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Hypo IgM: Phenotypic characterization of B cells in patiens with Selective IgM deficiency (SigMD)
Portugal ( PorMSIC) - Nova University of Lisbon, Lisbon
Type of Research Project
- Clinical Project without Laboratory work
What is the background of the project?
Selective IgM Deficiency (SIgMD) was only recently considered under the classification of primary immunodeficiency. Different diagnosis criteria have been described: absent serum IgM (IUIS 2019); patients with either invasive or recurrent infections, usually bacterial, low IgM serum or plasma levels (with normal levels of IgG, IgG subclasses and IgA), normal IgG antibody response to all vaccinations, in the absence of any T-cell defect (ESID). Considering low IgM serum levels below 2SD of mean, diverse manifestations can be identified in SIgMD patients, which may be asymptomatic, or present with mild to severe infections, including sepsis and meningitis. Autoimmune and/or allergic manifestations are also recognized in these patients. Moreover, circulating B cells can also vary from absence, to decreased germinal center B cells only, or even normal numbers. Other Igs can also be affected, i.e., IgG subclass deficiency. Curiously, IgM is usually expressed as a membrane receptor, but not secreted. Considering the heterogeneous presentations of SIgMD, and understanding that other distinct predominantly antibody deficiencies (PAD) present defective B-cell patterns associated with unique clinical profiles, we believe patients with SIgMD should undergo a thorough immunological evaluation, assessed along with the patients’ clinical manifestations. The highlighted sentence belongs in the next section.
What is the aim of the project?
We aim the apply to SIgMD patients the B-cell characterization used by EUROFLOW consortium in the characterization of PADs, to better characterize the immune background of these patients, which can be an helpful tool both for diagnostic and therapeutic approaches. Thus, we aim the apply to SIgMD patients the B-cell characterization used by EUROFLOW consortium in the characterization of PADs, to better characterize the immune background of these patients, which can be a helpful tool both for diagnostic and therapeutic approaches.
What techniques and methods are used?
Flow Cytometry Briefly, peripheral blood samples from patients with SIgMD will be collected in Hospital D. Estafânia – Lisbon, and analyzed by Flow Cytometry in a BD FACS Canto II (eight color flow cytometer) at the Primary Immunodeficiency laboratory of NOVA Medical School. The panel of monoclonal antibodies will be then described by the Euroflow Consortium for B-cell analysis. Data analysis will be performed using the Infinicyt software, to identify and characterize pre- and post-germinal center B-cell subsets, and particularly analyze the sIg expression in each subset.
What is the role of the student?
- The student will observe the practical experiments but will be highly involved in the analysis of the results
What are the tasks expected to be accomplished by the student?
First, the student shall attend a small Biosafety course to be familiarized with the good laboratory practice guidelines implemented at the laboratory. For the project itself, the student is expected to develop a database to include all patient data (both clinical and laboratory variables), following the General Data Protection Regulation (GDPR) and Helsinki Declaration. Patients will be recruited from the Primary Immunodeficiency Department of Hospital D. Estefânia, and samples will be directly sent by the laboratory, where the student shall perform all the immunophenotyping protocols, data acquisition on the cytometer and data analysis with the analysis software. For this step of the project, about 20 patients are expected to be enrolled. After all patients are analyzed, the student shall perform the statistical analysis and present results to the laboratory team, for further discussion. A poster presentation and possible publication of data in a peer reviewed journal are also an objective of this internship. BD FACS DIVA and Cytognos Infinicyt software will be used.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
The student shall attend a small Biosafety course in the beginning of the traineeship, to be familiarized with the good laboratory practice guidelines implemented at the laboratory. For the project itself, depending on the Immunology background of the student, lectures will be provided weekly on Immunology and Primary Immunodeficiency topics. Also, bibliographic references will also be available for the student to deepen his/her knowledge. Professor Catarina Martins shall be responsible for these sessions.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a poster - The student will prepare a scientific report - The student’s name will be mentioned in a future publication
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Previous laboratory experience will be preferred. Experience in Flow Cytometry would be an advantage.
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
- Gupta S & Gupta A. Selective IgM Deficiency-An Underestimated Primary Immunodeficiency. Front Immunol. 2017 Sep 5;8:1056. doi: 10.3389/fimmu.2017.01056. eCollection 2017.
- Chovancova Z; et al. Selective IgM Deficiency: Clinical and Laboratory Features of 17 Patients and a Review of the Literature. J Clin Immunol. 2017 Aug;37(6):559-574. doi: 10.1007/s10875-017-0420-8. Epub 2017 Jul 21.
- Louis AG; et al. Analysis of subsets of B cells; Breg; CD4Treg and CD8Treg cells in adult patients with primary selective IgM deficiency. Am J Clin Exp Immunol. 2016 Mar 23;5(1):21-32. eCollection 2016.
- Blanco E; et al. EuroFlow PID group. Defects in memory B-cell and plasma cell subsets expressing different immunoglobulin-subclasses in patients with CVID and immunoglobulin subclass deficiencies. J Allergy Clin Immunol. 2019 Sep;144(3):809-824. doi: 10.1016/j.jaci.2019.02.017. Epub 2019 Feb 28.
- Lucuab-Fegurgur DL & Gupta S. Comprehensive clinical and immunological features of 62 adult patients with selective primary IgM deficiency. Am J Clin Exp Immunol. 2019; 8(6): 55–67.
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