SU1727077A1 - Method for determining individual sensitivity to immune response modifiers in peptic ulcer patients - Google Patents

Abstract

This invention relates to medicine and can be used to determine the individual sensitivity of patients with gastric ulcer to immunomodulators. The purpose of the invention is to improve the accuracy of the method. Blood is taken from the patient, the lymphocytes of the cells are isolated, after attachment to the glass they are incubated in the presence of an immunomodulator, and the stability index of lysosomal membranes is determined in them and, if it is increased 2 times or more compared to the control, individual sensitivity to this immunomodulator is determined. Compared to the prototype, the accuracy is increased by a factor of 2.4-5.7.

Description

The invention relates to medicine, specifically to clinical immunology, and can be used to determine the sensitivity of an organism to immunomodulators in order to conduct immunocorrective therapy of immunodeficiency states.

The purpose of the invention is to improve the accuracy of the method.

The method is carried out as follows.

Into a tube containing 0.5 ml of anticoagulant, venous blood is drawn to the 5 ml mark. Shake well. To another centrifuge tube, pour 1 ml of a ficoll (Uppsa / a) and layer the test blood, centrifuge for 20 minutes at 1500 rpm. After the formation of white rings, pipette the MQ non-nuclears ring with the lower plasma layer with a pipette and transfer it to another clean tube. With

cells are not washed with this, diluted with medium 199 at a ratio of: 1 cell count + 2.2 portion of medium, counted in a Gorev chamber, adjusted with medium 199 to the desired concentration (2 x 10 6 cells / ml), applied to a plate 0.25 ml in each well and incubated for 60 min (to ensure adhesion of mononuclear phagocytes). The cells are then washed three times with medium 199 and introduced into the wells with 0.2 ml of culture medium consisting of medium 199, comprising 2.5% fetal calf serum and 0.5% glutamic acid. In the first two cells, only the culture medium (control) is introduced, in the following, 0.03 ml of the studied immunomodulators at a dilution of 1: 5. Incubate for 5 hours. Then, 0.06 ml of liquid (containing secreted lysozyme) is taken from the wells and the cells are frozen and thawed four times. Next, evaluate lysozyme to Dorofatechuk,

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determine the stability index of lysomal membrane (PSLM) for each immunomodulator by the formula: PSLM 100%, where PSLM is the stability index of lysosomal membranes; Ls - secreted lysozyme, µg / ml, Л0 - total lysozyme, µg / ml. The same indicator is also determined in the control for immunocorrection, the immuno-modulator is selected, on contact with which the PSLM increases 2 or more times compared to the control.

Example. Patient G., 54 years old, diagnosis: gastric ulcer. Entered for surgical treatment. The disease began 25 years ago. He was periodically treated, first with a temporary improvement, then with no effect. Concomitant diseases: chronic tonsillitis, tonsillogenous intoxication, manifestation of pain in the joints, cardiopathy, frequent acute respiratory infections (up to 5 times a year). Allergic history is favorable. The study of the immune status: phagocytosis indicators are normal, a sharp decrease in T-ROCK (26% and absolute value - 0.455 thousand / mm), the level of complementary serum activity - up to 27.7 CH / 50 ml, high values of circulating immunological complexes (CIC) - 160 (m). In the study of subpopulations of T- and B-lymphoidites with the help of monoclonal antibodies (MCAT) SD1, SD4, SDz, B, VA, VM, a sharp decrease in T-helper cells to 21% was found, a slightly increased level of T-suppressors — 31%, according to the results of the test, the Rebuck Skin Window - a decrease in the functional activity of T-lymphocytes.

According to the results of the clinical and immunological examination of the patient along with the operative treatment of the underlying disease, a T-immunity immunity stimulator is recommended. In this case, thymalin has the greatest labilifying effect on the lysosomal membranes of phagocytes. In this case, the increase in PSLM increased by 3.5 times compared with the control. The patient received thymalin in a dose of 3 mg / kg, 1 time per day for 3 days before surgery and 2 days in the postoperative period.

A smooth postoperative course was noted. 7 days after the operation and the course of T-ROCK immunocorrection, the indices rose to normal (56%), CAS and CEC returned to normal (52.6 CH / 50 ml and 60 m, respectively). The level of T-helper cells increased to 52%, the total population of T-cells was 59%, and according to test data, the skin window was normalized (0.5).

When using the proposed method, the accuracy of the method is increased by a factor of 2.4-5.7. Thus, the accuracy of determining individual sensitivity to immunomodulators is 75-87% using the proposed method, and 13-36% according to the prototype.

Claims (1)

Invention Formula The method of determining individual sensitivity of patients with gastric ulcer disease to immunomodulators, including blood sampling, isolation of a leuco-suspension, cultivation of it, incubation of a sample with cells adhering to glass in the presence of an immunomodulator, and without it, in order to improve the accuracy of the method , in the cells of both samples, the content of total and secreted lysozyme is determined and the stability index of the cell membranes is calculated, and with an increase of more than 2 times in the cells of the sample with an immunomodulator m compared to another sample was determined by the sensitivity of patients to immunomodulation - torus.