Immune response of mice of different inbred lines to Clostridium oedematiens anatoxin

Mice belonging to a number of inbred strains were immunized intradermally with Cl. oedematiens alpha-toxoid. The immunization was repeated 30 days later. On the 20th and the 30th days after the first injection and on the 10th day after the second one the antibody level against the toxoid was determined in the blood of mice by the passive hemagglutination test. The maximum response to the primary immunization was observed in the mice of the C3H strain, and the minimum one--in mice of the DBA/2 strain; the difference was more than 30-fold. The rest of the strains used in the test (A,CBA, BALB/c, AKR, CC57BR) displayed an intermediate level of the immune response. The differences reduced after the repeated immunization. The immune response to this antigen in mice is supposed to be genetically controlled.     

The influence of postradiation chemical signals of mice on the humoral immune response in recipients in different time relative to antigenic stimulus

The influence of volatile urine chemosignals of irradiated (4 Gy) mice on the primary humoral immune response to sheep red blood cells in intact recipients was investigated. It was demonstrated that the direction of immunomodulatory effect is dependent upon the time at which the postradiation chemosignals was initially applied. The antibody response to antigen was markedly suppressed in mice that were exposed before antigen injection. When chemosignals applied immediately following inoculation of antigen the antibody response was unaffected. The immune response was increased when chemosignals was loadeded for 1-10 days after immunization. The possible mechanisms of immunomodulation are considered.     

Effect of sophora japonica total flavonoids on pancreas,kidney tissue morphology of streptozotocin-induced diabetic mice model

To observe effect of sophora japonica total flavonoids on pancreas, kidney tissue morphology of streptozotocin-induced diabetic mice model. Mice received tail vein injection of streptozotocin (60 mg/kg) for diabetes modeling. The model mice were divided into five groups, to be respectively fed with high, middle and small doses of sophora japonica total flavonoids solution, metformin solution and saline of the same volume. Another blank control group was set to be fed with saline of the same volume. The mice were administered once a day for 30 consecutive days, to be euthanatized after fasting blood glucose level testing on 30th day with pancreas, kidney taken out for pathological section and microscopic examination. The mice chain streptozotocin diabetes modeling was successful, with significant pathological changes (P < 0.01) in pancreas, kidney. Compared with model group, high, middle and small doses of sophora japonica total flavonoids could significantly alleviate streptozotocin-induced pancreas, kidney damage (P < 0.01). Conclusion: Sophora japonica total flavonoids can effectively alleviate pancreas, kidney injury of streptozotocin-induced diabetic mice model.     

Cyclic kinetics and mathematical expression of the primary immune response to soluble antigen

The conveyer hypothesis is based on the fact that because of clone predetermination, antibody production takes place in an organism without the presence of antigen as a result of natural cell differentiation. Soluble antigen is an analogue of a specific mitogen which gives rise to reproduction mainly of cells carrying on their surface the immunoglobulin receptors to the given antigen. The mathematical model of the conveyer hypothesis takes into account the initial conditions, among them the background level of antibody-producing cells before injection of a soluble antigen, migration of precursor cells in the draining lymphoid organ, and the rate of precursor differentiation, including the rate of the change of the immunoglobulin receptor number on the cell surface. Changes of antigen concentration in blood determine the intensity of precursor proliferation. Comparison of real experiments (intraperitoneal injection of capsular antigen ofPasteurella pestis into inbred mice) with calculations done on the basis of the developed mathematical model shows a definite qualitative resemblance with the kinetics of antibody-producing cells and free antibodies as well as with the decrease of free antigen concentration in blood. In spite of some differences between model experiments and real experiments the conveyer hypothesis and its mathematical model appear suitable for describing the primary immune response of mice immunized with low doses of capsular antigen ofPasteurella pestis.     

Effect of preliminary administration of allogenic cells on transplantation immunity in mice receiving cyclophosphamide

It was shown that injection of 1 X 10(8) spleen cells from C57Bl/6 mice to CBA mice one day before the injection of cyclophosphamide (CY) helped the take of 2 X 10(7) allogeneic or semiallogeneic cells (injected for the second time 3 to 6 hours after C)). Criterion of survival is the ability of the donor cells to produce antibodies to sheep red blood cells in the recipients tolerant of this antigen. Injection of 1 X 10(8) allogeneic cells two days before CY produces no protective effect. Killer-cells proved to appear on the second day after the immunization with allogeneic cells; their peak was reached on the 5th day. The data obtained suggest that CY eliminated the recipients' lymphocytes, which responded to the transplantation antigens, whereas the killer-cells already formed were stable to the CY action.     

Immunodepressant action of cyclophosphamide in different strains of mice

A study was made of the immunodepressive effect of cyclophosphamide (CP) on mice of 3 strains (BALB/c, CBA, and DBA/2) immunized with sheep red blood cells (SRBC). With the optimal immunizing dose of the antigen (5 X 10(8) SRBC) the most pronounced immunodepression was noted in DBA/2 mice, and with the high dose (6.2 X 10(9))--in DBA/2 and CBA mice. The CP action proved to depend on the dose of the antigen administered; in BALB/c mice a reduction in the number of the antibody-forming cells was the same with both SRBC doses, in DBA/2 mice an increase of the antigen dose led to reduction of immunode pression, and in CBA mice -- to its enhancement (with sufficiently high CP doses). Determination of the rate of oxidative CP hydroxylation by the liver microsomes of mice showed it to be comparatively low in DBA/2 and CBA mice, and much greater in BALB/c mice. It is supposed that the detected differences in the immunodepressive action of CP could be connected with different sensitivity of the target cells and (or) with the peculiarities of its metabolism in mice belonging to different strains.     

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.     

T-lymphocyte subpopulations and immune response in mice of the ASC strain with depressive-like behaviour

Analysis of the content of CD16/32+, CD4+ and CD8+ -cells was perfomed in the peripheral blood and spleen ofmice of the ASC strain with high predisposition to depressive-like state in comparison with mice of parent CBA strain having no depressive behaviour. In both cases, ASC mice showed a decrease in the percentage of CD16/32+ and CD4+-cells along with an increase CD8 cells and lowering of immunoreactivity index (CD4+/CD8+). Changes in cellular subpopulations found in intact ASC mice was accompanied with animals' low capacity to respond to T-dependent antigen: sheep red blood cells at the dose of 5 x 10(8). In contrast to CBA mice the percentage and absolute number of IgM-antibody-forming cells were significantly decreased in the spleen of ASC mice on the 4th and 5th days after immunization as well as the numbers of IgG-antibody-forming cells on the 6th day of the immune response. Possible mechanisms underlying the immune reactivity inhibition under depressive-like state are discussed.     

Immunological properties of Propionibacterium acnes. I. Potentiation and Suppression on antibody response to sheep and hamster erythrocytes in mice.

Adjuvant activity of phenol-treated cells of Propionibacterium acnes C-7 in antibody response was investigated in ICR mice. Simultaneous administration (day 0) of P. acnes (i.p.) and sheep red blood cells (SRBC) (i.v.) enhanced the formation of direct plaque-forming cells (PFC) on days 2, and the formation of indirect PFC response on day 7 and thereafter. Conversely, pretreatment from 11 to 14 days before antigen injection suppressed markedly the antibody response. The potentiation and the suppression of immune response depended on doses of antigen and of P. acnes, the timing of adjuvant injection and the time of assay. The two opposite phenomena caused by P. acnes were also confirmed in antibody response against hamster red blood cells (HRBC). Pretreatment with P. acnes 1 to 14 days before antigen injection suppressed markedly anti-HRBC antibody response, whereas P. acnes injected simultaneously with HRBC or one day after injection of the antigen induced prolongation of antibody response and the production of 2-mercaptoethanol-resistant antibody.     

The regulation of hematopoietic stem cell proliferation and differentiation (CFU-S) during antigenic exposure

Hemopoietic stem cell (CFUs) proliferation is controlled by regulatory activities (stimulator and inhibitor) produced by bone marrow macrophages. Previously it has been shown that antigen administration stimulates CFUs proliferation. The data obtained in this study show the possible mechanism of antigen-induced stimulation of CFUs proliferation. 3-4 days after antigen injection bone marrow cells of BDF1 mice cease to produce inhibitory activity in contrast to similar cells of control animals. Therefore, increased CFUs proliferation in immunized mice can be due to decreased production of inhibitory activity and resulting abundance of stimulating factors. In BAlB/c mice CFUs proliferation is not changed after antigen injection and their bone marrow cells continue to synthesize inhibitory substances. Differentiation of CFUs into committed blood precursor cells may depend on the proliferation level in CFUs population since activation of CFUs proliferation in immunized BDF1 mice is accompanied by a decreased number of CFU-GM and CFU-M but an increased number of BFU-E. It should be noted that intact BAlB/c mice show a high level of CFUs proliferation similar to that of immunized BDF1 mice.     

The carcino-embryonic antigen of experimental tumors of the intestine]

The occurrence of a tumour-specific antigen in the intestinal tumours (induced in 51 albino rats by subcutaneous injection of 1,2-dimethylhydrazine), as well as in the blood serum was studied by double immunodiffusion in agar gel. Intestinal tumours proved to contain a water-soluble antigen absent in other tissues, including the colonic mucosa of control animals. This antigen was found in 70 per cent of the tumour-bearing rats. The antigen is perchloric acid-solubilized and is also present in the embryonic tissues in toto on the 7th and the 9th days of rat gestation. The features of this antigen were analogous with those of the carcino-embryonic antigen in human digestive tract tumours.     

Effect of trypsin on the immune response in localized staphylococcal infection

The influence of trypsin on the formation of immune response induced by a thymus-dependent antigen at different periods of localized staphylococcal infection has been studied. A single intramuscular injection of bovine trypsin has been found to enhance immune response to sheep red blood cells (SRBC) in healthy mice and, to a still greater degree, in mice with localized staphylococcal infection. the development of localized staphylococcal infection has been shown to have no influence on the manifestation of the immuno-suppressive effect of splenocytes in SRBC-immunized mice or to enhance this effect. The injection of trypsin decreases the immunosuppressive effect of splenocytes in healthy or staphylococcus-infected hyperimmune mice.     

IgM and IgG response to sheep red blood cells in mouse hepatitis virus-infected nude mice.

In nude mice which originally had no ability to respond to sheep red blood cells, an enhanced response to the same antigen with IgM-IgG switching was demonstrated during subacute infection with mouse hepatitis virus. IgM antibody-producing cells in the spleen were detected at days 2 to 6 after the antigen injection and IgG antibody-producing cells appeared at day 6 or later. The secondary IgG response, though not remarkable, was recognized after reinjection of the antigen 10 days after the first injection.     

Evidence for Generation of Suppressor T Cells in Mycobacterium lepraemurium-Infected CBA/J Mice,a Mouse Strain Highly Susceptible to the Infection

Susceptibility to Mycobacterium lepraemurium (MLM) infection markedly differed between two mouse strains, CBA/J and C57BL/6. CBA/J mice showed high susceptibility to MLM infection and developed either very weak or no delayed-type hypersensitivity (DTH) to MLM antigen after the injection of MLM. In contrast, C57BL/6 mice, which were resistant to MLM infection, showed significant DTH reaction to MLM antigen after the injection. Treatment of CBA/J mice with cyclophosphamide (Cy) conferred significant resistance to MLM infection on the CBA/J mice, and the treated mice developed a strong anti-MLM DTH response after the MLM injection. When spleen cells from MLM-infected CBA/J mice were transferred to Cy-treated and MLM-infected syngeneic mice, the anti-MLM DTH reaction of the recipient mice was suppressed. Treatment of the spleen cells to be transferred with anti-Thy-1.2 antibody or anti-I-J^k antiserum plus complement abrogated the suppressive activity. Thus, it is suggested that the high susceptibility of CBA/J mice to MLM infection is due to the generation of Cy-sensitive, I-J^k-positive suppressor T cells after infection with MLM.     

Plasmid vector-linked maturation of natural killer (NK) cells is coupled to antigen-dependent NK cell activation during DNA-based immunization in mice

Plasmid DNA vaccines serve in a wide array of applications ranging from prophylactic vaccines to potential therapeutic tools against infectious diseases and cancer. In this study, we analyzed the mechanisms underlying the activation of natural killer (NK) cells and their potential role in adaptive immunity during DNA-based immunization against hepatitis B virus surface antigen in mice. We observed that the mature Mac-1(+) CD27(-) NK cell subset increased in the liver of mice early after DNA injection, whereas the number of the less mature Mac-1(+) CD27(+) NK cells in the liver and spleen was significantly reduced. This effect was attributed to bacterial sequences present in the plasmid backbone rather than to the encoded antigen and was not observed in immunized MyD88-deficient mice. The activation of NK cells by plasmid-DNA injection was associated with an increase in their effector functions that depended on the expressed antigen. Maturation of NK cells was abrogated in the absence of T cells, suggesting that cross talk exists between NK cells and antigen-specific T cells. Taken together, our data unravel the mechanics of plasmid vector-induced maturation of NK cells and plasmid-encoded antigen-dependent activation of NK cells required for a crucial role of NK cells in DNA vaccine-induced immunogenicity.     

Parallel formation of delayed-hypersensitivity effectors and T-suppressors in the intraperitoneal administration of a massive dose of ram erythrocytes

Injection of 6 X 10(9) sheep red blood cells (SRBC) to mice led to parallel formation, on days 4-5, of delayed hypersensitivity effector cells (the activity was tested in local transfer experiments) and delayed hypersensitivity T-suppressors preventing sensitization of syngeneic recipients. After massive injection of SRBC the activity of spleen suppressors gave 2 peaks: on days 5 and 14. Five days after massive antigen injection only T-cells capable of sorbing on a specific antigen manifested suppressor activity. On day 14 T-cells capable of sorbing on specific antibodies showed a specific activity, whereas T-cells capable of sorbing on a specific antigen retained only part of their activity. The mechanism of delayed hypersensitivity inhibition following massive antigen injection by suppressors obtained by day 5 is reviewed in terms of Germain and Benacerraf's theory postulating that delayed hypersensitivity is regulated by Ly 2+, I-J+ antiidiotypic suppressors capable of sorbing on specific antibodies and formed upon injection of Ly 1+, I-J+, Id+ inductor cells capable of sorbing on a specific antigen.     

An intracameral injection of antigen induces in situ chemokines and cytokines required for the generation of circulating immunoregulatory monocytes

Anterior Chamber-Associated Immune Deviation (ACAID) induced by an intracameral injection of antigen generates antigen-specific regulatory splenic T cells that suppress specifically cell-mediated immunity specific for the injected antigen. Circulating F4/80(+) cells recovered from mice receiving an intracameral injection of antigen are thought to be ocular in origin and induce the development of thymic and splenic regulatory T cells. We have shown previously that after the intracameral injection of antigen there is a CCR2/CCL2-dependent infiltration of circulating F4/80(+) cells into the anterior chamber associated with the generation of circulating, ACAID-inducing F4/80(+) monocytes. Here we tested the hypothesis that the intracameral injection of antigen induces events in the anterior chamber that are associated with the induction of circulating immunoregulatory monocytes that induce the suppression of cell-mediated immunity. The intracameral injection of antigen resulted in aqueous humor (i) a time- dependent increase of CCL2 and CCL7, (ii) a transient increase in TNF-α, and (iii) an infiltration of CD11b(hi), Gr1(hi) and F4/80(+) as well as F4/80(-) and Gr1(hi) peripheral blood cells into the anterior chamber. Further characterization of these F4/80(+) cells revealed that they are Ly 6C(hi), LY6G(lo) or negative, 7/4 (LY6B)(hi), CD115(+), CD45(+), CD49B(+), and CD62 L(+). Antibody-mediated neutralization of TGF-β in situ in the anterior chamber prevented the induction of circulating, ACAID-inducing monocytes and ACAID. These cells did not increase in the irides of ACAID-refractory CCR2-/- and CCL2-/- mice that received an intracameral injection of antigen. Our results extend our suggestion that ACAID is initiated as the result of a mild proinflammatory response to intracameral injection that results in the infiltration of a CCR2(+) subset of monocytes into the anterior chamber where there is a TGF-β-dependent induction of an immunosuppressive phenotype in the infiltrated monocytes that recirculate to induce antigen-specific regulatory T cells.     

Leukocyte mobilization in mice by polyanions. Decline of leukocyte mobilizing capacity after transplantation of lymphoma

The leukocyte mobilizing polyanions dextran sulphate (DS) and polymethacrylic acid (PMAA) were administered to AKR and (C57BL x CBA) F1 mice at various times after transplantation of syngeneic lymphoma cells. In nonleukaemic mice DS and PMAA increased the number of circulating leukocytes 3--4-fold. The extent of leukocyte mobilization in leukaemic mice depended on the interval between transplantation of the lymphoma cells and injection of the polyanion. During the development of leukaemia in AKR as well as in (C57BL x CBA) F1 mice the capacity to react upon injection of polyanions with leukocyte mobilization gradually decreased. For DS, this decrease started before the number of leukocytes increased in the peripheral blood. On the other hand, the capacity for PMAA-induced leukocyte mobilization was fully preserved for several more days. In heavily leukaemic mice neither DS nor PMAA could further increase the number of peripheral blood leukocytes. In such mice the distribution pattern of leukaemic blast cells, small lymphocytes, granulocytes and monocytes was also hardly or not affected by injection of the polyanion.     

Effects of deoxycoformycin in mice. I. Suppression and enhancement of in vivo antibody responses to thymus-dependent and -independent antigens

The effect of 2'-deoxycoformycin (DCF) on the PFC responses of AKR mice to SE, TNP-Ficoll, and TNP-B. abortus was examined. Subcutaneous injection of DCF 4 days before antigen caused suppression of all three responses by 70 to 78%. In contrast, injection of DCF 1 day after antigen caused enhancement of both the anti-SE and the anti-TNP-Ficoll responses. Although a single high dose of cortisone acetate injected 4 days before antigen caused a similar suppression, the effect of DCF was not mediated via a steroid release, inasmuch as DCF also suppressed the immune response in adrenalectomized mice. The response of BALB/c mice to TNP-Ficoll was also inhibited by DCF pretreatment and enhanced by injection of DCF after antigen. In contrast, in athymic mice DCF caused suppression of the anti-TNP-Ficoll PFC response, whether injected before or after antigen. These results are interpreted as suggesting that DCF causes suppression primarily via an effect on B cells. The enhancement seen in normal but not in athymic mice may possibly be ascribed to an effect on suppressor T cells. Apparently the enhancement of both TD and TI responses caused by DCF injected 1 day after antigen in normal mice is the net result of these two opposing effects. The results imply that helper T cells are resistant to DCF.     

Delayed-type hypersensitivity response in an isogenic murine model of paracoccidioidomycosis

The specific delayed-type hypersensitivity (DTH) response was evaluated in resistant (A/SN) and susceptible (B10.A) mice intraperitoneally infected with yeasts from a virulent (Pb18) or from a non-virulent (Pb265)Paracoccidioides brasiliensis isolates. Both strains of mice were footpad challenged with homologous antigens. Pb18 infected A/SN mice developed an evident and persistent DTH response late in the course of the disease (90th day on) whereas B10.A animals mounted a discrete and ephemeral DTH response at the 14th day post-infection. A/SN mice infected with Pb265 developed cellular immune responses whereas B10.A mice were almost always anergic. Histological analysis of the footpads of infected mice at 48 hours after challenge showed a mixed infiltrate consisting of predominantly mononuclear cells. Previous infection of resistant and susceptible mice with Pb18 did not alter their DTH responses against heterologous unrelated antigens (sheep red blood cells and dinitrofluorobenzene) indicating that the observed cellular anergy was antigen-specific. When fungal related antigens (candidin and histoplasmin) were tested in resistant mice, absence of cross-reactivity was noted. Thus, specific DTH responses againstP. brasiliensis depend on both the host's genetically determined resistance and the virulence of the fungal isolate.Abbreviations DTH delayed-type hypersensitivity - DNFB dinitrofluorobenzene - FN18 Fava Netto's antigen obtained from isolate Pb18 - FN265 Fava Netto's antigen obtained from isolate Pb265 - SRBC sheep red blood cells