ANTITOXINS (Greek anti-against + toxins) — the specific antibodies which are formed in a human body and animals under the influence of toxins (anatoxins) of microbes, poisons of plants and animals, having ability to neutralize their poisonous properties.
And. are one of factors of immunity (see) and carry out the leading protective role at toksinemichesky infections (tetanus, diphtheria, botulism, gas gangrene, some streptococcal and staphylococcal diseases, etc.).
In 1890 Bering and Kitasato (E. Behring, S. Kitasato) for the first time observed that serums of the animals who were repeatedly receiving nonlethal doses diphtheritic and a tetanin, gained ability to neutralize these toxins (see). At Pasterovsky institute in Paris of Ru (E. Roux) in 1894 the first anti-toxic antidiphtherial serum was received, to-ruyu it the first entered into broad practice. Anti-toxic serum against gas gangrene was received by Veynberg (M. Weinberg in 1915 immunization of animals the increasing doses of live culture. After opening by G. Ramon in 1923 of anatoxins (see) receiving any And. does not meet great difficulties.
In an organism under natural conditions And. are formed as a result of the postponed toksinemichesky infection or owing to a carriage of toxicogenic microorganisms, are found in blood serum and can provide immunity to toksinemichesky infections.
Anti-toxic immunity can be created and it is artificial: active immunization by anatoxin or administration of anti-toxic serum (oroimmunity). At primary immunization by anatoxin speed of education And. depends on sensitivity immuniziruyemy, on a dose and quality of anatoxin, on intervals and the speed of a resorption of antigen in an organism. At immunization by the sorbed or pretsipitirovanny anatoxins which are applied in a crust, time, emergence and accumulation And. in blood occurs more slowly, than at immunization by the same doses of not sorbed anatoxins, but credits And. much higher and are found longer time. After primary immunization «immunological memory» in an organism to education And. vaguely long time, up to 25 years remains, and it is possible — and all life. At a revaccination development And. in an organism occurs very quickly. For the 2nd day after a revaccination significant amounts are found And., which credits continue to accrue in the next 10 — 12 days. Bystry development And. at a revaccination has great practical value in prevention of tetanus and other toksinemichesky infections. For prevention of tetanus of newborns carry out immunization and a revaccination by tetanic anatoxin of pregnant women. Formed And. have ability to pass through a placenta in an organism of a fruit, and also to be transferred to the newborn with milk of mother.
Anti-toxic serums receive immunization of horses and cattle the increasing doses of anatoxins, and then and the corresponding toxins. Education And. at animals occurs more intensively in case of use of pretsipitirovanny antigens — 1% of Calcium chloratum or 0,5% of potassium-alum. For increase in a caption And. at horses producers apply various stimulators (see Adjuvants).
The Soviet scientists (O. A. Komkova, K. I. Matveev, 1943, 1959) developed a method of receiving polyvalent antigangrenous (Cl. perfrin-gens, Cl. oedematiens, Cl. septicum) and antibotulinic antitoxins of types A, B, C and E from one producer. In this case the horse is immunized small doses of several antigens. This method found broad application in practice of production of polyvalent antigangrenous and antibotulinic serums from one producer with satisfactory credits of all
A. A. of antidiphtherial and antitetanic horse serum generally contain in γ1-, γ2-, β2-фракциях globulins.
And. in applied medicine are applied to prevention and treatment of diphtheria, tetanus and botulism. With the help And. at people it is possible to create oroimmunity of such tension which protects from a disease in case of penetration into an organism of a contagium or toxin as it happens at botulism. To the children who had contact with the patient with diphtheria enter And. for the prevention of a disease of diphtheria. At an injury to children and adults, neimmunizirovanny against tetanus, enter antitetanic serum. At identification of cases of botulism to all persons eating the product which caused a disease enter polyvalent antibotulinic serum for prevention.
For receiving medical action early introduction is very important And., capable to neutralize the toxin circulating in blood. Therefore efficiency serotherapy (see) depends substantially on the term of use And. Results of treatment And. at different infections are not identical. At treatment of diphtheria at people good results are received; at treatment of tetanus and botulism the best results are received at introduction And. at the beginning of a disease. Treatment of staphylococcal sepsis homologous alpha and staphylococcal antitoxin is effective (S. V. Skurkovich, 1969). At gas gangrene medical action And. it is called in question though many doctors continue to apply it.
However introduction to people of heterological anti-toxic serums for prevention and treatment of infections sometimes is followed by complications. In rare instances at administration of horse serum at the person can develop acute anaphylaxis (see), sometimes from the death. The serum disease develops in 5 — 10% of cases (see). Therefore in the USSR and other countries at people instead of horse serum apply the homologous immunoglobulin from donor blood containing tetanic antitoksinony Homologous antitoxin seldom to prevention of tetanus causes undesirable reactions and is in an organism in a necessary caption up to 30 — 40 days (K. I. Matveev, S. V. Skurkovich and sotr., 1973).
For elimination of the complications observed from administration of heterological native anti-toxic serums various ways of cleaning are offered And. from ballast proteins: salting-out by neutral salts, fractionation by means of electrodialysis, digestion by means of enzymes. The best results were received by method of peptic digestion (I. A. Perfentyev, 1936). Purification of anti-toxic serums with method of proteolysis in the USSR was carried out in Ying-those epidemiology and microbiology of Η. F. Gamalei of the USSR Academy of Medical Sciences (A. V. Beylinson and sotr., 1945). Advantage of a method of proteolysis (diaferm-3) is that it gives big extent of cleaning by 2 — 4 times And., than other methods, but at the same time 30 — 50% are lost And. At proteolysis there is a profound change of a molecule A. and reduction of its anaphylactogenic properties. Methods of cleaning and concentration of Ampere-second by use of aluminum hydroxide, filtering through sefadeksa (molecular sieves) and use of ion exchange are developed. At t ° 37 ° during 20 days a caption And. in the purified serums decreases a little, then is stabilized and remains invariable up to 2 years and more. After lyophilic drying under vacuum at low temperatures a caption And. decreases by 2 — 25%. Dried up And. keep the physical. and specific properties can be also kept for a number of years.
And. are exposed to obligatory control on harmlessness on Guinea pigs and an apirogennost on rabbits.
Contents And. in anti-toxic serums it is expressed in the international units (ME) accepted by World Health Organization that corresponds to the minimum quantity of serum neutralizing the standard unit of toxin expressed in the minimum lethal, necrotic or reactive doses depending on a species of an animal and toxin. E.g., ME of antitetanic serum corresponds to its minimum quantity neutralizing about 1000 minimum lethal doses (Dim) of standard toxin for a Guinea pig weighing 350 g; ME of antibotulinic antitoxin — the smallest amount of serum neutralizing 10 000 Dim of toxin for mice weighing 18 — 20 g; ME of standard antidiphtherial serum corresponds to its minimum quantity neutralizing 100 Dim of standard toxin for a Guinea pig weighing 250 g.
For some serums which do not have the accepted international standards national standards are approved, and their activity is expressed in national units which are called the anti-toxic units (ATU).
At titration And. at first define conventional (pilot) unit of toxin. The pilot dose of toxin is designated by a Lt symbol (Limes tod) and is established in relation to the standard anti-toxic serum produced State. Scientific research institute of standardization and control of medical biological supplies of L. A. Tarasevich of M3 of the USSR. For definition of a pilot dose of toxin according to the level of titration (to 1/5, 1/10 or 1/50 ME) of 0,2 ml add the decreasing or increasing doses of toxin of 0,3 ml to a certain amount of standard serum. After keeping at the room temperature within 45 min. this mix is entered to intravenously white mice of 0,5 ml on each mouse. Watch 4 days animals. Take that minimum quantity of toxin for a pilot dose, a cut in mix with the accepted dose of standard serum causes death of 50% of the mice taken in experience.
Antibotulinic anti-toxic serums of types A, B, C, E and antigangrenous (Cl. perfringens) In, With titrate at the level of 1/5 ME. The pilot dose of toxin podtitrovyvatsya also to 1/5 ME standard serums. Antibotulinic serum like F and antigangrenous serums of types A, D, E, and also antitetanic serum are titrated at the level of 1/10 ME. The pilot dose of toxin podtitrovyvatsya surely to 1/10 ME standard serums. Antigangrenous serum (Cl. oedematiens) is titrated at the level of 1/50 ME. The pilot dose of toxin podtitrovyvatsya to 1/50 ME standard serums. Examinees of serum part depending on an estimated caption and of 0,2 ml add a pilot dose of toxin of 0,3 ml to various cultivations of serum (counting on 1 mouse), leave mix for connection at the room temperature within 45 min. and enter 0,5 ml to intravenously white mice. Antitetanic serum is titrated by hypodermic introduction of 0,4 ml of mix to a back pad of a mouse. On each dose take not less than two mice in experience, prepare mix from calculation not less than on 3 mice. At each titration of serum Comte a role of activity of a pilot dose of toxin with standard serum is surely put.
The principles of titration of diphtheritic antitoxin the same, as other serums, only cultivations of standard serum and a pilot dose of toxin are jointly entered vnutrikozhno to a Guinea pig (Remer's method). Previously with standard serum the so-called necrotic dose vytitrovyvatsya — limes necrosis (Ln) of diphtheritic toxin representing that smallest amount of toxin, a cut at intradermal introduction to a Guinea pig (of 0,05 ml) in mix from 1/50 ME standard antidiphtherial serums causes to 4 — to the 5th day formation of a necrosis. Titration of diphtheritic antitoxin by Ramón's (flocculation test) method is made by means of toxin or anatoxin, in Krom previously define the maintenance of the antigenic units (AU) in 1 ml. One antigenic unit of toxin designated as a threshold of flocculation — limes flocculationis (Lf) is neutralized by one unit of diphtheritic antitoxin. For titration of small amounts diphtheritic And. also the intradermal method of Janszen on rabbits is applied.
And. are widely applied to prevention and therapy of toksinemichesky infections. Besides, they are used for neutralization of poisons of snakes, spiders and poisons of a plant origin.
Bibliography: Ramone G. is forty years old of research work, the lane with fr., M., 1962; Rezepov F. F. and d river. Definition of harmlessness and specific activity of immune serums and globulins, in book: Metodich. the management on laborat. to evaluation test bakt. and virusn. drugs, under the editorship of S. G. Dzagurov, page 235, M., 1972; Toxins-anatoxins and anti-toxic serums. M, 1969; Behring and. To i t and in a t oh, of Über das Zustandekommen der Diphterie-Immunität und der Tetanus-Immunität bei Tieren, Dtsch. med. Wschr., S. 1113, 1890; Kuhns W. J. a. Pappenheimer A. M. Immunochemical studies of antitoxin produced in normal and allergic individuals hyperimmunized with diphtheria toxoid, J. exp. Med., v. 95, p. 375, 1952; Miller J. F. A. P. a. o. Interaction between lymphocytes in immune responses, Cell. Immunol., v. 2, p. 469, 1971, bibliogr.; White R. G. The relation of the cellular responses in germinal or lymphocytopoietic centers of lymph nodes to the production of antibody, in book: Mechanism. antibody formation, p. 25, Prague, 1960.
K. I. Matveev.