Unit of Molecular Immunopathology and Histocompatibility

Contact Phone Numbers
Head: Dimitrios Degiannis 210 94 93 023
Scientific Personnel: Antigoni Chaidaroglou 210 94 93 024
Polikseni Gourzi 210 94 93 024
Eleni Pantou 210 94 93 019
Technician: Antonios Kapsalis
Fax 210 94 93 018

 

 

Diagnostics

The Molecular Immunopathology and Histocompatibility Unit (MIHU) provides Clinical-laboratory work and consultation to OCSC’s doctors and patients and outpatients.

MIHU supports preoperatively and postoperatively the Onassis Cardiac Surgery Center Transplant Program. The Unit also, covers a wide range of immunological tests exploring systemic immune-mediated diseases, and meets the needs of a wide range of molecular tests to investigate coagulation-hemostasis syndromes. Moreover, it has developed extensive activity in the field of Molecular Microbiology for septic patients hospitalized in the ICU, and also, in the field of Molecular Virology in the investigatory framework of patients with myocarditis or dilated cardiomyopathy. Additionally, pharmacogenetic tests on the metabolism and sensitivity to coumarins (Sintrom) and antiplatelet drugs (Plavix) are conducted.

The Laboratory participates in the External Quality Control programs of the British UK Neqas for the methodologies of donor-recipient intersection with flow cytometry, detection and identification of anti-HLA antibodies by Luminex and level of immunosuppressive drugs (cyclosporine, tactrolimus, everolimus).

 

In MIHU the following segments/modules operate:

  1. Histocompatibility – Transplantation Immunology

In the MIHU laboratory the pre-transplant examination of the candidates for heart transplantation and the immune monitoring of the transplanted patients of the Transplant Unit are conducted.

The Laboratory is on-call daily for thoracic organs transplantation.

 

The tests carried out are:

  • Immunophenotyping of lymphocytes by flow cytometry (CD3 +, CD19 +, CD4 +, CD8 +, CD16 / 56 +)
  • Intersection T and B lymphocyte donor with recipient serum at thoracic organs transplantation by flow cytometry (XMatch)
  • Detection of specific antibodies against HLA class I and II antigens by Luminex
  • Identification of antibody specificity against HLA class I and II antigens by method Luminex
  • Genotypic analysis of low resolution of HLA – A, B, DRB1, DQB1 genes by PCR-SSP (Sequence Specific Primers)
  • Qualitative and quantitative CMV and EBV detection with real-time PCR
  • Levels of immunosuppressive medication (Neoral, Tacrolimus, Certican)

 

  1. General Immunology

In the MIHU laboratory the serological determination of autoantibodies, immunoglobulin, complement and cardiolipin is carried out.

The Laboratory receives samples daily, except Saturday and Sunday, from 08:00 to 16:00.

The results are delivered within 5 working days.

Specifically, the exams carried out are:

Exams for autoimmune diseases – Control autoantibodies

  • Antinuclear antibodies (ANA)
  • Antibodies to smooth muscle (ASMA)
  • Anti ds-DNA antibodies
  • ENA (anti-SS-A, SS-A 52, SS-A 60, SS-B, Sm / RNP, RNP-70, Scl-70, Jo-1)
  • Rheumatoid factor
  • Antibodies against CCP (cyclic citrulinated peptide)
  • Antibodies to basement membrane (GBM)
  • Antibodies against cytoplasm of neutrophils / PR3 (c-ANCA)
  • Antibodies against myeloperoxidase neutrophils / MPO (p-ANCA)
  • Antibodies to parietal cells
  • Antibodies to mitochondria
  • Antibodies against thyroglobulin (Tg)
  • Thyroid microsomal antibodies (TPO)
  • Antimyocardiac antibodies (ACA)
  • Antibodies to cardiolipin IgG, IgM
  • Antibodies against beta 2 glycoprotein IgG, IgM
  • Antibodies to phosphatidylserine IgG, IgM
  • Antibodies to phosphatidic acid IgG, IgM
  • Antibodies against phosphatidylinositol IgG, IgM

Control Immunoglobulins-Complement

  • Serum Immunofixation
  • Quantitative determination of immunoglobulins IgG – IgA – IgM – IgE
  • Complement Control (factor C3, factor C4)

 

  1. Molecular Virology

It involves the molecular detection of viruses that are primarily involved in the pathogenesis of myocarditis.

The Laboratory receives samples daily, except Saturday and Sunday, from 08:00 to 16:00.

The results are delivered within 3 working days.

Suitable samples: Whole blood, tissue particles in biopsy material, cerebrospinal fluid (CSF) samples from respiratory.

Methodology: Real-Time PCR.

Pathogens detected

  • Cytomegalovirus (CMV)
  • Epstein-Barr (EBV)
  • Herpes Simplex Virus (HSV) Type 1/2
  • Adenoviruses
  • Influenza A / B
  • Parvovirus
  • Enteroviruses (Echo- / Coxsackie- / Polio- / Phino-)
  • Human Herpes Virus (HHV) 6,7,8

 

  1. Molecular Microbiology

It involves the simultaneous molecular detection of the 25 most common pathogens associated with sepsis.

The Laboratory receives samples daily, except Saturday and Sunday, from 08:00 to 11:00.

The results are delivered the same day.

Samples taken from 11:00 to 16:30 are analyzed the next working day.

Reception and analysis of samples passed operating hours can be made only after consultation with the Laboratory and only for OCSC’s inpatients.

Suitable samples: Whole blood in EDTA

The exam can be performed on other body fluids (cerebrospinal, pericardial fluid, bronchial lavage fluid, synovial fluid etc.) after consultation with the Laboratory.

Methodology: Multiplex Real-Time PCR (SeptiFast)

Bacteria and fungi detected:

Gram-positive: Staphylococcus aureus, Coagulase Negative Staphylococci, Streptococcus pneumoniae, Streptococcus spp., Enterococcus faecium, E. faecalis

Gram-negative: Escherichia coli, Pseudomonas aeruginosa, Klebsiella (pneumoniae / oxytoca), Serratia marcescens, Enterobacter (cloacae / aerogenes), Proteus mirabilis, Acinetobacter baumannii, Stenotrophomonas maltophilia

Fungi: Candida albicans, C. parapsilosis, C. tropicalis, C. glabrata, C. krusei, Aspergillus fumigatus

 

  1. Genetics

Molecular Thrombophilia Testing

The Laboratory receives samples daily, except Saturday and Sunday, from 08:00 to 16:00.

The results are delivered within 5 working days.

Suitable samples: Whole blood, genomic DNA.

Methodology: Reverse hybridization (strip assay) and Real-Time PCR.

Mutations detected:

  • Detection of a point mutation G1691V in the factor V (FV) of coagulation (Leiden)
  • Detection of Cambridge mutation of the V factor of coagulation
  • Detection of the Hong Kong mutation of the V factor of coagulation
  • Detection of point mutation G20210A of the factor II of coagulation (Prothrombin)
  • Detection of point mutation C677T of the MTHFR factor
  • Detection of point mutation A1298C of the MTHFR factor
  • Detection of polymorphism 4G/5G in the promoter region of the PAI

Pharmacogenetics

The Laboratory receives samples daily, except Saturday and Sunday, from 08:00 to 16:00.

The results are delivered within 5 working days.

Suitable samples: Whole blood, genomic DNA.

Methodology: Real-Time PCR

Genes checked:

  • Detection of polymorphisms of the CYP2C9 gene and the VKORC1 gene (associated with the metabolism of anticoagulants, Sintrom)
  • Detection of polymorphisms of CYP2C19 gene (associated with metabolism klopidigrelis, Plavix)

 

Future activities in Genetics

During 2016, MIHU will introduce a new activity that will involve the congenital cardiovascular disease genetics. Cardiovascular diseases are a group of complex diseases of the heart and blood vessels that may be caused to a great extent by genetic factors and are inherited in offspring. Using the new technology of sequencing next generation’s DNA (next generation sequencing) our Laboratory     will soon offer tests that detect the genetic basis of most cardiovascular diseases. These include cardiomyopathies (hypertrophic, dilated), arrhythmogenic syndromes (LongQT, Brugada, ShortQT, ARVC), familial hypercholesterolemia, and vascular diseases.

 

Research work

MIHU, in collaboration with the Transplant Unit and the OCSC’s Cardiology Departments is specialized in research Immunological and Cardiovascular diseases at the cellular, serological and molecular level, with particular emphasis on the role of immunological and inflammatory parameters in predisposition, treatment and prognosis of coronary artery disease, cardiac insufficiency, cardiomyopathies, and in acute and chronic rejection of transplants chest.