Experimental study of the immunostimulating properties of tomitsid

Tomicide, a preparation with antibacterial properties, accumulates in the culture medium during the growth of one of streptococcal strains. The injection of the preparation into mice simultaneously with the antigen (sheep red blood cells) produces an adjuvant effect. Tomicide has been found capable of inducing the production of interferon detected in the serum of the animals. During experimental stress created by intensive exercise tomicide prevented a decrease in the normal (anamnestic) level of antibodies, produced an immunostimulating effect and, at the same time, enhanced the total physical endurance of the animals, manifested by an increased duration of swimming.     

Immunostimulating and protective effects of terrilytin preparations in staphylococcal infection

In mice infected with staphylococci there was observed less pronounced development of the immune response to sheep red blood cells (SRBC) than in intact animals subjected only to immunization. Administration of free terrilytin to the infected mice increased the immune response development induced by SRBC while the use of immobilized terrilytin normalized it. The supernatant liquid of the spleen cells (SLSC) from the mice treated with the free of immobilized terrilytins stimulated development of the immune response to SRBC in the infected animals and inhibited the function of the suppressor cells in the spleen. The immunomodulating and protective effects of the SLSC fractions isolated with column chromatography on Sephadex G-150 were investigated. Substances of the low molecular fraction of the SLSC proteins (molecular weight of 10-15 kD) from the mice treated with the terrilytins showed immunostimulating and protective properties. The factors inducing both the activity types included peptides and the ribonucleotide component playing a significant role in realization of the immunostimulating and protective effects of the free and immobilized proteases.     

Tolerance-like condition developing in mice under the effect of antigen of erythrocyte lysate]

Lysate of sheep red blood cells obtained by the treatment of these cells with distilled water and purified by ultracentrifugation in cold possessed a weak immunogenicity. Its administration to mice caused the state of hyporeactivity to sheep red blood cells (a reduction of the immune response level to 10-25% of control. The capacity of the mise spleen cells to respond by immune reaction to the red blood cells following adoptive transfer was not disturbed. At the early periods after the lysate administrations the mouse spleen cells possessed a weak supressive activity in case of their transfer to the intact animals. The blood serum of mice treated with the lysate possessed a blocking activity which disappeared after the serum absorption with sheep red blood cells. A conclusion was drawn that hyporeactivity originating in mice after the lysate administration was caused by the presence in the serum of antibodies inhibiting the immune response.     

Changes in the immunotropic activity of neutrophilokins in the course of experimental staphylococcal infection

The influence of the products secreted by activated neutrophils (neutrophilokins) of mice, both intact and infected with staphylococci, on the activity of mouse spleen cells in the graft-versus-host reaction, immune response to sheep red blood cells and the antigen-presenting function of peritoneal macrophages was studied. Neutrophilokins of intact mice stimulated the activity of immunocompetent cells. Neutrophilokins obtained from infected mice on day 3 after infection produced an immunosuppressing effect. On day 7 after infection the immunostimulating activity of neutrophils was restored and showed practically no difference from the normal level.     

Antibody-producing ability of splenic cells of the progeny of female rats with chronic lesions of the liver

Spleen antibody-forming function in the offspring of female rats with experimental chronic autoimmune liver affection has been studied for its specificities. The data obtained testify to that these animals are characterized by the increased immunoreactivity at all the stages of investigation, that is confirmed by an increase in the number of antibody-forming cells in their spleen. Besides, the splenocytes of immunized animals have an immunostimulating effect on the productive phase of antibodygenesis and has no such effect on the inductive phase.     

Study of immunoregulating properties of an interferon inducer in animals with various reactions to the antigen

Influence of dsRNA isolated from killer yeast of S. cerevisiae on humoral and cellular immune responses in mice CBA/CaY and C57Bl/6Y with opposing reaction to the antigen was studied. It was shown that after administration of the yeast dsRNA preparation to the animals simultaneously with the antigen there was an increase in the number of the antibody forming cells in the spleen and the titer of hemolytic antibodies in blood serum of the animals with high and low reactions to the antigen. After sensitization with different doses of sheep red blood cells (10(7) and 10(8)) the preparation had immunomodulating action on development of DTH in mice CBA/CaY. The effect of the dsRNA preparation on the immunity system depended on the preparation dose, antigen loading and animal genotype and was the most marked in mice CBA/CaY with interferon levels in blood serum 2-3 times higher than those in mice C57Bl/6Y.     

Estimation of cell surface associated protease activity and its application to lymphocytes

A new method has been developed to estimate proteolytic activity available at the cell surface. Radioiodinated protein substrates are covalently linked to modified polystyrene-divinylbenzene beads with various diameters. These beads are presented to viable cells. Secreted enzyme activity is estimated when no contact occurs between beads and cells. Surface associated proteolytic activity is estimated by the increased rate of iodinated peptide release due to a contact between beads and cells. This method was applied to various lymphocyte preparations. In the absence of serum, mouse spleen lymphocytes produce three- to fourfold higher proteolytic activity than lymph node cells. This activity is completely inhibited by serum diluted 1:10. Since the proteolysis is so marked in the case of spleen cells, one must conclude that lymphocytes removed from the serum and treated in buffered mediums at 37° C have enzymatically altered surface properties. Cell surface associated enzyme activity was measured using rat lymph node lymphocytes with less than 0.1% contamination by granulocytes. This predominantly thymus derived, T cell population had 30% increase in proteolysis due to contact between cells and solid-phase localized substrate of casein. The released enzymatic activity was inhibited by diisopropylfluorophosphate, but its effect on the surface associated enzyme activity remains questionable since it perturbs several membrane functions.     

Experimental screening of two BCG preparations produced according to different principles

Summary Two BCG preparations produced according to different principles for use as an adjunct to surgery in a comparative clinical trial in melanoma patients have been examined for immunostimulating, toxic, and antitumor activity in various murine and guinea pig models and for immunostimulation in vitro of human lymphocytes. One BCG preparation was prepared in the classic way, i.e., grown as a surface culture and subsequently homogenized in a ball mill, while the other preparation was grown as a dispersed culture.In general, a comparable effect on spleen weight of the proliferation of lymph node cells and activation of spleen macrophages was observed when a similar amount of culturable particles of either BCG preparation were administered. In parallel with these effects, an increase of the serum glutamic pyruvic transaminase was detected. Activation of natural killer cells and the induction of an immune reaction in lymph nodes appeared to be a function of the total amount of injected viable and dead micro-organisms.The synergistic effect of the BCG preparation grown as a dispersed culture on the stimulation of human lymphocytes by phytohaemagglutinin, lacking in the BCG preparation grown as surface culture and subsequently homogenized in a ball mill, was due to the stabilizers present in the former material.The effect of BCG treatment on the growth of an originally chemically induced murine fibrosarcoma depended on the treatment schedule. Growth enhancement, growth retardation, or no effect was observed, depending on the regimen used. Although differences between the two BCG preparations could be demonstrated in the various models the clinical relevance of these reported results can only be evaluated in conjunction with the outcome of the comparative clinical trial.Now Fellow of the Koningin Wilhelmina Fonds of the National Cancer League of The NederlandsNow Secretary of the Health Council, The Hague, The Netherlands     

Comparative study of the proteolytic products of fibronectins in the serum of adult and embryonic animals formed under the effects of proteinases of the spleen

The effects of two spleen proteinases, cathepsin D and thiol proteinase I active in neutral media--on the structural properties of fibronectins from blood plasma on adult animals and their embryos were investigated. Proteinase I and cathepsin D caused rapid fragmentation of all fibronectins under study. Fibronectin from calf embryonic serum was more sensitive to proteinase I than that from adult animal serum. The molecular weight and the correlation between the proteolytic products formed under the influence of each enzyme, on the embryonic and "adult" fibronectins, are very similar but not identical. Similar results were obtained in experiments with proteolytic products of chicken serum and embryo fibronectins. Fragmentation of embryonic fibronectin occurs more rapidly than that of chicken fibronectin; the fibronectin proteolytic products differ both qualitatively and quantitatively. However, the determination of structural differences between these fibronectins is considerably hampered by the presence of protein contaminations in chicken fibronectin preparations.     

Tissue producing the serum factor stimulating the release of cathepsin D from lysosomes in vitro

Pancreatectomy as well as thyroparathyroidectomy resulted in the quick disappearance of a serum factor (stimulating cathepsin D release from lysosomes in vitro) from the rat or mouse blood. Extirpation of other organs such as duodenum, stomach, spleen, kidney, submaxillary gland, testis, adrenal gland or hypophysis, showed no effect on the serum factor level. Glucagon (but not insulin or thyroxine) given to the pancreatectomized animals restored the serum factor level in a dose-dependent manner. The serum factor-like activity was detected only in the parathyroids (but not thyroid), and the release of activity from parathyroid-slices was stimulated by glucagon, suggesting that the parathyroid may produce and/or secrete the serum factor under the influence of glucagon.     

Allergenic properties of the substance produced by the Streptococcus strain sp. Thom-1606

The newly isolated biologically active agent "STP" (the substance produced by Streptococcus strain sp. TOM-1606) has been found to possess no allergenic properties. This new preparation has shown hyposensitizing activity on the level with the allergen. The morphological data obtained in the study of the organs of experimental animals have revealed the immunostimulating action of preparation "STP" on the body, that is confirmed by the characteristic transformation of the internal organs of experimental animals.     

Comparative study of the properties of a proteolytic enzyme isolated from the myelin of animals and from the biological fluids of disseminated sclerosis patients

A proteolytic enzyme with the activity of 8-26 U/mg protein was isolated from purified animal myelin preparation obtained by an original technique. The optimal pH of the enzyme was found to be 9.6-9.8. Its substrate specificity was studied. An enzyme with similar characteristics and identical electrophoretic mobility was isolated from the blood serum of patients with disseminated sclerosis and then purified. The major part of the enzyme activity in the blood and myelin was bound and was manifested only after special treatment. It is suggested that a similar proteolytic enzyme is present in human myelin, whose activation in demyelinating diseases may result in myelin destruction.     

The effect of a ribosomal Shigella sonnei vaccine on cellular immune reactions]

The influence of S. sonnei ribosomal vaccine on hematopoiesis, T- and B-cell-mediated immune reactions has been studied in the course of the development of experimental vaccinal process. The vaccine stimulated hematopoiesis, that was characterized by a dose-dependent increase in colony-forming units in the spleen (CFUs), a rise in CFUs in the blood and bone marrow and an increase in the pool of proliferating stem cells in bone marrow, shortly after injection. A pronounced immunostimulating effect of the vaccine on the formation of antibody-producing cells (APC) to heterologous antigen (sheep red blood cells) in the spleen has been established, and the vaccine has also been found to stimulate, though to a lesser extent, APC synthetizing specific antibodies to S. sonnei LPS. The injection of S. sonnei ribosomal vaccine influences the functional activity of effector T-cells; in its turn this phenomenon produces phasic changes in the migration activity of spleen cells in the presence of specific LPS and surface polysaccharide antigen of S. sonnei in phase I.     

STP (the substance produced by a strain of Streptococcus sp. Thom-1606) as a stimulant of the nonspecific resistance of the macroorganism

Preparation STP, a new immunostimulating agent, is a substance produced by Streptococcus strain sp. Thom-1606 and capable of enhancing the nonspecific activity of the body as shown in animal experiments. The optimal dose-time parameters of the administration of the immunostimulator have been established by the method of the mathematical planning of experiments. As a result, the survival of all animals used in the experiment has been achieved. Mouse peritoneal macrophages have been found to form the population of target cells whose phagocytic activity is enhanced under the effect of the immunostimulating agent STP.     

Characteristics of the cellular reactions of the T- and B-dependent areas of the regional lymph nodes in dogs to the administration of immunodepressants

Mesenteric, bifurcational, axillary and popliteal lymph nodes have been studied in 22 healthy mature male dogs. Amount of blast cells, small lymphocytes, plasma cells and macrophages has been taken into account in the paracortical zone, in the germinative centers and in the medullary cords. For two weeks to one group of the animals every day imuran in turn with aurantin (10 mg/kg and 25 mg/kg) are injected, or antilymphocytic serum (ALS) intraperitoneally every other day (0.1 ml/kg). The combined injection of imuran and aurantin produces a more pronounced toxic effect to the hemopoietic organs than ALS. ALS is more specific for T-dependent zones of the lymph nodes. In the dose and interval mentioned ALS is an immunostimulating preparation for the immunocompetent cells of the germinative centers of the lymph nodes. The reaction of the lymph nodes depends on their regional belonging.     

Relation Between Testosterone Levels in Serum and Proteolytic Activity in the Neck Muscles of the Norwegian Reindeer (Rangifer tarandus tarandus)

The characteristic hypertrophying of the male reindeer neck muscles during the rutting period, which coincides with the rise in serum testosterone levels, is found to be accompanied by a significant decrease in lysosomal proteolytic activity of the neck muscle cells. Conversely, during the period of neck muscle involution following the rapid decline in serum testosterone after the rut, a marked increase in neck muscle proteolytic activity is found. These changes in lysosomal activity do not parallel the changes in feed consumption and nutritional stage of the animals during the rutting period. The lysosomal proteolytic activity of the male reindeer neck muscles thus may be, at least partly, under the control of sex steroid hormones.     

Effect of splenic humoral factors on the cholinesterase activity of liver tissue and blood serum in experimental toxic hepatitis in rats

Activity of cholinesterase in the liver of rats with experimental toxic hepatitis decreases, while in the serum it remains unchanged. Introduction of splenin, a spleen preparation, normalizes indices of the cholinesterase activity, extract of muscles possessing no such property. Boiling-heated spleen preparation looses its activity. Activity of the studied enzyme remains unchanged both after splenectomy and splenin introduction to intact animals.     

The immune system response to antigen in cold- and warm-adapted rats

In rat experiments, it was shown that long-term adaptation to cold modified the response pattern of the immune system to antigen stimulus by variously changing the different components of the response. There was a considerable decrease in the number of antigen-binding cells in spleen and peritoneal exudate and also a marked decrease in general antibodies in blood. Conversely, the number of antibody-forming cells in spleen increased twofold. The shift in the thermal conditions after immunization caused changes in the immune response. After the placement of cold-adapted animals to the warm environments the number of antigen binds and antibody-forming cells in spleen, as well as hemagglutinin titers in serum normalized, became indistinguishable from those in the warm-adapted control group. The placement from warm to cold after immunization of warm-adapted rats also produced considerable shifts in parameters of the immune response. However, these shifts were not entirely consistent with those observed for long-term adapted to cold. The obtained data evidence that the thermal factor of the environment is of major importance to the function of immunocompetent cells.     

Differentiation of stem cells from different parts of the hematopoietic system of adult thymectomized CBA mice stimulated with thymarin or cortexine

Essential differences were detected in differentiation of GFUs from bone marrow and peripheral blood. It was shown that as a result of thymectomy of adult animals the ability of bone marrow CFUs to form granulocytic colonies decreased and that of splenic CFUs to form erythroid colonies increased. The immunostimulating low-molecular-weight polypeptides, thymarin and cortexine , normalized the differentiation of CFUs from bone marrow and spleen but interfered with the formation of erythroid colonies from CFUs of peripheral blood of thymectomized mice.     

Mechanism of action of serum globulin preparations]

Experiments on mice demonstrated that serum homologous globulin preparations containing normal autoantibodies caused induction of autoantibodies in subcutaneous injection to intact animals; this was recorded on the basis of increase of the blood autoantibody level and of the plaque-forming cells content in the spleen tissue. This activation was seen as soon as the first hours--24 hours after the administration of the preparation and failed to disappear in 7 days, persisting in repeated administration of the preparation.