SERUMS immune (synonym antiserums) — blood preparations of animals or person, antibody-containing; are used for diagnosis, treatment and prevention of various diseases.
Studying of immune serums is closely connected with development of the humoral direction in immunology developed from the middle of the 80th of 19 century. In 1890 E. Bering and S. Kitasato found in blood serum of rabbits, immunizirovanny a tetanin, the antitoxins neutralizing this poison. In 1891 P. Ehrlich opened antibodies (see) to toxins of a plant origin. Thanks to works E. Ra and G. N. Gabrichevsky antidiphtherial serum was widely used at treatment diphtherias (see). In 1894 Pfeyffer and V. I. Isaev described a lysis of cholera vibrioes in an organism of an animal under the influence of specific immune serum (see. Isaeva — Pfeyffera a phenomenon ).
Zh. Borde, studying effect of anticholeraic serum in I. I. Mechnikov's laboratory, established that the cholera vibrio under its influence loses the mobility. In 189(5 g Gruber and Durham (M. of Gruber, H. E. Durham) showed that as a result of immunization of animals of serum of their blood gained properties to agglutinate the corresponding microbes. Antibodies — agglutinins were so open. In 1897 R. Kraus established that blood sera of immunizirovanny animals affect filtrates of the corresponding bouillon cultures: in a transparent filtrate the deposit (precipitated calcium superphosphate) drops out. In 1899 F. Ya. Chistovich the first received pretsipitina to proteins of animal origin.
In 1898 Zh. Borde opened at Guinea pigs, immunizirovanny rabbit erythrocytes, antibodies — hemolysins and hemagglutinins. In 1900. I. I. Mechnikov, injecting Guinea pigs an emulsion of a spleen or mesenteric limf, nodes of a rabbit, received specific serums — antileukocytic serums. Serums against other cells of an organism were soon emitted. Thus, receiving immune serums is based on property of antigens (see) to cause in an organism antibody formation (see).
Immune serums receive from immunizirovanny animals or the person, and also from convalescents, blood to-rykh contains antibodies after postponed inf. diseases (serum of convalescents). G. may contain so-called normal antibodies, napr, the alloantibodies, or the isoantibodies which are formed at the person or an animal during a zhkzna and not depending on artificial immunization (see). As a result of repeated immunization receive S., antibody-containing in high credits — hyperimmune serums. Distinguish diagnostic and treatment-and-prophylactic serums.
- 1 Diagnostic serums
- 2 Treatment-and-prophylactic serums
- 3 Table 1. The LIST of the MAIN DIAGNOSTIC IMMUNE SERUMS, THEIR RECEIVING, USES IN CLINICAL IMMUNOLOGY AND the FORM of RELEASE
- 4 Table 2. LIST of the MAIN DRUGS of IMMUNOGLOBULINS AND ANTI-TOXIC SERUMS, SOURCE of THEIR RECEIVING, USE AND WAY of INTRODUCTION
Diagnostic serums, as a rule, contain antibodies in higher caption, than treatment-and-prophylactic. They are received as from small a lab. animals (rabbits, Guinea pigs), immunizirovanny the killed microorganisms, antigens, anatoxins, and from large (goats, horses, rams) if a large amount of serum is necessary. Diagnostic S. apply in various immunol. reactions for establishment of a look, subspecies or a serotype (serovar) of the activator inf. diseases, definitions of various antigens in biol. liquids. At the same time use S., antibody-containing, called depending on character immunol. reactions. So, in an agglutination test — agglutinins, in a precipitation test — pretsipitina, in reaction of binding complement — a complement to a vyazyvayushchpa of an antibody, in an immunofluorescence — fluorescent antibodies, in an immunoradiomet-richesky method (see. A radio immuno-logical method) — antibodies, marked radioisotope, in an immunoenzyme method (see. Enzyme-immunological method ) — antibodies, marked enzyme, in an electronic and microscopic immunohistochemistry — antibodies, marked enzyme or ferritin, etc. Therefore depending on character immunol. reactions distinguish agglutinating, precipitant, fluorescent, hemolitic, marked isotopes, enzymes, and other diagnostic serums.
Among diagnostic S. distinguish antimicrobic, to the Crimea antibacterial and anti-virus, and also anti-toxic Pages belong.
Antibacterial serums usually use as agglutinating. On degree of specificity they are divided on not adsorbed (native) and adsorbed (polyvalent, antibody-containing to several antigens or several determinant groups of antigen, and monoreceptor). Not adsorbed serums contain cross reacting antibodies since various microbes can have similar antigens. The serums intended for detection of nek-ry salmonellas (e.g., S. typhi, S. paratyphi, S. typhimurium, S. enteritidis, S. anatum, S. newport, S. choleraesuis), shigellas and escherichias concern to them. Not adsorbed agglutinating serums use for statement of the developed agglutination test on purpose identifications of microbes (see). At the same time for exact with-erol. identifications of the activator (see. Serological researches ) it is important to consider that reaction went to a caption or a half of a caption of serum. Not adsorbed serums because of rather low specificity find smaller application, than adsorbed.
The adsorbed polyvalent serums are capable to agglutinate several related bacteria having the general antigen. The adsorbed monoreceptor serums contain antibodies only against a certain antigen, i.e. they have high specificity (diagnostic type-specific serums). They are received preferential adsorption of antibodies (see. Castellani method ). The adsorbed serums use in an agglutination test (see) on a slide plate, a thicket for identification of the activators relating to this. Enterobacteriaceae, napr, to identification of enteropathogenic escherichias apply polyvalent and standard OK - serums, to definition of belonging of the activator to a genus of salmonellas — the agglutinating adsorbed polyvalent salmonellezny O-serum (groups A, B, C, D, E), to definition serol. group and a serotype of the activator — separate adsorbed About - and then N-serums. For definition in the studied material of antigens, napr, botulinum toxins or
O-typroid antigen, use the corresponding antibodies of serum occluded on erythrocytes — so-called antitelny erythrocyte diagnosticums (see. O-agglutination ).
Anti-virus diagnostic S. use in various reactions of immunity, napr, hemagglutination-inhibition reactions (see. Hemagglutination ), reactions of binding complement (see), reactions of an immunofluorescence (see), etc. Special requirements are imposed to anti-virus serums depending on a type of a virus. A variety of sources of anti-virus serums is connected with availability in them of nonspecific inhibitors, to-rye can suppress and mask receptors of viruses. Anti-virus serums clear of thermolabile inhibitors warming up at t ° 56 ° within 30 min., and of thermostable inhibitors fermenting, influence by carbonic acid, etc. depending on the nature of a virus. Besides, define availability of nonspecific agglutinins. For the purpose of suppression of bacteriophages apply to the antifago-Vyya S. V a lab. to practice they are added to mediums for release of bacteria from their bacteriophages.
From anti-toxic diagnostic serums most often use serums for detection of botulinum toxins in a neutralization test of in vivo (see. Botulism, laboratory diagnosis ) and identifications of toxin of the causative agent of diphtheria in a precipitation test (see. Diphtheria ).
Precipitant serums use in precipitation tests (see) for definition of soluble antigens of microbic, plant or animal origin, definition of autoantibodies, S-reaktiv-nogo a squirrel, for diagnosis inf. diseases, identification of specific accessory of blood protein, detection of certain substances in products at suspicion on falsification.
Fluorescent (lyumineetsp-ruyushchy) serums represent globulinovy fraction of immune blood serum of animals, marked fluorescent dyes — flyuorokhroma (see). They are applied to detection of microbes by method of a direct immunofluorescence (see) in patol. material and at pilot studies. From a similar tselyo use also marked flyuorokhromam antiglobulinovy and an anticomplement rny serums at an indirect immunofluorescence.
In a wedge, immunology widely apply diagnostic serums to blood typing (see), carrying out fabric typing, to the characteristic immunol. status of an organism. Blood groups on the AVO system define by the standard hemagglutinating serums prepared from blood of the person. They are specific since they contain certain alloantibodies — alpha, beta or an alpha 4 beta antibodies. Blood serum of the AV (IV) group of these antibodies does not contain.
For definition a Rhesus factor accessory of blood of nek-ry patients, pregnant women and initially inspected donors use standard serum anti-Bh0(D), and also the serum for Koombs's test (see Koombs reaction) prepared from blood of animals (rabbits, goats, rams), immunizirovanny a serum protein of the person. Apply serums anti-rh to an additional blood analysis of the donor' (C) and anti-g" (E), the serums containing two antibodies — anti-V0 (With - f-D) or anti-Rho (D - f-E) or three antibodies — анти-Rh^ (With + D + E).
For fabric typing at allogenic transplantations and hemotransfusions (see Immunity a trance plantatsionny, antigens of gistos of a mestimost, t. 20, additional materials) so-called test HLA-and NA-, NB reactants use. The HLA reactant is received from persons, sensibilized the corresponding antigen (e.g., blood serum of women, sensibilized during pregnancy, or blood serum of immunizirovanny donors). Antibodies define by limfotsitotoksichesky test (see. Leukocytic tests) or reaction of leukoagglutinating (see Agglutination).
For definition immunol. the status of an organism apply monospetsi-fichesky serums to immunoglobulins of the person of the classes G, A, M, to-rye receive from rabbits or sheep, immunizirovanny cleared by IgG, IgA, IgM of the person. These serums for removal of geterolo-gichny antibodies «exhaust» the corresponding adsorbents. Therefore S. contain only antibodies against immunoglobulins of the corresponding class, i.e. are monospetsifi-chesky. With their help define type and amount of immunoglobulins for assessment of nek-ry immunodeficiency (see. Immunological insufficiency), quality of drug of immunoglobulin (see), and also the monovalent antibodies appearing at nek-ry inf. diseases. These serums use in reaction of radial immunodiffusion according to Mancini (see. Immunodiffusion ).
In table 1 the list of the main diagnostic immune serums, their receiving, use in clinical immunology, a form of release is given.
concern To them anti-toxic, antibacterial, anti-virus serums, and also immunoglobulins. Anti-toxic serums receive by method of hyper-immunization of large animals (usually horses) by parenteral administration of the accruing doses of anatoxins, then the corresponding exotoxins (see Antitoxins). Anti-toxic serums apply to passive immunization (see).
Anti-toxic serums use for treatment and prevention of toksinemichesky infections, to-rykh action on an organism of exotoxins of bacteria — causative agents of tetanus, botulism, diphtheria, gas gangrene, staphylococcal infections is the cornerstone. Anti-toxic serums are also serums, antibody-containing against poisons of snakes, spiders, poisons of a plant origin. Antibodies of anti-toxic serums neutralize effect of the corresponding toxins, providing thereby to lay down. - the prof. effect.
The anti-toxic serums received from giperimmunizirovanny animals clear and concentrate (for the purpose of removal of inactive ballast proteins); control their physical properties, sterility, an apirogennost, harmlessness, specific activity, amount of protein, a pH value and availability of residual ammonium sulfate. Specific activity of anti-toxic serums is expressed in the international units (ME). Take the minimum quantity of serum for unit, a cut neutralizes standard unit of toxin. For serums, yet not hsheyushchy international standards, national standards — the anti-toxic units (ATU) are approved.
Definition of specific activity of anti-toxic serums is made two-stage titration. At the first stage establish a pilot dose of toxin by means of standard anti-toxic serum. At the second stage define activity of ispytuyemy anti-toxic serum by means of a pilot dose of toxin (see. Antitoxins ).
Standard samples of anti-toxic serums undergo control in the State scientific research institute of standardization and control of medical biological supplies of L. A. Tarasevich and are dispatched to the manufacturing enterprises producing serums. Standard serums conform to the accepted international standards that facilitates a possibility of comparison of properties of the serums made in the different countries.
Antibacterial serums receive from blood of horses or oxen, giperimmunizirovanny the corresponding killed bacteria or their antigens. They did not find broad application with the medical or preventive purposes because of existence of more effective antimicrobic remedies. From antibacterial serums sometimes apply heterogeneous immunoglobulins (antianthracic gamma-globulin and protivoleptospirozny gamma-globulin).
Anti-virus serums (heterogeneous immunoglobulins) receive from blood of animals, immunizirovanny vaccinal strains of viruses or the corresponding viruses. They are cleared by method of spirit sedimentation at a low temperature. Most gamma-globulin against a tick-borne encephalitis, antirabic gamma-globulin, etc. was widely used (see. Immunoglobulins ).
The immunoglobulins received from blood of the person (homologous immunoglobulins) are, except for normal immunoglobulin, immunoglobulins of the directed action. Advantage of homologous immunoglobulins before heterological is weak reactogenicity and longer circulation of antibodies in an organism (within 30 — 40 days). Receive homologous immunoglobulins by fractionation by alcohol at a temperature below zero (by Kohn's method). In a wedge, practice immunoglobulins with the most reduced anti-complementary activity have the greatest value, to-rye use for intravenous administration. Such drug is immunoglobulin normal human for intravenous administration. In immunoglobin drugs for intravenous administration aim to remove ability of Fc-area of molecules of immunoglobulins to activate system of a complement, and also to prevent aggregation of molecules of immunoglobulins. It is applied at various forms of immunological insufficiency (see).
Among various immunoglobulins of the directed action, or gamma-globulins of the directed action, allocate an anti-Rhesus factor-ny immunoglobulin — aHTn-Rh0(D), to-ry use for immunoprevention of a hemolitic disease of newborns (see). It is entered to primapara Rh-negative women into a puerperal period or after abortion; at repeated pregnancies at such women healthy Rh-positive children are born. Hyperimmune plasma of donors, and also plasma of women, sensibilized Rh-Hr antigen in the natural way to time of repeated pregnancies can be a source of receiving drug.
Immunoglobulin of the directed action is also anti-lymphocytic (heterogeneous) immunoglobulin which is a part of An of the tilimfotsitarny serum (see) containing various antibodies to lymphoid cells is preferential to lymphocytes. The anti-lymphocytic serum received by immunization of animals by cells of a thymus gland and lymphocytes chest limf, a channel has the greatest immunodepressive activity. It is widely used at transplantation of vitals, first of all kidneys.
Antireticular cytotoxic serum (serum of the Pilgrim) is received from blood serum of a horse, immunizirovanny tissue of a spleen or marrow of the person. It contains antibodies cytotoxins, active in relation to cells of these bodies. Use of antireticular cytotoxic serum especially reasonablly at is long not healing ulcers and wounds, and also in the presence of slowly resolving inflammatory centers.
In tab. 2 the list of the main drugs of immunoglobulins and anti-toxic serums, a source of their receiving, use and a way of introduction is given.
Receiving immune serums includes: preparation of the corresponding immunogen, i.e. the substance stimulating products of antibodies; immunization of animals; control of the received serums. For receiving serums against such haptens, as pharmakol. drugs or low-molecular hormones, apply as immunogen, or the immunizing drug, conjugates hapten carrier. Degree of purity of proteinaceous immunogens is checked by means of immunochemical and physical and chemical researches. Immunization is one of the most variable processes during the receiving serums since the dose of immunogen can influence properties (e.g., specificity) formed anti-* bodies. Small doses of the entered antigen allow to save a consumption of immu-nogenny drugs, induce emergence of avidny antibodies in a high caption. At small doses of antigen small amounts of foreign impurity therefore collateral antibodies are not formed also are entered into an organism. For stimulation of antibodyformation immunogens mix with adjuvants (see), napr, a Freund's adjuvant. In addition to the stimulating function, adjuvants perform also the depositing function concerning immunogen. Except a dose of antigen, it is also important to consider degree of its purity, a way, terms and the sequence of introduction. The choice of an animal depends on estimated amount of serum. Receiving serum from rabbits, being good producers of antibodies, demands the maintenance of a large number of such animals. In a large number serum can be received from horses or other large animals.
For definition in serums of quantity of antibodies and for the characteristic of activity of diagnostic serums, napr, the agglutinating immune serums, make their titration. The caption of the agglutinating immune serums is defined as limit cultivation of blood serum, at Krom the agglutination test is found. Usually prepare the cultivations of serum increasing in a geometrical progression (double). For the characteristic of activity of blood serum, as a rule, consider result of reaction in each cultivation or totally in all cultivations by quantity of conventional positive units.
Table 1. The LIST of the MAIN DIAGNOSTIC IMMUNE SERUMS, THEIR RECEIVING, USES IN CLINICAL IMMUNOLOGY AND the FORM of RELEASE
Table 2. LIST of the MAIN DRUGS of IMMUNOGLOBULINS AND ANTI-TOXIC SERUMS, SOURCE of THEIR RECEIVING, USE AND WAY of INTRODUCTION
Bibliography: Immunological methods, under the editorship of X. Frimelya, the lane with it., M., 1979; Kovalyov I. E. and Field O. Yu. Antibodies to physiologically active connections, M., 1981; Methods of researches in immunology, under the editorship of. I. Lefkovit-s and B. Pernis, lane with English, M., 1981; The Guide to vaccinal and serumal business, under the editorship of P. N. Burgasov, JM., 1978; Smirnov V. V., etc. Scientific bases of production of diagnostic drugs (serum for identification of bacteria), Kiev, 1980, bibliogr.; The reference book on use of bacteritic and virus drugs, under the editorship of S. G. Dzaguro-va and F. F. Rezepova, M., 1975; Sh l I - x about in E. N. Immunologiya, an immunodiagnosis, immunoprevention of infectious diseases, Chisinau, 1977; In e h-ú i n g E. Die Blutserumtherapie, Bd 1, Lpz., 1892.
A. S. Bulls, E. P. Pashkov.