology (X-ray crystallography) and biochemistry (production and isolation of soluble proteins) as well as several important techniques including circular dichroism and surface plasmon resonance.

Besides the state of the art crystallography facilities within the Center, we also have full access to X-ray robotic facilities in the Structural Genomic Consortium (SGC) at Karolinska Institutet. Sampling of diffraction data from protein crystals at the highest achievable resolution is performed on a routine basis in several European synchrotrons.

The ambition is to further develop the technical capacities of the Center in structural biology and biochemistry through the acquisition of additional state-of-the-art equipment, including novel Äkta Purifier with a specific refrigerator as well as temperature-controlled bacteria shaker, that allows for high performance purification, isolation and characterization of proteins.

We are planning to purchase additional instruments for dynamic light scattering, circular dichroism spectroscopy as well as a Surface Plasmon Resonance Biacore 3000 system that will allow for analyzes of protein binding affinity and thermodynamics.

For more information please contact:

Adnane Achour

Imaging at the cellular and tissue level

To perform complete studies of immunological and pathogenic mechanisms, it is essential to have advanced equipment for visualization and quantification of immune cell functions.

At the Center, researchers have established an advanced visualization facility that provides researchers with front-line imaging technology, technical support and imaging-related expert advice to researchers. Specific acquired computerized image analyses programs for quantification of biological and immunological mediators have been developed in the research environment.

The Center also hold advanced equipment for fluorescence microscopy analyses, including a Nikon A1R confocal microscope, which is an excellent tool for studies of immune cell functions, viral and bacterial entry into host cells, as well as intracellular vesicular transport and diffusion mechanisms. It allows studies of single molecule dynamics, surface structures, exocytic and endocytic trafficking, receptor/ligand and cell-cell interactions in living and fixed specimens. The Nikon A1R is equipped with the total internal reflection fluorescence (TIRF) application, which is an ideal tool for live imaging investigating both the mechanisms and dynamics of protein interactions at the cell membrane surface.

As a joint effort with S. Strömblad and R. Toftgård at the Novum Research Center in Huddinge, the Center has established a state-of-the art imaging facility. The new facility offers a wide range of microscopes for advanced fluorescence microscopy and includes a multiphoton confocal microscope Zeiss 710, which allows extended time lapse experiments on live cells. This includes four-dimensional (4D, i.e. 3D over time) imaging of dynamic cellular events and quantitative modeling of molecular mechanisms regulating immune cell behavior in more complex settings such as, e.g., organotypic cultures, tissue explants, and whole body organs. This microscope allows simultaneous analyses of a range of fluorophores and also provides a cutting edge technology for imaging of live human immune cells and host pathogen interactions under more physiological conditions.

For more information please contact:

Anette Hofmann

Tissue model systems

At the Center, researchers have access to unique biopsies from patient cohorts.

Analyses of sections or isolated cells from biopsies at the cellular or molecular level provide information of high clinical relevance that are further strengthened by detailed analyses in appropriate human cell and tissue model systems.

From simple co-culture models of human tissue cells and, ultimately, to the generation of whole organs or representations of whole organs in the laboratory, scientists in the research environment develop human tissue model systems in vitro that can complement essential work currently achievable only in vivo.

Our commitment to this development is essential. It includes specialized cultures of tissue cells as well as ex vivo explant models. Several approaches have been adapted to study immune cell interactions with tissue specific cells in monolayer-based cultures. Although these culture systems have provided important information with respect to tissue-specific influences on immune cell function, moving from cell monolayers to three-dimensional (3D) cultures is motivated by the need to work with cellular models that mimic the functions of living tissues. Scientists in the research environment engineer three-dimensional (3D) tissue models, so-called organotypic cultures that also contain immune cells. Our approach of developing and using 3D tissue models that mimic real tissues provides unique tools to study human immune cell functions that are present in live tissue and often missed in monolayer-based cell cultures.

In 3D cultures, tissue specific cells acquire a polarized phenotype and a large number of cell-cell contacts occur, which are likely to affect immune cell function and responses to external stimuli, such as pathogens. In addition, the 3D tissue model allows performance of live imaging of immune cells within tissue using 4D, i.e. 3D over time fluorescence imaging techniques. For this purpose we generate tissue models with fluorescent cells, e.g. epithelial cells, fibroblasts, dendritic cells and monocytes. Thus, creating tissue models with immune cells provide us with a technological platform to increase our understanding of human immune cell responses, migration and positioning in tissue, in addition to interactions with pathogens in more physiological models.

For more information please contact:

Mattias Svensson

In vivo imaging

To increase our understanding of infectious diseases it is crucial to have sophisticated models for studying immune responses and disease progressions in vivo.

Non-invasive in vivo biophotonic imaging is a new technology that allows the detection of light-producing biological reactions in living animals in real time. Recently, a system has been acquired allowing combined bioluminescence and fluorescence imaging, as well as 3D reconstruction of signal localization in organs. This new system allows the assessment of up to 12 parameters simultaneously.

Ongoing and projected areas of application include assessment of disease progression in infection models with luciferase or fluorophore-tagged parasitic, bacterial and viral pathogens. Also, by adoptive transfer of luciferase-expressing or fluorophore-tagged immune cells, the involvement of these cells in various infection models can be monitored in vivo in settings of acute and chronic infection.

For more information please contact:

Antonio Barragan

Advanced flow cytometry

Researchers at the Center have been at the leading edge since 2003 in the field of flow cytometry technology.

The advanced flow cytometry lab at CIM is equipped with two BD LSR Fortessa instruments with four lasers that allows assessment of up to 18 fluorescence parameters simultaneously. This technology allows characterization of immune cell subsets at high resolution in exquisite detail, and the value of rare patient samples can be maximized. The instruments are currently used by many groups in the environment in research related to human immune cell function and infectious diseases.

For more information please contact:

Hernan Concha
Monika Enquist
Martin Ivarsson
David Malone

Large-scale isolations of human cells

To study functional aspects of human immune cells it is crucial to have efficient methods of obtaining primary human cells.

Researchers at the Center have developed several sophisticated methods for isolating large numbers of specific primary cells. As an example, our researchers now isolate human hematopoietic progenitor cells (HPC) from bone marrow and cord blood.

A number of other cell isolation systems have been established within the Center for isolation of large numbers of differentiated cells. For example, researchers have developed unique sorting procedures for isolation of distinct dendritic cell (DC) populations directly from blood. DC isolation is managed in collaboration with the Department of Transfusion Medicine at the Karolinska University Hospital. In a similar way, sorting procedures for isolation of NK cells directly from blood has been established. In addition, a protocol for isolation of DC directly from healthy skin (i.e., Langerhans cell DC and dermal DC) has been established. Other important sources of human immune cells are lymphoid tissues, e.g., tonsils, lymph nodes and gut-associated lymphoid tissue.

Many studies also involve isolation of parenchymal cells, including, e.g., hepatocytes and pancreatic islet cells. A method for isolating lymphocytes from the human liver was recently been devised in the research environment in collaboration with the Department of Transplantation Surgery. Human pancreatic islet cells are obtained from the Nordic Network for Clinical Islet Transplantation.

For more information please contact:

Karin Loré
Kalle Malmberg

In vivo models

Experimental models constitute an important complement to in vitro experiments and studies on patient material.

Researchers at the CIM use experimental models with an aim to gain increased understanding for complex host-microbe interactions, disease mechanisms and for testing new vaccine- and drug candidates. The Center is also actively involved in the generation of needs-driven, novel in vivo models for studies within these and related research areas.

Large scale GMP-production unit

Tailored cell therapies may become the treatment of choice in some infectious and tumor diseases as well as transplantation medicine.

Several phase I/II clinical trials are currently planned or ongoing at the Center. To minimize the risk during the cell processing procedures and storage of cells intended for treatment, the EU and EMEA have developed guidelines for handling of cells.

At the Karolinska University Hospital, the Vecura unit is a GMP contract manufacturer of gene and cell therapy products. Researchers at the Center have a close collaboration with Vecura, where cells are now produced in large scale for cell-based immunotherapies. This includes isolation of NK cells for adoptive transfer to patients with hematological malignancies.

Vecura unit, Karolinska University Hospital