
The overall goal of the Einstein-Montefiore CFAR Immunology and Pathology Core is to enable CFAR investigators to perform functional and anatomical analyses of immunological and pathological cells and tissues infected with HIV and other pathogens, and provide translational investigators with a wide array of immunological assays and consultations to further their research studying HIV, its clinical manifestations, and comorbidities.
Core Mission
- To provide investigators with the research infrastructure and optimized technologies and protocols to perform state-of-the art immunologic and biochemical assays.
- To provide investigators with an appropriate biohazard containment facility that is capable of examining the effects of infection with HIV and AIDS-associated pathogens on tissues by characterizing morphological, molecular and phenotypical changes using histological, immunohistochemical, in situ hybridization analyses and live cell imaging.
- To provide for investigators with support for the isolation and culture of primary human and rodent cells including endothelial cells, astrocytes, microglia and neurons for research studies.
- To train investigators in the performance of techniques that are extant in the Core and to assist investigators in the development of new immunological assays and pathological techniques that are required for the accomplishment of their research goals.
Core Facilities
- Fluorescent AxioImager D1 Semi-motorized microscope for video capture
- Cryostat and vibratome for cutting infected samples
- Single cell microinjector (Axoporator)
- ELISA reader and a multiplex Luminex reader for cytokine assays
- MSD SECTOR Imager 2400 plate reader
Core Services
- Quantification of cytokines, chemokines and other mediators by ELISA, Luminex or flow cytometry.
- Laser-capture microdissection of infected cells
- Assistance in characterization and differentiation of cultured cells.
- Extraction and analysis of RNA from uninfected and infected tissues or cells.
- In situ hybridization analyses of tissues for microscopic evaluation of cell type-specific expression of cytokines, chemokines and other mediators.
- Tissue processing and preparation for histopathology analysis of HIV-1 or opportunistic pathogen-infected human clinical samples and animal tissues.
- Immunocytochemistry for HIV-1 antigens or nucleic acids, other pathogen-specific antigen detections, leukocyte surface markers, cell-specific markers and mediators such as cytokines and chemokines.
- Interpretation and photographic documentation of the histopathology and immunocytochemistry findings.
Core Training Programs
- Performance of pathology, immunocytochemistry, photography, and quantitative image analysis.
- Techniques for performing a wide range of cell trafficking and live imaging studies.
- Use of ELISA, Luminex or flow cytometry to quantify production of cytokines, chemokines and other mediators.
- Techniques for primary cellular isolation and culture from primary human or animal tissue.
Recent Core Publications
Buckner CM, Calderon TM, Willams DW, Belbin TJ, Berman JW. Characterization of monocyte maturation/differentiation that facilitates their transmigration across the blood-brain barrier and infection by HIV: implications for NeuroAIDS. Cell Immunol. 2011;267(2): 109-23. PMID: 21292246.
Cosenza-Nashat M, Zhao ML, Suh HS, Morgan J, Natividad R, Morgello S, Lee SC. Expression of the translocator protein of 18 kDa by microglia, macrophages and astrocytes based on immunohistochemical localization in abnormal human brain. Neuropathol AppI Neurobiol. 2009;35(3):306-28. PMID: 19077109.
Eugenin EA, Clements JE, Zink MC, Berman JW. *Human immunodeficiency virus infection of human astrocytes disrupts blood-brain barrier integrity by a gap junction-dependent mechanism. J Neurosci. 2011;31:9456-65. PMID: 21715610; PMCID: PMC3132881.
Floris-Moore M, Fayad ZA, Berman JW, Mani V, Schoenbaum EE, Klein RS, Weinshelbaum KB, Fuster V, Howard AA, Lo Y, Schecter AD. *Association of HIV viral load with monocyte chemoattractant protein-1 and atherosclerosis burden measured by magnetic resonance imaging. AIDS. 2009;23:941-9. PMID: 19318907.
To access Core services please contact Dr. Berman or Dr. Lee.
Charge-back
fees
RNA
extraction and Q-RT-PCR, $50/sample for analysis of up to 5 primer pairs.
ELISA: if
paired antibodies are available, $7/sample/assay; if paired antibodies are not
available, the user will purchase the appropriate ELISA kits, and will pay
$6/sample.
Immunocytochemistry, immunofluorescence,
histopathology and consultation: $100/antibody, including controls, for up to 5 slides. Bulk immunohistochemistry: $10/slide per
antibody.
In Situ Hybridization and consultation: $50/slide per probe including controls.
Tissue culture and cell isolation: Processing fee of $100/preparation.
Live cell imaging: $50 per hour.