Immunomodulatory activity of biopolymeric fraction RLJ-NE-205 from Picrorhiza kurroa

In the last three decades, numerous biopolymeric fractions have been isolated from medicinal plants and used as a source of therapeutic agents. The most promising biopharmacological activities of these biopolymers are their immunomodulatory effects. The biopolymeric fraction RLJ-NE-205 was isolated and purified from the rhizomes of Picrorhiza kurroa. We evaluated the effects of biopolymeric fraction RLJ-NE-205 from P. kurroa on the in vivo immune function of the mouse. Balb/c mice were treated with the biopolymeric fraction RLJ-NE-205 (12.5, 25 and 50 mg/kg body weight) for 14 days with sheep red blood cells (SRBC) as an antigen. Haemagglutination antibody (HA) titre, plaque forming cell (PFC) assay, delayed type hypersensitivity (DTH) reaction, phagocytic index, proliferation of lymphocytes, analysis of cytokines in serum and CD4/CD8 population in spleen (determined by flowcytometry) were studied. At the dose of 50 mg/kg, significant increases in the proliferation of lymphocytes (p<0.001) and cytokine levels (IL-4 and IFN-gamma) in serum (p<0.001) were observed. A dose dependent increase was demonstrated in HA titre (p<0.05), DTH (p<0.01), PFC (p<0.05), phagocytic index (p<0.05) and CD4/CD8 (p<0.01) population. This suggests that the biopolymeric fraction RLJ-NE-205 improves the immune system and might be regarded as a biological response modifier.     

The effect of salazosulfapyridine on the in vitro antibody production in murine spleen cells

The effect of salazosulfapyridine (SASP) on the antibody response of murine spleen cells in vitro was studied. SASP inhibited the response to sheep red blood cells (SRBC), a T-cell-dependent (TD) antigen, dose-dependently and was most effective at a dose of 2 x 10(-4) M without cell toxicity. No remarkable inhibition was seen with the main metabolites of SASP, 5-aminosalicylic acid (5-ASA) and sulfapyridine (SP). SASP failed to inhibit antibody production to T-cell-independent antigens such as dinitrophenyl-Ficoll or trinitrophenyl (TNP)-lipopolysaccharides, although the response to TNP-keyhole limpet hemocyanin, another TD antigen like SRBC, was inhibited. Further, this drug did not show any depression of the anti-SRBC plaque-forming cell (PFC) response in spleen cells treated with anti-Thy1.2 antibody plus complement. The inhibition of anti-SRBC PFC response by SASP was accompanied by a reduction of interleukin 2 (IL-2) secretion. Our results suggest that SASP may act on T cell populations and may inhibit the T-cell-dependent antibody response partly through a depression of IL-2 production. The active compound appears to be SASP itself, rather than its metabolites.     

Antibody responses in hyperthyroid rats

This study evaluated antibody production against sheep red blood cells (SRBC) in hyperthyroid rats during treatment with triiodothyronine (T(3)). The immune response was evaluated by measuring plaque forming cells (PFC) in the spleen and by enzyme-linked immunosorbent assay (ELISA) in serum of male Wistar rats (180+/-10 g) treated with 25 mug/day of triiodotironine (T(3)) during 7-12 days and immunized with SRBC at the 8th day of treatment. The results showed that anti-SRBC antibody production was significantly decreased in animals treated for 12 days when compared to normal rats immunized with the same antigen, as evaluated by the two assays. These results show that in this experimental model hyperthyroidism decreases antibody response. We previously observed the opposite effect, that is, an increase in this response in hypothyroid rats resulting from the treatment with propylthyouracil, a blocker of thyroid hormone biosynthesis. It is suggested that antibody production is affected by thyroid hormone levels.     

903中药复方煎剂对小鼠体液免疫的增强作用

903中药复方煎剂对小鼠脾细胞的PFC数、血清溶菌酶活性和抗SRBC的溶血性抗体水平均有明显增强,尤其是PEC数比正常对照组的增强更为明显(P<0.01)。此外,本煎剂能对抗环磷酰胺(CY)对小鼠免疫功能的抑制作用。提示903中药复方煎剂对小鼠的体液免疫有增强作用。     

Reduced T-helper cell function in mice exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD).

Antibody production to the T-dependent antigen SRBC is highly sensitive to suppression by polychlorinated dibenzo-p-dioxins. The present study provides evidence for a defect in T-helper (TH) cells in TCDD-exposed mice. Because spleen cells from nonimmune TCDD-exposed mice did not show suppressed antibody responses when adoptively transferred to irradiated hosts, we used a hapten-carrier (TNP-SRBC) system with cell separation/reconstitution techniques to determine the effects of TCDD on carrier-specific TH cells. In vitro cultures of spleen cells from SRBC-primed TCDD-treated (5 micrograms/kg) mice produced fewer anti-TNP plaque-forming cells (PFC) when immunized with TNP-SRBC, as compared to cells from primed vehicle-treated controls. A reduced anti-TNP PFC response was also observed in experiments where non-immune B-cells were induced to produce anti-TNP PFC by TH-cells obtained from carrier-primed TCDD-exposed mice, as compared to carrier-primed vehicle-exposed mice. Removal of Lyt-2+ (suppressor) T-cells in these experiments did not alter the anti-TNP PFC response. These results provide direct evidence for reduced activity of TH-cells after exposure to TCDD.     

SA3443, a novel cyclic disulfide compound, depresses anti-SRBC antibody-forming cell responses in the mouse through inhibition of antigen-presenting cell activities.

(4R)-Hexahydro-7,7-dimethyl-6-oxo-1,2,5-dithiazocine-4-carboxylic acid (SA3443) is a newly synthesized cyclic disulfide compound which has potential hepatoprotective properties. The effect of SA3443 on the induction of anti-sheep red blood cell (SRBC) plaque (antibody) forming cell (PFC) responses was investigated in vivo and in vitro. SA3443 (approximately 3 mg/kg/day) remarkably decreased the number of anti-SRBC PFC in the spleens of mice immunized with a high dose of SRBC in vivo. The addition of SA3443 (approximately 1 x 10(-7) M) at the initiation of mouse spleen cell cultures in vitro also exerted a significant inhibitory effect on subsequent PFC response to SRBC, and removal of SA3443 after 24 h did not reverse its inhibitory effect. Pre-incubation of isolated adherent spleen cells with SRBC and SA3443 resulted in a similar inhibition of subsequent PFC response, but a pre-incubation of macrophage-depleted cells with SRBC and SA3443 or a pre-incubation of the unseparated spleen cells with SA3443 in the absence of SRBC had no effect. These findings have suggested that SA3443 may depress antibody response through inhibition of macrophage antigen-presenting cell activity.     

Effect of traxanox on antibody formation in C57BL/6 mice

C57BL/6 mice, lower responders to sheep red blood cells (SRBC), were intraperitoneally immunized with 5 X 10(8) SRBC on day 0. Traxanox (10 and 30 mg/kg) administered orally on days 0 and 1 potentiated the production of spleen- and thymus-rosette forming cells (RFC) assessed on day 7. The production of hemolytic plaque forming cells (HPFC) to SRBC in the spleen of the syngeneic recipient mice assessed on day 4 was inhibited by the transfer of spleen-RFC obtained from the vehicle-treated donor mice, but not by that obtained from the traxanox (30 mg/kg)-treated donor mice. The same results were obtained in the thymectomized-recipient mice. The activity of the spleen-RFC obtained from the vehicle-treated donor mice was abolished by treatment with anti-Thy 1.2 or anti-Lyt 2.2 antibody and complement. On the other hand, the activity of the spleen-RFC obtained from the traxanox-treated donor mice was abolished by treatment with anti-Lyt 1.2 antiserum and complement. Traxanox (3 and 30 mg/kg) also caused the induction of the Thy 1.2-positive RFC in the spleen of the thymectomized mice. These results suggest that traxanox has a capacity to potentiate the immune responses to SRBC in C57BL/6 mice by the induction of Lyt 1.2-positive cells (helper T cells).     

Suppression of primary antibody response by a single exposure to cadmium in mice

Suppression of primary antibody response to sheep red blood cells (SRBC) by a single intraperitoneal (i.p.) injection of cadmium into mice was investigated by the methods of in vitro plaque-forming cell (PFC) assay. BALB/c mice were given 1.8 mg cadmium/kg body weight, and 1, 2 or 7 days later, spleen cells from exposed and control mice were cultured with SRBC. PFC responses of all exposed groups were significantly suppressed compared to those of control groups. Addition of control adherent cells to spleen cells from exposed mice failed to recover control level. In the cell-reconstitution experiments, the activity of B-cell function from the exposed group was suppressed more by cadmium than that of T-cell function. These results suggest that the suppression of primary PFC response by cadmium exposure may be caused by the inactivation of B-cells.     

Effect of diisopropylfluorophosphate on the antibody response

The impact of the in-vivo administration of diisopropylfluorophosphate (DFP) on the antibody response in mice was examined. The antigen-specific plaque-forming cell (PFC) response of mice injected with DFP (4 mg/kg) at the time of immunization with a macrophage-dependent T-cell antigen, trinitrophenylated keyhole limpet hemocyanin (TNP-KLH) or sheep red blood cells (SRBC), was significantly depressed. In contrast, the injection of a similar dose of DFP had no effect on the PFC response of mice immunized with a macrophage-independent B-cell antigen, lipopolysaccharide (LPS). This differential effect suggests a possible detrimental effect of DFP on cells involved in antigen processing and/or presentation which are required for the antibody response to SRBC and TNP-KLH. These results, however, do not exclude T cells as a possible target and the cellular targets of DFP action remain to be established. The DFP-mediated depression of the PFC response to SRBC was still evident when DFP was given as early as 1 day before, but not 1 day after, the injection of antigen, suggesting that DFP may well affect an early event of the antibody response. For the secondary TNP-specific IgM PFC response, a significant depression was observed only when DFP was administered at the time of antigen challenge. The injection of DFP at the time of antigen priming, however, was effective in depressing the IgG, but not the IgM, PFC response. Therefore, DFP may also interfere with the generation of antigen-specific memory cells for the secondary IgG response.     

Anti-inflammatory action of Cudrania tricuspidata on spleen cell and T lymphocyte proliferation

This study examined whether an extract of Cudrania tricuspidata shows anti-proliferative effects in anti-CD3/CD28-mediated spleen and CD4+CD25- T cells and decreases the production of the proinflammatory cytokines interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) in anti-CD3/CD28-mediated CD4+CD25- T cells. The proliferation of anti-CD3/CD28-mediated spleen cells and CD4+CD25- T cells was effectively suppressed by C. tricuspidata. This extract, however, did not show cytotoxicity in spleen cells under conditions where the antigen was not stimulated using CCK-8 analysis. C. tricuspidata also decreased the production of the pro-inflammatory cytokines IL-2 and IFN-gamma by selective inhibition of this extract on proliferating cells in anti-CD3/CD28-mediated CD4+CD25- T cells. These results suggest that C. tricuspidata may be useful in the treatment of autoimmune diseases and organ transplantation through the inhibitory action of T cells in inflammation.     

Effects of an orally applied aqueous-ethanolic extract of a mixture of Thujae occidentalis herba, Baptisiae tinctoriae radix, Echinaceae purpureae radix and Echinaceae pallidae radix on antibody response against sheep red blood cells in mice

The influence of the oral administration of an aqueous-ethanolic extract of a mixture of Thujae occidentalis herba, Baptisiae tinctoriae radix, Echinaceae purpureae radix and Echinaceae pallidae radix, on the immune response in mice was investigated. The data show that the extract significantly enhances the antibody response to sheep red blood cells (SRBC), induces an increase in the numbers of splenic plaque forming cells (PFC) and an increase in the titer of specific antibodies in the sera of treated animals. The long-term application of the extract over several months also stimulated the PFC-response without affecting spleen weight, total cell yield per spleen or white blood cell count in mice.     

Analysis of lead effects on in vivo antibody-mediated immunity in several mouse strains

The effect of administration of lead acetate (10 mM in the drinking water) for 8 weeks on the in vivo sheep red blood cell (SRBC) specific plaque-forming cell (PFC) responses of inbred A, BALB/c, C57Bl/6, DBA/1, SJL, and NZW/NZB F1 mice and outbred CFW mice was examined to determine if lead was immunomodulatory in a genetically related manner. Lead did not suppress the SRBC-specific PFC/10(6) splenocytes or PFC/spleen response in any mouse strain when compared to the responses of strain-matched control mice. In addition, 10 mM lead-treated BALB/c mice manifested augmented PFC/10(6) splenocytes (17%; p less than .05) but unchanged PFC/spleen responses. Correspondingly, serum concentrations of SRBC-specific antibody (measured by radioimmunoassay) and serum immunoglobulin G, M, or A isotypes were also unchanged by lead acetate treatment in all tested mouse strains. There were no observable lead-related histopathological changes or deposition of immune complexes or antibasement membrane antibody in the kidneys of treated mice. Further, splenocytes from lead-treated, SRBC-immunized mice cultured with T-independent antigens (TNP-LPS, TNP-Ficoll) or with a T-dependent antigen (SRBC) exhibited direct and indirect specific PFC responses that were unchanged from those of control mice. The H-2K/D haplotypes of the outbred CFW mice were determined by microcytotoxicity to include r, q, u, and s. These results suggest that lead acetate (10 mM) administered po for 8 weeks does not suppress the primary direct humoral immune response to SRBC in inbred and outbred mice of several H-2 haplotypes (k/d; d; b; q; d,z; s; r; and u).     

Anti-CD40 Treatment of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-exposed C57Bl/6 mice induces activation of antigen presenting cells yet fails to overcome TCDD-induced suppression of allograft immunity

We have previously demonstrated that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) suppressed the induction of the costimulatory molecule CD86 (B7-2) on B220+ and Mac-1+ spleen cells following the injection of allogeneic P815 tumor cells. In this study, TCDD exposure was shown to suppress CD54 and major histocompatibility complex (MHC) class II expression on B220+, Mac-1+, and CD11c+ splenic antigen presenting cells (APC). Furthermore, interleukin-12 (IL-12) production by spleen cells from P815-immunized mice was significantly decreased following exposure to TCDD. To determine if exogenous costimulation could enhance the activation of APC, vehicle- and TCDD-treated mice were injected with an agonistic antibody to murine CD40. Stimulation with anti-CD40 increased the expression of CD86, CD54, and MHC class II on splenic APC and greatly enhanced the production of interleukin-12. TCDD treatment had minimal effects on the anti-CD40-induced expression of accessory molecules on splenic APC. TCDD exposure had no effect on anti-CD40-induced IL-12 in the plasma but suppressed its production from cultured spleen cells. Surprisingly, although stimulation via CD40 increased the activation of APC, allograft effector functions were not restored in TCDD-treated mice, perhaps due to persistent defects in antigen processing and presentation, cytokine production, T cell function, or CD40-independent pathways of APC activation.     

Selective immunosuppression of prodigiosin 25-C and FK506 in the murine immune system.

The immunosuppressive effects of prodigiosin 25-C were studied in comparison with FK506. Both prodigiosin 25-C and FK506 suppressed T cell proliferation in response to concanavalin A (con A) or phytohemagglutinin (PHA) more significantly than that to lipopolysaccharide. However, prodigiosin 25-C inhibited con A-mediated mitogenic response more strongly than PHA-mediated one. FK506 showed no selectivity among those responses. In addition, when higher concentration of con A was used an inhibitory effect of prodigiosin 25-C became more evident whereas that of FK506 became less evident. Furthermore, prodigiosin 25-C affected neither interleukin-2 (IL-2) production nor IL-2 receptor (IL-2R) and transferrin receptor (TF-R) expression in vitro, though FK506 extensively inhibited IL-2 production and significantly suppressed IL-2R and TF-R expression. When comparing the effects of prodigiosin 25-C and FK506 in vivo by injecting antigens of different nature to a mouse, prodigiosin 25-C selectively inhibited cytotoxic T lymphocyte (CTL) activity induced by an allogenic mastocytoma, P815, without affecting production of antibody against a thymus dependent (TD) antigen, sheep red blood cell (SRBC). On the contrary, FK506 significantly inhibited both CTL induction and the antibody production. When Brucella abortus, a thymus independent (TI) antigen, and SRBC were simultaneously challenged to a mouse, neither prodigiosin 25-C nor FK506 affected antibody production against the TI antigen while the effect on the TD antigen were the same as described above. The present results revealed the unique immunosuppressive property of prodigiosin 25-C which was different from that of FK506.     

CHARACTERIZATION OF AN APPROACH TO DEVELOPMENTAL IMMUNOTOXICOLOGY ASSESSMENT IN THE RAT USING SRBC AS THE ANTIGEN

Although developmental immunotoxicology is an area of emerging importance, few data exist that compare profiles of activity in neonatal and adult rodents. One of the factors contributing to the paucity of comparative results is that it is unclear whether immunotoxicological test procedures optimized to evaluate the immune system of adults can be integrated into studies of younger animals. Therefore,the following objectives were addressed in this study for both male and female Crl:CD(SD)BR rat pups and weanlings: (1) to quantify baseline values for absolute and relative splenic lymphocyte populations using flow cytometry in nonimmunized animals; (2) to optimize and establish baseline values for the primary humoral immune response to sheep red blood cells (SRBC) using either an enzyme-linked immunosorbant assay (ELISA) or the plaque-forming cell (PFC) assay; and (3) to determine the impact of SRBC immunization on the histology of the spleen. Responses for each age group were compared both withinandbetween litters. Spleencell counts and absolute andrelative (to body weight) spleen and thymus weights were also obtained. Analysis by flow cytometry indicated the following overall mean absolute and relative (%) numbers of splenic lymphocytes in nonimmunized 10-day-old rats, respectively: 0.18 ± 0.08 × 10 8 (13 ± 5) CD45 + ; 0.13 ± 0.04 × 10 8 (9 ± 3) OX12 + ; 0.07 ± 0.01 × 10 8 (5 ± 1)W3/25 + CD3 + ;0.03 ± 0.01 × 10 8 (2 ± 0.5)OX8 + CD3 + cells. Because of the difficulty in administering SRBC by tail vein injection, the pups received SRBC intraperitoneally but were still unable to generate a primary IgM response. Histologically germinal centers were observed in rat pups immunized with SRBC. Taken together,these results suggest that the spleens of 10-day-old rats consist predominantly of functionally immature lymphocytes. Analysis by flow cytometry indicated the following mean absolute and relative(%)numbers,respectively, for nonimmunized 21-day-old rats: 0.96 ± 0.25 × 10 8 (48 ± 5) CD45 + ; 0.77 ± 0.16 × 10 8 (39 ± 3) OX12 + ; 0.23 ± 0.02 × 10 8 (12 ± 2) W3/25 + CD3 + ; 0.14 ± 0.02 × 10 8 (7 ± 1)OX8 + CD3 + cells. Results when using the ELISA indicated that the SRBC-specific IgM antibody log 2 titer for rat weanlings was considerably less than the titer obtained for young adult rats, whereas results with the PFC assay indicated a response in weanlings that was within the historical range of responses for adult rats. The latter results were consistent with the histological analysis in that prominent germinal centers were observed in weanlings immunized with SRBC. Based on our flow cytometric, ELISA, and PFC assay data, a litter can be used as the unit of comparison for developmental immunotoxicity. Our results with SRBC suggest that it may not bepossible toevaluatea humoral immune functional parameter in rat pups due to the apparent immature status of their immune cells; however, additional antigens must be examined. Results in weanlings suggest that an antibody response to SRBC of sufficient magnitude can be demonstrated with the PFC assay,but that a high background may preclude the use of the SRBC-specific IgM ELISA.     

Immunopharmacologic studies of D-penicillamine-L-cysteine disulfide

Effects of D-penicillamine-L-cysteine disulfide (P-C) on some immunological parameters were examined in normal and immunity-impaired mice and rats. P-C enhanced the DNA synthesis in concanavalin A-stimulated mouse spleen cell cultures in vitro. In vivo, administration of P-C produced either enhancement or depression of plaque forming cell (PFC) response and delayed type hypersensitivity (DTH) to sheep red blood cells (SRBC) in low responder mice to SRBC, depending on the dose of P-C. P-C restored the impaired PFC response in hydrocortisone-pretreated mice. The enhancing effect of P-C was not shown in high responder mice to SRBC, but an inhibiting effect was observed. P-C inhibited the suppressor cell induction on PFC response in mice immunized with a supraoptimum dose of antigen. In adjuvant arthritic rats, P-C induced severe arthritis by eliminating the suppressor cells regulating this disease process. The relevance of these findings and mode of action of D-penicillamine in rheumatoid arthritis is discussed.     

Immune alterations in rats following subacute exposure to tributyltin oxide

Adult male Fischer 344 rats were dosed by oral gavage with bis(tri-n-butyltin)oxide (TBTO) in peanut oil for 10 consecutive days, at dosages ranging from 1.25 to 15 mg/kg/day. Other groups of rats were dosed daily for 10 days by oral gavage with cyclophosphamide (CY) at dosages ranging from 0.75 to 6 mg/kg/day. These rats served as positive controls for the immune assays employed. The immune function parameters examined included the following: delayed-type hypersensitivity (DTH) and antibody responses to bovine serum albumin (BSA), primary antibody responses to sheep red blood cells (SRBC) and trinitrophenyl lipopolysaccharide (TNP-LPS) and enumeration of splenic lymphocyte populations. The DTH and antibody responses to BSA were not affected by TBTO exposure; however these responses were suppressed in rats dosed with CY at 6 mg/kg/day. The plaque forming cell (PFC) response to the T cell-dependent antigen SRBC was enhanced in rats dosed with TBTO at from 5 to 15 mg/kg/day. On the other hand, the PFC response to the T cell-independent antigen TNP-LPS was unaffected by TBTO exposure. Rats dosed with CY had suppressed PFC responses to SRBC and TNP-LPS at dosages of 3 and 6 mg/kg/day, respectively. Enumeration of splenic lymphocyte populations from TBTO-exposed rats revealed a reduction in OX8- but not W3/25- or IgG-positive cells. These results, as well as results from an earlier study from this lab, suggest that T lymphocytes are a primary target for TBTO-induced immune alterations and that the enhancement of the PFC response to SRBC in TBTO-exposed rats may be mediated by alterations in the suppressor (OX8-positive) T lymphocyte population.     

Effects of single exposure to cadmium on the primary humoral antibody response

Effects of a single intraperitoneal (i.p.) injection of cadmium (Cd) on the primary humoral antibody responses against sheep red blood cells (SRBC) in mice were studied by assaying splenic plaque forming cells (PFC). PFC responses in mice were suppressed when exposed to Cd 2 days after immunization, and inconsistently stimulated when exposed before immunization. Dose-response relationships were observed in the suppressive effect of Cd exposure 2 days after immunization, but not consistently in the stimulative effect of Cd exposure before immunization. Thymus weights and cell numbers decreased markedly 4 days after Cd exposure with or without the antigenic stimulus. Splenic weights increased 2 days after Cd exposure, while the number of spleen cells was dramatically decreased 1 days after Cd exposure and still remained below normal 2 days after exposure.     

Modification of immune response by cinnarizine

Effects of cinnarizine on immune response in mice were investigated. Mice were orally administered with cinnarizine and were immunized with sheep red blood cells (SRBC) intravenously. Numbers of plaque forming cells (PFC) to SRBC in spleen of these mice were assayed and delayed-type hypersensitivity (DTH) response to SRBC was measured. 1) PFC response in immunization with 5 X 10(6) cells/mouse of SRBC was enhanced by administration of 25 mg/kg of cinnarizine, while the response in immunization with 5 X 10(8) cells/mouse was suppressed by 25 to 200 mg/kg of cinnarizine. 2) From study on timing of administration, suppression of PFC response by 6.25 to 200 mg/kg of cinnarizine was observed at 24 hr. after the immunization. 3) 12.5 to 200 mg/kg of cinnarizine suppressed polyclonal B cell activation induced by lipopolysaccharide (LPS). 4) Colchicine induced suppressor T cell inactivation was prevented by administration of 50 mg/kg of cinnarizine and it was suggested that cinnarizine may induce suppressor T cells from the study of adoptive cell transfer system. 5) 50 mg/kg of cinnarizine showed the suppression of DTH response in expression phase, but not in induction phase. It was concluded that immune responses in mice were modified by cinnarizine.     

Immune alterations in mice exposed to the herbicide simazine

Simazine, a triazine herbicide, was investigated for its in vivo immunomodulatory properties. Male C57Bl/6 mice were treated with vehicle or 300 or 600 mg/kg body weight (bw) simazine daily orally for 4 wk. The immune system was evaluated by the antibody response to sheep red blood cells (SRBC; plaque assay and serum immunoglobulin G), natural killer (NK) and macro-phage activities, lymphocyte subpopulations in the spleen and thymus, and concanavalin A (Con A)- and lipopolysaccharide (LPS)-stimulated lymphocyte proliferation using splenocytes. Body weight and spleen and thymus weight decreased generally in simazine-treated mice, while the weight of adrenal glands was higher than in the control. Simazine treatment (600 mg/kg) induced an increase in the percentage of CD4(+) cells in spleen and CD8 + in thymus. Simazine inhibited the IgM plaque-forming cell numbers and lowered the level of IgG and the proliferation of mitogen-stimulated B cells and T cells. In addition, splenic NK and peritoneal macrophage activities in exposed mice were significantly decreased. Exposure to simazine also decreased cytokine production by macrophages, such as interleukin-1 (IL-1), IL-6, and tumor necrosis factor-alpha (TNF-alpha). Taken together, data indicate that the immune system was suppressed by oral simazine exposure.