A study of the effect of analogs of nucleic acid bases on immunogenesis using Jerne's method

Summary A study was made in dynamics on the number of spleen cells producing antibodies following immunization of mice with erythrocytes. It was found that shortly after immunization the number of these cells was extremely small and subsequently grew rapidly. In animals given injections of 6-mercaptopurine or 6-thioguanine during immunization the number of antibody forming cells reduced considerably. Upon use of 6-thioguanine this effect was the most pronounced. The findings obtained are considered in the light of the clonal-selection theory.Department of General Immunology and Oncology (Chief Active Member USSR Academy of Medical Sciences Prof. L. A. Zil'ber), N. F. Gamalei Institute of Epidemiology and Microbiology (Director-Corresponding Member USSR Academy of Medical Sciences Prof. O. V. Baroyan)(Presented by Active Member USSR Academy of Medical Sciences L. A. Zil'ber) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 60, No. 8, pp. 85–89, August, 1965.     

Changes in the number of antibody-forming and stem cells in the spleen of unilaterally nephrectomized mice

A suspension of spleen cells from intact and unilaterally nephrectomized mice, obtained in the latter case 19–21 h after the operation, was injected into lethally irradiated (CBA×C57BL/6) F₁ mice 24 h after irradiation. On the 8th day after injection of the cells the number of colony-forming and plaque-forming cells in the spleen of the irradiated recipients was determined. To determine the number of plaque-forming cells, a mixture of spleen cells and sheep's red cells was injected into the irradiated recipients. The number of colonies in the recipients' spleen 19–21 h after the operation either was unchanged or was significantly reduced. Stimulation of the production of antibody-forming cells was observed at these same times, and it coincided in time with the period of manifestation of the ability of the splenic lymphoid cells of the unilaterally nephrectomized mice to induce proliferation in the kidney of the intact recipients.Laboratory of Experimental Genetics, Institute of Medical Genetics, Academy of Medical Sciences of the USSR. Laboratory of Growth and Development, Institute of Human Morphology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR, N. A. Fedorov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 78, No. 9, pp. 69–72, September, 1974.     

Effect of regeneration of hematopoietic organs on the number and type of splenic colonies

Two thirds of the spleen was resected or the bone marrow from one tibia was removed in CBA mice. On the 8th day after irradiation and injection of a suspension of intact spleen cells into the animals hematopoietic colonies were obtained and examined microscopically. In the animals of the experimental groups some increase was observed in the number of colonies compared with the control, and the number of granulocytic colonies was significantly higher although the number of colonies belonging to other hematopoietic series remained unchanged. The authors suggest that these changes may be due both to the local effect of the proliferating stroma of the spleen and to the action of a factor secreted by the regenerating stroma of the hematopoietic organs.Laboratory of Growth and Development, Institute of Human Morphology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 85, No. 2, pp. 219–221, February, 1978.     

Effect of regeneration of the spleen and bone marrow on number of hematopoietic colonies in the mouse spleen

Two thirds of the spleen (group 1) or the bone marrow from the right tibia (group 2) was removed from sexually mature male CBA mice. On the eighth day after lethal irradiation and injection of 1·10⁶ nucleated cells from the intact spleen the number of hematopoietic splenic colonies was counted. A significant increase in the number of colonies was observed in the animals of both experimental groups compared with the control intact mice. the authors suggest that this increase may have been caused both by the local effect of the regenerating splenic stroma and by a certain stimulating factor secreted by the regenerating hematopoietic tissue.Laboratory of Growth and Development, Institute of Human Morphology, Academy of Medical Sciences of the USSR, Moscow. Department of Histology, Biological Faculty, Moscow State University. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 82, No. 11, pp. 1375–1376, November, 1976.     

Effect of chronic pyrogenal stress on the mitotic regime and number of DNA-synthesizing cells in the corneal and lingual epithelium of albino rats

The effect of prolonged stress on the mitotic regime and number of DNA-synthesizing cells in the corneal and lingual epithelium was studied in response to pyrogenal. Injection of pyrogenal for 5 days caused a decrease of 41% in the number of mitoses in the corneal and lingual epithelium. The decrease in the number of dividing cells did not correlate with changes in the rate of mitosis. The number of pathological mitoses in the corneal epithelium of intact rats remained unchanged during stress. The index of labeled nuclei in the corneal and lingual epithelium of the control rats was 12.6 and 10.8 respectively, which did not differ significantly from their values in the experimental animals (12.2 and 12.2).Central Research Laboratory, Khabarovsk Medical Institute. Laboratory of Cytology, Institute of Human Morphology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 8, pp. 248–250, August, 1978.     

Origin of cells reducing normal hemolysins in guinea pigs

Cells producing normal antibodies against sheep's erythrocytes were revealed by local hemolysis in agar in the spleen of embryos in the late stage of development and of newborn guinea pigs. Preliminary immunization of the pregnant animals with sheep's red cells did not affect the number of hemolysin-producing cells in the spleen of the embryos or newborn guinea pigs. The number of normal antibody-forming cells in the spleen of sterile adult guinea pigs was the same as in animals of the same species kept under ordinary conditions. The results suggest that normal hemolysin-producing cells arise in guinea pigs not by antigenic stimulation through cross-reacting antigens of the external environment, but by physiological differentiation of predetermined precursor cells. This hypothesis corresponds to one of the postulates of the clonal selection theory of immunogenesis.Laboratory of Immunologic Tolerance, N. F. Gamaleya Institute of Epidemiology and Microbiology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR G. V. Vygodchikov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 78, No. 12, pp. 51–54, December, 1974.     

Ability of cells of the regenerating spleen to effect the graft versus host reaction

The ability of cells of the regenerating (after single or twice repeated resection) and intact spleen of mice to induce the graft versus host reaction was studied by two methods. The regenerating spleen was shown to be less capable than the intact of bringing about this reaction.Laboratory of Growth and Development, Institute of Human Morphology, Academy of Medical Sciences of the USSR. I. M. Sechenov Second Moscow Medical Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 81, No. 3, pp. 373–374, March, 1976.     

Effect of gnotobiotic conditions on the histophysiology of liver and spleen tissues

The liver and spleen of gnotobiotic Wistar rats were studied by histochemical methods and the liver electron-microscopically. Under germfree conditions of existence of the animal the succinate dehydrogenase and nonspecific esterase activity in the liver decreased, fatty infiltration of the cytoplasm of the hepatocytes and Kupffer cells increased, and some of the cells developed fatty degeneration. Meanwhile acid phosphatase activity and the number of lysosomes increased in the biliary poles of the hepatocytes, whereas in the spleen destruction of erythrocytes and the liberation of free iron and pigments, which stimulate the excretion of bile in germfree animals, were increased.Laboratory of Embryonic Histogenesis, Research Institute of Human Morphology, Academy of Medical Sciences of the USSR. Research Laboratory of Experimental Biological Models, Academy of Medical Science of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 85, No. 1, pp. 81–84, January, 1978.     

Cytological analysis of the intact and regenerating liver of various strains of rats

A comparative cytological study was made of the control and regenerating liver in two strains of rats: August and cotton-tail. Two-thirds of the liver was removed from both groups of experimental animals and the intact organ served as the control. The animals were killed five or six at a time 30 h and 3, 8, 42, and 120 days after partial hepatectomy. The number of binuclear cells, the dimensions of the mononuclear hepatocytes and of their nuclei, and the mitotic activity and ploidy of the cells were studied. With the exception of mitotic activity, the regenerating and intact liver of the August rats differed from the regenerating and intact liver of the cotton-tail rats in all the above-mentioned cytological parameters. It is concluded that differences in the cytological parameters between the two different strains are due to the genotype of the particular strain.Laboratory of Growth and Development, Institute of Human Morphology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 82, No. 8, pp. 987–989, August, 1976.     

Comparison of ribonuclease isolated from the liver and pancreas of burned and intact rats

Alkaline RNase was isolated from the pancreas and liver of intact and burned rats and purified 1400 and 2000 times, respectively. The pH-optimum of enzyme activity was 7.6–7.8. Biosynthesis of RNase under normal conditions and after burns was studied by incorporation of labeled amino acids. Inhibition of synthesis of alkaline RNase was found in both organs of the burned animals. The half-life of RNase from the liver of intact rats was 65 min and from the liver of burned animals 100.8 min. Enzymic activity of preparations of alkaline RNase from the liver and pancreas of intact and burned animals did not differ significantly.A. A. Vishnevskii Institute of Surgery, Academy of Medical Sciences of the USSR. Research Institute of Child and Adolescent Physiology, Academy of Pedagogic Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. A. Vishnevskii.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 81, No. 3, pp. 305–307, March, 1976.     

Morphological and functional changes in the immune system during reparative regeneration of the liver

During reparative regeneration in the liver functional activity of the immune system is increased. Resection of the liver is accompanied by sharp changes in the structure of the thymus and spleen. The thymus-dependent parts of the spleen are particularly reactive. Resection of the liver in rats is accompanied by an increase in the number of stem cells in the bone marrow, as determined by the splenic colonies method. The number of colony-forming cells in the spleen of recipients of lymphocytes taken from the hepatectomized animals is significantly greater than in the spleen of recipients of lymphocytes from intact rats. If the limb is screened, ability to form endogenous colonies also is increased in partially hepatectomized rats compared with intact animals. Resection of the liver in rabbits is accompanied by a significant increase in immunological reactivity on the first to third days after the operation. The reactions of leukergia and leukocytolysis are sharply intensified during this period.Department of General Biology and Central Research Laboratory, S. M. Kirov Gor'kii Medical Institute, (Presented by Academician of the Academy of Medical Sciences of the USSR, A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 10, pp. 480–484, October, 1977.     

A state resembling tolerance arising in mice under the influence of antigen of lysed erythrocytes

A lysate obtained by treating sheep's erythrocytes with distilled water, purified by ultracentri-fugation in the cold, was shown to possess weak immunogenicity. Its injection into mice caused a state of hyporeactivity to sheep's erythrocytes (the level of the immune response was depressed to 10–25% of the control). The ability of mouse spleen cells to give an immune response to erythrocytes was undisturbed after adaptive transfer. In the early stages after injection of the lysate the spleen cells of the mice had weak suppressive activity when transplanted into intact animals. Blood serum of mice treated with the lysate possessed weak blocking activity which disappeared after absorption of the serum with sheep's erythrocytes. It is concluded that the hyporeactivity arising in the mice after injection of the lysate is due to the presence of antibodies inhibiting the immune response in the serum.Laboratory of Immunogenetics, Institute of Medical Genetics, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR N. N. Zhukov-Verezhnikov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 85, No. 3, pp. 310–314, March, 1978.     

Specific suppression of the immune response in the adoptive transfer system

Injection of spleen cells (SC) of syngeneic animals immunized with large doses of sheep's red cells (SRBC) into intact mice led to marked specific suppression of the immune response of the recipients. The highest suppressive activity was shown by SC taken from donors on the 14th day after intraperitoneal injection of SRBC. The SC of intact animals and of mice receiving a preliminary injection of rat red cells did not affect the immune response of the intact recipients on immunization with SRBC. Treatment of the immune SC with anti-T serum (ATS) or anti-B globulin (ABG) and complement considerably reduced or completely abolished the suppressive activity. Injection of a mixture of two suspensions of immune SC, one treated with ATS and the other with ABG, into intact recipients did not lead to suppression of the immune response. It is postulated that the suppressor cells in this particular model are T lymphocytes, expressing common antigens or antigens cross-reacting with B cells.Laboratory of Immunogenetics, Institute of Medical Genetics, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR N. N. Zhukov-Verezhnikov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 83, No. 5, pp. 571–573, May, 1977.     

Dependence of the number of antibody-forming cells in the spleen of rats immunized with typhoid antigen Salmonella typhi on the number of cells spontaneously producing antibodies against this antigen

It was shown by the method of local passive hemolysis in gel that the number of antibody-forming cells (AFC) in the spleen of five batches of rats obtained at different times from the nursery correlates after immunization withSalmonella typhi O antigen with the number of spontaneous AFC against that same antigen in animals of the same batch. This may indicate that the number of spontaneous AFC reflects the number of B lymphocytes immunocompetent toward the particular antigen and that it can be used to some degree to assess immunologic reactivity to that antigen before immunization.Personal Group of Academicians of the Academy of Medical Sciences of the USSR V. K. Navrotskii, Ukrainian Postgraduate Medical Institute, Khar'kov. (Presented by Academician of the Academy of Medical Sciences of the USSR V. K. Navrotskii.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 81, No. 2, pp. 201–202, February, 1976.     

Mechanism of action of aspirin on platelet function

Inhibition of platelet aggregation and of the reaction of liberation of platelet factor 3 under the influence of aspirin was shown to be due to the action of the drug not only on the platelets, but also on plasma cofactors: In experimentsin vitro the blood plasma of rats receiving aspirin reduced the aggregating power of the platelets of intact animals; blood plasma of intact rats increased the aggregating power and accessibility of factor 3 of the platelets of animals receiving aspirin.Department of Radiation Pathological Physiology, Scientific-Research Institute of Medical Radiology, Academy of Medical Sciences of the USSR, Obninsk. (Presented by Academician of the Academy of Medical Sciences of the USSR N. A. Fedorov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 83, No. 4, pp. 434–435, April, 1977.     

Effect of a highly purified factor from the thymus on cellular and humoral indices of immunity in thymectomized mice

In experiments on thymectomized adult CBA mice the effect of a homogeneous factor of polypeptide nature from the thymus, with mol. wt. about 5000 (thymarin-III) on the cellular and humoral indices of immunity was studied in animals. Thymectomy in animals was shown to sharply reduce the number of T-cells in the spleen. Correspondingly, the ability of the mice to produce both IgM- and IgG-antibody-forming cells and humoral antibodies against a thymus-dependent antigen (sheep's red blood cells) was sharply inhibited in the mice. Subcutaneous injection of thymarin-III in a dose of 1 g/kg into the animals daily for 7 days completely restored the T-cell population of the spleen and restored the normal immunologic reactivity of the animals.Department of Microbiology and Immunology, Institute of Experimental Medicine, Academy of Medical Sciences of the USSR, Leningrad. (Presented by Academician of the Academy of Medical Sciences of the USSR V. I. Ioffe.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 7, pp. 51–53, July, 1978.     

Kinetics of hematopoietic stem cells in the bone marrow of hepatectomized mice

Two thirds of the liver was removed from (CBA×C57BL/6j)F₁ female mice. On the 5th day after the operation a significant increase was observed in the number of endogenous colonies in the spleen of the partially hepatectomized animals. This increase was not connected with a change in the number of stem cells in the bone marrow, for partial hepatectomy at different times after the operation did not affect the number of colony-forming units in the bone marrow.Laboratory of Virology, Oncologic Scientific Center, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR N. A. Kraevskii.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 85, No. 2, pp. 218–219, February, 1978.     

Properties and kinetics of the B lymphocyte population of rats during development

Cells carrying surface immunoglobulins (Ig⁺ cells) detected by the indirect immunofluorescence method, and cells forming rosettes with sheep's red blood cells (SRBC) with antibodies adsorbed on their surface and with complement (RFC), were found in the liver and spleen of rat fetuses at the 15th and 20th days of development. The relative percentage of Ig⁺ cells and RFC in the liver remained low and about the same level in rats on different days of postnatal development. In the spleen and bone marrow the number of Ig⁺ lymphocytes and RFC increased during the first month of the rat's life, to reach a maximum in animals aged 30 days, and fell sharply in old rats. No Ig⁺ cells or RFC were present in the thymus or they were found in very small numbers at certain times of investigation. Ig⁺ lymphocytes with caps of fluorescence on their surface appeared in the spleen and bone marrow on the fifth and 10th days of life of the rat and their number rose considerably by the age of 30 days and in adult rats. No such cells were present in the lymphoid organs of old (40 months) animals.Laboratory of Embryonic Histogenesis, Institute of Human Morphology, Academy of Medical Sciences of the USSR. Institute of Molecular Biology, Academy of Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 10, pp. 491–494, October, 1977.     

Effect of azathioprene on DNA synthesis,mitotic activity,and hypertrophy of cells in the regenerating liver and intact spleen of rats

Two-thirds of the liver was removed from August rats, and a suspension of azathioprene in 1% carboxymethylcellulose was injected in a dose of 40 mg/kg. The level of DNA synthesis was determined after 22, 48, 72 h with the aid of thymidine-H³, and the mitotic activity and the dimensions of the cells and their nuclei, were determined in the regenerating liver and intact spleen. Injection of azathioprene led to a decrease in the number of DNA-synthesizing cells in the regenerating liver, and spleen and depressed mitotic activity of the hepatocytes. Considerable development of hypertrophy of the cells and nuclei under the influence of azathioprene also were observed in the hepatocytes of the regenerating liver and the small lymphocytes of the white pulp of the spleen, i.e., the weight of the regenerating liver of the rats receiving azathioprene was restored by hypertrophy as well as mitotic division of the cells.Laboratory of Growth and Development, Institute of Human Morphology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 78, No. 11, pp. 102–105, November, 1974.     

Rosette formation in a system of adoptive transfer of cell populations from donors with an altered serotonin level

The formation of the primary immune response to injection of cells of various immunocompetent organs of nonimmune intact donors and also of animals receiving serotonin into sublethally irradiated recipients was studied. Considerable changes were found in the development of the immune response, mainly on account of IgM-rosette-forming cells, both after transfer of spleen cells alone and after combined injection of spleen cells with bone marrow and thymus cells from the same donors. The results are evidence of a possible effect of serotonin on changes in the relative proportions and migration of T- and B-cells in different organs and also on the action of serotonin on T-and B-suppressor cells.Laboratory of Physiology of Immunity, Institute of Physiology, Siberian Branch, Academy of Medical Sciences of the USSR, Novosibirsk. (Presented by Academician of the Academy of Medical Sciences of the USSR V. P. Kaznacheev.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 88, No. 12, pp 688–691, December, 1979.