Effect of Activated Immunocompetent Cells on the Content of Multipotent Stromal Cells in Splenic Transplants of CBA and CBA/N Mice Administered with Polyvinylpyrrolidone.

One day after intraperitoneal injection of polyvinylpyrrolidone (PVP) to recipient CBA and CBA/N mice, the count of multipotent stromal cells (MSC) in the 4-month-old splenic transplants was minimum in CBA/N→CBA/N group in comparison with the transplants of intact recipients (0.6 from the control level), but increased by 2.3, 3.2, and 3.7 times in CBA/N→CBA, CBA→CBA, and CBA→CBA/N groups, respectively. In the blood serum of recipient CBA/N mice with 4-month splenic transplants of CBA donors, the levels of some cytokines (IL-5, TNFα, and IL-2) was significantly increased 1 and 24 h after PVP injection in contrast to mice with bone marrow transplants, which attests to activation of the innate immunity mechanisms in this (splenic) transplantation variant. Probably, this phenomenon can be explained by the fact that the splenic transplants contain a sufficient number of CD+B-1a lymphocytes that can restore the response of recipient CBA/N mice to PVP. Thus, similar to bone marrow transplants [5], MSC count in splenic transplants increased only in groups, where the recipients were capable of responding to PVP. In other words, after injection of PVP to recipient mice, MSC counts in the spleen and bone marrow at this moment are determined by availability of activated immunocompetent cells. Overall, the novel data attest to close relationships between the stromal tissue of hematopoietic and lymphoid organs, on the one hand, and immune system, on the other.

Simultaneous Administration of NOD-2 (MDP) and TLP-4 (LPS) Ligands to Bone Marrow Donors 24 h before Transplantation Increases the Content of Multipotent Stromal Cells (MSCs) in Bone Marrow Grafts in CBA Mice Compared to the Total Result of Their Isolated Administration.

In 3-month bone marrow transplants of CBA mice from bone marrow donors receiving single injections of TLR-4 ligand (LPS) or NOD-2 ligand (muramyl dipeptide, MDP) 24 h before transplantation, an increase in the total number of MSCs (by 2.6 and 1.9 times, respectively), as well as a slight increase in the number of nuclear cells and the mass of bone capsules (by 1.3 and 1.2 times) were observed. After combined administration of MDР and LPS to donors, the total content of MSCs in the grafts was higher by 1.6 times in comparison with the total result of their isolated administration (and by 7.2 times in comparison with the control). At the same time, the concentration of osteogenic MSCs in the grafts of all groups was almost the same and corresponded to the control level. The number of nuclear cells and the mass of bone capsules of the grafts after combined administration of LPS and MDP were close (~80%) to the sum of the results of their isolated administration. These findings suggest that activation of the stromal tissue and the success of bone marrow transplantation depend on the intensity of innate immune responses. These data can be useful for the development of optimal methods of tissue transplantation.

Involvement of Bone Marrow Multipotent Stromal Cells in the Processes Presumably Provoking Vascular Calcification.

During serial transplantation of bone marrow derived from young and aged donor CBA mice to 5-month-old recipients, the counts of multipotent stromal cells (MSC) in transplants from young donors assessed at each passage surpassed those of aged donors by 3.2, 7.8, 3.0, and 2.2 times attesting to the age-related decrease of active pool of bone marrow MSC. The medullary curettage in mouse femur increased the total number of MSC and the number of osteogenic MSC both in the contralateral femur and in the bone marrow transplants attesting to spread of the effects of osteogenic factors after bone injury onto the bone tissue of the body even if this tissue if not topographically related to the skeleton. Combined and simultaneous administration of antigenic complex of S. typhimurium (or LPS) with BMP-2 markedly increased the count of osteogenic medullary MSC by 3.6 or 4.6 times in comparison with intact control or by 2.1 and 2.7 times in comparison with administration of BMP-2 alone, which probably resulted from enlargement of the pool of osteogenesis-inducible MSC due to inflammation. Addition of BMP-2 to the culture of splenic stromal cells where osteogenesis does not occur under normal conditions provoked appearance of MSC colonies with alkaline phosphatase activity attesting to involvement of inducible osteogenic MSC in vascular calcification. It can be hypothesized that the reaction to the age-related changes in the bone tissue and osteoporosis is similar to the reaction to bone marrow injury and includes initiation of systemic inflammation and elevation of blood BMP-2, both of which are prerequisite for vascular calcification.

Effect of Activated Immunocompetent Cells on the Number of Multipotent Stromal Cells in Bone Marrow Transplants of CBA and CBA/N Mice in a Short Time after Polyvinylpyrrolidone Administration to Animals.

One hour after polyvinylpyrrolidone administration, the content of multipotent stromal cells in the spleen of CBA and CBA/N mice increased almost equally (by 2.5 and 2.9 times, respectively), but in 24 h, the effectiveness of multipotent stromal cell cloning in the spleen of CBA/N mice decreased almost to the control level, whereas in CBA mice, the number of multipotent stromal cells continued to increase. Serum concentration of IL-5, TNFα, and IL-2 in both lines was elevated in 1 h after polyvinylpyrrolidone administration, which is likely to reflect activation of the innate immunity. One day after polyvinylpyrrolidone administration, the number of multipotent stromal cells in bone marrow transplants in the CBA/N→CBA/N and CBA→CBA/N groups remained practically unchanged, while in groups CBA→CBA and CBA/N→CBA it was equally increased (by 3.6 and 3.4 times, respectively). Thus, the number of multipotent stromal cells in bone marrow transplants after 1 day was increased only in groups where recipients (CBA mice) were capable of responding to polyvinylpyrrolidone administration, i.e. the number of stromal cells by this term, was apparently determined by the presence of activated immunocompetent cells. These findings also indicate that activation of the stromal tissue dur ing immune response can have a two-phasic pattern: the first phase (1 h after antigen adminis tration) can be determined by activation of innate immunity receptors (in multipotent stromal cells or other cells) observed in CBA and CBA/N mice, and the second phase occurs during further development of the immune response (that was observed in CBA mice, but not in CBA/N mice due to absence of CD+B-1a lymphocytes). The findings attest to close interactions between the stromal tissue and the immune system.

Pretreatment with Antiviral Preparation Kagocel Normalizes the Content of Bone Marrow Multipotent Stromal Cells and TNFα in Blood Serum of CBA Mice Disturbed by Administration of S. typhimurium Antigen Complex In Vivo and Maintains High Concentration of IL-10 and Th1 Cytokines.

Pretreatment with the active substance of antiviral preparation Kagocel, inductor of type I endogenous IFN, in a daily therapeutic dose (30 µg/mouse) 3 h prior to administration of S. typhimurium antigens to CBA mice reduced the number of bone marrow multipotent stromal cell (significantly increased by 3.2 times on the next day after antigen injection) to the initial level. Thus, activation of the stromal tissue induced by administration of the bacterial antigen was blocked. In addition, preliminary administration of Kagocel modulated the cytokine profile of the blood serum affected by S. typhimurium antigens: reduced 1.6-fold elevated concentration a proinflammatory cytokine TNFα to the control level (in 4 h after antigen injection) and maintained this level in 20 h after antigen administration. Kagocel also maintained the concentration of anti-inflammatory cytokine IL-10 at the level surpassing the normal by 1.6 times and high concentrations of Th1 cytokines (IL-2, IFNγ, and IL-12). These results suggest that Kagocel can reduce the immune response to bacterial antigens (similar to type I IFN [7]), which can contribute to its therapeutic and preventive effects in addition to its well documented antiviral activity and then this preparation can be used for the therapy of diseases accompanied by excessive or chronic inflammation.

[Cryoglobulinemia in a case of multiple myeloma associated with viral hepatitis B].

Cryoglobulinemia is a biochemical disorder accompanying many infectious, systemic, and lymphoproliferative conditions. Cryoglobulins are proteins capable of reversible precipitation or gel formation at temperatures below 37 degrees C. There are 3 types of cryoglobulins depending on the type of immunoglobulins in their composition; hence, three types of cryoglobulinemia occurring in different diseases. Clinical manifestations of cryoglobulinemia are sometimes inapparent, various combinations of the symptoms of the disease frequently occur. An example of atypical acute cryoglobulinemic vasculitis in a patient with multiple myeloma and viral hepatitis B is presented. Methods of diagnostics of this disease and its treatment are described.

[Iron deficiency anemia. Modern diagnostic and treatment strategy. Criteria for therapeutic efficacy].

Modern views of epidemiology, etiology and pathogenesis of iron deficiency anemia are considered. Mechanisms of iron metabolism regulation are described based on the most important literature data and the results of the authors' research. The authors present their own and literature clinical experience of using iron-containing drugs with reference to the existing recommendations on the treatment of iron deficiency anemia. Causes of low treatment efficiency are discussed and the ways to address this problem are proposed based on the published results of clinical research.

[Syphilis as a chronic systemic infection].

During the last STDs'epidemic in Russia (1994 - 2004) over 2.7 min people have been infected with syphilis. At present the structure of syphilis morbidity is characterized by 37% of latent forms, including 1.5% late latency. The increased level of late latency may result from: an ever-growing number of those with asymptomatic syphilis; disorderly and self-dependent usage of antibacterial preparations; spread of the virus chronic infections (herpes, hepatitis B, C) altering the macroorganism immune response; alcohol and drug abuse which decreases the efficacy of specific therapy. In general, chronization of the syphilitic infection may be caused by antigenically inert treponemal cell surface; paucity of outer membrane protein; residence of treponemas within an immunoprotective niche; uncompleted phagocytosis of treponemes with macrophages. Syphilis remains an "infectio magna" and demands thorough attention to all diagnostic and therapeutic procedures on every stage of the disease.

Use of transcranial magnetic stimulation with measurement of motor evoked potentials in the acute period of hemispheric ischemic stroke.

Functional derangements in the brain during the acute period of ischemic hemispheric stroke (IS) were assessed in terms of the severity of the motor neurological deficit in the acute period of IS and neurophysiological measures of motor evoked potentials (MEP) using transcranial magnetic stimulation (TCMS). A total of 52 patients (23 women, 29 men, mean age 58.5 +/- 8.7 years) were studied. Patients were divided into two subgroups: group 1 consisted of 29 patients with good functional outcomes from the acute period; group 2 consisted of 23 patients with poor functional outcomes. The use of TCMS for recording MEP demonstrated increases in the latency of the M response both after stimulation of the projection of the motor area of the cortex of the lesioned hemisphere and after stimulation of the spinal cord. There were increases in the central motor conduction time (CMCT) in the lesioned hemisphere of the brain and a negative correlation was seen between the severity of the neurological defect and CMCT on the one hand (r = -0.65 to -0.78; p < 0.001) and, on the other, the latency of the M response in TCMS of the motor zone of the cortex on the side of the hemispheric stroke (r = -0.65 to -0.79; p < 0.001). The increases in the latency of the M response and CMCT have prognostic significance for early assessment of the outcome of IS.

[Production and characterization of monoclonal antibodies to group-specific antigenic determinant of group A streptococcal polysaccharides]. / Poluchenie i kharakteristika monoklonal'nykh antitel k gruppospetsificheskoi antigennoi determinante polisakharida streptokokkov gruppy A.

Hybridomas producing monoclonal antibodies (McAb) to group A streptococcal polysaccharide (A-PS) were obtained. Of these, 3 clones were selected: 2 clones producing IgG3, precipitating McAb and 1 clone producing IgM nonprecipitating McAb. The results of the competitive inhibition in the enzyme immunoassay suggested that precipitating and nonprecipitating McAb reacted with nonidentical epitopes of A-PS, though determinants, specifically reacting with the given McAb, had a common site which included N-acetyl-D-glucosamine. On the surface of bacteria, in addition to protein M, the presence of the given determinants of A-PS was established in the direct immunofluorescence test. The newly developed method of direct immunofluorescence with the use of specially selected precipitating McAb was the basis for the development of rapid diagnosticum, permitting the identification of group A streptococci.

[Specificity of a panel of monoclonal antibodies to group A Streptococcus polysaccharide and search for cross reactions with mammalian tissues]. / Izuchenie spetsifichnosti paneli monoklonal'nykh antitel k polisakharidu streptokokkov gruppy A i poisk perekrestnykh reaktsii s tkaniiami mlekopitaiushchikh.

A panel of monoclonal antibodies (McAb) to different determinants of group A Streptococcus polysaccharide (A-PS) has been studied. As revealed in this study, A-PS contains at least 4 determinants, common with different epidermal antigens. McAb, cross-reacting (CR) with different mammalian tissue antigens, have not been found to be group-specific. Experiments on the inhibition of the immunoenzyme reaction of McAb with A-PS indicate that CR determinants include rhamnose and beta-N-acetyl-D-glucosamine of rhamnose alone. In addition, we have confirmed our earlier suggestion that group-specific high-affinity precipitating antibodies acted on different sites of A-PS. Contrary to earlier opinions, antibodies to group-specific determinants of A-PS have been found not to react with epidermal antigens.

[A trial of the use of low-molecular-weight heparin (fraksiparin) in myocardial infarct]. / Opyt primeneniia nizkomolekuliarnogo geparina (fraksiparina) pri infarkte miokarda.

Efficacy and safety were studied of nonfractionated versus low-molecular heparines (NFH, LMH) in acute myocardial infarction (AMI). A total of 99 AMI patients entered the trial including those after thrombolytic therapy. The highest preventive effect in respect to both venous and arterial thrombosis was achieved at subcutaneous administration of NFH in twice daily dose 12500 U and LMH in a single daily dose 25000 U. High doses of LFH promoted hemorrhagic complications which were absent in administration of LMH. NFH had a proaggregant action on the platelets. It is concluded that LMH is an effective and safe modality in the treatment of AMI patients acting through inhibition of activity of blood coagulation factor X, antiaggregant effect on platelets, improving microrheology of red cells and activation of fibrinolysis.

[The enhancement of the immunogenicity of streptococcal group-A M protein by conjugation with a synthetic polyelectrolyte]. / Povyshenie immunogennosti M-belka streptokokkov gruppy A pri kon''iugatsii s sinteticheskim poliélektrolitom.

Immunization with the polypeptide fragment of group A streptococcal protein M conjugated with the copolymer of acrylic acid and N-vinylpyrrolidone in complete Freund's adjuvant has been found to lead to a sharp increase in the level of antibodies to the type-specific determinants of protein M, detected in the enzyme immunoassay (EIA). The possibility of the application of such sera to preliminary typing of streptococci in EIA with the use of whole microbial cells as antigens has been shown. The data on high activity of the sera thus obtained in the bactericidal test with streptococci of the homologous type are presented. Recommendations on the use of sera obtained by the above method for highly precise typing of the virulent cultures of group A streptococci in the bactericidal test are given.

Immunogenicity of a polypeptide fragment of the streptococcal group A M protein conjugated with an acrylic acid N-vinylpyrrolidone copolymer.

Immunization of animals with a polypeptide fragment of the streptococcal group A M protein (pep M) conjugated with a synthetic copolymer caused a significant increase in the titres of bactericidal type-specific antibodies. In some cases bactericidal antibodies to streptococci of other types were also induced.