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NK CELLS (FLOW CYTOMETRY)

Categoría: Immunological .
Días hábiles de entrega: 5
 

EDTA blood.

Store samples at temperatures between 10-25 ° C. (Maximum 24h of outlet); refrigerant gels or ice pack ice can be used for shipping.

The sample must be obtained by blood puncture (venous blood) with EDTA anticoagulant (Tube purple lid), strictly respecting the blood-anticoagulant relationship. A cell suspension, whole blood plus fluorescently labeled antibodies directed against specific cellular proteins, resulting in an antigen-antibody complex labeled with the fluorochrome, generating fluorescence signals of the antigen-antibody complex, which are detected by the flow cytometer.

When the reagent is added to the sample, the antibodies labeled with fluorochromes present in the reagent bind specifically to the antigens they are directed against, allowing the detection by flow cytometry of the different lymphocyte subpopulations. A lysis is then performed using a red cell lysing solution.

05 working days.

LEUKOCYTES (Cell / uL) xxx (V.R .: 4500-10500).

Lymphocyte counts.

TOTAL LYMPHOCYTES (Cells x mm3) x (%).

LYMPHOCYTES T CD3 (Cellulasxmm3) x (%) (V.R .: 700-2100).

LYMPHOCYTES T CD16 + CD56 (Cellulasxmm3) x (%) (V.R .: 745-553).

The count of the lymphocyte subpopulations is generally expressed as number of cells labeled by microliter μL (absolute count) or as percentage of labeled cells relative to the total lymphocytes or leukocytes present in the sample which in turn can be determined by Flow Cytometry on a base to its characteristic FSC / SSC pattern (size / granularity or internal complexity). The absolute count of each of the lymphocyte subpopulations (CD3 / CD16 + CD56), is calculated from the count of absolute lymphocytes, obtained from the hemogram performed the first 6 hours, once the sample was extracted.

NK and NKT cells are important as part of the cellular immune response. These cells represent other types of lymphocytes other than B or T lymphocytes. Their participation in immunopathogenesis of some diseases makes them an important part of the clinical laboratory cellular immune analysis, among other applications. The NK cells of the maternal-fetal interface have a primordial role in the pathogenesis of preeclampsia, a specific disease of pregnancy that is an important cause of maternal-fetal morbidity and mortality. In infectious diseases, it has been shown, for example, that the expansion of this population in patients infected with human immunodeficiency virus (HIV), precedes the development of adaptive immunity by CD8 + T cells during acute infection. The study of this population seems to be an indicator of monitoring the progression of the disease. NKTs are multifunctional cells, which can enhance immunity against microorganisms, eliminate tumors, suppress autoimmune diseases, and promote tolerance. Part of its functions depend on the recognition of lipids presented by the CD1 molecule, in addition to its interaction with dendritic cells. NKT activity may be increased in the pathogenesis of some autoimmune diseases, allergies and atherosclerosis. Additionally, the knowledge of NK and NKT cells in different pathological conditions has allowed proposing the use of these cells alone or in combination with other modalities of treatment of some types of cancer.

Alter G, Teigen N, Ahern R, Streeck H, Meier A, Rosenberg ES, et al. Evolution of innate and adaptive effector cell functions during acute HIV-1 infection. J Infect Dis 2007; 195: 1452-60.

Godfrey DI, Berzins SP. Control points in NKT-cell development. Nat Rev Immunol 2007; 7: 505-18.

Ljunggren HG, Malmberg KJ. Prospects for the use of NK cells in immunotherapy of human cancer. Nat Rev Immunol 2007; 7: 329-39.

Sentman CL, Barber MA, Barber A, Zhang T. NK cell receptors as tools in cancer immunotherapy. Adv Cancer Res 2006; 95: 249-92.

Van Kaer L. NKT cells: T lymphocytes with innate effector functions. Curr Opin Immunol 2007; 19: 354-64.



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