[INTERCONNECTION BETWEEN CELL MICROVESICULAR TRANSPORT AND PATHOGENS PERSISTENCE IN VITRO AND IN VIVO].

This review presents an information and proof evidence toward to the role of microvesicles, originating from the different sources pro- and eucaryotes in the initiation and development of persistence of several human and animal pathogens. Also an information about another properties of microvesicles, as well as the reference of role in the different somatic pathology, intercellular interaction and in the intracellular transport of biologically active macromolecules as well as life origin and evolutionary events.

Transmissible retrovirus in Epstein-Barr virus-producer B95-8 cells.

Epstein-Barr virus (EBV) released from the B95-8 marmoset cell line has served as a prototype for biologic and biochemical studies of EBV. Here we identify and characterize a retrovirus carried by many cultures of B95-8 cells. The experiments were stimulated by the isolation of a cDNA clone from B95-8 cells in which sequences from the EBV large internal repeat were linked to gag sequences similar to those of squirrel monkey retrovirus, human isolate, SMRV-H. However, among 413 amino acids predicted from the nucleotide sequence of the gag region of the B95-8 SMRV isolate there were 48 amino acid changes that distinguished this virus from SMRV-H originally isolated from a human lymphoid cell line by Oda et al. (1988, Virology 167, 468-476). Nucleic acid and antibody probes were developed for the B95-8 isolate of SMRV. Using such probes, we found that SMRV-B95-8 was readily transmissible, independent of EBV, as an infectious virus to human B and T cell lines. SMRV-B95-8 was highly fusogenic in the presence or absence of EBV. The ultrastructural appearance of the B95-8 retrovirus was characteristic of a type D retrovirus. Cells dually infected with EBV and SMRV-B95-8 did not demonstrate increased levels of lytic EB viral replication. SMRV-B95-8 did not by itself cause lymphocyte immortalization or enhance immortalization by EBV. Thus SMRV-B95-8 does not contribute to the major biologic properties of the B95-8 strain of EBV.

Platelet-mediated cytotoxicity and its enhancement by platelet activating factor.

Platelet cytotoxicity was assessed in 70 cancer patients with various tumor localizations and in 30 normal donors. The data presented reveal that the ACL cell line displays the highest sensitivity to platelet cytotoxicity. Using the ACL cells, we discovered that platelets from oncological patients and normal donors display comparable cytotoxicity. The level of platelet lytic activity is irrelevant to tumor localisation; however, it appears to be dependent on the stage of tumor growth. Incubation of platelets, both from donors and patients, with PAF (concentration range 10 pM to 10 nM) results in a significant rise of the killing activity of platelets. PAF induces greater cytotoxicity enhancement for platelets with lower initial activity, this pattern appearing to be the specific feature of the PAF mediated effect. Hence, platelets can be considered as effector cells relevant to antitumor immunity; PAF-mediated enhancement of platelet cytotoxicity can appear to be useful in the search for new immunotherapeutic drugs.

Intracellular type A retrovirus movement associated with an intact microtubule system.

Intracytoplasmic type A particles known to be precursors to type B retroviruses in murine, hamster and marsupial cells are closely associated with microtubules and microtubule organizing centres. In this publication, the active participation of microtubules in the intracellular transport of the particles to the cell surface has been examined in NIH 3T3 cells infected with M432 virus using vincristine sulphate (VCR) as inhibitor of microtubule polymerization. The release of virus at different times after exposure to VCR was quantified by reverse transcriptase determinations of cell supernatants and by electron microscopic quantification of the number of virions at the cell surface using freeze-dried whole cell replicas. These studies indicate that VCR inhibits both microtubule polymerization and virus release, and thus suggest that intact cytoplasmic microtubules are necessary for intracellular transport and release of virus.

In vitro studies of a co-carcinogenic effect of vaccinia and herpes group viruses.

The results of studies of a co-carcinogenic effect of two human infectious viruses in tissue culture are reported here. Viable vaccinia virus actively replicating in the cells of primary BALB/c tissue culture and in a number of continuous murine cell lines has been shown to induce in them expression of major structural p30 protein of murine retroviruses. Vaccinia virus has been also shown to cause biochemical transformation of murine cells. Evidence for the capacity of herpes simplex virus type 2 to induce malignant transformation of BALB/3T3 murine cell line has been obtained and confirmed by transplantation to mice. Transformed cell clones did not contain complete infectious herpes simplex virus but were resistant to superinfection with this virus. N-tropic endogenous murine retrovirus of C type with buoyant density in the saccharose density gradient of 1.18 g/cm3 and a reverse transcriptase activity was expressed in the transformed cells. In the virion structure six proteins typical of these viruses with the prevalence of p30 have been demonstrated. Competitive radioimmunoassay revealed a very high level of virus production: p30 level reached 7500 ng p30 R-MuLV per mg of viral protein. Specificity of this results was shown in control experiments.

The ultrastructure of the tubular complex in the cytoplasm of cytolytic T lymphocytes.

The DNA synthesis, the cytolytic activity, and the ultrastructure of cytolytic T lymphocytes (CTL) derived from mixed mouse thymocyte culture on day 5 were studied. CTL of thymic origin were absorbed by centrifugation on the surface of target cell (TC) monolayers. At different time intervals after absorption, single tubular structures (TS) and complex of tubular structures (CTS) linked with ergastoplasmic reticulum, secretory granules, Golgi apparatus, coated vesicles, and multivesicular bodies were detected in the CTL cytoplasm. The linkage of CTS with ergastoplasmic reticulum, ribosomes, and secretory lymphocyte mechanism suggests the possibility of a secretory-receptor mechanism of TC cytolysis. The release of numerous secretory vacuoles was accompanied by the enlargement of the cytoplasmatic lymphocyte membrane surface, which seemed to cause its shedding. 'Membranosomas' were observed on CTL membrane; their role is still obscure and awaits further study.

Influenza in the USSR in 1977: recurrence of influenzavirus A subtype H1N1.

Early in November 1977, several outbreaks of influenza were reported in the far eastern region of the USSR. The epidemic spread rapidly throughout the country affecting mainly people under the age of 20 years. Most of the strains of virus isolated were found to be influenza A subtype H1N1. The serological characterization of the strains is described in this paper.

Mason-Pfizer virus characterization: a similar virus in a human amniotic cell line.

Mason-Pfizer monkey virus (MP-MV) is a RNA virus with an RNA-instructed DNA polymerase first isolated from a rhesus monkey mammary adenocarcinoma in 1970. Until recently, there have been no other isolates. A continuous human amnion cell line, AO, was found to be producing a virus indistinguishable or closely related to the Mason-Pfizer virus as measured by morphological, immunological, and biochemical methods. By thin-section electron microscopy, the extracellular virus particle in AO line is 115 to 130 nm in diameter and has a preformed nucleoid (80 to 90 nm) before budding, properties which are also characteristic of MP-MV. Two proteins of the virus from the AO line were studied. By immunodiffusion, sera which react specifically with MP-MV give a line of identity with virus from the AO line. The AO viral RNA-instructed DNA polymerase purified by phosphocellulose chromatography was specifically inhibited by anti-MP-MV polymerase sera, and the AO cells contained both DNA and RNA sequences related to MP-MV (3)H-DNA. Viruses thus far indistinguishable from MP-MV have also recently been found by others in different human lines, raising again the question of the species of origin of MP-MV. Because the virus in the AO cells cannot be differentiated from MP-MV, we attempted to determine the origin of MP-MV virus by measuring DNA sequences related to MP-MV (3)H-DNA in uninfected human and rhesus monkey cells. The quantity of MP-MV-like DNA sequences in uninfected primate tissues was found to be much lower than the amount of DNA sequences of murine type-B or type-C viruses in uninfected murine tissues. Thus, it was not possible to determine whether the virus produced by AO cells or MP-MV was of human or monkey origin, or both.