ANTI-LYMPHOCYTIC SERUM (ALS, ALS; Greek anti-against + a lymphocyte[s]) — the immune serum containing different antibodies to lymphoid cells is preferential to lymphocytes.
The name ALS is followed by the name of a species of an animal, from to-rogo serum, and also the name of a look is received, against fabric to-rogo antibodies are developed, e.g.: rabbit anti-rat, horse anti-human, etc.
ALS is for the first time received by I. I. Mechnikov in 1899 at immunization of a Guinea pig splenic tkanyo rats. A. M. Bezredka in 1900 showed cytotoxic action of ALS on lymphoid cells of in vitro; later leucio-and limfopenichesky action of ALS in vivo, and also an inhibiting effect on development of reactions of hypersensitivity of the slowed-down type was shown. The ALS last properties drew attention of transplantologists as there was a possibility of impact on reactions of transplant immunity which are mediated through lymphoid cells. Intensive studying and use of ALS began since 1963 — 1964 when Woodruff and Anderson (M. of Woodruff, N. Anderson) achieved considerable lengthening of terms of survival of an allogenic skin rag from rats during the use of rabbit anti-rat ALS.
ALS receive receiving by immunization of animals of one look (producer) lymphoid cells of other look (donor). As producers can serve: mice, rats, Guinea pigs, dogs, etc. For receiving ALS in the medical purposes in quality of producers use rabbits, horses, sometimes monkeys, donkeys.
For immunization use cells of a spleen, limf, nodes, a thymus gland, the lymphocytes received it is intravital by drainage chest limf, a channel (ductus thoracicus); there is an experience of receiving ALS by immunization by fabric of almonds of the person. ALS received by immunization by cells of a thymus gland and lymphocytes chest limf, a channel have the greatest immunodepressive activity.
Immunization provides various schemes; introduction can be intravenous, intraperitoneal, intramuscular, hypodermic; at two last antigen is entered in a Freund's adjuvant; use the combined schemes — intravenous with the subsequent hypodermic administration of antigen; quantity of the lymphoid cells entered for one injection — 0,2 — 50 billion depending on a type of a producer and its sizes.
Cleaning and processing
In the serums received by way of bloodletting are defined hemagglutinins (and hemolysins) in significant amounts which removal is made one-double (for the experimental purposes) or repeated (for the medical purposes) adsorption, using erythrocytes of the donor (20 — 30% of the total amount of serum). Presence of hemagglutinins and hemolysins is admissible in credits less than 1: 16. Antithrombocytic antibodies are admissible in the credits which are not exceeding 1: 8 in reaction of binding complement. ALS used in the medical purposes is inactivated warming up in the water bath at t ° 56 ° within 20 — 30 min.
Processing of ALS can be finished with allocation of the active beginning — an anti-lymphocytic γ-globu-lin, a cut make by ethanol fractionation or fractional extraction of 40% ammonium sulfate with the subsequent dialysis against a distilled water. Further processing with use of Sephadex G-200 shows that the most part of anti-lymphocytic activity is localized in IgG-fraction.
Activity of ALS is characterized on a complex of tests
1. Increase in duration of survival (prolongation) in a skin rag serves as an indicator of actually immunodepressive activity. This test is carried out on laboratory animals taking into account survival of an allogenic skin rag in the group receiving ALS and in control group. Normal terms of rejection of an allogenic skin rag of 7 — 8 or 10 — 13 days depending on a species of a laboratory animal. Active ALS increases terms of life of allotransplant from several days to several weeks.
2. Cytotoxic action on lymphoid cells: in vitro by Gorer's method — O'Gormana (1957) or to his modifications on lymphocytes of a donor animal with use of intravital dye tripanovy blue is carried out.
3. Oppression of primary immune response on fabric, a proteinaceous or bacterial antigen; it is carried out on small laboratory animals at immunization by erythrocytes of a ram, a bull seralbumin or culture of bacteria; considerable decrease in antiserum capacities at the animal processed by ALS in comparison with antibodyformation at a normal animal serves as an indicator of activity of ALS. Possesses an inhibiting effect anti-lymphocytic γ-glo-bulin as normal IgG does not cause such effect.
4. The transforming activity is studied by in vitro on ability to cause a blasttransformation of lymphocytes (see. Immunity transplant ). Are compared not stimulated and stimulated under the influence of ALS 2 — 3-day cultures of lymphocytes; the accounting of reaction is made by a morphological research of cultures or on inclusion of radioactive N 3 - thymidine or With 14 - glycine in DNA or RNA of the sharing lymphocytes.
5. Braking of rosetting: lymphocytes of the person or mice being are mixed in saline solution with erythrocytes of a ram «sockets» owing to immune sticking to their cover of erythrocytes of a ram can create in vitro. The active ALS entered to the person or a mouse inhibits spontaneous «rosetting» at an incubation of test lymphocytes with erythrocytes of a ram (90 min., t ° 37 °). Sensitivity of reaction of an ingibition increases in presence of a complement of a Guinea pig. The serums intended for clinical use are tested using the called tests on primacies. Primary selection is made on the lowest monkeys (macaques Rhesus factors, baboons) and final — on a chimpanzee.
In an experiment is shown that ALS is capable to increase terms of survival of an allogenic skin rag at mice, rats, rabbits, monkeys. The areactivity created by ALS in relation to allotransplant extends to a secondary transplant and to heterograft. ALS extends also terms of survival of a kidney and liver, replaced to rats and dogs.
Clinical use is carried out in two directions so far. Cases of use of ALS at diseases in which pathogeny there is an autoimmune component are known (acute recurrent rheumatism, rhematoid polyarthritis, etc.). At the same time improvement of a state, normalization of the indicators characterizing a disease is observed (delay of ROE, disappearance of joint pains, disappearance of an exudate in joint bags).
In recent years ALS and anti-lymphocytic globulin widely use at transplantation of vitals, first of all kidneys. Since 1971 the majority of transplantations of a kidney is carried out during the use of ALS as the immunodepressive agent.
The clinic of transplantations of ALS uses always in a complex with other immunodepressive substances (see). Inclusion of ALS in a complex of immunodepressive therapy increases duration of survival of the renal transplants taken from living related donors. When use of ALS in the general scheme of an immunosuppression significantly did not extend terms of life of a renal transplant, it nevertheless was useful since allowed to reduce a dose of corticosteroids and, therefore, to reduce the frequency of emergence of infectious complications.
To the person of ALS enter intramusculary, subcutaneously, intravenously. Test across Bezredka on hypersensitivity shall precede the first injection of ALS. Intramuscular and hypodermic introduction of ALS sometimes is followed by burning sensation and pain in an injection site, a swelling, reddening, raising of t ° to 38 — 39 °. Intravenous administration (slow, a drop method) without serious consequences, however is to some extent risky because of a possibility of development of an acute anaphylaxis or toxic changes in a kidney. Single introduction of ALS causes the lymphopenia noticeable in 5 min. after an injection. The number of lymphocytes during 4 — 12 hours after an injection decreases by 20 — 80%; normalization occurs in several days; parallel to a lymphopenia the leukopenia sometimes develops. At long introduction of ALS the number of lymphocytes is determined at the certain level and can return to initial figures.
At transplantation of a kidney to the recipient enter 3 — 10 ml of anti-lymphocytic γ-globulin a day (respectively in terms of protein). Schemes of use are various, following is most usable: daily within two weeks before transplantation and then each two days within a month after renal transplantation, it is obligatory with azathioprine (an imuran and corticosteroids); introduction of the last stops from 3rd week. Use of ALS can be extended depending on character of a transplant (related, from a corpse etc.) and conditions of the recipient.
Side effects and complications
1. Long introduction of ALS can cause pathological changes in parenchymatous bodies, and also owing to trombopenichesky action to be followed by internal hemorrhages. Before use of ALS in the medical purposes carry out tests on the toxic and hemorrhagic effects consisting in repeated introduction (7 — 8 days) of ALS to a small laboratory animal with the subsequent histologic inspection of parenchymatous bodies and taking into account internal hemorrhages.
2. Administration of heterological globulin (in the form of ALS or anti-lymphocytic γ-globulin)))))))))) is followed by emergence of the pretsipitin directed to proteins of an animal producer. This process accelerates elimination of ALS from an organism that was proved with use of a radioactive label. Besides, precipitated calcium superphosphates of a cell-bound immune complex, being besieged in a transplanted organ (kidney), can be the reason damage of kidneys.
3. The ALS some parties possess the pyrogenic action (raising of t ° to 38 — 39 °), cause a swelling, puffiness and a hyperemia in the place of an injection. See also Cytotoxins .
Bibliography: Kraskina H. A., Holchev N. V. and Lopatin T. K. The characteristic of depressive action antilimfo cytic serum and antilimfo-cytic globulin on products of antibodies at mice, Zhurn. mikr., epid, and immun., JVa 2, page 83, 1971; Medical Avar P. B. Biological effect heterological anti-limfotsntarnykh syvorotok.v book. - Organ transplantation and fabrics at the person, iod of an edition of F. Rapaport and Zh. Dosse, the lane with English, with 411, M., 197 3, bibliogr.; In and 1 η e N. In vivo evaluation of antihuman lymphocyte s; 'ra, Fed. Proc., v. 29, p. 117, 1970; James K. Anti-lymphocytic antibody, €lin. exp. Immunol., v. 2, *p. 615, 1967, bibliogr.; Metchnikoff E. Etude sur la r6sorption des cellules, Ann. Inst. Pasteur, 1.13, p. 737, 1899; S t and of z 1 T.E.Heterologous antilymphocyte globulin, histoincompatibi-lity matching, and human renal homotransplantation, Surg. Gynec. Obstet., v. 126, p. 1023, 1968, bibliogr.; Traeger J. €. Utilisation chez l'homme de globulines antilymphocytaires, Minerva nefrol., t. 15, p. 96, 1968, bibliogr.; Woodruff M. F. A. Immunosuppression by antilymphocytic serum, in book: Organ transplantation today, ed. by N. A. Mitchison a. o., p. 90, Amsterdam, 1969; Woodruff M. F. A. a. Anderson N. F. The effect of lymphocyte depletion by thoracic duct fistula and administration of antilym-phocytic serum on the survival of skin homografts in rats, Ann. N. Y. Acad. Sci., v. 120, p. 119, 1964.
Yu. M. Zaretskaya.