RU2340348C1 - Method of treatment of atopic dermatitis - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: for treatment of atopic dermatitis, use the autologous T-lymphocytes, activated with the help of anti CD3 antibodies and interleukin-2. The activated T-lymphocytes are administered subcutaneously once a week within 4 weeks and further once a month within one year. Simultaneously perform basic therapy by antihistamine preparations and topical glucocorticosteroids.

EFFECT: depression of clinical implications of disease at the expense of influence on the changed immune response through an induction of the antiergotypical response.

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Description

The present invention relates to medicine, in particular to clinical immunology and allergology, and relates to a method for the treatment of atopic dermatitis.

Atopic dermatitis (AD) is one of the most frequent and problematic dermatoses, the incidence of which in the world, according to various sources, in adults can be from 5% to 16.8% of the population. Beginning in early childhood, in about half of cases, it can occur for many years, exacerbating, then subsiding with a characteristic seasonal dependence [1, 2, 4].

Atopic dermatitis is characterized by allergic sensitization with overproduction of immunoglobulin E against various environmental antigens. In allergic sensitization, activation of Th2 cells plays a large role, since these cells can mediate IgE synthesis through IL-4. Current data indicate that the Th-2 response is normally suppressed by T-regulatory cells (Treg), while in atopic patients this suppression is weakened [4, 9, 10, 12].

Regulatory T cells are an important component of the homeostasis of the immune system and control its functioning through interaction with lymphocytes, monocytes, NK cells and dendritic cells in an activated state. Moreover, according to recent studies, Treg cells actively migrate to the foci of inflammation and have a negative effect on target cells through contact-dependent (perforin / granzyme mediated killing) and contact-independent (production of anti-inflammatory cytokines - TGF-β and IL10) mechanisms [5 , 6, 8, 11, 14].

The standard methods of treatment currently used, including the use of systemic and topical drugs, are not effective enough, which served as the basis for the search for new areas of systemic therapy for this disease, the purpose of which is to activate the mechanisms of suppression of activated cells.

For many years, allergen-specific immunotherapy has been used in the complex treatment of atopic dermatitis - subcutaneous administration of low doses of allergen in increasing concentrations [15]. This method of treatment is taken as a prototype. According to recent studies, such treatment leads to the induction of T-regulatory cells that suppress the Th2 response to the allergen and switch antibody production from IgE to IgG4 [9, 10].

It was shown that in the process of vaccination with antigen-specific cells, regulatory types of T-cells of 2 types are activated: anti-idiotypic and anti-ergotypic. Moreover, anti-ergotypic T cells, unlike anti-idiotypic ones, recognize effector T cells regardless of their TCR affiliation. Currently, molecules such as TCR, CD25, CD122, and HSP60 epitopes are considered as ergotopes [5]. In experimental studies, it was shown that regulatory T cells modulate the Th2 immune response characteristic of allergic diseases, reducing the production of proinflammatory cytokines by CD4 + Th2 cells [10, 12].

However, allergen-specific immunotherapy requires the use of specific allergens, which is not always possible, because not in all cases it is possible to establish a relationship of a particular allergen with the development of the disease, in addition, most patients have polyvalent sensitization.

The aim of the invention is to provide a method for the treatment of atopic dermatitis, using the effect on the altered immune response characteristic of patients with atopic dermatitis, through the induction of an anti-ergotypic response.

This is achieved through the use of T-lymphocytes, which are isolated from the patient’s peripheral blood and then activated using anti-CD3 antibodies and IL-2. Activated autologous T-lymphocytes are administered to the patient subcutaneously 1 time per week for 4 weeks and then 1 time per month for 1 year against the background of basic therapy with antihistamines and topical glucocorticosteroids.

Immunotherapy of atopic dermatitis using autologous activated T-lymphocytes has advantages compared to the prototype because it is based on the use of the patient’s own cells, therefore, the proposed method is more immunogenic and significantly less allergenic. The method allows to achieve a significant clinical effect and even complete recovery of patients.

The method is as follows.

To 200 ml of peripheral blood stabilized with heparin, add 10-20 ml of 10% gelatin (Promix) and incubate for 45 minutes in an incubator at 37 °. The resulting leukoprecipitate is collected, diluted with buffered saline with EDTA (PBS-EDTA) containing 100 ml of a solution of 2 mg of thienam (Merck), 125 μl of gentamicin (KRKA) and 500 μl of FBS (Fetal Bovine Serum, HyClone); then centrifuged in a gradient ficoll-verographin (Sigma, Sweden) for 35 minutes at 1.2 thousand rpm in an OS-6 centrifuge. The resulting ring of mononuclear cells is collected and washed three times with PBS-EDTA at 1.2 thousand rpm for 10 minutes. The cell pellet is resuspended in 10 ml of complete RPMI 1640 medium (FSUE SSC WB Vector) (with the addition of 2 mg of thienam, 125 μl of gentamicin and 30 mg of L-glutamine (FSUE SSC WB Vector) per 100 ml of medium) and the number of cells in the Goryaev chamber is counted . All obtained MNCs are put into culture mattresses (NUNC, Italy) at the rate of 2 million cells per 1 ml of complete medium with 10% FBS and activators (1 μl / ml anti-CD3 antibodies (SPC MedBioSpektr) and 50 units / ml IL-2 (Biotech LLC) ) On the 5-6th day (according to the state of culture), 70% of the medium is replaced with a new one with the same composition. On days 10-14, cells are harvested, precipitated at 1.2 thousand rpm for 10 minutes, resuspended in 10 ml of FBS and the number of cells is counted. FBS and DMSO (Riedel-deHaën, Germany) are added so that as a result, 30 million cells per 1 ml of FBS with 10% DMSO are obtained and packaged in 1 ml cryovials. Store cells in a freezer at -80 ° C.

Before administration, 10 ml of physiological saline is added to the cells and precipitated for 5 minutes at 1 thousand rpm in a medical laboratory centrifuge. The precipitate is resuspended in 2 ml of physiological saline and is administered to patients in this form.

This treatment has been tested in a clinical setting. Clinical trials were carried out on the basis of the Department of Allergology, Clinic of Immunopathology, State Research Institute of Medicine, Siberian Branch of the Russian Academy of Medical Sciences, Novosibirsk. It was used to treat patients with atopic dermatitis.

The study included 18 patients (7 men and 11 women). The age of the patients ranged from 17 to 49 years (mean age 24.5 ± 1.8 years). Patients received basic therapy with antihistamines and topical glucocorticosteroids.

Inclusion criteria

1. Men and women aged 18 to 65 years inclusive.

2. The diagnosis of blood pressure was verified in accordance with ICD-10 at least 6 months before the patient was included in the study.

3. The presence of moderate blood pressure activity.

4. The associated dose of basic drugs should be stable during the 1-3 months preceding the screening.

5. Men and women of childbearing age should not plan childbirth for the period of the study, and when included in the study should use adequate contraceptive measures (except for oral contraceptives).

6. Patients must be sufficiently mobile to follow the schedule of visits.

7. Patients should be able to give informed consent, and this consent must be obtained before any screening procedure begins.

Exclusion criteria

1. Simultaneous participation in another clinical trial.

2. Inconsistency with inclusion criteria.

3. The presence of serious infections such as hepatitis, pneumonia, pyelonephritis, herpes zoster in the 3 months prior to screening.

4. Severe somatic condition caused by progressive diseases of the liver, cardiovascular system, kidneys, respiratory system, blood system, as well as endocrine or cerebral origin.

5. Any known malignant disease or history of cancer in the previous 5 years.

6. The presence of active alcoholism or signs of alcoholic liver damage.

7. The presence of acute infectious disease.

8. Any vaccination in the previous 3 months.

The research methods used to identify the clinical efficacy of activated autologous T cells included:

clinical characteristics on a SCORAD scale:

A) the prevalence of the process (%);

B) the intensity of the manifestations, including the presence of erythema, edema / papule formation, weeping / crusting, excoriation, lichenification, dryness (expressed in points from 0 to 3);

C) subjective symptoms: itching and sleep disturbance (expressed in points from 0 to 10);

clinical characteristics on the DCSS scale:

erythema, edema, weeping, lichenification, papules, dryness, peeling, cracks, itching (expressed in points from 0 to 3);

DLQI quality of life assessment.

The severity was also evaluated by the number of relapses per year, the average duration of relapses, the prevalence of skin lesions [3, 7].

The occurrence of local and general reactions to cell administration was monitored.

Statistical analysis was performed using the method of comparing two dependent Viscoxon values.

Intervention. The subjects received treatment with activated autologous T-lymphocytes in combination with standard therapy. Standard therapy: antihistamines (Zirtek 0.01, 1 t. 1 time per day), topical glucocorticosteroids (Elokom cream).

Activated autologous T-lymphocytes were injected subcutaneously with a frequency of 1 time per week No. 4, then 1 time per month, a total of 10 injections.

Assessment of the condition of patients was carried out initially, on the 5th and 10th administration of activated autologous T cells.

In 4 patients local adverse reactions were noted in the form of hyperemia, swelling, pain at the injection site, in 7 patients - common in the form of exacerbation of atopic dermatitis, an increase in body temperature to subfebrile numbers (1 patient).

By the end of treatment (10 introduction), 3 patients (16.7%) achieved clinical recovery, 5 (27.8%) significant improvement (symptom reduction by 85-100%), and improvement (symptom reduction by 50-85%) - 8 (44.4%), a slight improvement (reduction of symptoms by less than 50%) - 1 (5.55%), unchanged - 1 patient (5.55%). Thus, a good effect of therapy (symptom reduction by more than 50%) was observed in 16 people (88.9%), and a pronounced effect was not observed in 2 patients (11.1%). The average value of the SCORAD and DCSS indices was initially 52.2 ± 4.8 and 11.9 ± 1.3 points, respectively, both groups were identical. By 5 injections in patients with a good response to treatment, the indices were 22.8 ± 4.5 and 6.7 ± 1.1, without effect 83 ± 13 and 19 ± 4, and 10 injections in patients with a good response were 10 , 2 ± 1.9 and 3.1 ± 0.5, without effect - 60.5 ± 35.5 and 14.5 ± 8.5 points, respectively.

The DLQI quality of life index was initially 10.8 ± 1.7 points, by 5 injections in the group of patients with a good effect - 7.1 ± 1.9, without an effect - 29 ± 1, and by the end of treatment in patients with a good effect - 2 , 9 ± 0.8, without effect - 21.5 ± 8.5 points.

Below are the results that explain this method of treatment of atopic dermatitis.

Example 1. Patient C., 20 l. Diagnosis: atopic dermatitis, a common form, of moderate severity. The area of damage to the skin was widespread: on the skin of the front surface of the chest, arms, legs, rashes of an erythematous-papular nature, excoriation, hemorrhagic crusts. Upon admission, she complained of skin rashes accompanied by intense skin itching, worse at night, dry skin, peeling, and sleep disturbance. From 3 years old suffers from atopic dermatitis. The course of the disease is year-round with exacerbation in the autumn-winter period. Exacerbations are provoked by psycho-emotional stress, eating chocolate, citrus fruits. The severity of clinical manifestations on the SCORAD and DCSS scales was initially 45 and 12, respectively, the DLQI quality of life index was 5. The patient in combination with standard therapy with antihistamines (Zyrtec 0.01 at 1 t per day) and topical glucocorticosteroids (Elokom cream) therapy was carried out with activated autologous T-lymphocytes according to the scheme. After 2 injections, adverse reactions were noted in the form of muscle soreness at the injection site, an increase in body temperature to 37.6 ° C during the day. After 3 and 4 injections, some pain was also noted at the injection site, but less pronounced. After 4 injections, an exacerbation of the disease was noted in the form of the appearance of a small number of new rashes in the area of the elbow bends, increased itching at night. Starting from the 6th injection, there was a tendency to a decrease in the severity of clinical manifestations in the form of a decrease in the number of rashes, a decrease in skin hyperemia, a decrease in the intensity of itching, and an improvement in sleep. After the course of therapy, a significant improvement in the patient's condition was noted: the prevalence of skin rashes significantly decreased, the rashes did not recur, the itching of the skin did not bother, and sleep was normal. The severity of clinical manifestations by 5 administration of activated autologous T-lymphocytes on the SCORAD and DCSS scales was 54 and 15, respectively, the DLQI quality of life index was 18. The severity of clinical manifestations after a course of treatment on the SCORAD and DCSS scales was 11 and 3, respectively, the index quality of life DLQI became 4. Thus, the patient noted a significant improvement.

Example 2. Patient M., 19 l. Diagnosis: atopic dermatitis, a common form, of moderate severity. The area of damage to the skin was widespread: erythematous-squamous eruptions on the skin of the trunk, elbow, knee bends, single papules in the face, hyperemia, swelling, peeling of the eyelids, serous crusts on the scalp. Upon admission, she complained of rashes accompanied by intense skin itching, worse at night, swelling and itching of the eyelids, dry skin, peeling, and sleep disturbance. The course of the disease is year-round without a clear seasonality. Exacerbations are provoked by psycho-emotional stress. The severity of clinical manifestations on the SCORAD and DCSS scales was initially 43 and 11, respectively, the quality of life index DLQI was 22. The patient in combination with standard therapy with antihistamines (Zirtek 0.01 to 1 t. Per day) and topical glucocorticosteroids (Elokom cream) therapy was carried out with activated autologous T-lymphocytes according to the scheme. After 1 injection, adverse reactions were noted in the form of hyperemia, swelling, pain at the injection site. By the 5th injection, an exacerbation of the disease was noted in the form of the appearance of new rashes in the face, trunk, and increased dry skin. Starting from the 6th injection, there was a clear tendency to reduce the severity of clinical manifestations in the form of a decrease in the number of rashes, a decrease in hyperemia, dry skin, a decrease in the intensity of itching, improved sleep, and rash did not recur. By 9 injections, the skin was completely cleansed, the itching did not bother, sleep was normal. After the course of therapy, a persistent clinical recovery was noted: the skin was clean, there was no itching, sleep was normal. The severity of clinical manifestations for 5 administration of activated autologous T-lymphocytes on the SCORAD and DCSS scales was 50 and 13, respectively, the DLQI quality of life index was 24. The severity of clinical manifestations after a course of treatment on the SCORAD and DCSS scales was 0 and 0, respectively, the index quality of life DLQI became 0. Thus, the patient has a complete clinical recovery.

Therefore, the inclusion of activated autologous T-lymphocytes in standard treatment for patients with atopic dermatitis is characterized by clinical efficacy in 88.9% of cases, and the severity of clinical manifestations of the disease decreases by an average of 80.5% / 74% according to SCORAD / DCSS indices, respectively. the quality of life improves by 73.2%, which makes it possible to widely use this method in clinical practice for the treatment of patients with atopic dermatitis.

Claims (1)

A method of treating atopic dermatitis using vaccine therapy, characterized in that they use autologous T-lymphocytes isolated from the patient’s peripheral blood and activated with anti CD3 antibodies and interleukin-2 (IL-2), which are administered subcutaneously to the patient once a week for 4 weeks and then 1 time per month for 1 year against the background of basic therapy with antihistamines and topical glucocorticosteroids.