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INVENTION
Patent of the Russian Federation RU2136002
METHOD FOR DIAGNOSTICS OF NEST ALOPECY
The name of the inventor: Dyakov Valery Mikhailovich; Parfenov German Ivanovich
The name of the patent holder: Dyakov Valery Mikhailovich; Parfenov German Ivanovich
Address for correspondence: 111402, Moscow, Alley Zhemchugova, d1, ap.141, Zhilin A.S.
Date of commencement of the patent: 1999.01.26
The invention relates to the field of medicine, in particular to dermatology, and can be used in the diagnosis and treatment of alopecia, in particular alopecia areata. Visually assess the condition of the skin and hair, additionally perform an immunological analysis of the blood and the violation of the cellular system of the immune system, mainly by the number of natural killer-NK (CD 16), exceeding the norm, determine the immune-reactive phase of the disease. The method ensures an increase in the accuracy of the diagnosis.
DESCRIPTION OF THE INVENTION
The invention relates to medicine, in particular to dermatology, and can be used in the diagnosis and treatment of alopecia areata.
Various theories of the occurrence of alopecia are known, for example, trophoneurotic, neuroendocrine, infectious and the theory of autoantibodies [BME, Ch. Ed. B.V. Petrovsky, vol.1, ed. "Soviet Encyclopedia", M., 1974].
The etiology and pathogenesis of alopecia areata are not known. Hair loss is associated with such causes as general intoxication, nervous stress, bacterial and fungal lesions, lack of vitamins and trace elements, etc.
However, none of the above factors can independently cause sudden and rapidly progressing alopecia against the background of recent anamnestic well-being.
It is established that some negative factors, called "trigger" triggers, can potentiate the appearance of the first centers of alopecia.
Such factors include acute and chronic stress, ovarian dysfunction, scalp damage (hair pulling, skin prick, chemical action, etc.), craniocerebral trauma, general anesthesia, some oral contraceptives, allergic and atopic reactions, various Kind of intoxication, hypovitaminosis, etc.
In clinical practice, in the detection of the causes of the disease, as a rule, tests that are typical not for alopecia are prescribed, but to determine the concomitant pathology: general blood and urine tests, biochemical analyzes, hormones in the blood, bacteriological and virological studies.
It is known that the diagnosis of alopecia are performed mainly by visual examination of the skin and hair condition [VVVladimirov, "Diagnosis and treatment of skin diseases", ed. LLP Medtehturservis, 1995;
V. Ya. Arutyunov, "The Problem of Alopecia", Bulletin of Dermatology and Venereology, No. 12, 1971].
This diagnosis does not reveal the root causes of alopecia and does not determine effective ways of treating it.
The invention is aimed at improving the quality of the diagnosis of nest alopecia ("rapid alopecia").
At the same time, a well-known method for diagnosing nested alopecia characterized by visual examination of the skin and hair condition is supplemented by carrying out an immunological analysis of the blood and breaking the cellular part of the immune system, mainly by the number of natural killers - NK (CD16), exceeding the norm in absolute and relative indices, Determine the immunoreactive phase in the development of alopecia.
Studies carried out by the authors in 1992-1998. In the medical center "Blagovest" for the treatment of alopecia (Moscow) showed that the development of alopecia are associated primarily with a change in the immunological status of the organism. In this case, damage to the cells of the hair follicle is determined not only by humoral and nervous factors, but primarily by strictly differentiated cells of the immune system.
In the pathogenesis of alopecia is a genetic predisposition. The genetic anomaly of the immune system does not appear at birth, it only potentiates the possibility of the development of the disease.
Immunodeficiency is not universal, but shows a strong specificity for a specific antigen. The introduction of an antigen is characterized by the absence at this stage of an inflammatory process in the paraxial region of the hair follicle, which indicates a possible defect in the recognition of a foreign agent by antigen-presenting cells.
Two etiological factors were examined: random invasion of the virus with low virulence (avirulence) and congenital specific deficit of antiviral protection. Damaged "trichovirus" cells of the hair follicle are target cells, which after infection do not stop their vital cycles and do not interfere with the natural growth of hair.
Langerhans cells (antigen presenting skin macrophages) play a crucial role in initiating the primary immune response. Normally, they are the ones who are obliged to accept information from the trichovirus and express trichantigen on its surface, thus launching the entire complex of immune responses. With alopecia, this does not happen, and the target cells of the hair follicle become antigen-presenting cells, which are not capable of utilizing the virus.
Trichovirus changes the DNA fragment of the sixth chromosome responsible for the expression of antigens of the main histocompatibility complex. First of all it concerns antigens of the first class of histocompatibility HLA-A9, B12, BW18. These antigens are represented by abnormal receptors on the surface of a matted cell. Cell mutation is point-like and does not interfere with normal hair growth, does not activate cytotoxic T-lymphocytes (CD8), and does not trigger the whole complex of the immune response with the formation of antibodies at the onset of the disease. It only makes recognizable lesions for natural killers -NK (CD16).
Studies have shown that in the immunoreactive phase, lymphocytosis is caused by an increase in the number of natural killers-NK (CD16), which make contact with mutated cells expressing tropic receptors. Such exposure leads killers to a state of excitation with the manifestation of cytotoxicity. The lymphotoxins released by the killer violate the membrane permeability of target cells and induce apoptosis (self-death of the cell). The confirmation of apoptosis is the absence of inflammation and necrotic process in the centers of alopecia with Alopecia areata ("Rapid alopecia").
It is known to conduct immunological studies for the diagnosis of various diseases. For example, monoclonal antibodies are used to determine the subpopulations and the number of immune cells, the method of preparation of which was developed by Ts. Milstein and G. Keller in 1975
Immunophenotyping (determining) subpopulations of lymphocytes is used by immunofluorescence analysis (ELISA) of blood in which monoclonal antibodies labeled with fluorochrome bind to membrane antigens of living cells when they are mixed. Then, this slurry under pressure enters an optoelectronic cytofluorimeter (for example, from Becton Dickinson, Becton Dickinson, USA).
The principle of phenotypic analysis of cells is to determine the size of cells and to identify certain monoclonal antibodies bound to cells, which is reflected in the immunogram
["Lymphocytes, Methods," J. Claus, Moscow. "The World", 1990].
The main lymphocytes detected by monoclonal antibodies are: T-lymphocytes (CD3), B-lymphocytes (CD19), T-helpers (CD4), T-suppressors (CD8), natural killers - NK (CD16), etc. ., The number of which and the mutual ratio may vary depending on the initial state of the immune system and its response as a result of the disease.
The amounts of these lymphocytes taken as the norm are shown in the table.
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As shown by the authors' studies, immunophenotypic analysis of the blood of patients with alopecia areata reveals deviations in the immunogram from the norm. The disturbance of the cellular part of the immune system in alopecia primarily affects the natural killer-NK (CD16). Exceeding their number from the norm to 100% or more in absolute amounts, and an increase in the relative amount to 70% of the upper limit of the norm indicates excessive proliferation of natural killers -NK (CD16) in alopecia areata. The decrease in the number of T-lymphocytes (CD3) from the lower limit of the norm ranges from 0-15% in relative amounts and 0-31% in absolute values. From the above, it can be concluded that alopecia (focal) alopecia is an autoimmune reaction of the organism directed towards mutated cells of the hair follicle. In this case, the autoimmune response is realized through the action of natural NK killers (CD16) on the hair follicle target cells, in which the mechanism of programmed cell death (apoptosis) is triggered. |
Below are examples of clinical examinations of patients with alopecia areata, conducted in 1998 in the medical center "Blagovest" on the basis of the Research Institute of Immunology (Moscow) using immunoassay.
Example 1 . Patient M., 15 years old, revealing the first focus of alopecia for 2 months before the examination. At visual inspection the continuation of the process of hair loss is noted. In the general blood test, there is a small lympho- and monocytosis, biochemical and hormonal indices within the norm. Diagnosis: alopecia areata, subtotal stage, 30% of the presence of hair.
Immunological examination of cytofluorimeter (3-color flow laser cytometry, reagents and equipment from Becton Dickinson-Becton Dickinson, USA) revealed the CD3 + T-lymphocyte content in absolute values of 1258 cells / μl and in relative 57%. In this case, the content of natural killers -NK (CD16) was in absolute and relative amounts, respectively, 717 cells / μl and 33%.
Conclusion: there was an excess over the norm of absolute and relative indices for natural killers and a decrease in T-lymphocytes, which shows a pronounced immunoreactive phase of alopecia areata.
Example 2 . Patient Zh., 28 years old, the first foci of alopecia were recorded at the age of 7, after 1 year the head hair was completely restored. Recurrence of the disease began 5 months before the examination, with the hair on the head lost completely. In the general blood test, minor lymphopenia was noted. Respiratory diseases rarely get sick, he considers himself healthy. Diagnosis: alopecia areata, total stage.
Immunological examination on a cytofluorimeter (3-color flow cytometry, reagents and equipment from Becton Dickinson, USA) revealed the CD3 + T-lymphocyte count in absolute values of 842 cells / mm3 and in relative 59%. In this case, the content of natural killers -NK (CD16) was 463 cells / mm3 and 33%, respectively, in absolute and relative amounts.
Conclusion : there was an excess over the norm of absolute and relative indices for natural killers and a decrease in T-lymphocytes, which indicates the immunoreactive phase of the disease.
A total of 78 patients with different stages of alopecia areata (focal, subtotal, total, universal), including 42 children from 5 to 13 years, 23 teenagers from 13 to 17 years and 13 adults. All of them were at different stages of hair loss and in different phases of immunological activity (immunoreactive phase, immune-indifferent phase).
A common feature for most patients was that an increase in the number of natural killer-NK (CD16) and a decrease in the number of T-lymphocytes (CD3) accompanied a rapid process of objective hair loss. This proves the participation of natural killers in the pathogenesis of nesting ("rapid") alopecia.
The number of patients with impaired immunological status during the period of active hair loss was 68.5% for natural killers and 39.6% for T-lymphocytes (CD3), which indicates the immunoreactive phase in the acute period of the disease.
The results of the conducted studies confirmed the possibility of reliable identification of the immunoreactive phase of alopecia areata, which allows to form a new understanding of the etiology and pathogenesis of the disease.
CLAIM
A method for diagnosing nested alopecia by performing a visual examination of the skin and hair condition, characterized in that an additional immunological analysis of the blood is carried out, and in violation of the cellular part of the immune system, mainly by the number of natural killer NK-cells (CD 16) exceeding the norm, the immunoreactive phase of development is determined Disease.
print version
Date of publication 01.04.2007гг
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