To isolate B cells from the total lymphocyte population using a modification of the rosette (by which in the last issue, we evaluated the distribution of T-cells). This method is called the EAC-rosette.
The method consists in the following. On the surface of B-lymphocytes have a receptor for complement component C3. Complement – a group of serum proteins that serves to kill bacteria. To identify a receptor for C3 complement component studied lymphocytes mixed with specially treated bovine red blood cells (which are first treated red cell antibody, and then fresh frozen mouse serum – a source of complement). After co-incubation of leukocytes and erythrocytes in the cells form figures outlets.
Norm: the relative content of 10 – 30%
absolute content of 100 – 900 cells / mm or 0.1 – 0.9? 10 9 / L
The reasons for increasing the content of B-lymphocytes:
The reasons for reducing the content of B-lymphocytes:
O-lymphocytes or null lymphocytes – cells is without a receptor specific for B-and T-lymphocytes. Among them there are NK-cells, young (immature) cells.
The study of the functional state of lymphocytes.
To determine the functional state of the cells examined the ability of these cells proliferate in response to the mitogens (substances that stimulate the reproduction), as well as the synthesis of antibodies.
Phytohemagglutinin (PHA) – is one of those substances that have on the white blood cells of mammals mitogenic. Receiving PHA of bean seeds.
To conduct the study leukocytes isolated from the blood by centrifugation (at Spin blood cells settle to the bottom of its layers depending on their weight), cultured (grown) in the presence of PHA for 72 hours.
Next, from cell culture prepare smears, fixed them in methanol (methyl alcohol), and stained in the same way as blood smears. In the light microscope with immersion system (such a microscope usually look swabs tissue slices) determine the percentage of blasts (young cells) in relation to the total number of lymphocytes.
Depending on the quality of reagents used, and the conditions of cultivation results of the response may be different. Of great importance is the quality of the serum added to the culture medium. Therefore, physicians need to find out in the laboratory in which research was conducted, what numbers are the norm.
Reduction of the proliferative response to PHA indicates the presence of immune deficiency, but the reasons for it may be different.
Increase the proliferative response to PHA may indicate the presence of infection or some autoimmune disorders.
Evaluation of the functional state of phagocytes.
Phagocytes (in this case – neutrophils) feed microorganisms. Phagocytic activity is assessed using the method to determine the fraction of cells able to form within itself “digestive bubble” – phagosome. “Eats” material may be particles of latex, red blood cells, test cultures of bacteria (eg, E. coli).
To estimate the “digesting” the ability of neutrophils using NBT-test (NST – it NBT). The absorption of microbes by phagocytes accompanied by an increase in oxygen consumption by the cells and the formation of hydrogen peroxide and oxygen free radicals (they both can be grouped under the name “reactive oxygen species” – ROS). The essence of the reaction is that the NBT in the presence of reactive oxygen species is colored in blue, and in their absence remains colorless.
NBT-test rate of 10-30%
The reasons for increasing the phagocytic activity of leukocytes:
acute bacterial infection.
Reasons for the decrease of the phagocytic activity of leukocytes:
acquired immune deficiency syndrome
The rate for Ig A: 0,6 – 4,5 g / l
The reasons for increasing the content of Ig A:
autoimmune diseases with the localization process in the lungs or in the intestines
Ig A nephropathy
The reasons for reduction of Ig A:
chronic illness (especially respiratory and gastrointestinal tract)
The rate for Ig M: 0,4 – 2,4 g / l
The reasons for increasing the content Ig M:
infection (acute exacerbation or chronic)
autoimmune diseases, especially rheumatoid arthritis.
The reasons for reduction of Ig M:
primary and secondary immunodeficiencies.
The rate for Ig G: 6,0 – 20,0 g / l
The reasons for increasing the content Ig G:
carried forward in the recent past infection.
The reasons for reduction of Ig G:
primary and secondary immunodeficiencies.
The rate for Ig E: 20 – 100 Re / l
Ig E content increased in various types of allergies.
PS: all is normal for adults!