Discipline - Immunology and Transplant Immunology - Fundeni Clinical Institute

Professor Constantinescu Ileana, PhD tenured
Professor Tanase Alina-Daniela, PhD tenured
Associate Professor Calenic Bogdan, PhD tenured
Lecturer Maruntelu Ion, PhD tenured
Assistant Professor Tizu Maria, PhD fixed term
Assistant Professor Ursu Andreea Mirela, PhD fixed term

The Immunogenetics and Virology Center of Fundeni Hospital, in which the discipline conducts its activities, is the first center of molecular biology for transplant immunology in Romania, being accredited by the European Federation of Immunogenetics (EFI) since 2006.

Since 2006, our laboratory has been the Reference Center for Transplant Immunology in Romania, as well as a methodological forum, organizing international workshops, round tables and courses in the domain of histocompatibility, immunogenetics and virology, congresses and other national and international scientific events.

The main activity of the Immunogenetics and Virology Center involves running all immunologic and virological investigations related to the results of a transplantation procedure, for both solid organs (kidney, heart, liver, etc) and hematopoietic stem cells.

These investigations are conducted for both transplant recipients and donors (alive or brain dead) and include: indirect viral diagnosis (serologic screening) and direct diagnosis (determination of viral load) for hepatic viruses (HCV, HBV), HIV-1,2, Herpes viruses (EBV, CMV, HSV-1,2), parvovirus B19, BK polyomavirus, adenoviruses, HTLV-1,2, toxoplasma gondii; tumoral markers; screening and identification of cytotoxic antibodies; HLA genotyping and the determination of donor-recipient compatibility; crossmatch test; Chimerism.

Likewise, the Immunogenetics and Virology Center runs the necessary investigations for monitoring the transplant recipient during the post-transplant period: evaluation for viral infections and tumoral markers in the context of immunosuppressive treatment; rejection immunology (anti-HLA antibodies); dosage of blood levels of immunosuppressants (tacrolimus, sirolimus, cyclosporine);

The other sections of the Fundeni Clinical Institute, which do not conduct transplant activities, the laboratory performs immunologic and virological diagnostic tests, direct and indirect, as well as tumoral immunology and specific HLA genotyping, associated with certain diseases. The Immunogenetics Center is also implicated in the coordination, recruitment, donation and virological and immunologic testing for the National Register of Voluntary Donors of Hematopoietic Stem Cells.

The Immunogenetics and Virology Center is also a Learning and Medical Education Center in the domain of Immunology and Transplant Immunology for all the Immunology centers in the country: Iasi, Targu Mures, Timisoara and Cluj.

The goal is to establish standard operating procedures (SOP) in the molecular genetics of HLA, standardize the methods used regarding HLA genotyping, screening and identification tests of cytotoxic antibodies in the serum of receiving patients and the crossmatch test.

Likewise, we have proposed to standardize the algorithms used in the domain of molecular virology and for monitoring immunosuppressive therapy (Cyclosporin, Tacrolimus, Sirolimus) in post-transplant patients

Research directions:

  • New findings in the diagnosis and treatment of autoimmune diseases and the impact of autoimmune diseases on the transplant of solid organs
  • Transplant immunobiology of solid organs (kidney, liver, heart)
  • Molecular genetics and immunologic diagnosis and its impact on the evolution of bone marrow transplantation
  • Virtual crossmatch, clinical and immunogenetic aspects and prognostic factors in kidney transplantation
  • Molecular biology of acute subclinical and clinical rejection in the transplantation of solid organs
  • Molecular biology of GVHD post-transplant of peripheral hematopoietic cells
  • The role of tumoral immunogenetics in the diagnosis and monitoring of different types of post-transplant cancers
  • Oncogenes and the expression of HLA alleles in different types of cancers (renal, urinary bladder, and prostate cancers). The efficiency of cellular immunity stimulation in different types of cancer.
  • HLA and its prognostic value in oncology (urologic cancers)
  • Immunotherapy for cancers of the urinary bladder
  • Maybe the appearance of post-transplant cancers can be prevented?
  • What is the role of molecular diagnosis in the prevention of the appearance of post-transplant renal neoplasms?
  • Applied pharmacogenetics for the prediction of post-transplant immunosuppressive treatment response
  • Molecular genetics diagnosis for renal, prostate and urinary bladder cancers
  • The role of vaccine therapy in cancer treatment
  • How anti-HLA antibodies influence the selection of donor-receiver in renal transplantation and what is their impact on post-transplant evolution?

Immunology course, 3rd year (romanian and English language modules)

Thematic

  • Definition of general aspects of the immune response
  • The origin, destination and structure of the cells and tissues belonging to the immune system
  • Antigens
  • Antibodies
  • B cell development
  • T cell development
  • Structure and role of the major histocompatibility complex (MHC)
  • The magic of antigen processing and presentation
  • T cell receptors: structure and function in the activaiton of T cells
  • Humoral immunity
  • Inflammation
  • Immediate hypersensitivity reaction
  • Delayed hypersensitivity reaction (DTR)
  • Lysis mediated by complement vs cell-mediated lysis
  • Microbial immunity and immunoparasitology
  • Autoimmunity
  • Transplantation
  • Immunopathology: when the immune system responds incorrectly
  • Secondary immunodeficiency
  • Allergic diseases
  • Connective tissue diseases
  • Vasculitis syndrome
  • Diseases with mixed determinism
  • Psycho-neuro-immunology and neuro-endocrino-immunology
  • Cancer and the immune system
  • Immunochemistry
  • Cellular investigations
  • Qualitative and management issues

Bibliography

  1. Imunologia transplantului, Ileana Constantinescu, ISBN: 978-973-708-431-6, 2009
  2. Basic concepts in immunology, John Clancy, JR, ISBN 0-07-011371-8, 1998
  3. Immunology-Problem based, Reginald M. Gorczynski, Jacqline Stanley, ISBN: 9781416024163 25.99, 2006
  4. How the immune system works, third edition, Lauren Sompayrac, ISBN: 978-1-4051-6221-, 2008
  5. Immunology, Stephen Boag & Amy Sadler, ISBN: 978-0-340-92558-4, 2007
  6. Oxford handbook of clinical immunology, Gavin Spickett, ISBN: 0-19-262721-x, 1999
  7. Roitt’s Essential Immunology, Peter J., Delves I., Seamus J., Martin Dennis R., Burton Ivan M. Roitt, 12th edition, 2011.
  8. Human Leucocyte antigen: The major Histocompatibility Complex of Man, Clinical Diagnosis and management by Laboratory Methods, J. Bernard Henry, M.D., twentieth ed., 2001.
  9. Genetics and molecular genetics of the MHC, reviews in Immunogenetics, Rhodes, D.A., Trowsdale, J., 1999.
  10. HLA Beyond tears. De novo, Rodey Glenn E., 2000
  11. Molecular Diagnostics a training and study guide, Tsongalis Gregory J, Coleman William B., 2002.
  12. Transplantul renal, Sinescu I., Manu M.A., harza M., 2007.
  13. Oncogeneza virala-aspecte de etio-patogeneza, Petre Calistru, Radu Iftimovici, Ileana Constantinescu, Petru Adrian Mutiu in colaborare cu Vasile Nastase si Andrei Antoniu Dinu, cap.8 Virusuri cu potential oncogen pentru om, pag.363-449, editura Academiei Romane, 2015, ISBN 978-973-27-2512-2
  14. Tratat de Urologie, I.Sinescu, Editura Universitara „Carol Davila", Bucuresti, 2011, ISBN: 978-973-708-558-0

Practical Lessons for the Immunology course (3rd year)

  1. Cellular investigations – detection of specific cellular products
  2. EIA technique
  3. ELISA technique
  4. Electrophoresis of serum proteins
  5. Immunogram
  6. Cell cultures
  7. Cellular proliferation analysis
  8. Apoptosis tests
  9. Adhesion markers
  10. Flow cytometry – its importance in the immunologic diagnosis of various conditions
  11. NK cells
  12. Toll-like receptors
  13. Molecular immunogenetics techniques – PCR techniques (HLA, chimerism, viremia)
  14. DNA extraction
  15. RNA extraction
  16. HLA genotyping in transplantation
  17. Major histocompatibility complex
  18. Immunobiology of renal transplantation
  19. Immunobiology of hematopoietic stem cell transplantation
  20. National register of honorific donors of hematopoietic stem cells
  21. Immunology of heart transplantation
  22. Detection of KIR gene polymorphism through SSP
  23. Quantitative detection of serum IL2R (enzymatically enhanced chemoluminescence)
  24. Detection of cytokine gene polymorphisms through SSP
  25. Quantitative detection of serum neopterin (quantitative ELISA): interpretation in the clinical context

Optional course: Transplant Immunology, 4th year students (English module and Romanian module)

Topics covered in the course

  1. Immunogenetics and transplantation: genetic basis for transplantation, introductive notions, basic elements of genetics (DNA, genes and chromosomes, loci, alleles and the segregation phenomenon)
  2. Major histocompatibility complex; MHC structure, genes for MHC class I and class II. HLA nomenclature. "Linkage disequilibrium" phenomenon. Ethnic variations.
  3. HLA class I molecules: structure, organization, function. Regulation of class I gene expression. Other class I genes.
  4. HLA class II molecules: structure, organization, function. Regulation of class II gene expression. Other class II genes.
  5. Minor histocompatibility antigens. Genes of the immune response (KIR genes). Polymorphism of cytokine genes and their implication in transplantation
  6. Evaluation of the humoral and cellular immune response in transplant patients
  7. Genotyping and HLA donor matching – receptors in different types of transplantation
  8. Immunological basis for the acute and chronic transplant rejection phenomenon in different types of transplantations. Preventing the acute and chronic transplant rejection phenomenon in solid organ and cellular transplant patients.
  9. Immunology of renal and pancreatic transplantation – kidneys
  10. Immunology of liver transplantation
  11. Immunology of hematopoietic stem cell transplantation
  12. Immunology of heart transplantation
  13. Immunology of hand, skin and corneal transplantation
  14. Immunologic evaluation protocols accredited by EFI (European Federation for Immunogenetics) before the transplant of solid organs or cells.
  15. Complex immunologic problems related to viral, bacterial, fungal and parasitic infections in the transplant population
  16. Immunologic monitoring post-transplant of solid organs or stem cells
  17. Virological monitoring post-transplant of solid organs or cells
  18. Management of waiting lists
  19. Management of Immunogenetics and Virology Centers: standardization of methods, quality assurance
  20. National register of honorific donors of hematopoietic stem cells

LP Themes for the optional course "Transplant Immunology" for 4th year students

  1. HLA genotyping using the SSOP technique; interpretation in the context of the respective transplant
  2. HLA genotyping using the SSP technique; interpretation in the context of the respective transplant
  3. Crossmatch testing using CDC: interpretation for different types of transplants
  4. Crossmatch testing using ELISA: interpretation
  5. ELISA testing for the detection of anti-HLA antibodies: interpretation and clinical significance
  6. ELISA testing for the identification of anti-HLA antibodies: interpretation and clinical significance
  7. Detection of cytotoxic antibodies using the Luminex technique
  8. Electrophoresis of serum proteins
  9. Genetic amplification reaction used for the quantitative detection of HCV RNA: interpretation and clinical significance for transplantation
  10. PCR reaction for the quantitative detection of HBV DNA
  11. PCR reaction for the detection of herpes viruses: interpretation and clinical significance
  12. Quantitative detection of cyclosporine in whole blood
  13. Quantitative detection of tacrolimus in whole blood
  14. Quantitative detection of sirolimus in whole blood
  15. SBT technique for resolving HLA allele ambiguities

For the discipline of immunology and transplant immunology, we propose the following themes for professional development courses that may be conducted at the level of the immunology and transplant immunology discipline of IC Fudeni:

Post-graduate courses:

  1. Molecular Biology in Laboratory Medicine: the advantages, as well as benefits and problems with the interpretation of results.
  2. Current issues in transplant immunology.
  3. Infections with hepatic viruses HBV, HDV and HCV: specific aspects of diagnosis and treatment.
  4. Immunologic and virology monitoring after renal transplant
  • immunology
  • molecular immunology
  • molecular biology
  • molecular virology
  • transplantation
  • molecular genetics
  • oncology
  1. Project code: POSDRU 86/1.2/S/57748

Title: Strategic program to promote innovation in education services through open, continuous (INSEED)

Beneficiary: University Politehnica Bucharest

Partner: Carol Davila University

  1. Project code: 59497

Project title: Training and Organizational Immunology Laboratories employees by implementing advanced technologies and quality management – IMUNOLAB

Beneficiary: Romanian Society of Immunology

  1. Project code: POSDRU/89/1.5/S/64153

Project title: Support postdoctoral research in the field of reconstructive surgery transplant

Beneficiary: Carol Davila University

  1. Title: „Development of the National Strategy on Environmental Research, Technological Development and Innovation for 2014-2020”

Beneficiary: Executive Agency for Higher Education, Research, Development and Innovation

Contract Financing for Projects Sector Plan no. 6S / 20.12.2012, Health Panel

  1. Title: Specializare pentru performanta, eficacitate si raspuns in domeniul transplantului de organe – S.P.E.R.

Beneficiary: Project co-financed by the European Social Fund Operational Programme Human Resources

Development 2007-2013, Invest in the people, POSDRU 81 / 3.2 / S / 59 616

  1. Clinical and therapeutical importance of HBV genotyping in Romania, Constantinescu, I; Nedelcu, F; Toader, MA; Vasile, D, Journal of medicine and life, 2008 Apr-Jun, Vol.1, Issue 2
  2. Primary immunodeficiencies of the B lymphocyte, Moise, A; Nedelcu, F D; Toader, MA; Sora, SM; Tica, A; Ferastraoaru, DE; Constantinescu, I, Journal of medicine and life, 2010 Jan-Mar, Vol.3, Issue 1
  3. Cytotoxic antibodies–valuable prognostic factor for long term kidney allograft survival, Moise, A; Nedelcu, D; Toader, A; Sora, M; Tica, A; Ferastraoaru, DE; Constantinescu, I, Journal of medicine and life, 2010 Oct.-Dec., Vol.3, Issue 4
  4. Cytokine gene polymorphisms support diagnostic monitoring of Romanian multiple myeloma patients, Banu, C; Moise, A; Arion, C V; Coriu, D; Tanase, A; Constantinescu, I, Journal of medicine and life, 2011-Aug-15, vol. 4, Issue 3
  5. Quantification of Cyclosporine, Tacrolimus and Sirolimus Concentrations from Whole Blood of Bone Marrow and Renal Transplantated Patients – Benefits for Long Term Outcome and Survival, Constantinescu, I; Istrate, G., Revista de Chimie, 2013 Nov. ,Vol.64, Issue 11
  6. Cytotoxic antibodies screening in kidney transplantation: clinical relevance and challenges,Constantinescu Ileana, Moise A, Nedelcu D, Toader A, Petre D, I Sinescu,27nd Annual EFI European Immunogenetics and Histocompatibility Conference, Maastricht, Netherlands, 11-14 May 2013,Tissue Antigens – Immune response genetics, vol. 77, issue 5, pag.433-434; ISSN: 0001-2815 print,. Impact factor=2,753
  7. Quantification of Cyclosporine, Tacrolimus and Sirolimus, Concentration from Whole Blood in Bone Marrow and Renal Transplantation – articol– Ileana Constantinescu, Gabriela Istrate, Revista de Chimie, Bucuresti, noiembrie, 11, 2013, ISSN-0034-7752
  8. Hepatitis B Virus Core Promoter Mutations in Patients With Chronic Hepatitis B and Hepatocellular Carcinoma in Bucharest, Romania, Constantinescu, I; Dinu, AA; Boscaiu, V; Niculescu, M, Hepatitis Monthly, 2014 Oct., Vol.14, Issue 10
  9. Complex immunology issue in renal transplanted chronic HBV/HCV carriers,Nastase V, Dinu AA, Moise A, Nedelcu FD, Toader MA, Mircescu G, Sinescu I, Constantinescu Ileana,28nd EFI – European Immunogenetics and Histocompatibility Conference,25-28 May  2014, Stockholm, Sweden,Tissue Antigens, vol. 84, issue 1, pag.84-85. ISSN: 0001-2815, Impact factor=2,350
  10. The frequency of HLA alleles in the Romanian population, Constantinescu, I; Boscaiu, V; Cianga, P; Dinu, AA; Gai, E; Melinte, M; Moise, A, Immunogenetics, 2016 Mar., Vol.68, Issue 3

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