The advantage with this technology is that analysis can be performed on at least one sample with very high sensitivity and precision. With this method, we can investigate independently patients and we can suggest targeted therapies according to the following tests:
1. Immunological Phenotype
– Subpopulations of Lymphocytes are investigated such as the absolute number of NK cells, T-cells, B-cells, T cytotoxic or NK like cells and T helper cells in all biological fluids (eg. CSF, blood, BAL). By this way, we can detect the nature of the disease.
In the above methodology, usually the populations of cells are written with a (+) or (-) sign, which indicate the presence or absence of specific surface antigen.
Applications of Immunophenotype in Clinical Laboratorial Medicine
- Infertility, miscarriages, failed IVF trials,
- Cancer,
- HIV,
- Hematological malignancies (leukemias, lymphomas),
- Autoimmune diseases (Hashimoto’s multiple sclerosis etc.),
- Septic syndrome (viral Hepatic cirrhosis),
- Lung diseases,
- Inflammations general,
- Investigation of therapeutic efficacy and follow ups,
- Frequent sickness due to low immunogical response.
2. Detection of 13 cytokines Th1/Th2 important of balance of immune system
The investigation of immune response of lymphocytes type I(Th1) and II (Th2) is very important for the good function of the immune system because:
- Ø TH1 produce IL-2, IFN-γ, IL-8, IL-12, TNF-β
- Ø TH2 produce IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-13
- Ø TH17 produce IL-17
- Ø TH23 produce IL-23
The imbalance of Th1/Th2 is observed in the following diseases:
Irritable bowel syndrome (IBS), cystic fibrosis, chronic gastritis, tuberculosis, thromocytopenia, aging, cancer, autoimmune diseases, and others.
ΤΗ17 lymphocytes participate in regulating autoimmune reactions like chronic asthma, rheumatoid arthritis, inflammatory bowel disease, and multiple sclerosis.
ΤΗ23 λεμφοκύτταρα lymphocytes are characterized by most autoimmune diseases and the symptoms for the patient are insomnia, pain to the whole body, fibromyalgia, stiffness, and depression.
3. Detection of platelet functionality (platelet receptors)
The platelet receptors to be investigated have:
Specificity for specific additive substances, which are modulated during their activation expressing different epitopes.
Flow cytometry applies in the following ways in platelets:
To investigate without addition of stimulator, the status of activation of platelet (in vivo),
To detect the membrane of glycoproteins of platelets,
To estimate the capability of stimulation of platelets after the addition of agonist (in vitro), in order to be investigated the aggregation of leucocytes-platelets. All these can help doctors to estimate precisely the therapy with drugs such as agonists with receptors GPIIb-IIIa. Also, this testing of receptors can help in diagnosis in most thromboembolic diseases such as Bernard-Soulier or thrombo diseae Glanzmann. Flow cytometry can also help doctors from selecting the ideal drug for their patients just by testing their platelet receptors and test drugs on their own platelets.
Investigation of functionality of platelet via the following antibodies (PAC-1, CD41, CD42a, CD42b, CD61, CD62P) with main purpose the recognition of surface antibodies as shown below:
- CD41 – Glycoprotein IIb
- CD42a – Glycoprotein IX
- CD42b – Glycoprotein Ib
- CD61 – Glycoprotein IIIa
The above platelet diagnostic tests are useful for the following diseases:
- Major injury, fractures,
- Pregnancy,
- Malignancy,
- Birth control or hormonal restoration,
- Hypertension,
- Anti-phospholipid syndrome,
- Paroxysmal nocturnal hemoglobinuria (PNH),
- Inflammatory bowel disease,
- Nephrotic syndrome,
- Emboli,
- Myelodisplastic syndrome,
- Estimation of surgical outcome, (orthopaedics)
4. Testing – Detection of allergic factors with definition of degranulated basophils
The marker of degranulation of basophils is used for the study of pharmacological or biological supplement for the therapy in allergic reactions, for the study and comparison of allergic response with natural allergens, in chemical modifying substances, and in recombinants allergens, even in diseases like allergic rhinitis, atopic dermatitis, bronchial asthma etc. for the evaluation of the progress of therapy.
5. More investigating tests with flow cytometry
* (QUANTITATION of SURFACE RECEPTORS):
CD38, CD20
*DNA ANALYSIS with detection of cells in different multiplication phases:
1. RELAXATION (G0/G1) 2. SYNTHESIS DNA (S 3. MITOSIS (G2/M), DNA INDEX, apoptosis, and detection of alterations in cellular membrane:
*Caspases detection if requested specifically for any project.
NOTE: Our center in the near future will present new innovative tests with new antibodies for most diseases, which will guide potentially for the more targeted and individualized diagnostic and therapeutic approach.
