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Molecular and biological characterization of human memory NK cell in vitro expansion.
Universita degli Studi di Roma 'La Sapienza'
Experimental Medicine Department Dipartimento di Medicina Sperimentale Sapienza Universita' di Roma Viale Regina Elena, 324 00161 Roma, Italy
Type of Research Project
- Basic science
What is the background of the project?
Memory NK are a recently discovered subset of Natural Killer cells that display some peculiar characteristics that don’t belong to innate immunity, such as long half-life and antigen specificity (in the mouse). Memory NK development is not completely understood, but it is associated to CMV infection in both humans and mice. Among the peculiar characteristics of this subset, there’s an increased responsivity to antibody stimulation; the majority of NK cells can recognize IgG-opsonized target cells through their Fc-receptor CD16, however, memory NK cells respond better to this stimulation than normal NK cells. Linked to CD16 there’s also a phenotypic characteristic that is used to identify the memory subset among NK cells; indeed, memory NK cells lack the FcεRIγ chain, that, together with TCR ζ, is responsible for CD16 signaling ability. Their ability to respond better to antibody-stimulation and their long life make memory NK cells an interesting subset as a potential tool to exploit in clinical therapies that rely on the action of monoclonal antibodies, such as hematological malignancies like B cell leukemias and lymphomas.
What is the aim of the project?
- Setting up a method for memory NK cells expansion in vitro through CD16 stimulation; - Testing feasibility of their use in combination with mAb-based clinical therapies, in the context of B cell tumors treatment. - Better describe the biology of memory NK cells, by investigating receptor expression pattern, response to cytokine stimulation and synergy of CD16 activation.
What techniques and methods are used?
For the major part, memory NK cell biology will be investigated through multiparametric cytofluorimetric analysis. Through the use of fluorochrome-conjugated specific antibodies for surface and intracellular markers, together with fluorescent dyes, we are able to identify memory NK cells and to evaluate their phenotypic and functional characteristics. Moreover, many different techniques of cell isolation, culture and preparation will be used for obtaining memory NK cells from healthy donors and to specifically cultivate memory NK cells.
What is the role of the student?
- The student will mainly observe
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- If the project includes “lab work”
- the student will take active part in the practical aspect of the project
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
The student, at the end of the internship, will be able to: - set up cell culture manipulation, under sterile conditions - perform mononuclear cell separation from peripheral blood samples - perform immunostaining of cell suspensions for immunocytofluorimetric analysis - evaluate cell concentration by microscopic cell counting (Thoma chamber) - set up lymphocyte polyclonal stimulation - analyze cytofluorimetric data
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- No specific outcome is expected
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Subjects passed: Cellular Biology; Biochemistry; Human Physiology; Microbiology; Immunology.
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months) - Pre-Medical students from the American-British system
- 1. Trends Immunol. 2016 Dec;37(12):877-888. doi: 10.1016/j.it.2016.09.005. Epub 2016 Oct 21. Harnessing NK Cell Memory for Cancer Immunotherapy. Fehniger TA1; Cooper MA2.
- 2. Front Immunol. 2017 Sep 13;8:1143. doi: 10.3389/fimmu.2017.01143. eCollection 2017. Natural Killer Cell Memory: Progress and Implications. Peng H1; Tian Z1;2.
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