Immunomodulatory effects of alcbholic and hydroalcoholic extracts of Moringa olifera Lam leaves

Effects of 50, 100 and 200 mg/kg body weight of the alcoholic and hydro-alcoholic extract of leaves of M. olifera were studied on various immune paradigms like delayed type hypersensitivity reaction using SRBC as an antigen, determination of antibody titer, neutrophil adhesion test as an indicator for neutrophil index, total leucocyte count in cyclophosphamide induced immunosuppressed animals and carbon clearance assay as a measure of phagocytic activity. Hydro-alcoholic extract of M. olifera substantially enhanced cellular immune response, humoral immune response, neutrophil index and phagoctic activity in doses of 100 and 200 mg/kg body weight. The ethanolic extract (200 mg/kg body weight) was efficient in improving immune response. The results suggest that M. olifera has a significant role to play as an immune stimulator.     

The effects of diabetes with hyperlipidemia on P450 expression in APA hamster livers

The effect of chronic hyperglycemia and hyperlipidemia induced by streptozotocin (SZ) on the expression of P450 in the liver of APA hamsters was studied in this experiment. No effect on the total activity of P450 was seen in SZ-induced diabetic hamsters throughout the experimental period. At 1 and 6 months after SZ-injection, the levels of CYP1A, 2C6, and 3A of SZ-injected hamsters were much lower than those of age-matched control hamsters. CYP2B expression tended to decrease and CYP2E1 and 4A expression tended to increase in SZ-injected hamsters, although the results were not significant. At 3 months after SZ-injection, however, no significant difference between SZ-injected and normal hamsters was seen in these P450 isozymes. On the other hand, CYP2C11 expression was slightly depressed in SZ1M and SZ6M, and almost equivalent to control hamsters in SZ3M. Immunohistochemistry by the use of each isozyme antibody revealed that SZ-induced diabetes affected the localization of CYP2C6, 3A, and 4A in the hepatic acinus. The expression of CYP2C6 and 3A was depressed mainly in the periportal region of the acinus, and CYP4A expression was induced mainly in the perivenous region by SZ-induced diabetes. On the other hand, the expression pattern of CYP1A, 2B, 2C11, and 2E1 were not affected. These results demonstrate that the effects of SZ-induced diabetes on hepatic P450 differ for each isozyme in APA hamsters and also differ from those of other experimental diabetic animals, including golden hamsters.     

Effects of different levels of selenium on growth performance and immunocompetence of broilers under heat stress

An experiment was undertaken to evaluate the effect of dietary selenium (Se) levels on growth performance and immune competence of broilers under heat stress. Birds were raised in either a thermoneutral (TN, 23.9°C constant) or heat stress conditions (HS, 23.9°C to 38°C cycling) and were fed a corn-soybean meal basal diet supplemented with Se at 0, 0.2 and 0.4 mg/kg. A total of 240 one-day-old male broiler chicks were randomly assigned to six groups; each group had four replicates of 10 birds. Body weight and feed intake were not influenced by dietary Se, while feed conversion was significantly improved by a Se-supplementation of 0.2 mg/kg. HS significantly reduced body weight, feed intake and feed conversion. Numbers of abdominal exudate cells (AEC), percentage of macrophages in AEC, phagocytic macrophages, internalized opsonised and unopsonised sheep red blood cells (SRBC) were significantly increased by dietary Se. Both primary and secondary antibody responses were characterised by increasing titres of antibody to SRBC by dietary Se when birds were exposed to HS (p < 0.05). Lymphoid organ weights, antibody responses, incidence of macrophages in AEC, and phagocytic ability of macrophages were also significantly reduced under HS. These results indicated that HS severely reduced growth performance and immunocompetence of broilers, whereas the immune response of broilers improved by dietary Se supplementation under HS.     

Dietary vitamin A can improve immune function in heat-stressed broilers

This experiment was undertaken to evaluate the effect of dietary vitamin A on the performance and immune competence of broilers under heat stress (HS). A total of 180 birds, at 22 days of age, were randomly assigned to be reared either at 24°C (thermoneutral, TN, 24°C, constant) or 24°C to 38°C (heat stress, HS, cycling) until the age of 42 days. Birds were then supplemented with vitamin A at 750, 1500, 15 000 IU/kg. Each of the 2 × 3 factorially arranged treatments were replicated in six cages, each containing five birds. Humoral immunity was assessed by intravenous injection of 7% sheep red blood cells (SRBC) followed by evaluation of serum for antibody titers in primary and secondary responses. Cell-mediated immunity was assessed by using a Sephadax stimulation method to recruit abdominal exudate cells (AEC) to evaluate macrophage phagocytic ability. Body weight (BW) and feed conversion were significantly affected by dietary vitamin A (P < 0.05). HS significantly reduced BW, feed intake and feed conversion (P < 0.05). Numbers of AEC, percentage of macrophages in AEC, phagocytic macrophages, internalized opsonized and unopsonized SRBC were increased by dietary vitamin A (P < 0.05). Both primary and secondary antibody responses were characterized by increasing titers of antibody to SRBC by dietary vitamin A when birds were exposed to HS (P < 0.05). Lymphoid organ weights, antibody responses, incidence of macrophages in AEC and phagocytic ability of macrophages were all significantly reduced under HS. These results indicated that HS severely reduced performance and immunocompetence of broilers, whereas the immune response of broilers improved by dietary vitamin A supplementation under HS.     

Immunomodulatory effects of two sapogenins 1 and 2 isolated from Luffa cylindrica in Balb/C mice

Two Triterpenoids (sapogenins 1 and 2) isolated from Luffa cylindrica were subjected to immunomodulatory activity in male Balb/c mice. Mice were treated with three doses of sapogenins 1 and 2 (10, 30 and 100 mg/kg) and levamisole (2.5 mg/kg) used as a standard reference drug for 15 days. Immune responses to T-dependent antigen SRBCs were observed using parameters like HA, PFC, DTH, lymphocyte proliferation and phagocytosis. As regards these parameters, sapogenins 1 and 2 elicited a significant increase in the HA, PFC and DTH response at dose 10 mg/kg (P<0.01) and 100 mg/kg (P<0.001), respectively. Sapogenins 1 and 2 also showed significant dose-dependent decrease and increase in lymphocyte proliferation assay and phagocytic activity of macrophages. Overall, sapogenins 1 and 2 showed dose relative immunostimulatory effect on in vivo immune functions in mice.     

Effect of modified low density lipoproteins on humoral immune response and functional activity of macrophages in mice

Intravenous injection of acetylated low density lipoproteins (acLDL) in mice in a dose of 0.5 mg per mouse decreased the intensity of humoral immune response to sheep red blood cells (SRBC) by 35%. The addition of acLDL to mouse peritoneal macrophages in vitro resulted in inhibition of Fc-dependent phagocytosis of SRBC and fourfold increased secretion of prostaglandins E2 by macrophages. Fc-dependent phagocytosis of SRBC was also found to be inhibited by oxysterols (25-hydroxycholesterol and 7-ketocholesterol), added to the incubation medium of macrophages in vitro in doses of 0.5-5 mg/ml. The conclusion was made that oxidative metabolism of cholesterol and arachidonic acid, contained in LDL, may mediate the immunomodulating effects of modified LDL.     

Supressor activity of spleen cells during medically induced immunologic tolerance]

SRBC tolerance was induced in mice (CBA X C57BL/6) F1 by single intraperitoneal injection of 6 X 10(9) SRBC and of cyclophosphamide (100-200 mg/kg) in 44-46 hours. Spleen cells of tolerant mice obtained at various periods after the tolerance induction (in 12-26 days) failed to decrease their immune response to SRBC after administration to intact syngeneic recipients. Contrary to intact mice, tolerant animals were incapable of producing suppressor cells after a single SRBC immunization. Only when 3 additional injections of high SRBC doses (6 X 10(9)) were given to tolerant mice the spleen cells in them acquired the capacity to inhibit the immune response after administration to normal mice. It is supposed that the absence of suppressor cells in induction of the immunological tolerance by means of cyclophosphane was caused by the processes of clone elimination. Suppressor cells can originate in tolerant animals under the effect of intensive antigenic stimulation, this leading to enhancement of the tolerance state as a result of additional SRBC injections.     

Immune response in experimental animals immunized with Burkholderia pseudomallei surface antigens

The influence of the chromatographic fractions of B. pseudomallei surface antigenic complex (C, C1, D, H) on immune response in white rats and white mice was under study. These antigenic complexes were noted to produce perceptible stimulating effect on the immune system of white rats, in contrast to that of white mice. The immunization of the mice the above-mentioned fractions suppressed the phagocytic activity of peritoneal macrophages (PM) and slightly enhanced cell-mediated immunity. In experiments on white rats, fraction C induced the growth of specific antibody titers and stimulated the phagocytic activity of PM, as well as the indices of delayed hypersensitivity (DH). Fraction D showed a lower level of the induction of the phagocytic activity of PM and was inactive in the manifestation of cell-mediated immunity, but induced a high level of humoral immunity. Antigenic complexes C1 and H increased the phagocytic activity of PM and DH characteristics with a low level of antibody production. The studied fractions of the causative agent of melioidosis decreased the content of bactericidal cationic proteins (BCP) in rat blood neutrophils, and in mice a decreased content of BCP in phagocytes was registered. The fractions increased the activity of myeloperoxidase in blood neutrophils in mice and rats. As revealed with the use of immunoelectrophoresis, SDS PAAG electrophoresis and immunoblotting, the surface antigenic complex contained proteins of 18, 22, 39 kD and glycoproteins 42, 55, 90 kD. The latter glycoprotein was found in all the fractions under study, having protective properties.     

Target cells for the activity of a synthetic adjuvant: muramyl dipeptide.

Muramyl dipeptide (MDP), a synthetic adjuvant, increased the primary response of CBA mice to sheep red blood cells (SRBC). In reconstituted irradiated recipients, cooperation between T and B lymphocytes was required for the expression of adjuvant activity and MDP increased the efficiency of SRBC-educated T cells. The role of T-derived lymphocytes in mediating the MDP adjuvant activity was also demonstrated in irradiated mice and in mice reconstituted with various splenic cellular types of donors which had received SRBC and MDP 24 hr earlier. In our experiments, the macrophage did not seem to be involved, since MDP did not increase the phagocytic capacity of peritoneal exudate cells and MDP- and SRBC-pretreated macrophages had no increased ability to induce an anti-SRBC immune response. These results demonstrate the importance of T lymphocytes as mediators of the adjuvant activity of MDP.     

The effect of vitamin B6 deficiency on cytotoxic immune responses of T cells, antibodies, and natural killer cells, and phagocytosis by macrophages

The effect of vitamin B6 on cytotoxic immune responses of T cells, natural killer (NK) cells, cytotoxic antibody production, and macrophage phagocytosis was assessed in 5-week-old female C57B1/6 mice. Mice were fed 20% casein diets with pyridoxine (PN) added at 7, 1, 0.1, or 0 mg/kg diet, which represents 700, 100, 10, and 0% of requirement, respectively. Compared to mice fed 7 or 1 mg PN diet, animals fed 0 or 0.1 mg PN diet showed significantly reduced primary splenic and peritoneal T-cell-mediated cytotoxicity (CMC). Animals fed 0 mg PN diet also showed significantly depressed secondary T CMC of splenic and peritoneal lymphocytes against P815 tumor cells. Complement-dependent antibody-mediated cytotoxicity against P815 cells, phagocytosis of SRBC by macrophages, and native and interferon-induced NK cell activities against YAC cells were not affected by the level of vitamin B6 intake. The percentage of macrophages present in the peritoneal exudate cells was increased in animals fed the 0 mg PN diet. The immune responses were not enhanced or altered by the excess intake of vitamin B6 (7 mg PN). It appears that vitamin B6 is an essential nutrient for maintenance of normal T-cell function in vivo.     

Macrophages of athymic nude mice: Fc receptors, C receptors, phagocytic and pinocytic activities

Expression of Fc receptors (FcR) for IgG1, IgG2A, IgG2B, IgM, IgA and IgE, binding of C3 and C5 complement components and phagocytic and pinocytic activities were determined in peritoneal and omental macrophages of nu/nu, nu/+ and +/+ Balb/c mice. nu/nu mice showed a higher proportion of FcR and complement receptor-bearing peritoneal macrophages along with a significantly higher phagocytic activity of peritoneal macrophages both in vitro and in vivo. Tests of pinocytic activity in these cells and phagocytic activity in omental phagocytes yielded similar results. We conclude that athymic mice compensate their immune defects by a higher phagocytic activity of their professional phagocytes and a higher expression of receptors mediating this process.     

Immunomodulatory activity of Sulforaphane, a naturally occurring isothiocyanate from broccoli (Brassica oleracea)

The effect of Sulforaphane on the immune system was studied using BALB/c mice. Intraperitoneal administration of five doses of Sulforaphane (500 microg/dose/animal/day) was found to enhance the total WBC count (12,950 cells/mm3) on 9th day. Bone marrow cellularity (23 x 10(6) cells/femur) and number of alpha-esterase positive cells (1346.66/4000 cells) were also increased by the administration of Sulforaphane. Treatment with Sulforaphane along with the antigen, sheep red blood cells (SRBC), produced an enhancement in the circulating antibody titre and the number of plaque forming cells (PFC) in the spleen. Maximum number of PFC (315.83 PFC/10(6) spleen cells) was obtained on the 6th day. Administration of Sulforaphane also showed an enhancement in the phagocytic activity of peritoneal macrophages. Moreover administration of Sulforaphane significantly reduced the elevated level of TNF-alpha production by LPS stimulated macrophages. These results indicate the immunomodulatory activity of Sulforaphane.     

Modulation of the immune response by anaphylatoxin in the microenvironment of the interacting cells

It was shown that the anaphylatoxins C3a and C5a can modulate in vitro immunological reactivities. C3a suppresses both the in vitro polyclonal antibody response and the specific antibody response to sheep red blood cells (SRBC) of both mouse spleen cells and human peripheral blood cells. The target cell in the mouse for C3a appears to be an Lyt-1+2- suppressor-inducer cell and macrophages appear not to be required. In contrast to C3a, C5a enhances in vitro responses of mice. Both the response to SRBC and the mixed lymphocyte reaction are enhanced by C5a. This enhancement appears to be through an Ia- macrophage that contains receptors for C5a. It appears that enhancement may be brought about by interleukin 1, which is released when Ia- macrophages are pulsed with C5a. It is suggested that these anaphylatoxins, when present in high concentrations in the microenvironment of the interacting cells of the immune system, play a dynamic role in the regulation of the immune response. Peptide fragments cleaved from the Fc portion of antibody, complexed with antigen in this microenvironment, may have a similar regulating role.     

Changes in the phagocytic activity of macrophages under the action of different doses of antibiotics]

Penicillin, streptomycin and gentamicin, when introduced into mice in a single injection in doses of 0.5-500,000 U/kg, produced different changes in the phagocytic activity of mouse peritoneal macrophages. Bactericidal doses of these antibiotics either suppressed the activity of macrophages or left it unchanged. Their sub-bactericidal doses enhanced the phagocytic activity of macrophages. Combined use of penicillin and streptomycin produced a synergic effect.     

Antigenic markers on mouse lymphoid cells: origin of cells mediating immunologic memory

Specific antisera were used for the purification of thymus dependent and thymus independent or bursa equivalent lymphoid cells in the mouse. Spleen cells from mice immune to sheep erythrocytes, a thymus dependent antigen, or to E. coli 055:B5 lipopolysaccharide, a thymus independent antigen, were treated with anti-θ (C3H) serum or anti-MBLA serum and complement prior to their adoptive transfer into lethally irradiated syngeneic recipients. Syngeneic thymocytes, bone marrow cells, or spleen cells from nonimmune donors were appropriately added to antiserum treated cells prior to transfer. The secondary response to these antigens was assayed in recipient spleens six days after cell transfer. The kinetics of the primary response to SRBC was investigated as to its effect on origin of specific hyper-reactive T or B lymphoid cells.The adoptive response to CPS originated in the B lymphoid cell population. Immunologic memory to CPS was demonstrated in recipients of immune cells, compared to recipients of normal cells, by a five fold increase in antibody forming cells.The IgM and IgG adoptive immune response to high doses of SRBC depended upon an increased number of specifically hyper-reactive T-lymphoid cells to facilitate cooperation between T and B lymphocytes. High doses of SRBC initially stimulated T cell memory but at 42 days after priming an increased number of specifically hyper-reactive B lymphoid cells were present.     

Role of antigen-specific B cells in the induction of SRBC-specific T cell proliferation

In this study, we ask whether antigen presentation can be effected by antigen-activated B cells. Antigen-dependent in vitro proliferation of T cells from mice primed with SRBC or HoRBC occurs in the presence of B cells primed to the relevant antigen. B cells prepared from lymph nodes of mice primed with irrelevant antigens are not effective antigen-presenting cells for RBC-specific T cell proliferation over a wide range of SRBC doses. This is true even when both RBC and the antigen to which the B cells are primed are included in the culture. In contrast, B cells specific for a hapten determinant coupled to SRBC are able to support proliferation of T cells specific for SRBC determinants. We conclude from these data that antigen-specific B cells play a role in the induction of T cell proliferative responses to SRBC and HoRBC antigens. Two models are proposed: either B cells, upon antigen interaction with surface antibody, are able to act as accessory cells to induce Ia-dependent proliferation of immune T cells; or B cells augment the T cell proliferative response by secretion of antibody, leading to opsonization of the antigen for macrophage uptake and presentation.     

Splenopentin--modulator of immunological and behavioral reactions in secondary immunodeficiency state induced by experimental alcoholism

Effect of splenopentin on some patterns of immunity was studied in mice with chronic alcoholic intoxication. Splenopentin was administrated into animals once intraperitoneally (250 micrograms/kg). Administration of splenopentin was found to normalize several immunological patterns in animals with chronic alcoholic intoxication: the immune response to the thymus-dependent antigen sheep red blood cells and phagocytic activity of peritoneal macrophages. Also observations over C57B1/6 mice characterized by high level of alcoholic motivation showed that alcohol consumption in mice decreased after administration of splenopentin at a dose of 250 micrograms/kg during two weeks.     

凝结芽胞杆菌对免疫功能影响的实验研究

目的 研究凝结芽胞杆菌（TQ33）对实验动物免疫功能模型的影响。方法 以鸡红细胞混悬液对小鼠腹腔注射,观察TQ33对小鼠腹腔巨噬细胞吞噬功能的影响;制备绵羊红细胞（SRBC）,观察血球凝集程度,测定血清溶血素;以靶细胞（YAC-1细胞）与脾细胞（效应细胞）的反应检测TQ33。对小鼠NK细胞活性的影响;采用淋巴细胞转化法观察TQ33对细胞免疫的影响;计算小鼠胸腺／体重及脾脏／体重为脏器系数观察TQ33对小鼠脏器的影响。结果 与对照组比较,TQ33显著增加小鼠腹腔巨噬细胞吞噬功能;对小鼠体液免疫功能有一定的增强作用;可增强小鼠NK细胞活性;对ConA诱导下的小鼠淋巴细胞转化有增强作用;对脏器系数没有显著的影响。结论 凝结芽胞杆菌具有明显的免疫调节作用,预示有良好的应用前景。     

Trypanosoma cruzi: Correlation of resistance and susceptibility in infected inbred mice with the in vivo primary antibody response to sheep red blood cells

Nonspecific immune responses during the course of murine Trypanosoma cruzi infection were examined in mouse strains genetically resistant or susceptible to the Brazil strain of T. cruzi. Spleen cells from infected susceptible (C3H) or resistant [C57 B1/10 and FI (C3H × C57)]mice at various points during the course of infection exhibited a reduced response to concanavalin A and lipopolysaccharide in vitro. Since this reduced response occurred in both susceptible and resistant mice, it was not predictive of resistance or susceptibility in vivo. We next examined the kinetics of in vivo primary antibody response to sheep red blood cells (SRBC) in infected C3H and C57 mice. C3H mice exhibited inhibition of the direct plaque-forming cell assay (d-PFC) which persisted until death. In contrast C57 mice exhibited no inhibition of the response at Day 5 and subsequently a markedly augmented response was observed. Other strains of mice were similarly investigated: all the susceptible mice examined (A/J, BALB/c) showed inhibition or depression of the primary antibody response and resistant mice [B10Br, C57B1/10, SJL, F1 (C3H × C57)]demonstrated either no inhibition or considerable augmentation of this response. CS7 mice resistant to the Brazil strain were susceptible to the Tulahuén strain. The mice in this latter group exhibited a markedly significant inhibition of the in vivo primary antibody response to SRBC. Culture forms of the Brazil strain protected C3H mice from a virulent challenge. This immunization resulted in a markedly augmented antibody response. The data reported herein are consistent with the notion that inhibition of the primary antibody response to SRBC correlates with susceptibility whereas no inhibition or, indeed, augmentation of the response correlates with natural as well as acquired resistance.     

The suppressive effect of peritoneal exudate macrophages on production of antibody to sheep erythrocytes in vitro

The effects of peritoneal exudate macrophages on antibody response to sheep erythrocytes (SRBC) were investigated in mice. Peritoneal exudate macrophages obtained from mice injected intraperitoneally with proteose peptone or Corynebacterium parvum 4 days earlier had stronger ability to phagocytize and degrade SRBC than normal resident macrophages. These macrophages suppressed antibody formation to SRBC in vitro as well as in vivo. This suppression was overridden by increasing the amount of SRBC and diminished completely by pretreatment of the macrophages with iodoacetate and partly by pretreatment with 2-deoxyglucose, both known to be inhibitors of phagocytosis, but not by addition of indomethacin to the in vitro culture. These results suggest that the suppression of antibody response by peritoneal exudate macrophages was due to the increased activity of these cells as scavenger cells, resulting in a reduced amount of effective antigenic stimulation, and that it was not mediated by a prostaglandin-dependent mechanism. The scavenger function of these macrophages may be due to Ia-negative macrophages.