Ultrastructure of Chlamydia pneumoniae in cell culture.

The electron microscopic appearance of Chlamydia pneumoniae elementary bodies with pear-shaped, loose outer membrane has been suggested as one criterion of its classification as a new chlamydial species. The study of the original strain TW 183 in LCL 929 and HL cells and a low-passage isolate of Kajaani-6 isolate in HL cells revealed spherical compact elementary bodies common to other chlamydia.

Immunological activity against tick-borne encephalitis virus in fractions of sera from goats of different ages.

A comparative analysis of the antihaemagglutinating, virus-neutralizing and complement-fixing activities of 19 S, 7 S, 4S and 1.5 S fractions of blood sera from goats of different ages pastured in a tick-borne encephalitis focus was carried out. Changes in the immunological activity of different fractions in relation to the age of the animals are discussed.

Comparison of the ultrastructure of small dense forms of chlamydiae and Coxiella burnetii.

In the course of passaging of Coxiella burnetii (C.b.) in Alveonasus lahorensis ticks, the haemocytes contained cell forms with electrondense cytoplasm, intracytoplasmic lamellar membranes, and a peculiar limiting membrane--25 to 30 nm thick "envelope complex". Similar small forms occurred when C.b. had been cultured in the yolk sack of chick embryos. The dense forms of C.b. were similar to those of Rickettsiella cells. Dense forms (elementary bodies) surrounded by an "envelope complex" were found also in some chlamydiae cultured in yolk sacs of chick embryos.

[Morphology of Halprowia (Chlamydia) isolated in Reiter's syndrome]. / Morfologiia gal'provii (khlamidii), vydelennykh pri sindrome Reitera

Morphology and ultrastructure of Halprowia arthritidis, strain SR-1 (HSR), isolated from the synovial fluid of a patient with Reiter's syndrome, was studied in the membranes of the yolk sacs of the developing chick embryos and the L-cell culture. In acridine orange staining for light and fluorescent microscopy there was revealed intracellular cytoplasmic inclusions containing HSR structures at various stages of its reproduction characteristic of halprowia (chlamydia). The direct immunofluorescent method demonstrated the presence of a characteristic HSR antigen not only in the developed inclusions, but also at the early stages of infection, when the morphological HSR structures could not be found by light microscopy. The ultrastructure of the HSR inclusions and forms in the cycle of development (of the initial and elementary bodies) of the SR-1 strain was typical of other halprowia. A peculiar structure of a complex of cell wall and cytoplasmic membrane of the elementary body was described. Taking into consideration the biological characteristics of HSR revealed earlier it can be considered to be a typical representative of Halprowiales s. Chlamydiales. The data obtained on other halprowia, pointing out the fact that criteria of compactness and diffuseness of inclusions, the presence of absence of glycogen in the inclusions could not serve as taxonomic signs in classification of halrpowia, were confirmed on a model of the SR-1 studied.

[Electron microscopic determination of the localization of the group-specific antigen of Halprowise (Chlamydiae) by the direct immunoperoxidase method]. / Elektronno-mikroskopichekoe opredelenie lokalizatisii gruppospetsificheskogo antigena gal'provii (khlamidii) priamym immunoperoksidaznym metodom

The direct immunoperoxidase technique was applied to the study of the localization of a group-specific antigen of Halprowiae (Chlamydiae) in the causative agents of meningopneumonia (strain MP) and of paratrachoma (strain LB-I) at the individual stages of their developmental cycle in the L-cell (16, 24 and 48 hours after infection of the culture). The product of reaction to peroxidase pointing to the localization of the group-specific antigen was localized not only on the surface, but in the whole thickness of the cell wall of the initial bodies in the form of an even electron dense layer, 200--280 A in thickness. A weak positive reaction was also noted on the outer layer of the sytoplasmic membrane. The periplasmic space remained free of the reaction product. Localization of the reaction product in the intermediate and elementary bodies remained unchanged. At the initial stages of the developmental cycle (16 hours after the infection) the reaction) the reaction product was revealed not in the whole cell wall, butin some of its areas only. There were found no qualitative differences in the localization of the group-specific antigen in the Malprowiae strains under study.

[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.

[Improvement in the method of cultivating Halprowiae (Chlamydiae) in cell cultures]. / Usovershenstvovanie metoda kul'tivnirovaniia gal'provii (khlamidii) v kul'ture kletok.

Comparative evaluation of several methods increasing the effectiveness of cultivating Halprovia (7 strains of different origins) in L-929 and McCoy cell cultures has shown that the maximum effectiveness can be achieved by supplementing the standard method for the cultivation of obligate intracellular parasites with the method of forced adsorption of Halprovia on the monolayer and the treatment of cells with DEAE dextran or cycloheximide. The peculiarities of detecting cytoplasmic halprovian inclusions in infected cells by staining the specimens with fluorescent antibodies, with Lugol's solution, and according to May-Grünwald-Giemsa are described.

[Indication of halprowia (chlamydia) antigens by the direct immunoperoxidase method]. / Indikatsiia antigenov gal'provii (khlamidii) priamym immunoperoksidaznym metodom

The authors modified and approbated the direct immunoperoxidase method for the indication in the infected L cells of the halprowia (chlamydia) antigens--etiological agents of trachoma, ocular and urogenital forms of paratrachoma, arthritis, meningopneumonia and enzootic abortion of sheep. Different types of localization of the group-specific halprowia antigen were revealed in the cytoplasm of the affected cells by light microscopy; electron microscopy demonstrated localization of this antigen in the membrane of the cell wall of the elementary and initial bodies. Specificity of the test-system used, no lesser sensitivity than the sensitivity of direct immunofluorescent method of indication of the same microbies, with a possibility of detection of stably stained antigens by widely accessible light microscopy underly practical usefulness of direct immunoperoxidase method of the diagnosis of halprowiosis (chlamydiosis) of man and animals.

[The immune status of patients with acute inflammatory diseases of adnexa uteri associated with different combinations of microorganisms]. / Issledovanie immunnogo statusa u bol'nykh ostrymi vospalitel'nymi zabolevaniiami pridatkov matki, assotsiirovannykh s raznymi kombinatsiiami mikroorganizmov.

The immune status of 130 patients with acute inflammatory diseases of uterine appendages was studied. As etiologically associated infective agents detected in these patients were opportunistic microorganisms (group 1), Neisseria gonorrhoeae in combination with opportunistic microorganisms (group 2) and Chlamydia trachomatis in combination with opportunistic microorganisms (group 3). In all three groups of patients the response of T lymphocytes to PHA and hyperactivation of the B-cell element of immunity (an increase in the relative number of B-lymphocytes and the content of IgA and IgM in the serum) was found to be suppressed. In addition, in groups 1 and 3 an increase, and in group 2 a decrease in IgG were detected. In the mucus of the cervical canal in the patients of groups 1, 2 and 3 IgM was detected, while in patients of groups 2 and 3 a decrease in sigA was established.

[Unusual small forms in the growth cycle of halprowia (chlamydia) and their possible relation to manifestations of L-transformation]. / Neobychnye melkie formy v tsikle rezvitiia gal'provii (khlamidii) i ikh vozmozhnaia sviaz' s iavleniiami L-transformatsii

The authors carried out a comparative electron-microscopic study of the ultrastructure and the developmental cycle of 15 Halprowiae (chlamydiae) strains isolated in different pathology from man and animals. Halprowiae were cultivated in the yolk sac epithelium of the developing chick embryos and in the L-cell monolayer cultures without any special action of the antibiotics and medicines. Formation of minute round structures ("minute forms"), 50--120 nm in diameter, separating from the protoplast into the periplasmic space and surrounded with cytoplasmic membrane by the vegetative forms of a number of strains (MP, AP-23, 25-SM, LB-1, MyB), is described. Along with minute forms, Halprowiae formed polymorphous giant bodies with long protrusions into the inclusion cavity; there was also a separation of vesicles of the cell wall membrane. This apparently indicated a structural derangement of the cell wall in these vegetative forms of Halprowiae. On the basis of similarity of the mentioned structures detected in Halprowiae reproduction to the known morphological characteristics of the L-form bacteria a possibility of L-transformation in Halprowiae during the normal developmental cycle as a peculiar compensatory adaptation to the action of host cell is supposed.

[Ultrastructure of the initial stages of interaction between halprowiae (Chlamydiae) and host cells]. / Ul'trastruktura nachal'nykh étapov vzaimodeistviia gal'provii (chlamidii) s kletkoi-khoziainom.

The initial stages of the interaction of halprowiae (strain CP-1), isolated in Reiter's syndrome, with a monolayer culture of cells L-929 were studied. The electron-microscopic examination of the cells every 2 hours during the first 24 hours after the inoculation allowed to follow the processes of adsorption, endocytosis of elementary bodies and their transformation into reticulate bodies by the "decondensation" of nucleoid, which indicated the beginning of the vegetative (reproductive) stage of the developmental cycle. During all the periods of observation halprowiae were enclosed into the membranes of phagocytic vacuoles formed by host cells. Besides, in other vacuoles (probably, phagolysosomes) halprowiae at various stages of destruction could be observed. 8--10 hours after inoculation the phagosomes containing reticulate bodies began to merge, thus forming an inoculation the phagosomes containing reticulate bodies began to merge, thus forming an intracytoplasmic inclusion. In this inclusion a new generation of the agent was formed by binary-like fission. At the beginning of the vegetative stage halprowiae seem to be highly sensitive to the action of exo- and endogenic factors which may cause their L-transformation. Depending on the reaction of the host cell at the initial stages of interaction with halprowiae, 3 ways of such interaction are seemingly possible: developmental cycle, destruction in phagolysosomes. L-transformation.

[The ultrastructure of a new species of Chlamydia--Chlamydia pneumoniae]. / Ul'trastruktura novogo vida khlamidii--Chlamydia penumoniae.

The dynamic study of a new Chlamydia species, C. pneumoniae (strain TWAR, isolate TW-480), inoculated into the monolayer culture of cells L-929 was made 24, 48, 72 and 96 hours after inoculation. When compared with C. trachomatis and C. psittaci, C. pneumoniae were found to stand between these two species with respect to the morphology of their intracytoplasmic microcolonies (inclusions): they were round, almost bubble-like, but more densely packed with chlamydiae, surrounded by an undulate membrane, preserving its integrity until the late stages of their development cycle. In cells L-929 C. pneumoniae had a typical development cycle accompanied by the formation of vegetative and spore-like cells, reticular and elementary bodies, as well as intermediate cells, though this process was slower than in C. trachomatis and C. psittaci. Besides normal elementary bodies, many altered ones were formed in the process of the development of C. pneumoniae in cells L-929. Most of these alterations were similar to the process of bacterial L-transformation and could be regarded as the manifestation of chlamydial pathology related to the adaptation to new host cells.