Facilities:

Receipt of samples. Common area with the rest of the laboratories. Located in the general laboratory (ground floor, research building).

Blood drawing. Common area with the rest of laboratory blood draws. Located on the mezzanine floor.

Immunology laboratory area: Located on floor B1, research building, distributed in 3 main sub-areas:

  • Haematological, Cellular and Immunodeficiency Diagnosis Laboratory (Mixed: Haematology + Immunology).
  • Autoimmunity and Immunoallergy Laboratory.
  • Biochemistry, molecular biology and HLA and disease laboratory.

Outpatient area: Located on Floor 1 of the hospital (office 6 of the Haematology area). Clinical Immunology and Adult Immunodeficiency Consultation) and on the ground floor (Blood Bank. Gestational Pathology Immunology Consultation). It has the following resources:

  • 1 office for consultations, shared with the Haematology Service.
  • 1 waiting room shared with the Haematology Service.
  • 1 office for the Haematology/Oncology/Immunology Administration Desk.
  • 1 Oncohaematology Day Hospital. Shared with other Services.

The service does not have a Hospitalisation Unit, although it does offer hospitalised patient interconsultations.


Techniques and procedures

Haematological, Cellular and Immunodeficiency Diagnosis Laboratory

Fundación Jiménez Díaz's flow cytometry laboratory is focused on haematological and immunological diagnostics. It is multidisciplinary in nature, and includes specialists in Immunology and Haematology.

Equipment

The flow cytometry laboratory has the following flow cytometers (analysers):

  • Two FACSCantoTM II cytometers: three lasers (405 nm violet laser, 488 nm solid-state blue laser, and 633 nm He-Ne red laser) to determine 8 parameters at the same time, in addition to 2 scattering parameters (FSC and SSC).
  • An AQUIOSTM CL cytometer: automated and continuous loading. A 488 nm solid-state blue laser to work with 4 fluorescences at the same time, 2 scattering parameters (FSC and SSC), and electronic volume (EV).
  • CytoFlex: three lasers (405 nm violet laser, 488 nm blue laser, and 638 nm red laser) to determine 13 parameters at the same time, in addition to 2 scattering parameters (FSC and SSC).

Techniques and procedures

Flow cytometry is an objective, specific, sensitive, fast technology for multiparameter study of cells. Any sample can be studied, as long as a cell suspension is obtained. Fluorochrome-labelled monoclonal antibodies are used in preparing the samples. These bind specifically to the cellular, nuclear, cytoplasmic and/or surface molecules to be studied. The cells in suspension are acquired individually in the flow cytometer, such that the laser light emitted by the equipment is focused on each of them, generating:

  • Laser light scattering: can be frontal or lateral, reporting the size and complexity of the cell, respectively.
  • Excitation of antibody fluorochromes: Antibodies are specific to certain molecules. If the molecule under study is found in the cell, the antibody recognises it and the laser light excites its fluorochrome, releasing fluorescence. There will be no fluorescence emission if the molecule is not in the cell. It is therefore possible to identify whether a molecule is present or not, and, if present, in what amount.
  • The light signals are detected and transformed by the cytometer's electronic system, which generates accessible data that can be interpreted by the cytometrist using analysis software (InfinicytTM).

This technique is mostly applied in Fundación Jiménez Díaz's flow cytometry laboratory for:

  • Immunophenotypic study of cells in suspected haematological diseases: lymphomas, leukemias, multiple myeloma, myelodysplasias, etc. Diagnosis and minimal residual disease.
  • Paroxysmal Nocturnal Haemoglobinuria diagnostics.
  • Determination of foetal haemoglobin in maternal blood.
  • Study of hereditary spherocytosis.
  • Study of platelet antigens.
  • Determination of CD34 cells in haematopoietic progenitor transplantation.
  • Post-transplant immune reconstitution study.
  • Cellular viability study.
  • B-cell quantification in the evaluation of follow-up in anti-CD20 therapies.
  • Study of intraepithelial lymphocytes (IELs) in patients with suspected coeliac disease.
  • Determination of lymphocyte subpopulations in the follow-up of HIV patients.
  • Determination of oxidative capacity of neutrophils.
  • Study of lymphocyte subpopulations under suspicion of primary immunodeficiencies.
  • Initial screening in the HLA-B27 antigen study.

Biochemistry, molecular biology and HLA and disease laboratory.

Equipment

The laboratory has thermocyclers and real-time PCR equipment, an HLA typing unit for SSO, three automated units for immunoelectrophoresis, an immunoelectrofocusing unit for oligoclonal bands, a chemiluminescence analyser, and an immunoassay unit for HIV and HCV confirmations.

Techniques and procedures:

  • HLA study for diagnosis of Coeliac Disease HLA DQ2/DQ8.
  • Determination of disease-associated alleles HLA B57, HLA B51, HLA B27 and HLA DQ6.
  • Determination of IgG and IgD subclasses and monoclonal components and identification of cryoglobulins.
  • Identification of Oligoclonal Bands in CSF for the diagnosis of Multiple Sclerosis.
  • Determination of KL6, for diagnosis and follow-up of interstitial lung disease.
  • CSF determination of Alzheimer's disease biomarkers.
  • Determination of serology in relation to Hepatitis Virus (A, B and C), Rubella, Cytomegalovirus, Toxoplasma.
  • Determination of Hepatitis B and C Viral Load.
  • Identification of Hepatitis C genotypes.
  • Tests for diagnosis and follow-up of HIV patients: determination of antibodies against HIV1/2 and P24 antigen by 4th generation ELISA, viral load, CD4 T lymphocyte count and study of antiretroviral drug resistance mutations.

Autoimmunity and Immunoallergy Laboratory

Equipment

The Allergy Laboratory has 2 Phadia IC1000 (Thermo®) sample processing units. These instruments are configured to work in high-throughput laboratories, adjusting to the growing demand for in vitro allergy testing.

The Autoimmunity Laboratory has a wide variety of automated equipment to determine autoantibodies using different immunoassay platforms: ELISAs, chemiluminescence, fluoroenzyme immunoassay, multiplex techniques, strip immunoassay.

It has 2 fluorescence microscopes, an automatic reading system for slides, and software for full traceability of the patient and to generate an image library.

The service also uses a Laboratory Information System to ensure comprehensive management of results, enabling seamless integration and connectivity between the section's analysers.

Techniques and procedures:

  • Allergy Laboratory:
    • Wide range of tests focused on studying in vitro allergies. Screening techniques, specific identification of allergens, and study of specific IgE at molecular level are all performed. This gives a detailed picture of the patient's IgE profile, assessing the risk of developing serious clinical events, explaining the symptoms due to cross-reactivity, and reporting the potential effect of immunotherapy.
  • Autoimmunity Laboratory:
    • The presence of autoantibodies is part of the classification criteria in numerous autoimmune pathologies, and determining them is key in diagnosis.
  • Study of Organ-Specific Autoimmune Diseases:
    • Autoimmune Thyroid Disease.
    • Autoimmune Gastritis.
    • Autoimmune Hepatitis.
    • Primary Biliary Cholangitis.
    • Inflammatory Bowel Disease.
    • Diabetes Mellitus.
    • Autoimmune Nephropathies.
    • Autoimmune Blistering Diseases.
    • Autoimmune Polyglandular Syndromes.
  • Study of Systemic Autoimmune Diseases:
  • Rheumatoid Arthritis.
  • Systemic Lupus Erythaematosus.
  • Systemic Sclerosis.
  • Inflammatory Myopathies (Polymyositis/Dermatomyositis/Inclusion Body Myopathy).
  • Mixed Connective Tissue Disease/Overlap Syndromes.
  • Anti-phospholipid syndrome.
  • ANCA-positive vasculitis.
  • Coeliac disease
  • Biological Drug Monitoring: Infliximab, Adalimumab.
  • Determination of Interleukin 6.