Laboratory assessment of innate immune disorders

Functional assessment of B lymphocytes

Determining the concentration of immunoglobulins in blood serum and other biological fluids is one of the most important integral indicators of the functional usefulness of B-lymphocytes and the entire system of regulation of humoral adaptive immunity.

Traditionally, the concentration of serum immunoglobulins was determined by radial immunodiffusion in gel according to Mancini. Currently, this method is still in use, with preference given to monoclonal antibodies against certain antigenic determinants(epitopes) characteristic of certain classes of immunoglobulins.

The principle of the Mancini method consists in the fact that samples of the test sera are placed in agar wells that contain antibodies against IgM, IgG, IgA in a standard concentration. Immunoglobulins contained in the test serum diffuse into the agar and, when interacting with the corresponding antibodies, form precipitation rings. As long as excess antigen remains in the well, a gradual increase in the diameter of the precipitation ring occurs. The final size of the precipitation ring depends on the concentration of the corresponding immunoglobulins. The Mancini method allows you to determine the concentration of immunoglobulins equal to 10 μg/ml. The method error is 10%.

To diagnose innate immune disorders, the phagocytic and metabolic activity of neutrophils is usually determined and the state of the complement system is assessed.

The most informative for assessing the phagocytic system should be considered the phagocytic index, phagocytic number (reflects the absorption capacity of neutrophils) and the bactericidal index (reflects the digestive ability of a neutrophil).

Phagocytic index(PI) – the percentage of cells that entered phagocytosis out of their total number.

Phagocytic number(NF) is the average number of bacteria or latex particles ingested per neutrophil.

Also held Spontaneous and stimulated phagocytosis tests using latex particles. Phagocytosis with latex is based on the uptake of latex particles by neutrophils (adhesion, capture and complete uptake).

In a spontaneous test, leukocytes isolated from peripheral blood are mixed with a suspension of latex particles; When conducting an induced phagocytosis test, a pyrogenal solution is added to the resulting mixture. After incubation at 37 0 C for 30 minutes, smears are prepared, and then the percentage of neutrophils that have absorbed latex is calculated in fixed and stained Romanovsky-Giemsa smears.

Nitro blue tetrazolium reduction test–NST test allows you to evaluate the metabolic activity of blood granulocytes, reveals the reserve capabilities of the intracellular systems of phagocytes. The NBT test is based on the reduction of a soluble dye, nitroblue tetrazolium, absorbed by blood leukocytes into insoluble black diformazan (intranuclear inclusions - diformazan clumps) under the influence of superoxide anion formed in the NADP-H oxidase reaction, which initiates the process of stimulating phagocytosis.

Setting up a reaction. 0.02 ml of a 0.9% sodium chloride solution, 0.02 ml of a neutrophil suspension and 0.02 ml of a 0.75% NBT (nitro blue tetrazolium) solution are applied to a fat-free glass slide.

Commercial lipopolysaccharide from gram-negative bacteria E. coli or a solution of pyrogenal (lipopolysaccharide from S. typhi) at a dilution of 1:10 is used as a load. 0.02 ml of LPS, 0.02 ml of neutrophil suspension and 0.02 ml of 0.75% NBT solution are applied to another defatted glass. Place in a humid chamber for 30 minutes. Smears are prepared, dried, and fixed. Stain with 0.1% neutral red solution. Neutrophils are counted by calculating the percentage of leukocytes with reduced NBT granules (black diformazan).

Phagocytic activity of leukocytes– this is the determination of the content of neutrophils and monocytes capable of binding on their surface, absorbing and digesting a microbial test culture (labeled bacteria).

Indications for laboratory testing:

• Frequent recurrent infectious diseases,
• recurrent purulent inflammatory processes,
• long-term non-healing wounds,
• frequent postoperative complications,
• suspicion of autoimmune diseases,
• dynamic monitoring of such patients, assessment of the activity and effectiveness of therapy for collagenosis and rheumatic diseases.

Reasons for increased phagocytic activity of leukocytes:

• acute bacterial infections.

Reasons for decreased phagocytic activity of leukocytes:

• congenital immunodeficiencies
• chronic infections
• autoimmune diseases
• allergic diseases
• viral infections
• human acquired immunodeficiency syndrome

No special preparation is required for the study. Must be followed general rules preparation for research.

GENERAL RULES FOR PREPARATION FOR RESEARCH:

1. For most studies, it is recommended to donate blood in the morning, from 8 to 11 o’clock, on an empty stomach (at least 8 hours must pass between the last meal and blood collection, you can drink water as usual), on the eve of the study, a light dinner with a restriction eating fatty foods. For tests for infections and emergency studies, it is acceptable to donate blood 4-6 hours after the last meal.

2. ATTENTION! Special preparation rules for a number of tests: strictly on an empty stomach, after a 12-14 hour fast, you should donate blood for gastrin-17, lipid profile (total cholesterol, HDL cholesterol, LDL cholesterol, VLDL cholesterol, triglycerides, lipoprotein (a), apolipo-protene A1, apolipoprotein B); glucose tolerance test performed in the morning on an empty stomach after 12-16 hours of fasting.

3. On the eve of the study (within 24 hours), eliminate alcohol, intense physical activity, and medicines(in consultation with the doctor).

4. 1-2 hours before donating blood, refrain from smoking, do not drink juice, tea, coffee, you can drink still water. Avoid physical stress (running, quickly climbing stairs), emotional excitement. It is recommended to rest and calm down 15 minutes before donating blood.

5. You should not donate blood for laboratory testing immediately after physiotherapeutic procedures, instrumental examination, X-ray and ultrasound examinations, massage and other medical procedures.

6. When monitoring laboratory parameters over time, it is recommended to conduct repeated tests under the same conditions - in the same laboratory, donate blood at the same time of day, etc.

7. Blood for research must be donated before starting to take medications or no earlier than 10–14 days after they are discontinued. To assess the control of the effectiveness of treatment with any drugs, a study should be conducted 7-14 days after the last dose of the drug.

If you are taking medications, be sure to notify your doctor.

12. Cellular protective factors. Phagocytosis, stages, characteristics. Methods for determining phagocytic activity, phagocytic index, phagocytosis index.

For the occurrence of infection, along with the properties of the pathogen, a complex of factors and mechanisms of MK (sensitivity or resistance to infection) is important.

SKIN AND MUCOUS MEMBRANES

a mechanical barrier for most microorganisms - prevents penetration into the body. Constant desquamation upper layers epithelium, gland secretions help remove microorganisms from the surface.

bactericidal properties due to the action of lactic and fatty acids, various enzymes secreted by skin glands, lysozyme of tear fluid, saliva and other secretions.

NORMAL MICROFLORA

promotes maturation immune system,

plays a role in the nonspecific protection of the areas of the gastrointestinal tract, DP and MPT inhabited by them, because microbes living in certain biotopes prevent adhesion and colonization of mucous membranes by pathogenic microbes (pathogen antagonists).

But some representatives of N μF can cause diseases in cases of penetration in large numbers from one biotopes to others (with dysbacteriosis and immunodeficiencies).

HUMORAL FACTORS: lysozyme, complement, interferons.

PHAGOCYTING CELLS(I. I. Mechnikov in 1883). All phagocytic  are divided into: microphages(PMN: neutrophils, eosinophils and basophils) and macrophages various tissues of the body (connective tissue, liver, lungs, etc.). Macrophages, together with blood monocytes and precursors (promonocytes and monoblasts), are united in the system of mononuclear phagocytes (MPF). SMF is phylogenetically more ancient compared to immune.

Micro- and macrophages have a common myeloid origin (from PSC). Peripheral blood contains more granulocytes (mature cells, 60–70% of all white blood cells) than monocytes (1–6%). Monocytes, leaving the bloodstream, mature into tissue macrophages. The liver, spleen, and lungs are especially rich in them.

The membrane of all phagocytes is folded and carries many specific receptors and antigenic markers that are constantly updated. The lysosomal apparatus is well developed; lysosomes can fuse with the membranes of phagosomes or with the outer membrane. In the latter case, cell degranulation occurs and concomitant secretion of lysosomal enzymes into the extracellular space.

FUNCTIONS OF PHAGOCYTES:

Protective– cleaning  from infectious agents, tissue decay products, etc.

Representing– presentation of Ag epitopes on the phagocyte membrane

Secretory– secretion of lysosomal enzymes and other biologically active substances (monokines), which play an important role in immunogenesis.

STAGES OF PHAGOCYTOSIS:

Chemotaxis– targeted movement of phagocytes in the direction of the chemical gradient of chemoattractants (B! components, tissue degradation products, fractions C5a, C3a, lymphokines) is associated with the presence of specific receptors.

Adhesion– mediated by receptors, but nonspecific physical-chemical interactions can also occur. Adhesion immediately precedes endocytosis (uptake).

Endocytosis= phagocytosis (particles >0.1 µm) and pinocytosis. Phagocytic cells are capable of capturing inert particles (coal, latex), flowing around them with pseudopodia WITHOUT PARTICIPATION SPECIFIC RECEPTORS, unlike bacteria, Candida and other mk. The most effective phagocytosis is mediated by Fc receptors and receptors for C3 - IMMUNE. As a result of endocytosis, a phagosome is formed.

Only some bacteria (noncapsular strains of pneumococcus, strains of streptococcus lacking hyaluronic acid and M-protein) are phagocytosed directly. Most bacteria are phagocytosed only after they are opsonized by complement and/or antibodies.

Digestion- occurs in phagolysosomes, μ die as a result of the action of oxygen-dependent (“oxidative explosion”) and oxygen-independent mechanisms (cationic proteins and enzymes (including lysozyme)).

Incomplete phagocytosis– many virulent B! often do not die and persist for a long time inside phagocytes, thanks to various mechanisms (impaired fusion of lysosomes with phagosomes - toxoplasma, tbc; resistance to lysosomal enzymes - gono-, staphylo-, group A streptococci, etc.; exit from the phagosome - rickettsia, etc.).

REPRESENTING FUNCTION OF Macrophages consists of fixing antigenic epitopes of mk on the outer membrane. In this form they are presented for specific recognition by T lymphocytes.

SECRETORY FUNCTION consists in the secretion of biologically active substances (monokines - substances that regulate the proliferation, differentiation and functions of phagocytes, lymphocytes, fibroblasts and other cells). A special place among them occupies IL-1, k/y activates many functions of T-lymphocytes, incl. IL-2 production. IL-1 also has the properties of an endogenous pyrogen (acting on the nuclei of the anterior hypothalamus). Macrophages produce and secrete prostaglandins, leukotrienes, cyclic nucleotides, oxygen radicals (0 2, H 2 0 2), complement components, lysozyme and other lysosomal enzymes, interferon. Due to these factors, phagocytes can kill bacteria not only in phagolysosomes, but also outside cells, in the immediate microenvironment.

METHODS FOR DETERMINING PHAGOCYTIC ACTIVITY

The phagocytosis reaction is based on opsonization of the pathogen.

A fraction of phagocytes is isolated from the blood, gonococci and serum of the patient being examined are added to them (Am + C). After a certain time, the smears are examined and at least 100 phagocytes are counted. From these, the %  of captured microbes is determined. In N PHAGOCYTIC INDICATOR = 40-80%.

PHAGOCYTIC NUMBER - count the number of captured microbial cells, sum and divide by the number of phagocytes, obtain the number of microbial  absorbed by one phagocyte. In N PF = 1-5.

Study of phagocytic activity of leukocytes- a blood test that is aimed at determining reserve capabilities neutrophils and monocytes to perform their main function - the absorption and processing of foreign agents. The test is performed as part of an immunogram complex. It is indicated for patients with recurrent and chronic infections, acquired and genetic immunodeficiency conditions, autoimmune and oncological diseases, who have undergone complex operations, including organ transplantation. Whole blood is analyzed. The study is based on assessing the phagocytosis of fluorescently tagged bacteria. Normally, phagocytic granulocytes make up from 82 to 90% of the total, phagocytic monocytes – from 75 to 85%. Readiness of results – up to 8 days.

The phagocytic activity of leukocytes is a laboratory indicator that reflects the percentage of neutrophils and monocytes capable of binding to pathogenic microflora and digesting it. Phagocytes are cells that protect the body from the development of infections. They are considered a component of innate immunity; in the blood they are represented by two types of leukocytes - monocytes and neutrophils. Monocytes are large cells - macrophages. They have a pronounced ability to absorb, process large-sized cells and organic compounds. At the site of inflammation, they phagocytose bacteria, leukocyte mass, and affected cells. As a result, the tissues are cleansed and prepared for regeneration. Neutrophils are microphages; unlike monocytes, they absorb only small cells and organic components. After processing the agents, neutrophils die, release substances that damage bacteria and fungi, and increase the flow of immune cells to the site of inflammation.

A blood test for the phagocytic activity of leukocytes allows you to assess the reserve of monocytes and neutrophils for digesting foreign agents. Changes in the characteristics of phagocytes reflect not only the immunological reactivity of the body, but also the characteristics of some other processes - protein and carbohydrate metabolism, the presence of intoxication and exhaustion of the body, the activity of recovery after diseases, etc. Thus, the analysis is used not only in immunology and infectious diseases, but and in rheumatology, oncology, surgery. The results of the study are displayed as the percentage of active phagocytes to their total number. Active neutrophils and monocytes are detected using bacteria with fluorescent tags. The biomaterial for the study is whole blood with heparin.

Indications

A study of the phagocytic activity of leukocytes is indicated if congenital or acquired immunodeficiency is suspected. It is prescribed for prolonged, chronic and recurrent infectious diseases – characteristic feature decreased immunity. Most often, patients with pneumonia, sinusitis, otitis, enterocolitis, candidiasis, and cystitis are sent for analysis. Also, long-term non-healing wounds and complications after operations may indicate a lack of immune defense. Therefore, the analysis is performed in preparation for surgical intervention and in complicated cases postoperative period, during long-term recovery after injuries and burns. Other indications for this study include allergic, autoimmune and oncological diseases. The results allow us to evaluate the activity of immune defense (phagocytosis) and its role in the development of the disease.

A blood test for the phagocytic activity of leukocytes makes it possible to determine the actual readiness of the body to resist infections. However, it is worth remembering that this indicator changes under the influence of many factors. Thus, the activity of monocytes and neutrophils decreases after physical activity and with mental fatigue, and after eating high-calorie food it increases. Another limitation of the analysis is that the research procedure takes up to 8 working days, the results obtained reflect the state of the week ago.

Preparation for analysis and collection of material

To study the phagocytic activity of leukocytes, blood is taken from a vein. The day before the test, you need to eliminate alcohol from your diet, cancel sports training and other intense physical activity, and avoid stressful situations. It is also necessary to consult with your doctor about the effect of the medications you are taking on the test result; it is possible that some medications will be temporarily discontinued. The blood sampling procedure is usually performed in the morning, after an overnight period of fasting or 4 hours after eating.

Blood is taken from the ulnar vein using a puncture; to study the phagocytic activity of leukocytes, the method of assessing the phagocytosis of bacteria with a fluorescent label is most often used. Monocytes and neutrophils, the test material, are isolated from the blood by centrifugation and washing. Then a culture of luminescent bacteria is introduced into the sample, the mixture is resuspended and incubated, and the number of leukocytes that have phagocytosed the bacteria is determined by the intensity of luminescence. Analysis results are prepared within 7-8 business days. The phagocytic activity of monocytes and neutrophils can be determined by other methods, for example, by staining phagocytosed cells (Romanovsky-Giemsa method), by the activity of lysosomal enzymes, the production of cytokines, and the presence of cationic proteins.

Normal values

The result of a blood test for the phagocytic activity of leukocytes is expressed as a percentage of phagocytic cells from their total number. Normal values ​​for granulocytes are from 82 to 90%, for monocytes – from 75 to 85%. These rates are the same for patients of all ages and both sexes. A physiological decrease in phagocytosis can be detected during pregnancy, after physical activity that does not correspond to the level of training, and after emotional stress.

Increasing and decreasing the indicator

An increase in the phagocytic activity of leukocytes has no diagnostic significance; acute infections may be the cause. The activity of monocytes and neutrophils increases relative to the initial level.

Analysis of the phagocytic activity of leukocytes is an immunological research method. Its indicators make it possible to determine the reserve abilities of blood cells to absorb and digest infectious agents, that is, the body’s readiness to resist the development of the disease. If the test results are below normal, you should consult your doctor - an immunologist, an infectious disease specialist, a surgeon, a rheumatologist, or an oncologist. Physiological decline can be corrected correct selection physical activity, stress prevention.

PHAGOCYTIC ACTIVITY OF HUMAN BLOOD NEUTROPHILS IN HYPOTONIC ENVIRONMENTS IN THE PRESENCE OF ANTIBIOTICS

PHAGOCYTIC ACTIVITY OF NEUTROPHILS OF HUMAN BLOOD IN HYPOTONIC MEDIUM BY ANTIBIOTIC ACTION

A.A. Mishchenko, E.M. Savelyeva

A.A. Mischenko, E.M. Savelyeva

The phagocytic activity of human blood neutrophils in a hypotonic environment and in the presence of a number of antibiotics was studied. A decrease in tonicity by 1.5 -2.0 times causes an increase in phagocytosis parameters by 16%. In the presence of furosemide, the effect of hypotension is not observed. Various antibiotics cause strong inhibition of phagocytosis.

Phagocytic reaction of human neutrophils in the hypotonic medium and at the presence of antibiotics was investigated. Tonicity decrease of medium in 1.5 - 2 times causes increase of phagocytosis parameters on 16%. At furosemide presence action of a hypotonia is not shown. Various antibiotics caused strong inhibition of phagocytosis.

Keywords: neutrophil, phagocytosis, phagocytic activity, hypotension.

Key words: neutrophil, phagocytosis, phagocytic activity, hypotonycity.

Introduction

The main barrier to infection entering the body is the mucous membranes. Being multicomponent systems, they take part in many body reactions, including immune ones. Normally, the mucous membrane contains immunoglobulins and a small number of neutrophils and macrophages. It is these cells that are the first to come into contact with pathogens, but if the latter penetrate into the thickness of the tissues, lymphoid accumulations in the thickness of the mucous membranes become a barrier.

Since the surfaces of the mucous membranes are not isotonic with blood plasma, to assess the functioning of immune system cells under such conditions, it is advisable to conduct experiments in an anisotonic environment in vitro. Thus, it has been shown that in hypotonic solutions oxygen explosion and metabolism are activated in leukocytes arachidonic acid, the concentration of calcium ions increases. In our work, we conducted studies of the phagocytic activity of blood neutrophils during modeling of hypotonic conditions. Since in the case of an inflammatory process in the body, doctors often prescribe antibiotic therapy, the effect of antibiotics of various classes on the phagocytic activity of blood neutrophils was also studied.

Objects and Methods

The experiments used venous blood from male donors obtained from the Republican Blood Transfusion Station (Syktyvkar). 500 μl of blood was placed into the wells of the plate for immunological reactions. A suspension of yeast cells (Saf-Neva LLC), pre-washed three times with a 0.9% NaCl solution, was added to each well. The number of yeast cells averaged 30 thousand/1 μl of blood.

To reduce tonicity by 2.0 and 1.5 times, distilled water (pH 7.4) was added to the wells. In a number of experiments, furosemide was also added to wells with iso- and hypotonic media at a concentration of 1*10-5 Mol/l.

In experiments with antibiotics, lincomycin, ceftriaxone, amoxiclav and gentamicin were added to the wells at a concentration of 30 mg/l.

The samples were incubated in a thermostat at 370C for 20 minutes. Then the plate was placed on ice to stop the phagocytosis reaction, and 3 smears were prepared from each well. After drying and fixing, the smears were stained according to Giemsa-Romanovsky and viewed under a microscope at immersion magnification 15*90.

We calculated: 1) phagocytic activity - the number of active neutrophils out of 100 encountered during viewing; 2) phagocytic index - the average number of yeast cells absorbed by one neutrophil. The results were processed using the method of paired comparisons; the significance of differences between samples was assessed using the Wilcoxon test.

Results and discussion

In the control, the phagocytic activity of human blood leukocytes was 49.5±5%, the phagocytic index was 1.64±0.1 (n=20). The data are consistent with the results obtained from studying the phagocytosis of pathogenic Candida crusei. The phagocytic activity of neutrophils under these conditions is stimulated by P-glucans in the yeast cell wall, for which there are receptors on the surface of phagocytes, as well as by the opsonizing effect of the components of the complement system C3bi and IgG immunoglobulins present in the blood plasma.

When the tonicity of the medium decreased by 1.5 and 2.0 times (n=20), phagocytic activity increased by an average of 16.5%, amounting to 58.1±9.1% (p<0.05). Увеличился также фагоцитарный индекс до 1.97±0.06 и 2.14±0.58 при снижении тоничности в 1.5 и 2.0 раза соответственно. Таким образом, гипотония вызвала активацию фагоцитоза, что отразилось в увеличении как доли активных клеток, так и скорости поглощения фагоцитами дрожжей. Одним из механизмов такого

The effects of hypotension may be an increase in the concentration of intracellular calcium, as a result of which the cytoskeleton of cells, their motility and phagocytic activity changes. Additionally, cell activity under these conditions may change as a result of triggering the regulatory volume depletion response, RVD, in response to cell swelling. In order to exclude the influence of the latter, furosemide inhibited K+,Cl" - cotransport, the activation of which leads to RVD. Since the substance changes the chemotactic activity of cells, its effect was previously studied in an isotonic environment. However, furosemide did not change the phagocytic activity of neutrophils (n = 20 ), which is consistent with published data... Against the background of a hypotonic environment, furosemide did not change phagocytic activity compared to the control and reduced it compared to the results in a hypotonic environment in the absence of the substance (p<0.05). В частности, в присутствии фуросемида фагоцитарная активность и фагоцитарный индекс составили 45.9±6.7%; 1.83±0.1 и 50.5±5.6%; 1.7±0.1 соответственно при

hypotension 1.5 and 2.0. Therefore, by blocking the RVD response, furosemide thereby prevented cell activation under hypotensive conditions.

Under the influence of antibiotics, the indicators of phagocytic activity decreased. When exposed to ceftriaxone, phagocytic activity decreased by 78% to 10±2.3% (p<0.02), амоксиклав - на 70% до 17±3.9% (р<0.02), линкомицин - на 65% до 16±4.9% (р<0.02), гентамицин - на 76% до 11±3.6% (р<0.02). Фагоцитарный индекс в экспериментах с антибиотиками практически оставался неизменным, следовательно, данные препараты не влияют на скорость поглощения клеток.

An increase in phagocytic activity under the influence of antibiotics has been noted in a number of studies. According to other data, the phagocytic activity of leukocytes is suppressed by the action of antibiotics such as aureomycin. Oxytstracycline, erythromycin, chloramphenicol, polymyxin B do not cause noticeable changes in the phagocytic activity of leukocytes.

The antibiotics used in the experiments belong to different groups according to their action. Amoxiclav and ceftriaxone act as bactericidal drugs (inhibit the development of the cell wall, and suppress the synthesis of peptidoglycan, murein, specific to the bacterial cell wall). Lincomycin and gentamicin act as bacteriostatics at low concentrations and bactericidal when the concentration increases (they inhibit protein synthesis by binding to the 50s and 30s ribosomal subunits). All antibiotics in our experiments had an inhibitory effect on neutrophils. This may be due to a change in structure

antibacterial drugs due to metabolic processes in the body, as a result of which metabolic products become toxic to

the phagocytes themselves. Antibiotics are one of the main causes of neutropenia and agranulocytosis. This effect was exerted by penicillins, cephalosporins and sulfonamides. The aminoglycoside gentamicin increases the formation of lysosomes containing various virulence factors. Representatives of P-lactam antibiotics, amoxiclav and ceftriaxone, can suppress the oxidative burst reaction.

Data on the effect of lincosamines, which include the used lincomycin, on phagocytic activity are contradictory. At different concentrations of the drug, the authors note both an increase in phagocytic activity, as well as a decrease or no change. It is possible that the concentration of antibiotics used in our experiments was toxic to cells.

1. In the control, phagocytic activity and phagocytic index were 49.5±5% and 1.64±0.1, respectively.

2. Replacing an isotonic medium with a hypotonic one causes an increase in both phagocytic activity and phagocytic index, respectively, by 16.5% and 1.5-2 times.

3. In the presence of furosemide, the effect of hypotension on the phagocytic activity of neutrophils is not manifested.

4. Antibiotics ceftriaxone, amoxiclav, lincomycin and gentamicin cause inhibition of the phagocyrate activity of neutrophils by 78%, 76%, 65% and 70%, respectively.

1. Aleshina E.N. Study of the effect of amorphous and crystalline penicillins on the plague microbe // Tr. Rostov-on-Don PChI. Rostov-on-Don, 1959. T. 15. Issue. 1. pp. 153-160.

2. Arefieva N.A., Aznabaeva L.F. Immune reactions of the nasal mucosa: cytological diagnosis, treatment methods // Consilium Medicum. 2009. T. 11. No. 11. pp. 30-33.

3. Israelson M.I., Shpolyansky B.I., Boevskaya G.I. The influence of penicillin on the functional ability of the reticuloendothelial system and the phagocytic activity of leukocytes // Journal. micro epid. and immune 1951. No. 3. P. 59-62.

4. Lakin G.F. Biometrics. M.: Higher. school, 1980. 291 p.

5. Finkel E.A., Lutskaya S.I. Autovaccine therapy for tuberculosis: Monograph. Kyrgyzstan, 1972. 154 p.

6. Bazzoni F., Cassatella M.A., Laudanna C., Rossi F. Phagocytosis of opsonized yeast nucleates tumor necrosis factor - alpha mRNA accumulation and protein release by human polymorphonuclear leucocytes // J. Leukoc. Biol. 1991. V. 50. No. 3. P. 223-228.

7. Czop J.K., Valiante N.H., Janusz M.J. Phagocytosis of particulate activators of the human alternative complement pathway thought monocyte beta-glucan receptors // Prog. Biol. Res. 1989. V. 297. P. 287-296.

8. De Weck A.L. Pharmacologic and immunochemical mechanisms of drug hypersensitivity // Immunol Allergy Clin North Am. 1991. No. 11. R. 461-474.

9. Gilliland B.C. Drug-induced autoimmune and hematologic disorders // Immunol Allergy Clin North Am. 1991. No. 11. R. 525-553.

10. Hiura M., Ozawa M., Othsuka T. a. o. Stimulation of superoxide anion production in guinea pig polymorphonuclear leucocytes by hypotonic condition with protein kinase C activators // Arch. Biochem. Biophys. 1991. C. 15. No. 291. P. 31-37.

11. Kishi K., Hirai K., Hiramatsu K., Yamasaki T., Nasu M. Clindamycin suppresses endotoxin released by ceftazidime-treated Escherichia coli O55: B5 and subsequent production of tumor necrosis factor alpha and interleukin-1 beta // Antimicrob Agents Chemother. 1999. V. 43. No. 3. R. 616-22.

12. Kovaks T., Stas I. et al. Volume regulatory mechanisms of human granulocytes in hipoosmotic media // Acta Biochim. Biophys. Hung. 1989. V. 24, No. 1-2. P. 142-147.

13. De Weck A.L. Pharmacologic and immunochemical mechanisms of drug hypersensitivity // Immunol Allergy Clin North Am. 1991. No. 11. R. 461-474.

14. Labro M.T. Pharmacology of spiramycin in comparison with other macrolides // Drug Invest 1993. 6. Suppl 1. R. 15-28.

15. Lachani M., Usmani S. et al. In vitro effect of furosemideon hemiluminescence of polymorphonuclear neutrophils in preterm infants // Biol.Neonate. 1997. V. 72. No. 3. P. 142147.

16. Muniz-Junqueira MI, Mota LM, Aires RB, Junqueira LF Jr. Digitalis inhibits and furosemide does not change the in vitro phagocytic function of neutrophils of healthy subjects // Int Immunopharmacol. 2003. V. 3. No. 10-11. P. 1439-1445.

17. Munoz J., Geister R. Inhibition of phagocytosis by aureomycin // Proc. Soc. Exp. Biol. Med. 1950. V. 75. No. 2. R. 367-370.

18. Richardson M.D., Donaldson F. Interaction of Candida crusei with human neutrophils in vitro // J. Med. Microbiol. 1994. V. 41, No. 6. P. 380-388.

19. Vetvica V., Thornton B.P., Ross C.P. Soluble beta-glucan polysaccharide binding to the lectin site of neutrophil or natural killer cell complement receptor type 3 (CD 11b/CD 18) generates a primed state of receptor capable of mediating citotoxicity of IC3b - opsonized target cells // J. Clin. Invest. 1996. V. 98. P. 50-61.