SU874063A1 - Method of reducing immunologic activity of lymphocytes - Google Patents

Description

(54) METHOD FOR REDUCING IMMUNOLOGICAL ACTIVITY. Lymphocytes

I

The invention relates to medicine, to the problem of transplantation of organs and tissues for the treatment of hematopoietic depression.

There is a method of reducing the immunological properties of lymphocytes by treating NHjCB hematopoietic tissue cells, followed by administering the suspension to the recipient r.

However, MSI is a toxic substance and its application in clinical practice is associated with known difficulties.

The purpose of the invention is to simplify and increase the efficiency of the method by ... reducing the homotransplantation activity of lymphocytes.

This goal is achieved by the method of reducing the immunological activity of lymphocytes by treating a suspension of hematopoietic tissue cells with organic matter followed by administering the suspension to a recipient, the suspension of cells is treated with 14.9-15% glycerin solution for 15-30 minutes.

The method is carried out as follows.

Prepare a suspension of spleen cells, lymph nodes, or bone marrow of experimental animals in a preservative solution, which includes sucrose, glucose, EDTUMAL, trisubstituted sodium citrate,

10 10% solution of gelatin,. To the resulting cell suspension, glycerol is added in 14.9-15% concentration. Cell suspension with glycerin is stored in a TFI refrigerator for 15-30 minutes.

In vivo methods of proving a decrease in the immunological activity of lymphocytes under the action of 14.9-15% glycerol were used.

accounting for the abolition of endogenous colony formation (Example 1); accounting for the inactivation of non-singular stem cells (Example 2 and 3). Example 1 1 & hybrids (CBA X C57B16) f are irradiated with an EGO-2 apparatus at a dose of 600 r. In order to cancel the endogenous formation columns after 20 h: after irradiation, mice were injected with a suspension of cells of the lymph nodes of CBA in different concentrations. The first group was administered a suspension of cells without glycerol, the second group - a suspension of joieTOK with 14.915% glycerin (at an exposure of 15 U1n): the third group - a suspension). . f cells with 14.9% glycerol, (at an exposure of 30 min); the fourth group serves as a radiation control. On day 9 after transplantation of cells, spleens were removed from recipients, fixed in Buen’s solution and 10% formalin-HDM solution. The number of macroscopically visible colonies in each spleen is counted. The inactivation index is calculated. It was found that the introduction of lymph node cells (without glycerol} at a dose of 1.0x10; 2, and 4.0x10 leads to 53.5; 82.2; and 93.1% inactivation of endogenous colony-forming units, respectively. After exposure of cells of lymph nodes with glycerin for 15-30 minutes, a significant decrease or complete absence of the inactivating effect of donor lymphocytes on the stem cells of the recipient is observed (Table 1.). Table 1 shows the changes in ingt activation of endogenous CFU under the influence of 15% glycerol during transplantation Intact cell lymph nodes The CBA mice were sub-irradiated (600 r) to the recipients (CBA X C57B1b) p4. When the cell suspension of transplanted intact lymphocytes was transplanted to the recipients, the cells were recalculated into the number of living (morphologically preserved) cells. A dose of 5.0x10 is added to the spleen cells of CBA mice in different concentrations, and a suspension of the spleen cells of the young CBA is prepared in three types of suspensions: freshly prepared cells, freshly prepared cells with 14.9-15% glycerol (at an exposure of 15%). min and svezhezag About 14.9–15% glycerol cells at an exposure of 30 min. 4 Mixed cell suspension is injected with nutrient hybrids (CBAx C57B16) F 20 hours after irradiation. Recipient mice are irradiated with EGO-2 J-rays. in the lethal dose of 880 r (with exogenous colonies). It was established that freshly prepared cells of the spleen of CBA mice at up to 4.0 X 10 inactivate stem cells of C57B16 murine. The inactivation index is 51.3%. Addition of glycerol to the spleen cells (with an exposure of 15 min) leads to a decrease in the inactivation index of non-syngeneic stem cells to 35.8% (with the same dose of the spleen cells of CBA mice - 4.0 x 10 X The increase in the equilibration time of the spleen cells with glycerol to 30 min results in the inactivating effect of the stem cells of the C57B16 murine lymphocytes of the spleen of the SVL strains (Table 2; table 2). Liciter cells of the spleen and CBA cells are lethal (880 r) to irradiated recipients (CBA X) F. Example 3. Freshly prepared bone marrow cells with C57B16 pulps in a dose of 5.0x10 are added to bone marrow cells of the CBA line in different concentrations. Condition the preparation of irradiation suspensions, cell transplantation, and studies of the spleens are the same as Example 2. It was established that during the interaction of bone marrow cells with different lines (CBA and C57B16), stem cells of the C57B16 are inactivated. At a dose of fresh bone marrow cells, the molar CBA is 0.5x10, the inactivation index ... is 25.0%, and at a dose of 4.0x10 57.0%. Addition of glycerol to the bone marrow of CBA mice (at an exposure of 15–30 min) leads to a significant decrease or cancellation of the effect of inactivation of stem cells of C57B16 hsp cells with lymphocytes of the bone marrow of CBA mice Stab. 3). Table 3 shows the inactivation of nonsingen CFU during the transplantation of a mixture of intact bone marrow with C57B16 with intact (without glycerol and with 15% glycerol) bone marrow cells with CBA from irradiated CBA X C5 / B16 recipients F).

The proposed method allows to significantly reduce the homotransplantation activity of hematopoietic lymphocytes.

Table 1

table 2

Claims (1)

Invention Formula A method of reducing the immunological activity of lymphocytes by treating a suspension of hematopoietic tissue cells with organic matter followed by administering the suspension to a recipient, characterized in that, in order to simplify and increase the efficiency of the method by weakening the lymphocyte homotransplantation activity, 14.9-15% of cells are processed Nym glycerol solution for 15-30 minutes. Sources of information taken into account in the examination I. O.M. Strong at all.Differential susceptib. 11 ty. of Murine T-and B-Lymphocytes to Frelz-Thaw and Hypotonic Shook, Criobiology, 1974, y.l, 12, p.127-138.