Mandelic acid condensation polymer: novel candidate microbicide for prevention of human immunodeficiency virus and herpes simplex virus entry.

Presently marketed vaginal barrier methods are cytotoxic and damaging to the vaginal epithelium and natural vaginal flora when used frequently. Novel noncytotoxic agents are needed to protect men and women from sexually transmitted diseases. One novel candidate is a mandelic acid condensation polymer, designated SAMMA. The spectrum and mechanism of antiviral activity were explored using clinical isolates and laboratory-adapted strains of human immunodeficiency virus (HIV) and herpes simplex virus (HSV). SAMMA is highly effective against all CCR5 and CXCR4 isolates of HIV in primary human macrophages and peripheral blood mononuclear cells. SAMMA also inhibits infection of cervical epithelial cells by HSV. Moreover, it exhibits little or no cytotoxicity and has an excellent selectivity index. SAMMA, although not a sulfonated or sulfated polymer, blocks the binding of HIV and HSV to cells by targeting the envelope glycoproteins gp120 and gB-2, respectively, and also inhibits HSV entry postattachment. SAMMA is an excellent, structurally novel candidate microbicide that warrants further preclinical evaluation.

A novel system for the production of fully deleted adenovirus vectors that does not require helper adenovirus.

Fully deleted adenovirus vectors (FD-AdVs) would appear to be promising tools for gene therapy. Since these vectors are deleted of all adenoviral genes, they require a helper adenovirus for their propagation. The contamination of the vector preparation by the helper limits the utility of currently existing FD-AdVs in gene therapy applications. We have developed an alternative system for the propagation of FD-AdVs, in which the adenoviral genes essential for replication and packaging of the vector are delivered into producer cells by a baculovirus-adenovirus hybrid. A hybrid baculovirus Bac-B4 was constructed to carry a Cre recombinase-excisable copy of the packaging-deficient adenovirus genome. Although the total size of the DNA insert in Bac-B4 was 38 kb, the genetic structure of this recombinant baculovirus was stable. Bac-B4 gave high yields in Sf9 insect cells, with titers of 5 x 10(8)p.f.u./ml before concentration. Transfection of 293-Cre cells with lacZ-expressing FD-AdV plasmid DNA followed by infection by Bac-B4 at a MOI of 2000 p.f.u./ml resulted in rescue of the helper-free vector. Subsequent passaging of the obtained FD-AdV using Bac-B4 as a helper resulted in approximately 100-fold increases of the vector titer at each passage. This resulting vector was completely free of helper virus and was able to transduce cultured 293 cells. However, scaling-up of FD-AdV production was prevented by the eventual emergence of replication-competent adenovirus (RCA). Experiments are underway to optimize this system for the large-scale production of helper virus-free FD-AdVs and to minimize the possibility of generation of replication-competent adenovirus (RCA) during vector production. This baculovirus-based system will be a very useful alternative to current methods for the production of FD-AdVs.

[Enhancement of epidermal regeneration by recombinant vaccinia virus growth factor]. / Usilenie épidermal'noi regeneratsii s pomoshch'iu rekombinantnogo faktora rosta virusa ospovaktsiny.

Examining the specific activity has showed that recombinant vaccinia virus growth factor binds to appropriate receptors on the A-431 cell surface and prompts the healing acceleration of degree III burns in rats. This recombinant factor did not demonstrate pyrogenicity or toxicogenicity in tests on rabbits, guinea-pits, noninbred albino mice.

[Recombinant vaccinia virus expressing Japanese encephalitis virus protein E]. / Rekombinantnyi virus ospovaktsiny, ékspressiruiushchii belok E virusa iaponskogo zéntsefalita.

Recombinant vaccinia virus expressing protein E of Japanese encephalitis virus has been constructed. Polyclonal antibodies to JE virus reacted with recombinant protein E in immunoblotting. Immunochemical analysis of the recombinant protein E with monoclonal antibodies showed that both group specific and receptor domains of the protein were intact.

[Expression of a synthetic gene for human angiogenin in a recombinant vaccinia virus]. / Ekspressiia sinteticheskogo gena angiogenina cheloveka v sostave virusa ospovaktsiny.

The gene for angiogenin was cloned into vaccinia virus genome. The recombinant virus expressing angiogenin was obtained. The level of protein synthesis directed by the recombinant virus was analyzed by immunoblotting using monoclonal antibodies against human angiogenin.

[Immunochemical analysis of Escherichia coli expression products of I3 and A2 genes of vaccinia virus strain L-IVL genome]. / Immunokhimicheskii analiz produktov ékspressii v Escherichia coli genov I3 i A2 genoma virusa ospovaktsiny shtamma L-IVL.

Genes I3 and A2 of the vaccinia virus strain L-IVP DNA were cloned into bacterial expressing vectors. The monospecific antisera to the expression products of these genes in E. coli were obtained. By means of immunochemical cross-analysis two polypeptides of equal electrophoretic mobility were found in the virion preparations in the band of the major envelope protein p35. The major of them is the product of gene I3, and the minor is encoded by gene A2.

[Mapping the genes of the vaccinia virus, coding membrane proteins p34 and p40, by mRNA hybridization selection]. / Kartirovanie genov virusa ospovaktsiny, kodiruiushchikh membrannye belki p34 i p40, metodom gibridizatsionnoi selektsii mRNK.

The HindIII--J HindIII-F fragments of the vaccinia virus DNA strain Lister have been analysed by the technique of mRNA hybridization selection with the subsequent translation in cell-free protein synthesizing system from the rabbit reticulocytes. The viral mRNA hybridizable with the HindIII--J fragment was shown to direct the synthesis of 30 kDa polypeptide in the cell-free system. This polypeptide was demonstrated to react specifically with antiserum to plasma membrane protein p34. The viral mRNA hybridizable with the HindIII-F fragment was shown to direct the synthesis of 37 kDa polypeptide in the cell-free system. This polypeptide reacts specifically with antiserum to major membrane protein p40.

[Isolation and characteristics of vaccinia virus proteins associated with cell membranes of infected cells]. / Vydelenie i kharakteristika belkov virusa ospovaktsiny, assotsirovannykh s plazmaticheskimi membranami infitsirovannykh kletok.

The proteins of vaccinia virus associated with plasma membrane infected cells BHK-21, p60, p45, p42, p40,p35,p34,p28,p23 were isolated from plasma membranes using affinity chromatography, gel-electrophoresis and passive elution. An immunochemical characterization was carried out using specific antiserum to these proteins. Investigation of temporal regulation of proteins synthesis in infected cells showed that proteins p60, p45, p42, p40, p28 were late, and p35, p34, p23--early-late proteins. Immunochemical analysis of vaccinia virus mRNA cell-free translational products was carried out using specific antiserum. The polypeptide-precursors of viral proteins were identified.

[Molecular-biological study of vaccinia virus genome. III. Identification of the late gene product protein 36K from Hind-III-P-fragment of vaccinia virus strain L-IVP]. / Molekuliarno-biologicheskoe izuchenie genoma virusa ospovaktsiny. III. Identifikatsiia produkta pozdnego gena belka 36K iz HindIII-P-fragmenta genoma virusa ospovaktsiny shtamma L-IVP.

Vaccinia virus gene encoding 36K protein was cloned in pUR290 bacterial expressing vector and resulted in the synthesis of a chimeric protein in E. coli. The chimeric protein consists of beta-galactosidase and virus protein in C-termini. It has virus antigen specificity. By monospecific antibody 36K protein of vaccinia virus was determined to be non-virion. It is localized in the cytoplasm of infected cells.