Effect of pertussis vaccine on the functional activity of the peritoneal and splenic macrophages in 2 mouse strains genetically differing by the intensity of their immune response

The immunostimulating effect of corpuscular pertussis vaccine on the antigen-presenting and bactericidal functions of peritoneal and splenic macrophages in CBA and C57BL/6 mice, differing in the intensity of immune response to sheep red blood cells and Salmonella typhimurium, has been studied. The study has revealed that the injection of pertussis vaccine alters the functional activity of the cells under study, the effect depending on the immunizing dose, the strain of mice and the time elapsed from the moment of immunization. Pertussis vaccine enhances the low capacity of macrophages for antigen presentation in C57BL/6 mice with low responsiveness and alters the resistance of peritoneal and splenic macrophages to the cytopathic action of salmonellae.     

Capacity of triterpene glycosides from holothurians to stimulate antibacterial resistance in a model of experimental murine salmonellosis

When injected intraperitoneally into mice in doses of 40.0-0.4 microgram, Cucumarioside, the preparation of triterpene glycosides obtained from sea cucumbers (Cusumaria japonica), enhanced the resistance of the animals to the subsequent challenge with Salmonella typhimurium. The study of the duration of the persistence of salmonellae in mice receiving the preparation in a dose of 0.001 microgram revealed a decrease in the contamination of their organs. The same dose of the preparation stimulated the phagocytic activity of peritoneal exudate cells with salmonellae showing decreased cytopathogenic action. This suggests that Cucumarioside enhances nonspecific protective factors, activates the macrophagal system and facilitates the development of complete phagocytosis.     

Effect of T-activin on peripheral immunity organs in intact and thymectomized mice

Morphometry of the spleen, axillary lymph nodes and cytological assay of the bone marrow and peripheral blood were performed in (CBA X C57BL)F1 mice 1, 5, 10 and 15 days after subcutaneous injection of 0.5 microgram T-activin to intact and thymectomized (when adult) mice 2 months after operation. It was demonstrated that in intact animals, injection of T-activin stimulated the whole system of immunogenesis. The time course of plasmatization and the response of the germinative centers differing from that seen during antigen administration suggests that T-activin is not immunogenous, acting as a stimulant of the previous immune responses. The permanent amount of the degenerating cells attests to the lack of the toxic drug effect.     

Functional macrophage activity in infection with virulent and avirulent strains of the dysentery microbe

The effect of S. flexneri virulent and avirulent (vaccine) strains 2a on the cytoplasmic membrane of mouse macrophages has been studied by evaluating the action of these bacteria on the activity of 5-nucleotidase. The dynamics of the activity of 5-nucleotidase after the introduction of both virulent and avirulent strains has a phasic character with alternating rises and falls in the activity of this enzyme in comparison with the control. S. flexneri vaccine strain produces mainly a stimulating effect on the functional activity of peritoneal macrophages in mice, which is confirmed by a decrease in the activity of 5-nucleotidase; on the contrary, S. flexneri virulent strain- has mainly an inhibiting effect on the functional activity of peritoneal macrophages, which is confirmed by an increase in the activity of 5-nucleotidase in these cells. The comparative study of changes in the activity of 5-nucleotidase, following the introduction of S. flexneri, in mice, previously immunized with smallpox vaccine, and in intact mice has shown that the use of animals immunized with smallpox vaccine in the study of metabolic characteristics may lead to distortions in the results of the experiment.     

Persistence of the Salmonella enteritidis recombinant strain with expressed HBc antigen in mice and macrophage culture

The behavior of S. enteritidis recombinant strain E-23/pKDNA(3.1) with HBc antigen expressed by the eukaryotic promoter (DNA vaccine) in the body of mice after their infection per rectum and in the culture of macrophages was studied. The expression of HBc was shown to lead to the loss of the capacity of salmonellae for persistence in parenteral lymphoid tissue and for inducing the formation of anti-HBc antibodies. The study of the interaction of salmonellae with the macrophage culture revealed that the synthesis of HBc antigen inhibited the intracellular multiplication of salmonellae.     

Effects of T-activin on the morphology of the thymus gland in experimental injury of the lower limbs in mice (anti-stress effect of T-activin)

The effect of T-activin on thymic involution under the experimental trauma of femur was studied. T-activin in a dose of 1.0 micrograms/mouse was injected into young male (CBA X C57BL)F1 mice weighing 17.5-19.0 g before (I injection) or immediately after the fracture of the femur during 3 days. Morphometric analysis of the thymus was made 1, 5, 10 and 15 days after the trauma. It was found that T-activin suppressed the involution of the thymus, induced by the trauma, during the first 5 days and accelerate the process of its regeneration. It is suggested, that T-activin displays protective anti-stress effect on the mouse thymic involution.     

The correction of the functional activity of alveolar macrophages in inducing a primary immune response in influenza]

The functional activity of alveolar macrophages obtained from mice, both healthy and infected with influenza virus A/Aichi 2/68 (H3N2), as manifested by their capacity to initiate the development of primary immune response to sheep red blood cells and Escherichia coli lipopolysaccharide after the transfer of these macrophages to intact syngeneic recipients was studied. The capacity of alveolar macrophages to perform antigen-presenting functions in the induction of humoral immune response was shown, and at the same time the development of experimental influenza infection was found to essentially decrease these properties. The injection of the immunomodulating agent diuciphon into experimental mice somewhat enhanced the immune response after the syngeneic transfer of alveolar macrophages from infected mice to intact recipients.     

High Susceptibility of Strain A Mice to Endotoxin and Endotoxin-Red Blood Cell Mixtures.

Heppner, Gloria (University of California, Berkeley), and David W. Weiss. High susceptibility of strain A mice to endotoxin and endotoxin-red blood cell mixtures. J. Bacteriol. 90:696-703. 1965.-Strain A mice were shown to be considerably more susceptible to lethal effects of endotoxin lipopolysaccharide (LPS) than mice of several other strains. Complexes of sublethal quantities of LPS and sheep red blood cells were synergistically toxic for strain A mice. Separate administration of sheep red blood cells and heat-killed salmonellae, in either order and as long as 24 hr apart, also proved to be synergistically lethal for strain A mice, but not for R(III) animals studied comparatively. Sheep red blood cell lysates possessed the ability of the intact cells in forming lethal combinations for strain A mice with killed salmonellae. Strain A red blood cell-killed salmonellae complexes were also lethal for strain A mice, but less so then complexes made with sheep red blood cells. A x R(III) F(1) hybrid animals showed the same resistance characteristics as the resistant R(III) parental strain. Possible explanations for these findings are suggested, and their relevance to an immunological mode of action of endotoxin lethality is discussed.     

Study of the action of immunomodulator T-activin on electric properties of plasma membranes of thymus cells by the use of fluorescent probes

T-activin alters the electric properties of plasmatic membranes of the T-activin treated suspension of mouse (CBAX X057B1) F1 thymic cells. These alterations were registered by means of negative charged fluorescent probe l-anilino-naphthalene-8-sulfonate (ANS) and positive charged 4-(p-dimethylaminostyryl)-1-methylpyridinium (DSM) by measurement of their fluorescence in the individual thymic cell. Apparently, T-activin leads to depolarization of transmembrane potential on plasmatic membrane of thymic cells. It seems that action of T-activin is similar to the effect of polycations, which alter cells ionic streams during lymphocyte activation. However, there is a version, that T-activin acts like neuraminidase, but it is activity less expressed.     

The use of chemiluminescence for assessing the physiological status of macrophages activated by pertussis preparations

The work deals with the comparative analysis of oxygen-dependent metabolic processes in the peritoneal macrophages of intact and immune, mice after their interaction with whole-cell pertussis vaccine (WCPV), dialyzed pertussis antigen (DPA) and fraction 2 of this antigen. The study revealed that WCPV, DPA and its fraction 2 modulated the production of active forms of oxygen by peritoneal macrophages, evaluated by means of luminol-dependent chemiluminescence (CL). The influence of WCPV of oxygen-dependent metabolic processes in macrophages after the first contacts with them had a dose-dependent character: low concentrations activated and higher concentrations suppressed the "respiratory" explosion. The immunization of animals abolished the effect of extinguishing CL on the contact of macrophages with high doses of WCPV and essentially increased the level of their CL response to all pertussis preparations in comparison with those for intact cells. DPA and its fraction 2 were not inferior to WCPV in their capacity for inducing the "respiratory" explosion, and in high doses they essentially surpassed WCPV doses in this capacity, especially on the first contact with macrophages.     

Immunomodulating properties of salmozan and its effect on the functional activity of macrophages

The effect of salmozan on the resistance of mice to Listeria monocytogenes infection, the formation of delayed hypersensitivity (DH) to sheep red blood cells in the animals, as well as changes in some functional activity characteristics of macrophages have been studied. The study has revealed that salmozan enhances anti-infectious resistance, suppresses the dermal manifestations of DH, and decreases the level of 5'-nucleotidase in peritoneal macrophages, stimulating their phagocytic activity. The intensity of the drug action depends on the time of its administration. The most pronounced immunomodulating action and maximal changes in the function of macrophages have been registered simultaneously after the treatment of the animals with salmozan.     

In vivo administration of anti-asialo-GM1 antibody enhances splenic clearance of Listeria monocytogenes

Resistance of mice to infection by Listeria monocytogenes involves a biphasic response. The first phase consists of the first 48 h after infection, during which there is multiplication of Listeria in the liver and spleen of infected mice. In these nonimmune mice, macrophages and polymorphonuclear leukocytes are the effector cells involved in controlling multiplication. In the second phase, cell-mediated immunity develops, beginning on day 2, during which multiplication of Listeria is prevented by macrophages possessing increased microbicidal activity that is mediated through the action of lymphokines released by immunologically committed T lymphocytes. The purpose of the present study was to define a role for natural killer (NK) cells in natural resistance to Listeria during the first 48 h after infection, prior to the development of specific immunity. Splenic NK cell activity was enhanced following a sublethal intravenous injection of viable Listeria as early as 24 h after injection and remained elevated throughout the nonimmune phase of infection. Interestingly, treatment of mice with anti-asialo-GM1 significantly enhanced the ability of mice to clear Listeria from the spleen relative to infected controls possessing intact NK cell populations. This was evidenced by 23-fold fewer bacteria obtained from the spleens of anti-asialo-GM1-treated mice. In addition, Percoll-enriched NK cell populations obtained from 48-hour Listeria-infected mice do not exhibit in vitro listericidal activity. These observations suggest a regulatory role of NK cells in resistance against Listeria and preclude a role for NK cells in direct cytolysis. Perhaps these cells modulate the immune response to Listeria by down-regulating the activity of the immune cells crucial to listerial resistance.     

Delayed hypersensitivity and nonspecific cellular immunity. The role of mono- and polynuclear phagocytes

Differences in the influence produced by sensitization with BCG vaccine and Staphylococcus aureus and by the reaction of delayed hypersensitivity (DH) induced, respectively, by the injection of old tuberculin and staphylococcal phagolysate on the phagocytic activity of peritoneal macrophages and blood leukocytes in different animals were experimentally demonstrated. A considerable activation of the bactericidal and ingesting functions of macrophages was observed in animals showing a pronounced DH reaction (rabbits, guinea pigs and mice), while in Wistar rats no such activation was noted. The latter showed no DH reaction after sensitization with BCG vaccine and the injection of the specific antigen. Among different strains of mice, the activation of macrophages occurred in the animals with the most pronounced DH reaction. Sensitization with BCG vaccine led to an insignificant sensitization of macrophages, and sensitization with S. aureus even suppressed the phagocytic activity of macrophages. The treatment of mice with antimacrophagal preparations (carrageenan, silica and trypan blue, but T-lymphocyte antiserum) before and after the injection of the specific antigen into the sensitized animals abolished the stimulation of anti-infection immunity.     

Enhanced accumulation of inflammatory neutrophils and macrophages mediated by transfer of T cells from mice immunized with Listeria monocytogenes

We have previously shown that listeria-immunized mice recruit more inflammatory neutrophils and macrophages to the peritoneal cavity after i.p. injection of a sterile irritant than do nonimmune mice. Because the inflammatory phagocytes that were obtained from listeria-immune and nonimmune mice did not differ in their ability to kill Listeria monocytogenes in vitro, this suggested that the rapid recruitment of listericidal inflammatory neutrophils and macrophages may be critically important for resistance to listeriosis. In this study we demonstrate that the transfer of listeria-immune T cells, which enhances recipient resistance to listeriosis, also increases the ability of recipients to mobilize inflammatory neutrophils and macrophages to the peritoneal cavity after the i.p. injection of dead listeria. The transfer of enhanced inflammatory responsiveness was blocked by pretreatment of the transferred cells with anti-Thy-1.2 plus complement, and the magnitude of the inflammatory cell accumulation was dependent on the number of listeria-immune T cells that were injected. Inflammatory neutrophils and macrophages that were obtained from the mice after the transfer of listeria-immune or nonimmune T cells (plus dead listeria) did not differ in their ability to kill L. monocytogenes in vitro. These data suggest that the elicitation of an inflammatory response may be an important event in T cell-mediated resistance to listeriosis.     

Effect of trypsin on suppressor cell formation and function in localized staphylococcal infection

The present study has been made on (CBA X C57BL)F1 mice immunized with sheep red blood cells (SRBC) and inoculated with staphylococci (M-SRBC-S). The injection of splenic lymphocytes from syngeneic M-SRBC-S into intact mice has been found to suppress immune response to SRBC in these mice. The injection of trypsin into M-SRBC-S decreases the suppressive action of their lymphocytes on SRBC-induced immune response in syngeneic recipients. The injection of trypsin into the recipients has been found to produce no effect on the immunosuppressive action of transplanted lymphocytes obtained from M-SRBC-S. The injection of trypsin into M-SRBC-S induces the release of the factor, inhibiting the formation and function of suppressor cells, by their splenocytes. Previously formed suppressor cells block the release of the immunostimulating factor by the splenocytes of the animals receiving the injections of trypsin.     

Action of plant extracts on the natural immunity indices of animals

The influence of extracts from oak bark, St. John's-wort leaves and pine buds on natural immunity characteristics of mice has been studied. The injection of these extracts into mice has been found to enhance their resistance to infection with Staphylococcus aureus and Bordetella pertussis virulent cultures, to decrease the enzymatic activity of 5'-nucleotidase in the peritoneal exudate macrophages of mice and to increase the level of lysozyme in their blood. The action of these extracts has proved to depend on their dosage and the time of observation.     

Effect of insulin and isoproterenol on macrophage phagocytic activity

To reveal the influence of cyclic adenosine monophosphate (cAMP) on the completion of the phagocytosis of salmonellae, the influence of insulin and isoproterenol on the phagocytic activity of peritoneal macrophages obtained from mice infected with S. typhimurium strains differing in virulence was studied in vitro. The study showed that isoproterenol, while increasing the intracellular content of cAMP, suppressed the bactericidal properties of macrophages with respect to salmonellae, whereas insulin decreased the level of cAMP in the cells and thus facilitated more rapid and complete digestion of ingested bacteria irrespective of their virulence.     

Characteristics of the immunocyte cooperation in 2 strains of mice opposite in their sensitivity to dermaphytoses during the immune response to antigens of the causative agent

C57BL/6 and BALB/c mice, opposite in their sensitivity to dermatophytic infections, show similar activity of phagocytes as regards their capacity for the destruction of injected conidia of dermatophytes, but differ in the changes of this activity after immunization with dermatophytic antigenic complexes (DAC). Shortly after the injection of the antigens, the lymphocyte-mediated suppression of the fungicidal activity of macrophages, caused by the interaction of DAC with intact T-lymphocytes, was detected in the animals of both strains. Later in C57BL/6 mice resistant to mycosis the formation of cell-mediated immunity to DAC occurs, with the simultaneous production of factor stimulating the fungicidal activity of macrophages. In BALB/c mice sensitive to mycosis the injection of DAC induces active antibody production, but not the formation of delayed hypersensitivity with the resulting stimulation of the fungicidal activity of phagocytes. The injection of DAC into mice of the above-mentioned strains induces changes, peculiar to each strain, in the mitogen-induced proliferation of spleen cells and in the character of immune response to sheep red blood cells. Differences in the influence of DAC on the induction of immune response in C57BL/6 and BALB/c mice are realized by cells belonging to the population of T-lymphocytes.     

Pathogenesis and immunity in murine salmonellosis.

Salmonella is traditionally described as a facultative intracellular parasite, and host macrophages are regarded as the primary effector cells in both native and acquired immunity in mouse typhoid. This concept has not been unanimously accepted in the literature. Based on cell culture experiments and electron microscopic examinations of infected tissues, we observed that virulent Salmonella typhimurium is killed within polymorphs and macrophages of guinea pigs and mice. In a systemic disease, the organism propagates primarily in the extracellular locations of sinusoids and tissue lesions and within hepatocytes. Hence, it is more likely to be an extracellular pathogen and its virulence is directly related to its antiphagocytic property. The conspicuous absence of macrophages in the primary lesions of murine salmonellosis disputes the likelihood of their significant role in native resistance to the disease. Acquired cellular immunity is expressed as an enhanced antibacterial activity of macrophages facilitated by cytophilic antibodies rather than as an altered antibacterial action of immune macrophages. It is proposed that acquired immunity in murine salmonellosis is a synergistic manifestation of the innate capacity of polymorphs and macrophages to destroy ingested salmonellae, the activated antibacterial functions of macrophages mediated by cytophilic antibodies, the opsonic and agglutinating actions of antiserum, and the accelerated inflammation associated with delayed hypersensitivity to bacterial antigens. Unlike live attenuated vaccines, nonviable vaccines offer a significant, though not a solid, protection against subsequent challenges.     

Effect of T-activin on thymus involution in mice

The effect of T-activin on thymus involution in mice was studied. T-activin in a dose of 1.0 micrograms/mouse was injected into young male (CBA X C57BL)F1 mice weighing 24.0 +/- 2.0 g daily for 20 days. Morphometric analysis of the thymus was made 6 months after the treatment with T-activin was completed. It was found that T-activin induced the suppression of physiological involution of the thymus together with the enhancement of the processes of thymocyte transformation and proliferation.