Lymphocyte subpopulations in sensitization and specific immunotherapy in an experiment. Lymphocyte subpopulations in sensitization to staphylococci, Artemisia pollen and their combinations

The distribution of T- and B-lymphocytes in the body of guinea pigs was studied in different groups of the animals. As shown in this study, in delayed hypersensitivity to staphylococci the number of PE- and E-rosette-forming cells increased in the blood, the spleen, and the lymph nodes and decreased in the thymus; the number of EA- and EAC-rosette-forming cells decreased in the bone marrow and the spleen, the number of T gamma-suppressors decreased in the bone marrow and the distant lymph node. Immediate hypersensitivity to tarragon pollen induced the general increase of the content of T- and B-lymphocytes; the number of T gamma-cells decreased in the thymus, the bone marrow, and the lymph nodes and increased in the spleen. The characteristic features of combined microbial-pollen sensitization were the high content of B-cells in all lymphoid organs (except the thymus), a low level of T-lymphocytes in the blood and the peripheral lymphoid organs, the decreased number of T gamma-cells in most of the immunogenetic organs.     

Lymphocyte subpopulations in sensitization and specific immunotherapy in an experiment. Lymphocyte subpopulations in the specific immunotherapy of microbial allergy and subsequent heterologous pollen sensitization

The influence exerted by the specific immunotherapy (SIT) of delayed hypersensitivity (DH) to staphylococci and subsequent sensitization with tarragon pollen on the level of immunocompetent cells in the blood and lymphoid organs of guinea pigs was studied. On the whole, SIT normalized the characteristics of T- and B-lymphocytes, altered as the result of experimentally induced DH: the content of T-cells in the peripheral blood and the lymph nodes increased, while the number of B-cells in the blood and T gamma-suppressors increased. The subsequent heterologous sensitization with pollen abolished the effect of SIT, inducing the general decrease of the level of T gamma-lymphocytes and enhancing the number of T-lymphocytes in the lymph nodes.     

Development of resistance in Fisher lymphadenosis cells to dipin and bruneomycin when these preparations are used separately and in combination]

It was shown that dipin and bruneomycin resistant tumor cells appeared in mice with transplanted lymphadenosis after 10 passages on single use of the drugs. When the drugs were used in combination, no lymphadenosis cells resistant either to bruneomycin, or to dipin and their combination were found. The combined use of the drugs prevented development of resistance to them in the lymphadenosis cells at least during 10 passages on mice. The data on the possible prevention of the resistance development in the mouse lymphadenosis cells by means of combined use of low doses of dipin and bruneomycin provided an assumption that it is expedient to test the combination in clinics.     

T- and B-lymphocyte content in the peripheral blood of healthy subjects]

It was revealed by the method of rosette-formation that the content T- and B-lymphocytes differed in human blood depending on the time of the day and the season. T-cells were at their maximum in the morning and B-cells -- in the evening. The percentage content of B-cells was greater in autumn than in winter.     

Development of bruneomycin and rubomycin resistance in mouse lymphadenosis tumor cells and Staphyloccus when these preparations are used separately or together

Two sublines of mouse lymphadenosis (L-5178) resistant to bruneomycin and rubomycin used alone, as well as a subline with induced resistance to the combination of these drugs were employed in the study. The studies showd that in separate use of rubomycin and bruneomycin the tumor cell resistance to the respective drug was evident at the 10th passage. After 30 passages neither bruneomycin nor rubomycin produced reliable inhibition of the tumor growth in the respective subline of lyphadenosis. When the antibiotics were used in combination, no significant decrease in sensitivity of the tumor cells to either of the drugs or their combination was observed. The experiment with Staph. aureus also showed that the rate of the resistance increase to the drug combination was lower than that to the drugs used alone. Therefore, it was shown that the combined use of bruneomycin and rubomycin provided a means for preventing to a significant extent of development of the resistance in lymphadenosis tumor cells and Staph. aureus. This may be considered as an indication for clinical trials of the above combination.     

Determination of glutathione in lymphocytes and possible association of redox state and proliferative capacity of lymphocytes.

The glutathione (GSH) content of mouse T- and B-cells was determined and compared with the GSH content of human peripheral blood lymphocytes and human erythrocytes. Owing to the difficulty of obtaining large numbers of purified lymphocytes, a technique was developed to measure picomolar quantities of GSH. By this technique, mouse T- and B-cells, as well as mouse peripheral-blood lymphocytes, were found to contain approx. 30% of the GSH found in human peripheral-blood lymphocytes. The concanavalin A response of human peripheral-blood lymphocytes and human spleen cells was insensitive to 2-mercaptoethanol as well as to culture in 17% O2, whereas mouse lymphocyte responses were altered by 2-mercaptoethanol and inhibited by 17% O2. The capacity of human peripheral-blood lymphocytes, human erythrocytes, mouse T-cells and mouse B-cells to regenerate GSH stores after chemical oxidation by diamide was tested, and it was found that mouse cells were less capable of regenerating GSH than human erythrocytes or human peripheral-blood lymphocytes. In addition, the latter lymphocytes were less sensitive to oxidation of GSH and to inhibition of proliferation by diamide.     

The effect that ammonium and a deficiency of calcium have on lymphocyte subpopulations in common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.)

The patterns of changes in the numbers of T-, B-, and D-subpopulations of lymphocytes were studied in common carp (Cyprinus carpio L.) exposed to waterborne NH₄⁺ (1.5 and 2.5 mM/l) for 45 days and maintained in calcium-free water (30 and 90 days). It was revealed that the fish respond to studied factors by a decrease in T- and D-lymphocytes. The number of B-cells did not change significantly.     

Myeloinhibitory effect of the action of some antitumor antibiotics and its comparative assessment]

General toxic and myeloinhibitory effects of some antitumor antibiotics, such as rubomycin, olivomycin, bruneomycin and karminomycin administered intraperitoneally in a single LD50 to mice were studied. It was found that the general toxicity of bruneomycin and karminomycin was almost the same and 5 to 8 times higher than that of rubomycin and olivomycin. The use of the above antibiotics resulted in definite shifts in the blood systems of healthy mice. The most significant suppression of hemopoesis accompanied by a pronounced depression of the number of the myelocariocytes was observed after the use of olivomycin. The effect of karminomycin was characterized by suppression of erythro-, myelo- and lymphopoesis and depression of the number of the granulocytes and lymphocytes of the blood. Bruneomycin and rubomycin had a short-time myeloinhibitory effect. The erythroid cords of the bone marrow proved to be most sensitive to the inhibitory effect of the antibiotics. However, inhibition of the erythropoesis accompanied by deep reticulocytopenia did not induce the respective depression of the erythrocyte number. The lymphoid cords was in the 2nd place by its sensitivity to the antibiotics and the myeloid and megocariocytal cords were in the 3rd and the 4th places respectively. Complete reduction of hemopoesis in the animals was observed by the 10th day of the drugs use.     

Interaction of blood lymphocyte Fc receptors with IgG during in vitro cultivation

Bovine blood lymphocytes taken from normal cows and those suffering from chronic lymphocytic leukemia were cultured in complete medium 199 with 10% of heat-inactivated fetal bovine serum. After a 48 hour culturing an enhanced quantity of the Fc-receptor bound fluoresceinated immunoglobulin G (IgG) was established. When lymphocyte fractions enriched with T- or B-cells were cultured, the binding capacity of Fc-receptors for IgG (48 hours after culture) increased in both the cell populations. A study of the kinetics of interactions of Fc-receptors with IgG showed that the increased number of Fc-receptors after culturing was followed by an enhanced affinity of Fc-receptors towards IgG. The affinity of Fc-receptors of blood lymphocytes of cows with chronic lymphocytic leukemia was lower than that of normal lymphocytes.     

Immunologic competence of T- and B-lymphocytes in tolerance induced by cyclophosphamide]

A study was made of immunological competence of T- and B-lymphocytes of mice subjected to tolerogenic treatment (administration of a massive dose of sheep erythrocytes and cyclophosphamide 7 days before the experiment). The capacity of lymphocytes of tolerant mice to influence the interaction of normal T- and B-lymphocytes was also investigated. This form of tolerance was caused not by T-suppressors, but by a true deficiency of T-cells-helpers (both in the thymus and in the spleen), and partially of B-cells (in the spleen). Some lack of B-cells in the bone marrow was connected with a nonspecific action of cyclophosphamide. Cyclophosphamide is supposed to selectively eliminate cells proliferating in response to the antigen.     

Ca++-dependent immunosuppressive effect of ethylenediaminetetraacetic acid (EDTA)]

Subcutaneous injection of a sum total dose of 6.44.10(-5) M of ethylenediaminetetraacetic acid (EDTA), which binds divalent cations, inhibits the process of accumulation of antibody-forming cells in the spleen of mice (CBA X C57BL) F1 hybrids, immunizated with sheep red blood cells. In a dose of 5.2.10(-5) M CaCl2 eliminated the immunodepressive effect of EDTA almost completely. It was assumed that EDTA action was produced through the Ca++-controlled regulation mechanism of one of the following processes: the interaction between T- and B-lymphocytes; proliferation and differentiation of T- and B-cells in the course of the immune response to sheep red blood cells. A more complex mechanism of EDTA action on the above-mentioned processes is not excluded, however.     

Relationship between T- and B-lymphocyte cooperation and their syngeneity]

Cooperation efficiency of (CBA x C57BL/6) F1 thymocytes and CBA bone marrow cells in immune response to SRBC was compared with the syngenic combination of the same cells. Selectivity of interaction of the T- and B-lymphocytes of different origin was studied in incomplete cyclophosphamines (CBA x C57BL/6) leads to CBA chimerae, where donors were primed with SRBC and the recipients were either intact or tolerant to the given antigen. F1 T-cells proved to interact with the CAB-B-cells 10-15 times less effectively than with the syngenic B-cells. It is suggested that similarity between the antigenic structure of the cell membrane of the T- and B-lymphocytes, aiding their physical contact, increased the action efficiency of the T-mediator on the B-cell.     

Analysis of certain mechanisms of antigen-specific suppression of the immune response

CBA mice were immunized with sheep red blood cells (SRBC) to obtain immune spleen cells (ISc) which were used to suppressor cells. Administration of ISC to intact syngeneic recipients on the immunization day led to a more powerful suppression of the immune response as compared to that seen one day after antigen injection. Four days after immunization the animals' immune response was not liable to be suppressed. ISC extract possessed similar effects with respect to the immune response of normal spleen cells which were transplanted to the cyclophosphamide-treated recipients. The immune response of spleen cells from mice immunized with SRBC in a dose of 10(6) was less liable to be suppressed. Hyperimmune spleen cells from donors immunized with SRC in a dose of 10(9) were insensitive to ISC or to the extract. Experiments with the use of adoptive transfer of a mixture of immune and intact T- and B-cells have disclosed that B-cells from hyperimmune donors were resistant to suppression. Therefore, B-lymphocytes are the most probable target cells exposed to T-suppressors in the given system. The mechanism is discussed of the selective effect of T-suppressors on B-cells in the course of the immune response development during immunization with high doses of antigen.     

Interaction and migration of T- and B-lymphocytes in immune responses during carcinogenesis induced by methylcholanthrene]

The (CBA X C57BL) F1 mice were injected intramuscularly with methylcholantrene (MCA) in a dose of 0.3 mg, and their T- and B-cells ability to cooperate in the immune response against sheep red blood cells, and also migration of these cells from the thymus and the bone marrow to the spleen were studied. The MCA immunosuppressive action proved to be associated with the inhibition of migration and cooperation of T- and B-lymphocytes in the immune response. A conclusion was drawn that the immunosuppressive effect developing during the carcinogenesis was complex and it was realized at various stages of immunogenesis.     

Pathomorphological picture of the liver in mice administered antitumor antibiotics intraperitoneally and its comparative assessment]

A single administration of carminomycin, ribomycin or olivomycin in LD50 or treatment of the experimental animals with these antibiotics for 10 days in the therapeutic doses equal to 10 per cent of the LD50 induced distrophic and necrobiotic changes in the liver. The use of bruneomycin in the equivalent doses induced sclerotic process in addition to the above doses resulted in a decrease in the colour intensity of DNA, RNA and protein as compared to the control, the content of glycogen and a marked increase in the amount of lipids in the hepatocyte cytoplasm. The most pronounced shifts were observed with the use of carminomycin, rubomycin and especially bruneomycin in single doses. With the use of olivomycin in a single dose the shifts were less pronounced. It should be noted that with the use of carminomycin and rubomycin the damages were of the same character by their intensity. The changes in the liver on the use of carminomycin, rubomycin and olivomycin in single doses or during the treatment course were reversible, while on the use of bruneomycin they preserved to the end of the experiment.     

Treatment of Late Stage Disease in a Model of Arenaviral Hemorrhagic Fever: T-705 Efficacy and Reduced Toxicity Suggests an Alternative to Ribavirin

A growing number of arenaviruses are known to cause viral hemorrhagic fever (HF), a severe and life-threatening syndrome characterized by fever, malaise, and increased vascular permeability. Ribavirin, the only licensed antiviral indicated for the treatment of certain arenaviral HFs, has had mixed success and significant toxicity. Since severe arenaviral infections initially do not present with distinguishing symptoms and are difficult to clinically diagnose at early stages, it is of utmost importance to identify antiviral therapies effective at later stages of infection. We have previously reported that T-705, a substituted pyrazine derivative currently under development as an anti-influenza drug, is highly active in hamsters infected with Pichinde virus when the drug is administered orally early during the course of infection. Here we demonstrate that T-705 offers significant protection against this lethal arenaviral infection in hamsters when treatment is begun after the animals are ill and the day before the animals begin to succumb to disease. Importantly, this coincides with the time when peak viral loads are present in most organs and considerable tissue damage is evident. We also show that T-705 is as effective as, and less toxic than, ribavirin, as infected T-705-treated hamsters on average maintain their weight better and recover more rapidly than animals treated with ribavirin. Further, there was no added benefit to combination therapy with T-705 and ribavirin. Finally, pharmacokinetic data indicate that plasma T-705 levels following oral administration are markedly reduced during the latter stages of disease, and may contribute to the reduced efficacy seen when treatment is withheld until day 7 of infection. Our findings support further pre-clinical development of T-705 for the treatment of severe arenaviral infections.     

Isolation of purified populations of T- and B-lymphocytes under clinical conditions]

The authors present comparative characteristics of the methods of isolation of T- and B-lymphocytes in patients with chronic renal insufficiency. T-lymphocytes were obtained on columns with neilon cotton wool. B-cells were fractionated by repeated centrifugation in the gradient of phycol-urotrast density after staging E-rosette formation with the total lymphocyte pool. Methodical peculiarities of single-stage and two-stage B-lymphocyte fractionation were revealed.     

Impact of bacterial lectin on functional activity of lymphocytes

Impact of Azospirillum brasilense Sp7 lectin with carbohydrate specificity to L-fucose on T- and B-lymphocytes production, activity of IL-1alpha, and TNF-alpha was studied in vitro and compared with influence of red kidney bean phytohemagglutinin (PHA) on the same parameters. Increase of growth of T- and B-cells subpopulations has been shown that point to stimulatory effect of the bacterial lectin. Bacterial lectin of A. brasilense also increased synthesis of IL-1alpha and TNF-alpha. It has been shown that the bacterial lectin has more pronounced stimulatory effect on functional activity of lymphocytes compared with PHA.     

Quantitative behavior of B-lymphocytes in infection susceptible children following a single therapeutic substitution with human gamma globulin]

In 15 children with a lowered gammaglobulin level a single substitution with HGG was made, with the impact of this substitution on the number of B-cells in the peripheral blood being examined by means of the direct fluorescence antibody technique. In 9 from 15 children the substitution had no influence on the number of B-cells. However, a significant increase of the number of B-cells could be observed in 6 from 15 children. 14 days after the substitution the number of B-cells lay within the normal range again.     

Immunohistochemical identification of T- and B-lymphocytes delineated by the unlabelled antibody enzyme method

Summary T-lymphocytes and B-lymphocytes are identified in tissue sections of human tonsils by applying the unlabelled antibody enzyme method. The epithelium of the tonsils contains a majority of immunoglobulin-positive cells and fewer T-lymphocytes. In the subepithelial zones, areas composed of B-cells predominate, however, regions containing T-lymphocytes are also present. The latter are mainly arranged in the lamina propria around high-endothelial venules and often include plasma cells containing immunoglobulin. Follicles containing germinal centres display a complex structure which changes during development. The lymphocytic cap consists of densely packed lymphocytes, labelled heavily by anti-IgM and anti-IGD, and of individual T-lymphocytes. Germinal centres show a framework of immunoglobulin-positive dendritic reticular cells; they contain some heavily labelled lymphoid cells and several cells weakly labelled by anti-IgM and anti-IgA, as well as a small number of T-lymphocytes. Furthermore, the total areas of T- and B-lymphocytes measured by planimetry may differ considerably between different tonsils. Especially total areas of germinal centres vary to a great extent. The quantitative data on amounts of T- and B-cells achieved by planimetry are comparable to those reported in cellular suspensions of tonsils.