[Biological properties of the Escherichia of serological group 06 isolated in acute intestinal diseases of undetermined etiology]. / Biologicheskie svoistva ésherikhii serologicheskoi gruppy 06, vydelennykh pri ostrykh kishechnykh zabolevaniiakh neustanovlennoi étiologii.

The authors describe new serological and biochemical types of escherichia belonging to serological group 06, isolated in group and sporadic cases of sickness, from the affected children. Circulation of seven serological types was revealed--partial composition of the O-antigen proved to be serologicaly nonhomogeneous. Escherichia of a single sero-anzymatic type (06a6b: K13: H1) were isolated in a group affection.

[Biochemical and serological characteristics of Escherichia belonging to the serological group 01]. / Biokhimicheskaia i serologicheskaia kharakteristika ésherikhii serogruppy 01.

A circulation at the territory of the country of various biochemical and serological variants of escherichia belonging to serological group O1, isolated in acute intestinal diseases of children and adults, was revealed. Nonhomogeneousness of the partial composition of the O-antigen was demonstrated; K-antigens were determined; new H-antigens were described. Of the 10 serological types of escherichia there proved to prevail O1 : K? : Hp and O1 : K1 : Hp; in group and sporadic acute intestinal diseases there were for the first time isolated O1 : K1 : H34, O1 : K1 : H20, O1 : K1 : Hp, O1 : K51 : H7, and O1 : K? : H20.

[Monoclonal antibodies to Mycoplasma pneumoniae antigens]. / Monoklonal'nye antitela k antigenam Mycoplasma pneumoniae]

Approaches to obtaining stable mouse hybridomas, capable of producing monoclonal antibodies (McAb) to M. pneumoniae key antigens, were developed. As the result of hybridization experiments, 7 clones were obtained; of these, 4 clones stably synthesized IgG McAb. Clones H1/H9 and H9/B2 synthesized antibodies to thermolabile, proteinase-sensitive K protein, produced by cytoplasmic membranes of M. pneumoniae cells. The molecular weight of this protein was found to be 90 kD. McAb of clone H1/H9, labeled with horse-radish peroxidase and fluorescein isothiocyanate, specifically reacted with M. pneumoniae antigens in the immunofluorescence test and the enzyme immunoassay (EIA). The sensitivity of EIA was 0.25 ng/ml of antigen protein. These data are prerequisites for the development of diagnostic test systems for the detection of M. pneumoniae antigens in different biological substances obtained from patients with respiratory pathology.

[Production of hybridomas synthesizing monoclonal antibodies to streptococcal group A polysaccharide]. / Poluchenie gibridom, sinteziruiushchikh monoklonal'nye antitela k polisakharidu streptokokka gruppy A.

Nine stable hybridomas synthetizing monoclonal antibodies to the antigenic determinants of polysaccharide A of group A streptococcus were obtained. Three monoclonal antibodies possessed precipitating properties. The formation of hybridomas was found to be influenced by the presence of immune splenocytes and the standard conditions of cell fusion. The highest yield of hybridomas was observed under the conditions ensuring the growth of cell in 80-100% of the wells. Rapid and specific screening was found to be an important stage in obtaining hybridomas.

[Trial of pertussis toxin activity in vitro on CHO cells]. / Ispytanie aktivnosti kokliushnogo toksina in vitro na CHO-kletkakh.

The activity of B. pertussis toxin has been tested in the continuous culture of CHO (Chinese hamster ovary) cells. The in vitro method of testing B. pertussis toxin is rapid, highly sensitive and specific. The unit of activity of B. pertussis toxin is higher than in mouse tests by several orders. The specificity of the action of B. pertussis toxin on CHO cells has been confirmed by the test of the neutralization of the toxicity effect with antiserum.

[Immunobiological properties of monoclonal antibodies to Mycoplasma hominis antigens]. / Immunolobiologicheskie svoistva monoklonal'nykh antitel k antigenam Mycoplasma hominis.

Approaches to obtaining stable mouse hybridomas synthesizing monoclonal antibodies (McAb) to M. hominis key antigens were developed. 4 clones capable of the stable synthesis of McAb of different IgG classes were obtained. Clones A3/2 and A5/D produced antibodies to the thermostable determinant to with a mol. wt. of 80-120 kD, sensitive to sodium periodate and resistant to potassium proteinase. Clone H9/B2 synthesized McAb which interacted with potassium proteinase-sensitive M. hominis thermolabile determinant with a mol. wt. of 80 kD. McAb of clone A3/2, labeled with fluorescein isothiocyanate and horse-radish peroxidase, specifically reacted with M. hominis antigens in the immunofluorescence test and the immunoenzyme assay (EIA). The sensitivity of EIA was 0.25 ng/ml of antigen protein. These data may serve as prerequisites for the development of diagnostic test systems aimed at the detection of M. hominis antigens in different clinical substances.

[Synthesis of monoclonal antibodies to tissue-specific antigens of human skin and thymus epithelium in polyclonal activation by pertussis toxin]. / Sintez monoklonal'nykh antitel k tkanespetsificheskim antigenam épiteliia kozhi i timusa cheloveka pri poliklonal'noi aktivatsii kokliushnym toksinom.

Following the immunization of BALB/c mice with B. pertussis toxin, monoclonal antibodies (McAb) to the antigens of human epithelium, both dermal (the basal, superbasal or all epidermis levels) and thymic (the epithelium of the medullary zone, the cortical and medullary epithelium around Hassall's corpuscles), have been obtained. McAb have been obtained as the result of the polyclonal activation of autoreactive B-cells with B. pertussis toxin. McAb thus obtained can be used for the determination of the corresponding antigens in the epithelial tissues of the thymus and other organs in man, as well as for the diagnosis of tumors, histogenetically related to integumentary tissues of the epidermal genesis.

[The immunochemical and biological properties of monoclonal antibodies to Chlamydia trachomatis]. / Immunokhimicheskie i biologicheskie svoistva monoklonal'nykh antitel k Chlamydia trachomatis.

In this work some properties of 11 monoclonal antibodies to C.trachomatis, obtained in our earlier investigations, were studied and the antigens recognized by these McAb were characterized. Some McAb reacted with a genus-specific thermostable antigen, sensitive to sodium metaperiiodate and having a mol. wt. of 10 - 12 kD. Other McAb interacted with C.trachomatis species- and subspecies-specific thermostable proteins with a mol. wt. of 40 kD. Two McAb reacted with thermolabile protein with a mol. wt. of 32 kD and some C.trachomatis unidentified protein. The in vitro study of two McAb, active against subspecies-specific proteins, revealed the neutralizing activity of these McAb. Thus McAb obtained in this investigation may be for the differential identification of different determinants and for further analysis of the antigenic structure of C.trachomatis, as well as for the development of modern immunodiagnostic preparations.

[Immunodiagnostic preparations based on monoclonal antibodies to Chlamydia trachomatis]. / Immunodiagnosticheskie preparaty na osnove monoklonal'nykh antitel k Chlamydia trachomatis.

Highly sensitive diagnostic preparations for the detection of C. trachomatis by direct immunofluorescent and enzyme immunoassay techniques were obtained with the use monoclonal antibodies to C. trachomatis genus-specific polysaccharide antigen. The enzyme immunoassay diagnostic preparation permitted the detection of C. trachomatis in experimental specimens in a dose of 4-14 ng protein. The results of the primary clinical trial of the immunofluorescent preparation revealed that its effectiveness was not lower than that of similar foreign commercial preparations. The use of this diagnosticum for the study of clinical material obtained from 1,603 patients with different venereal diseases showed the presence of chlamydial contamination in 40.2% of the examined patients. The data thus obtained made it possible to recommend the newly developed immunofluorescent preparation for diagnosing infections caused by C. trachomatis.

[Identification of serological group 0128:K67 Escherichia]. / Ob identifikatsii ésherikhii serologicheskoi gruppy 0128:K67

The authors present the results of studies of etiology of acute group intestinal diseases in neonates from whom escherichia of serological group 0128ac:K67 possessing the following characteristics were isolated: of the same (with the H12 antigen) serological and enzymatic type (nonfermenting sucrose and raffinose, fermenting dulcit and sorbit the first 24 hours, and slowly fermenting ramnose). All the cultures isolated were resistant to the majority of antibiotics used at present, and were only weakly sensitive to erythromycin. Difficulties (agglutination of live cultures with production sera in the absence of low agglutinability of heated cultures) in serological typing of the cultures were due to different partial O-antigen composition of the cultures isolated and of the production strain used in the preparation of commercial sera of the given serological group (0128ab:K67). Because circulation of escherichia of serological 0128ac variant was revealed in the USSR there occurred a necessity of their identification in practical laboratories; for this purpose organization of industrial production of the corresponding serum is necessary.

[Biological characteristics of enteropathogenic escherichia of serologic group 015:K]. / Biologicheskaia kharakteristika énteropatogennykh ésherikhii serologicheskoi gruppy 015:K

A study was made of 239 strains of enteropathogenic escherichia 0151:K-- isolated in various regions of the USSR from patients with the clinical diagnosis of dysentery, gastroenteritis, intestinal coli-infection: a standard strain of the international collection of escherichia belonging to the given serological group was also studied. There was shown an increase in the role of these microorganisms among the enteropathogenic escherichia recorded at the territory of the USSR; they occupied the third place by the frequency of isolation after the serological group 0124:K72 and 0111:K58. There was established a common nature of the enzymatic characteristics of escherichia 0151:K--with shigellae by the absence of lactose, sucrose, inosite, adonite fermentation, the presence of gasless, immobile variants containing no lysin decarboxylase, and a possibility of rapid differentiation from shigellae in the use of acetate medium. Among the escherichia 0151:K--there was revealed the presence of 5 biotypes by the capacity to gas-formation in glucose, arabinose, sorbit, dulcit fermentation, and decarboxylation of lysin and ornithin; three biotypes are described for the first time. Industrial issue of the agglutinating serum 0151:K--is necessary to provide the diagnosis of these microorganisms at the territory of the USSR.

[Multispecific monoclonal antibodies]. / Mul'tispetsificheskie monoklonal'nye antitela.

Immunization of BALB/c mice with Rickettsia prowazekii antigens, Bordetella pertussis toxin and Legionella pneumophila cytolysin induces the synthesis of IgM autoantibodies of different specificity. Among monoclonal antibodies, multispecific antibodies with a wide reactivity spectrum have been found to make up high percentage (30-80%). Monoclonal antibodies interact with different bacterial antigens and tissue substances. A hypothesis has been put forward that normally the injection of the antigen is followed by the appearance of antigen-nonspecific "immunological noise", including the synthesis of both tissue-specific and multispecific autoantibodies. Such antigen nonspecific "immunological noise" must have a certain threshold level which can be determined with the use of hybridoma techniques. This problem is particularly topical for bacterial antigens, as many of them are used in the development of vaccinal preparations, which may lead to an increase in the synthesis of autoantibodies and induce different autoimmune disturbances in the body.

[Hybridomas synthesizing monoclonal antibodies to Bordetella pertussis toxins]. / Gibridomy, sinteziruiuschchie monoklonal'nye antitela k nekotorym toksinam Bordetella pertussis.

Hybridomas synthetizing monoclonal antibodies (McAb) to B. pertussis toxin (BPT) and endotoxin, or lipopolysaccharide (LPS), were obtained. The specificity of McAb to BPT was confirmed in the leukocytosis-stimulating factor neutralization test. Two hybridomas synthetized McAb, seemingly active against the common determinant of BPT and LPS. The McAb of one hybridoma reacted with the crude extract of BPT.

[Monoclonal antibodies to Legionella pneumophila cytolysin]. / Monoklonal'nye antitela k tsitolizinu Legionella pneumophila.

The fusion of spleen cells, taken from BALB/c mice immunized with the purified preparation of L. pneumophila cytolysin, with cells Sp2/0 and NP has been carried out. As a result, hybridoma cells producing IgG1, IgG3 and IgM antibodies to this protein have been obtained. All monoclonal antibodies (McAb) thus obtained react with L. pneumophila strain lysates in the precipitation test, while IgG3 and IgM antibodies react with erythrocyte diagnostic agents prepared from the lysate of L. pneumophila cells in the hemagglutination test. In the Western blot assay, McAb react with the 37 KD protein (cytolysin) and a number of other proteins from L. pneumophila cultures and L. pneumophila cell lysate, but do not react with the species-specific protein with a molecular weight of 29 KD, contained in the outer membrane of L. pneumophila, as well as with other species: L. bozemanii, L. dumoffii, L. longbeachae, L. micdadei. The possibility of using these McAb conjugated with FITC and peroxidase for the rapid diagnosis of Legionella infection is shown.

[Immunomorphological (light and electron microscopic) diagnosis of human squamous cell cancer using monoclonal antibodies]. / Immunomorfologicheskaia (sveto- i élektronno-mikroskopicheskaia) diagnostika ploskokletochnogo raka cheloveka s pomoshch'iu monoklonal'ny kh antitel.

It is shown with fluorescent and peroxidase labels for antibodies that the antigen to which the monoclonal antibody A-6/2 is directed, is a marker for cells of the basal stratified epithelium (basal-cell antigen). This antigen persists in squamous cell carcinomas of various sites and degrees of differentiation, but does not occur in tumors of other origins. With electron immunocytochemistry, the antigen was found to be located in tonofilaments and desmosomes of human squamous cell carcinoma. The A-6/2 monoclonal and basal-cell antigen may be used when developing immunomorphologic (light- or electronmicroscopic) procedures for histogenetic diagnosis of these carcinomas.

[Monoclonal autoantibodies to the epithelial basement membrane cells of human skin and thymus obtained through immunization with Rickettsia prowazekii antigens]. / Monoklonal'nye autoantitela k kletkam bazal'noi membrany épiteliia kozhi i timusa cheloveka, poluchennye pri immunizatsii antigenami rikketsii Provacheka.

As the result of immunization of BALB/c mice with the commercial preparation of typhus vaccine and R. prowazekii corpuscular antigen, in 29.2% and 40.3% of cases (respectively) the appearance of hybridomas synthesizing monoclonal antibodies (McAb) to different autologous structures (skin and thymic epithelium, cell nuclei, conjunctive tissue structures and vascular endothelium) has been revealed. The McAb under test have proved to be IgM-autoantibodies. McAb M-6, active against the basal membrane of human skin and thymic epithelium, produce quite a definite picture of disturbances in the differentiation of epithelium and can be used for the diagnosis of dyskeratosis.

[The detection of Rickettsia prowazekii in the vectors of louse-born typhus Pediculus humanus by using monoclonal antibody-based preparations]. / Vyiavlenie Rickettsia prowazekii v perenoschikha sypnogo tifa Pediculus humanus s ispol'zovaniem preparatov na osnove monoklonal'nykh antitel.

The results of this investigation indicate that preparations obtained on the basis of monoclonal antibodies have proved to be suitable for the detection of R. prowazekii antigens in the natural carrier of typhus when used in all types of the enzyme immunoassay; of these, the assay made by the capture method has been found to possess the highest sensitivity. The testing of the sensitivity limits has shown that this method known as ELISA-mu-capture, i.e. ELISA carried out with the use of antibodies to the mu chain of human immunoglobulin, is capable of detecting the antigen in a dose of 0.5-1 ng. Preparations based on monoclonal antibodies to species-specific R. prowazekii antigen permit the identification of the causative agent of typhus in its natural carrier within 24 hours.

[A new variant, for the USSR, of enteropathogenic escherichia 0151:K--, isolated from acute intestinal diseases under the conditional name "Crimea"]. / O novoi dlia SSSR raznovidnosti énteropatogennykh ésherikhii 0151:K--, vydeliaemykh pri ostrykh kishechnykh zabolevaniiakh pod uslovnym oboznacheniem "Krym"

A study was made of enteropathogenic escherichia "Krym" isolated in the USSR from patients with acute enteric diseases; their antigenic structure was unknown. Also cultures of new serological groups officially recorded as standard test-strains were investigated. By its antigenic structure "Krym" escherichia corresponded to the standard strain of the international set 0151:K--. Along with the H10 antigen described earlier "Krym" escherichia proved to possess H11 antigen; these two serological types of escherichia were revealed in the USSR, i.e. 0151:K--: H10 and 0151:K--: H11. The statement on the presence of a new H50 antigen described in escherichia 880-67 with an antigenic formula of 0151:K--: H50 should be revised due to its complex identity with the 0151: K--: H10 strain. For proper serological identification the conditioned name "Krym" should be replaced by "escherichia of serological group 0151:K--".

[Data on the biological characteristics of serological group O4 Escherichia isolated in acute intestinal diseases]. / Materialy k biologicheskoikharakteristike ésherikhii serologicheskoi gruppy O4, vydelennykh pri ostrykh kishechnykh zabolevaniiakh

A study was made of 155 strains of E. coli of the O4 serological group isolated from sick children and adults during group and sporadic acute intestinal diseases), from persons who came in contact with them, and also from healthy persons during prophylactic examination; three standard cultures were examined as well. Along with strains with a typical enzymatic activity there were strains which produced retarded lactose fermentation and also gas-free, immobile and lysin-negative strains resembling Shigellae. Eight biochemical types were determined among E. coli 04. A study of the antigenic structure by cross agglutinin adsorption indicated identicity of the strains by O-antigen and their difference by the K- and H-antigens. Circulation of E. coli of serological types O4: K12(L): H1,O4: K3(L): H5,O4: K3(L): H12,04: K12(L): H40,04: K52(L): H4, and O4: K12(L) HII was revealed; the first two serological types prevailed. Serological types of O4: K3(L):H12, O4: K12(L): HI and O4: K12(L): H40, isolated in cases of group and sporadic acute intestinal diseases were described for the first time.

[Characteristics of serological group O144:K? Escherichia isolated in acute intestinal diseases]. / Kharakteristika ésherikhii serologicheskoi gruppy O144:K?, vydelennykh pri ostrykh kishechnykh zabolevaniiakh

A study of 14 strains of E. coli belonging to serological group O144: K? isolated in clinical dysentery (bacteriologically unconfirmed), from contacts in the foci of clinical dysentery and acute enteric disease of unknown etiology, and also in prophylactic examination, was made. Among these cultures there were strains with shigella characteristics. Along with the serological type O144: K?: H--described earlier there was determined serological type O144: K?: H4 differing in the enzymatic activity and the absence of the capacity to produce shigellae keratoconjunctivitis. The cultures of the given serological type were isolated in the focus of bacteriologically-undeciphered acute intestinal disease, this pointing to their possible etiological role. In the light of the aforesaid escherichia of the O144: K? serologically groups could be referred to the enteropathogenic ones; in this connection it is necessary to prepare preparations for their serological identification.