Functional Parameters of Physiological Systems of Laboratory Primates after Administration of Doxorubicin and Transplantation of Mesenchymal Stem Cells.

We studied physiological parameters of rhesus monkeys after administration of anthracycline antibiotic doxorubicin. Intravenous administration of the drug caused intoxication manifested in in an abrupt body weight loss, baldness, vomiting, and exicosis. Intoxication in monkeys determined by the damaging effects of doxorubicin on organs and tissues is also characterized by significant changes in the blood: leukopenia, thrombocytopenia, neutropenia, monocytopenia, lymphocytosis, and a sharp drop of CD20+ B cell content. The total protein and albumin content in the blood significantly decreased. A sharp increase in C-reactive protein was also accompanied by an increase in activity of proinflammatory cytokine IL-6. Transplantation of mesenchymal stem cells in some cases can significantly alleviate doxorubicin-induced damage to organs and maintain normal clinical status of monkeys after two injections of the drug. Late transplantation of stem cells does not have a protective effect and does not protect the animals from the damaging effects of doxorubicin. We found that the protective effect of mesenchymal stem cells depends on the dose of the drug, total number of cells, and the time of their transplantation. It should be noted that human and monkey mesenchymal stem cells produce similar regenerative effects, at least in the doxorubicin toxicity model.

Use of Mesenchymal Stem Cells for Possible Repair of Doxorubicin-Damaged Organs and Tissues in Experimental Monkeys.

Three injections of doxorubicin to rhesus macaques cause severe intoxication, characterized by anemia, cachexia, and degeneration of the viscera. The life span of monkeys injected with the drug and receiving after 24 h mesenchymal stem cell transplantation varied from 96 to 120 days in comparison with 50-74 days in animals receiving stem cells before doxorubicin. Controls received doxorubicin and saline; the lifespan of one monkey was 24 days, of the other - 1 year and 8 months. The increase in activity of proinflammatory cytokine IL-6 was paralleled by an increase in the level of C-reactive protein.

Transplantation of Simian Mesenchymal Stem Cells to Baboons with Experimentally Induced Myocardial Infarction.

Culture of mesenchymal stem cells isolated from the bone marrow of primates by their characteristics met the requirements of stem cells. It was shown that transplantation of allogeneic mesenchymal stem cells (2 million cells per 1 kg body weight) immediately after ligation of the left anterior descending coronary artery between the middle and upper thirds led to neovascularization and capillarization of the ischemic myocardium.

[Preclinical study of AV0012 early stage inhibitor of hepatitis C virus infection: I. In vitro ADME and pharmacokinetics].

In vitro immunohistochemical investigations on the human hepatoma cell line (Huh7) infected with hepatitis C virus (HCV) strain JFH-1 showed that AV0012 compound blocks the early stages of viral infection. AV0012 also blocked viral infection spread in tissue culture through the secreted virus and through tight cell-to-cell contact. AV0012 is a specific inhibitor of HCV but not of related pestivirus, flaviviruses and other RNA-containing viruses such as bovine diarrhea (BVDV), Venezuelan equine encephalitis (strain TC-83), dengue type 2 (New Guinea), yellow fever (strain 17D), west Nile fever, parainfluenza (type 3) virus, RSV (strain A2), and Rhinovirus (type 2 strain HGP). It is established that human serum does not significantly affect the antiviral activity of AV0012 in vitro. The drug combination studies with AV0012 and interferon alpha 2a in vitro showed that the two inhibitors act additively, which makes possible the use of this combination in clinical tests. AV0012 is highly soluble and stable in aqueous solutions and murine blood plasma, has limited metabolic stability, low binding to human plasma proteins, high permeability through biological membranes, and only interacts with isoenzymes 2D6 and 3A4 of human cytochrome P450. In animal pharmacokinetic studies, AV0012 was rapidly absorbed into the blood stream upon oral administration, showed sufficiently long half-elimination times, and had high oral bioavailability that reached 92% in monkeys. Further preclinical development of AV0012 is in progress.

[Efficiency of cycloferon against slow viral infection caused by SV-40 in monkeys and man].

The latent infection caused by Simian virus (SV-40) defeats monkeys and others animals. Currently there are not the prophylactic and therapeutic measures against this disease. We showed the infringement of immunity of M. mulatta infected by SV-40. The use of the preparation Cyclopheron for the treatment of this infection led to the normalization of the functions of immunity of monkeys and to disappearance of the virus from the organism. We suggested the new method for prophylaxis and treatment of latent SV-40 infection by Cyclopheron, may be used for the correction of the immunity.

Possible aspects of using cultured stem cell of laboratory primates in experimental medicine.

We describe the methods of isolation and culturing of mesenchymal stem cells from 3 monkey species Macaca mulatta, Papio hamadryas, and Macaca fascicularis. Flow cytofluorometry showed that the cells do not express CD34, CD45, and HLA-DR, but most of them (78-98%) express CD90 marker. The cardioprotective effects of cultured mesenchymal stem cells in cardiomyopathy induced by administration of antitumor anthracycline drugs (doxorubicin).

[Comparative immunophenotypic characterization of human and monkey permanent lymphoid culture cells].

The aim of the study was to define the comparative immunophenotypic characteristics ofwidely spread lymphoid cell cultures, derived from Burkett's lymphoma named as Raji and P3HR-1 in comparison with analogous monkey cultures. It has been shown that P3HR-1 culture consists of similar type cells - activated B-lymphocytes CD23 with k phenotype, which demonstrates its monoclonality. Raji culture includes cells with markers of immature B-lymphocytes CD10 and CD24, as well as elements expressing CD10 antigens. T-cell markers were found in none of the cultures. In contrast to human cells, monkey lymphoid culture expressed both B- and T-cell markers. Moreover, in one of them, obtained from a green monkey, T-cells of suppressor type (CD8) prevailed. The immunophenotypic characteristics of primate lymphoid cell cultures, revealed by the study, are of great importance for their proper application to medical and biological studies.

[The virogenic status of cultured continuous simian lymphoid cells and their immunophenotypical characteristics].

Immunophenotyping of cultured continuous simian lymphoid cultures using a panel containing monoclonal antibodies to human B- and T-cell antigens has been carried out, by employing enzyme immunoassay and flow cytofluorometry. The test cultures showed a wide variety of cells containing B- and T-cell antigens with their varying expression. The cultures were also found to comprise lymphoblasts simultaneously containing B- and T-cell markers. Simian lymphotropic viruses, such as EBV-like and STLV-1 retrovirus, detected apart or simultaneously, have been verified by polymerase chain reaction. Whether there is a possible relationship between the type of cells in the culture and the type of their replicating virus(es) is discussed in the paper.

[Antibodies to herpesvirus type 8 in Kaposi's sarcoma patients and controls in Russia]. / Antitela k gerpesvirusu tipa 8 u bol'nykh sarkomoi Kaposhi i lits kontrol'nykh grupp v Rossii.

The purpose of the present study was to investigate the HH-8 seroprevalence among patients with Kaposi's sarcoma (KS), melanoma and gastric carcinoma (GC) as well as among renal recipients and blood donors. The obtained data revealed a high percentage of seropositive KS patients, which ranged from 83.6% in the classical disease type to 68.8% and to 71.4% in the immunosuppressive and AIDS-associated disease types, respectively. On the whole, the positive humoral response to HHV-8 reliably correlated with the positive findings if the viral genetic information in tumor tissue samplings. An unexpectedly high percentage of seropositive persons was found among the GC patients (41.8%) and among the renal recipients (26%), which is apparently predetermined by the immunosuppressive condition of such patients. Seroprevalence was found only in 4% of blood donors. Thus, the obtained data make it possible to conclude that KS cases, as diagnosed in Russia, are tensely associated with HHV-8 in spite of a low virus spread among the healthy population. Patients with pathology concomitant with a pronounced immunosuppression are characterized by a high prevalence of HHV-8 and belong to the category of persons with a KS risk.

[Cytogenic characteristics and virogenic status of lymphoid cells from malignant baboon lymphomas]. / Tsitogeneticheskaia kharakteristika i virogennyi status limfoidnykh kletok iz zlokachestvennykh limfom pavianov.

Four lymphoid cell cultures obtained from Papio hamadryads with malignant lymphomas were studied. The cultured cells produced two lymphotropic EBV-like herpes virus of Papio (HVP) and T-lymphotropic monkey retrovirus STLV-1 (HTLV-1 family) or HVP alone. More than 100 subcultured cell lines were shown to remain mixed B- and T-cellular, the levels of T cells of different subpopulations were 15-50%. Cytogenetic investigations showed that one of the 4 cultures was tumorogenic and the others were derived from the normal cells of monkeys with malignant lymphomas. It is suggested that the presence of a cultural virus (viruses) may affect the karyotypes of cells.

[The cytogenetic characteristics of human B-lymphoblastoid cell lines]. / Tsitogeneticheskaia kharakteristika B-limfoblastoidnykh kletochnykh linii cheloveka.

Permanent suspension B-cell lines HSL-1, HSL-2, HSL-3, HSL-4, HSL-5, HSL-8, HSL-9 have been obtained spontaneously from lymphocytes and cells of hematopoietic tissues of patients with various forms of leukemia and tumours. All cell lines contained Epstein-Barr virus (EBV). At early stages of cultivation (6-39th passages) normal karyotypes were established for all the tested lines which testified their origin from normal B-lymphocytes. In the process of cultivation of HSL-1 cell line, which (up to the 63rd passage) contained primarily the normal karyotypes, a substitution for tetraploid ones (92, XX, YY) by the 132nd passage was established. In line HSL-4, a pathological clone of cells with structural chromosome rearrangements (46, XY, -2, -12, der (12) t (2; 12) (p12, p13); del2 (p12), was revealed at the 26th passage. During a prolonged cultivation (up to the 130th passage) this abnormal clone forced out completely the clone with normal karyotype. Substitution of normal clone cells by cells with pathological karyotypes may point to their selective advantage.

[The cytogenetic characteristics of B-lymphoid lines from Papio hamadryas baboons and Macaca arctoides brown macaques]. / Tsitogeneticheskaia kharakteristika B-limfoidnykh linii pavianov gamadrilov Papio hamadryas i makakov burykh Macaca arctoides.

A cytogenetic analysis of 9 permanent suspension cell lines was performed. Cell lines were obtained from Papio hamadryas and Macaca arctoides cells by different methods. Five cell lines originated from non-malignant parental cells. Lines SPG-5 and SPG-6 were analysed at early stages of cultivation, with normal karyotypes being identified. In cell lines PL-11, MAL-1 and MAL-4 pathological clones with structural and numerical chromosomal rearrangements appeared only following a long-term cultivation. This phenomenon may be due presumably to the influence of cultivation on karyotype variability. In cell line LUG-4 chromosome marker 17p+ was found at early stages of cultivation. In sublines E9-1, E1-1 and E5-1, established from line LUG-4 by cloning, 17p+ chromosomal marker was found persisting. The presence of the marker 17p+ in all the cells at early passages testifies that the cell material from Papio with lymphoma, which was used for this line, might have the same chromosomal damage in its initial karyotype.

[The isolation and characteristics of continuous virus-producing lymphoid suspension cultures of African green monkey cells]. / Poluchenie i kharakteristika perevivaemykh virusprodutsiruiushchikh limfoidnykh suspenzionnykh kul'tur kletok afrikanskikh zelenykh martyshek.

Five continuous lymphoid suspension cell cultures were established by cultivation of lymphocytes from African green monkeys. The immunological typing revealed the mixed pattern of cultures consisting of B and T cell types. Two types of viruses: herpesvirus and retrovirus C-type were demonstrated by electron microscopy. Indirect immunofluorescence and molecular hybridization studies showed one of the viruses to be an EBV-related herpesvirus of green monkeys designated HVGM and the other to belong to H (S) TLV-1 group.

[Comparative analysis of genomes of oncogenic B-lymphotropic herpesviruses reproducing in cell lines obtained from different monkey species]. / Sravnitel'nyi analiz genomov onkogennykh B-limfotropnykh virusov gerpesa, reprodutsiruiushchikhsia v kletochnykh liniiakh, poluchennykh ot obez'ian razlichnykh vidov.

The aim of this work was to study genomes of B-lymphotropic oncogenic herpesviruses of Macaca arctoides (HVMA), Papio hamadryas (HVP) and Cercopithecus aethiops (HVGM). Blot-hybridization detected inter- and intraspecific genome heterogeneity of investigated viruses in restriction sites EcoR1, Pst1 and Hind111. Virus-specific sequences in two-cell lines obtained from HVMA-induced rabbit tumours can prove HVMA a role in malignant tumour induction in rabbits.

[Analysis of B-lymphotropic oncogenic herpesvirus of Macaca arctoides]. / Analiz B-limfotropnogo onkogennogo virusa gerpesa makakov burykh.

Some physico-chemical and molecular-biological parameters of B-lymphotropic herpes virus of Macaca arctoides (HVMA) have been analyzed. The buoyant density of virions and nucleocapsids in a gradient of the percoll density is 1.10 and 1.14 g/ml, respectively, while that of DNA in CsCl gradient is 1.717 g/ml, which is typical for herpes viruses of primates. At the same time the homology of DNA HVMA with DNA EBV and DNA HVP is 30-45%. Differences in restrictions sites between DNA HVMA, DNA HVP and DNA EBV are determined. The intraspecific heterogeneity of HVMA strains has been detected according to restriction sites of DNA. The HVMA genome size is about 170 kb. The DNA HVMA is colinear to the EBV genome, which is also typical of B-lymphotropic herpes viruses of primates.

[A method of determining Campylobacter pylori in the gastric mucosa in peptic ulcer and chronic gastritis]. / Metod opredeleniia Campylobacter pylori v slizistoi obolochke zheludka pri iazvennoi bolezni i khronicheskom gastrite.

An original method is described for the detection of Campylobacter pylori in gastric mucosa biopsy specimens in chronic gastritis and gastroduodenal ulcers. The method is based on the detection (by a special test) of extremely high urease activity of C. pylori. The method has been employed in examinations of 211 patients. The results and prospects for the use of this technique for the detection of C. pylori in patients with gastroduodenal diseases are discussed.