Splenic Plaque-forming Cell Response Research Articles

Effects of selenium-chitosan on blood selenium concentration, antioxidation status, and cellular and humoral immunity in mice.

One hundred and eighty Kunming mice were allotted to three groups in a randomized complete block design, including two treatments and one control. Mice in group 1 were fed a basal diet as control, while mice in groups 2 and 3 were fed the basal diet supplemented with 0.2mg/kg selenium as sodium selenite (SS) or selenium-chitosan (SC), respectively. On day 28 of the experiment, blood selenium concentration, glutathione peroxidase (GPx) activity, plasma superoxide dismutase (SOD) activity, malondialdehyde (MDA) content, and Con A-induced splenocyte proliferation were determined, and plasma interleukin-2 (IL-2) and interferon-γ (IFN-γ) concentrations, splenic plaque-forming cell (PFC) responses, serum hemolysis level (HC50), and delayed-type hypersensitivity (DTH) responses were determined on day 15 of the experiment. The results showed that blood selenium concentration, GPx activity, splenic PFC response, and plasma IL-2 and IFN-γ concentrations in SC group were higher than those in the control and SS groups (P < 0.01 or P < 0.05), respectively. Plasma SOD activity, Serum hemolysis level, DTH responses, and Con A-induced splenocyte proliferation in SC group were higher than those in control (P < 0.01 or P < 0.05). Plasma SOD activity, serum hemolysis level, DTH responses, and Con A-induced splenocyte proliferation in SC group were also higher than those in SS group, while there was no significant difference between SC and SS groups (P > 0.05). Plasma MDA content in SC group was lower than those in the control and SS groups (P < 0.01 or P < 0.05). It is concluded that SC supplement can increase blood selenium concentration, antioxidation status, and cellular and humoral immunity, and SC has better biological activity than SS in mice.

Adenosine-deaminase-associated immunodeficiency. I. Differential sensitivities of lymphocyte subpopulations exposed to 2-deoxycoformycin in vivo.

In order to obtain a better understanding of the degree of immune dysfunctions caused by the absence of adenosine deaminase, we gave a single i.p. injection of 2'-deoxycoformycin (2-dcf), a potent inhibitor of the enzyme ADA at various doses into adult Syrian hamsters. These animals were examined for their ability to mount primary in vivo antibody responses to helper T cell dependent (Th-d) and helper T cell independent (Th-ind) antigens. Hamsters treated with 0.5 mg/kg of 2-dcf mounted enhanced splenic plaque-forming cell (PFC) responses to sheep erythrocytes, a Th-d antigen, and to pneumococcal polysaccharide type III (SIII), a Th-ind antigen. Treatment of animals with 1.0 mg/kg of 2-dcf resulted in a significantly depressed (P less than 0.001) PFC response to Th-d antigen, but a further enhanced response to Th-ind antigen. One mechanism which may be responsible for such a dichotomous response to these two types of antigens was selective dysfunction of T cell subpopulations. At higher doses (1.5-4.0 mg/kg), PFC responses to both types of antigens were significantly suppressed. Immunoenhancement at low doses of 2-def was attributed to an increased susceptibility of T suppressor cells to 2-dcf. This hypothesis was confirmed by priming the 2-dcf-treated animals with low-dose Th-ind antigens. These animals failed to induce low-dose tolerance by stimulation of antigen-specific suppressor T cell subsets. At low doses, B cells and T helper cell functions were found to be intact, as further confirmed by priming the animals with the carrier keyhole limpet haemocyanin (KLH) and challenging with trinitrophenyl-KLH. This dose-dependent selective susceptibility of various T cell subpopulations and B cells may explain the heterogeneity of clinical, biochemical and immunological parameters observed in children with ADA deficiency severe combined immunodeficiency.

Immunosuppressive activity of FK-506 in rats: flow cytometric analysis of lymphocyte populations in blood, spleen and thymus during treatment and following drug withdrawal

Rats were immunized systemically with sheep red blood cells (SRBC) and given either FK-506 (1 mg/kg) or drug vehicle by i.m. injection for 7 days. In animals receiving FK-506, there was suppression (87%) of the splenic plaque-forming cell response on day 4 and marked reductions in the serum antibody titre throughout the 3-week period following immunization. Sequential flow cytometric analyses of blood lymphocytes revealed statistically significant attenuation, by FK-506, of the increase in relative numbers of OX-12+ (B) cells between days 4 and 7. Following drug withdrawal, OX-12 values remained elevated, whereas in control animals a decline was observed. These changes were reflected in concomitant increases in the relative numbers of OX-19+ (CD3+), W3/25+ (CD4+) and OX-8+ (CD8+) T cells; however, due to an overall reduction in lymphocytes by day 7, absolute values were not significantly affected compared with controls. The pattern of changes in OX-6+ (MHC class II+) cells in blood was similar to that observed for B cells. FK-506 also suppressed increases in the small proportion of blood-borne OX-40+ (activated CD4+) cells and OX-39+ (interleukin-2 receptor+) cells in the 7 day period following immunization; thereafter values for activation marker expression between treatment and control groups were similar. In the spleen, there were fewer significant differences between FK-506 and control groups in the incidences of cells expressing the above markers. OX-8+ cells, however, were significantly higher in drug-treated animals on day 7, and there were also reductions in the small proportions of OX-39+ and OX-40+ cells when compared with controls. In the thymus, reversible medullary atrophy induced by FK-506 was accompanied on day 7 by increases in the incidence of CD4+ and CD8+ cells and by a concomitant reduction in OX-44+ mature, medullary thymocytes. Two weeks after drug withdrawal, the phenotypic marker expression profile had been restored to normal in blood, spleen and thymus. These data provide new information on the apparent capacity of FK-506 to interfere with T cell maturation and its influence on lymphocyte activation in vivo.

In vivo blockage of nitric oxide with aminoguanidine inhibits immunosuppression induced by an attenuated strain of Salmonella typhimurium, potentiates Salmonella infection, and inhibits macrophage and polymorphonuclear leukocyte influx into the spleen.

Our laboratory has previously shown that after immunization with a strain of Salmonella typhimurium, SL3235, made avirulent by a blockage in the pathway of aromatic synthesis, murine splenocytes were profoundly suppressed in their capacity to mount an in vitro antibody plaque-forming cell (PFC) response to sheep erythrocytes. Evidence indicated that suppression was mediated by nitric oxide (NO), since the in vitro addition of NG-monomethyl-L-arginine blocked suppression. The present studies examined the effect of blocking NO production on Salmonella-induced immunosuppression by in vivo administration of aminoguanidine hemisulfate (AG). AG was administered to C3HeB/FeJ mice in their drinking water (2.5% solution) for 7 days prior to intraperitoneal inoculation with SL3235. AG treatment inhibited the increase in nitrate and nitrite levels in plasma and nitrite levels in the spleen seen in immunized mice. Importantly, AG treatment completely blocked suppression of the splenic PFC response and markedly attenuated the suppression of the response to concanavalin A in immunized mice, providing further evidence that Salmonella-induced immunosuppression is mediated by NO. AG treatment also alleviated the majority of the splenomegaly associated with SL3235 inoculation, which correlated with a blockage of influx of neutrophils and macrophages into spleens, as assessed by flow cytometry. AG treatment unexpectedly resulted in 90% mortality in mice injected with the highly attenuated vaccine strain of Salmonella, SL3235. Increased mortality in AG-treated mice correlated with inability to clear organisms from the spleen by day 15 postinoculation and with persistent bacteremia, compared with control mice. Collectively, these in vivo results underscore the dual biological consequences of NO production following Salmonella infection, with NO being necessary for host defense, but also having the potentially adverse effect of immunosuppression. A unifying hypothesis to explain how these seemingly paradoxical effects could both result from NO production is presented.

Inhibition of 3,3′,4,4′,5-Pentachlorobiphenyl-induced fetal cleft palate and immunotoxicity in C57BL/6 mice by 2,2′,4,4′,5,5′-hexachlorobiphenyl

3,3′,4,4′,5-Pentachlorobiphenyl (pentaCB) caused a dose-dependent induction of fetal cleft palate in offspring from pregnant C57BL/6 mice exposed to a single dose (783 or 1044 μg/kg) of this compound on gestation and 10. In contrast 2,2′,4,4′,5,5′-hexaCB did not cause cleft palate at a dose 271 mg/kg and, in pregnant mice cotreated with 2,2′,4,4′,5,5′-hexaCB (271 mg/kg) plus 783 or 1044 μg/kg 3,3′,4,4′,5-pentaCB, fetal cleft plane formation was significantly inhibited. 3,3′,4,4′-PentaCB (6 μg/kg) also inhibited the splenic plaque-forming cell (PFC) response and serum IgM levels in C57BL/6 mice treated with the T cell-independent antigen trinitrophenyl-lipopolysaacharide. At doses as high as 72 mg/kg, 2,2′,4,4′,5,5′-hexaCB was not immunotoxic; however, in mice cotreated with a immunotoxic dose of 3,3′,4,4′,5-pentaCB plus different doses of 2,2′,4,4′,5,5′-hexaCB (18, 36 and 72 mg/kg), there was a dose-dependent inhibition of 3,3′,4,4′,5-pentaCB-induced immunotoxicity. These non-additive (antagonistic) interactions of prototypical polychlorinated biphenyl (PCB) congeners may be an important consideration in development of a toxic equivalency factor approach for hazard and risk assessment of PCB mixtures.

Stressor-induced alterations of the splenic plaque-forming cell response: Strain differences and modification by propranolol

The effects of stressor application on the splenic plaque-forming cell (PFC) response was assessed in two strains of mice: the BALB/cByJ strain, which is highly responsive to stressors; and the more hardy DBA/2J strain. Both strains exhibited a peak PFC response 120 h following administration of sheep red blood cells (SRBC; 5 × 106 cells). Stressor exposure reduced the immune response; however, the appearance of such an outcome was dependent upon the time at which the stressor was applied relative to SRBC inoculation. In DBA/2J mice, foot-shock applied either immediately after SRBC inoculation or at the time of the peak immune response (120 h) resulted in suppression of the PFC response. In BALB/cByJ mice, both stressor severities provoked an immunosuppression when applied 120 h after inoculation, but when applied 96 h after immunization only foot-shock reduced the PFC response. At other intervals, the stressors were without effect. Pretreatment with the β-norepinephrine antagonist propranolol precluded the immunosuppression elicited by a stressor applied 96 h after inoculation, but did not affect the reduction of the PFC response elicited by a stressor applied 120 h after inoculation. It is suggested that several factors may contribute to stressor-provoked alterations of the immune response, and that the contribution of these factors vary over the course of an immune response being mounted.

Immunosuppressive Activity of Polychlorinated Biphenyl Mixtures and Congeners: Nonadditive (Antagonistic) Interactions

Immunosuppressive Activity of Polychlorinated Biphenyl Mixtures and Congeners: Nonadditive (Antagonistic) Interactions. Harper, N., Connor, K., Steinberg, M., and Safe, S. (1995). Fundam. Appl. Toxicol. 27, 131-139. The dose-response inhibition of the splenic plaque-forming cell (PFC) response and serum IgM units to the antigen, trinitrophenyl-lipopolysaccharide, was determined for several polychlorinated biphenyl (PCB) mixtures and congeners in female B3C3F1 mice. The ED50 values for Aroclor 1260-, 1254-, 1248-, and 1242-induced immunotoxicity varied by less than twofold from 355 to 699 mg/kg. The range of ED50 values for 2,3,7,8-tetrachlorodibenzo- p-dioxin (TCDD), 3,3′,4,4′-tetrachlorobiphenyl, 3,3′,4,4′,5-pentaCB, 3,3′,4,4′,5,5′-hexaCB, 2,3,3′,4,4′-pentaCB, 2,3′,4,4′,5-pentaCB, 2,3,3′,4,4′,5-hexaCB, 2,3,3′,4,4′,5,5′-heptaCB, 2,2′,3,3′,4,4′,5-heptaCB, and 2,2′,3,4,4′,5,5′-heptaCB were 4.6 to 4.9, 134 to 245, 4.7 to 7.0, 6.9 to 11.1, 88,000 to 121,000, 122,000 to 132,000, 99,000 to 157,000, 89,000 to 129,000, 117,000 to 240,000, and 132,000 to 238,000 μg/kg, respectively. The immunotoxicity-derived toxic equivalency factors (TEFs) for these congeners could be calculated from the ED50 (TCDD)/ED50 (congener) ratios and the TEF values were within the range of those previously determined for other aryl hydrocarbon receptor-mediated responses. Based on the known concentrations of these congeners in the PCB mixtures, TCDD or toxic equivalents (TEQs) in the mixture were calculated [i.e., TEQ = Σ (PCB congener × TEF)] using the immunotoxicity-derived TEFs (plaque-forming cells/10 6 viable cells). TEQ values for Aroclors 1260, 1254, 1248, and 1242 were 16.0, 54.4, 260.4, and 197 ppm, respectively. Based on the ED50 value for the immunosuppressive activity of TCDD (4.8 μg/kg), the calculated ED50 values for immune suppression by Aroclors 1260, 1254, 1248, and 1242 were 300, 88, 18, and 24 mg/kg, respectively. The ED50 (observed)/ED50 (calculated) ratios were 1.2, 5.9, 21, and 22.0 for Aroclors 1260, 1254, 1248 and 1242, respectively. Thus, for Aroclors 1254, 1248, and 1242, the high ED50 (observed)/ED50 (calculated) ratios (i.e., 5.9 to 22.0) indicate that the TEF approach overestimates the toxicity of these mixtures due to nonadditive (antagonistic) interactions of the PCBs. In contrast, the TEF approach was useful in determining the immunotoxicity of the Aroclor 1260 mixture.

Halogenated aromatic hydrocarbon-induced suppression of the plaque-forming cell response in B6C3F1 splenocytes cultured with allogenic mouse serum: Ah receptor structure activity relationships

The immunsuppressive effects of halogenated aromatic hydrocarbons (HAHs) were investigated in B6C3F1 female mice and in mouse splenocytes cultured with allogenic mouse serum using the Mishell-Dutton model for in vitro immunization to trinitrophenyl-lipopolysaccharide (TNP-LPS). Exposure to 2,3,7,8-tetrachlorodibenzo- P-dioxin (TCDD), 2,3,4,7,8-pentachlorodibenzofuran (PeCDF), 1,2,3,7,8-PeCDF, 1,3,6,8-tetrachlorodibenzofuran (TCDF), 3,3′,4,4′,5-pentachlorobiphenyl (pentaCB), or 3,3′,4,4′,5,5′-hexaCB resulted in a dose-dependent suppression of the splenic plaque-forming cell (PFC) response both in vivo and in vitro. The effective dose required to decrease 50% (ED 50) of the response to 2,3,7,8-TCDD, 2,3,4,7,8-PeCDF, 1,2,3,7,8-PeCDF, 1,3,6,8-TCDF, 3,3′,4,4′,5-pentaCB, or 3,3′,4,4′,5,5′-hexaCB in vivo was 14.1, 5.5, 1695, 34800, 21, and 19 nmol/kg, respectively, and in vitro was 7.0, 10.6, 149, 2325, 9.1 and 9.1 nM, respectively. There was an excellent rank order and linear correlation between the in vivo versus in vitro activities for these HAHs ( r < 0.99) and the relative immunosuppressive potencies of these compounds paralleled their binding affinities for the aryl hydrocarbon (Ah) receptor. These results show that splenocytes cultured with allogenic mouse serum is an Ah-responsive in vitro assay which can be used for quantitating the immunosup-pressive effects of HAHS.

Immunosuppressive Activity of Polychiorinated Biphenyl Mixtures and Congeners: Nonadditive (Antagonistic) Interactions

The dose-response inhibition of the splenic plaque-forming cell (PFC) response and serum IgM units to the antigen, trinitrophenyl-lipopolysaccharide, was determined for several polychiorinated biphenyl (PCB) mixtures and congeners in female B3C3F1 mice. The ED50 values for Aroclor 1260-, 1254-, 1248-, and 1242-induced immunotoxicity varied by less than twofold from 355 to 699 mg/kg. The range of ED50 values for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 3,3′ ,4,4′-tetrachlorobiphenyl, 3,3′ ,4,4′,5-pentaCB, 3,3′,4,4′,5,5′-hexaCB, 2,3,3′,4,4′-pentaCB, 2,3′,4,4′,5-pentaCB, 2,3,3′,4,4′,5-hexaCB, 2,3,3′,4,4′,5,5′-heptaCB, 2,2′,3,3′,4,4′,5-heptaCB, and 2,2′,3,4,4′,5,5′-heptaCB were 4.6 to 4.9, 134 to 245, 4.7 to 7.0, 6.9 to 11.1, 88,000 to 121,000, 122,000 to 132,000, 99,000 to 157,000, 89,000 to 129,000, 117,000 to 240,000, and 132,000 to 238,000 μg/kg, respectively. The immunotoxicity-derived toxic equivalency factors (TEFs) for these congeners could be calculated from the ED50 (TCDD)/ED50 (congener) ratios and the TEF values were within the range of those previously determined for other aryl hydrocarbon receptor-mediated responses. Based on the known concentrations of these congeners in the PCB mixtures, TCDD or toxic equivalents (TEQs) in the mixture were calculated [i.e., TEQ = Σ(PCBcongener × TEF)] using the immunotoxicity-derived TEFs (plaque-forming cells/106 viable cells). TEQ values for Aroclors 1260, 1254, 1248, and 1242 were 16.0, 54.4, 260.4, and 197 ppm, respectively. Based on the ED50 value for the immunosuppressive activity of TCDD (4.8 μg/kg), the calculated ED50 values for immune suppression by Arodors 1260, 1254, 1248, and 1242 were 300, 88, 18, and 24 mg/kg, respectively. The ED50 (observed)/ED50 (calculated) ratios were 1.2, 5.9, 21, and 22.0 for Aroclors 1260, 1254, 1248 and 1242, respectively. Thus, for Aroclors 1254, 1248, and 1242, the high ED50 (observed)/ED50 (calculated) ratios (i.e., 5.9 to 22.0) indicate that the TEF approach overestimates the toxicity of these mixtures due to non-additive (antagonistic) interactions of the PCBs. In contrast, the TEF approach was useful in determining the immunotoxicity of the Aroclor 1260 mixture.

Alterations in central catecholamines associated with immune responding in adult and aged mice

Central catecholamine alterations associated with immune activity are similar to those seen following stressor exposure. Inasmuch as aged animals exhibit more pronounced stressor-provoked alterations of central amines relative to younger animals, it was of interest to determine whether immune challenge would similarly induce more pronounced central amine variations in older animals. Fifteen-month old CD-1 mice challenged with 10 7 sheep red blood cells (SRBC) revealed an equivalent peak splenic plaque-forming cell response (4 days after antigen challenge) to that of 3-month-old mice challenged with 10 6 cells. Neither plasma adrenocorticotropic hormone (ACTH) nor corticosterone levels varied over days following immunization, although ACTH levels were generally higher in the older mice. In both age groups reductions of hypothalamic and locus coeruleus norepinephrine (NE) and increased accumulation of the metabolite MHPG coincided with (or preceded by 24 h) the peak immune response. However, increased accumulation of MHPG in the hypothalamus was greater and occurred earlier in the locus coeruleus of the aged mice. Likewise, at or about the time of peak immune responses nucleus accumbens dopamine (DA) levels were reduced and metabolites elevated in both age groups, while in the prefrontal cortex only DA metabolite levels were elevated. These data are commensurate with previous findings showing that SRBC inoculation may influence central neurotransmitters and that such effects correspond with the time of the peak immune responses. Moreover, in so far as hypothalamic NE utilization is concerned, it seems that the effects of SRBC inoculation are more pronounced in aged animals.

Enhanced Activity of Polynuclear Aromatic Hydrocarbon Mixtures as aryl Hydrocarbon (Ah) Receptor Agonists

Abstract The relative potencies of benzo[a]pyrene and a complex mixture of polycyclic aromatic hydrocarbons (PAH) produced as by-products of manufactured gas plant (MGP) residues as inducers of hepatic microsomal ethoxyresorufin O-deethylase (EROD) were determined in the B6C3F1 mouse. The ED50 values for the induction response were 78 and 65 mg/kg for benzo[a]pyrene and the MGP-PAH mixture, respectively, although benzo[a]pyrene and other compounds containing four or more rings were only trace components of this mixture. A comparison of the EROD induction potencies of benzo[a]pyrene and the MGP-PAH mixture showed that the mixture was approximately 706 times more active than expected based on its benzo[a]pyrene content (0.17%). The enhanced activity of the MGP-PAH mixture was also observed for several other aryl hydrocarbon (Ah) receptor-mediated responses, including inhibition of the splenic plaque-forming cell (PFC) response to both T-cell-dependent and independent antigens in B6C3F1 mice. The nature of t...

Modulation of the in vivo immune response by selective depletion of neutrophils using a monoclonal antibody, RP-3. III. Enhancement by RP-3 treatment of the anti-sheep red blood cell plaque-forming cell response in rats.

Because recent work has shown that neutrophils produce various cytokines and are activated by these agents, this study was undertaken to examine neutrophil participation in immune networks. In our previous work, we demonstrated that delayed type hypersensitivity to SRBC and accompanying mononuclear leukocyte recruitment are inhibited in vivo by selective depletion of neutrophils using a mAb designated RP-3. To elucidate the function of these cells in Ab production, we assessed the splenic plaque-forming cell response to SRBC in a rat model through selective depletion of circulating neutrophils by RP-3. This depletion which occurred at the time of immunization resulted in an increase in the number of anti-SRBC Ab producing cells, as determined by the direct and indirect plaque-forming cell response. This phenomenon was observed only when the Ag was administered i.p. and not with i.v. immunization. These results suggest that neutrophils suppress Ab production in certain situations.

Influence of change from grouped to individual housing on a T-cell-dependent immune response in mice: Antagonism by diazepam

Transferring CD-1 mice from grouped to individual housing and then maintaining them individually resulted in a decline in the peak IgM plaque-forming cell (PFC) response to sheep red blood cells (SRBCs). However, the immunosuppression was dependent on the amount of time mice were maintained individually. In particular, individual housing for 5–10 days prior to SRBC inoculation and for 4 days following inoculation resulted in a suppression of the splenic PFC response and serum antibody titers. Shorter periods of individual housing (4 days following inoculation) did not provoke the immunosuppression. Likewise, following more protracted individual housing (15–30 days prior to inoculation) the immunosuppression was not evident. Inasmuch as daily treatment with an anxiolytic, diazepam (1.0 mg/kg), antagonized the suppression induced by 5 days of individual housing, it was suggested that the change from group to individual housing and then maintenance of animals individually acted much like a stressor to induce the immunosuppression.

Suppression of B-cell mediated immunity in juvenile chinook salmon (Oncorhynchus tshawytscha) after exposure to either a polycyclic aromatic hydrocarbon or to polychlorinated biphenyls.

Juvenile chinook salmon (Oncorhynchus tshawytscha) were injected intraperitoneally with either the polycyclic aromatic hydrocarbon, 7,12-dimethylbenz[a]anthracene (DMBA)1 or with the commercial polychlorinated biphenyl (PCB) mixture, Aroclor 1254, to assess effects on the B-cell mediated immune response. B-cell mediated immunity was assessed by examination of the primary and secondary plaque-forming cell (PFC) responses of anterior kidney and splenic leukocytes to a T-independent antigen, TNP-keyhole limpet hemocyanin (TNP-KLH). Salmon exposed to DMBA at dosages of 20% or 1% of the 96 hr LD50 (12.7 mg and 0.6 mg/kg of salmon, respectively) or to PCBs at a dosage of 20% of the 96 hr LD50 (54.0 mg/kg of salmon) exhibited a suppressed PFC response. The secondary PFC response of anterior kidney and splenic leukocytes to both antigens and the primary splenic PFC response to TNP-LPS were suppressed in salmon exposed to either DMBA or PCBs. However, only the primary PFC response of anterior kidney leukocytes to TNP-LPS was suppressed in salmon exposed to PCBs and no suppression of this response was observed in salmon exposed to DMBA. Neither anterior kidney or splenic leukocytes from salmon exposed to DMBA or PCBs showed an altered primary PFC response to the T-dependent antigen, TNP-KLH. These results suggest that B-cell mediated immunity in salmon is suppressed by known mammalian immunosuppressants and that suppression of the PFC response observed previously in salmon from an urban estuary may be due to contaminant exposure.

Immunosuppressive effects of highly chlorinated biphenyls and diphenyl ethers on T-cell dependent and independent antigens in mice

The dose-dependent effects of 2,2′,3,3′,4,4′,5,5′,6-nonachlorobiphenyl (nonaCB), 2,2′,3,3′,4,4′,5,6,6′-nonaCB, 2,2′,3,3′,4,5,5′6,6′-nonaCB and decaCB on the suppression of the splenic plaque-forming cell (PFC) response to the T-cell-dependent antigen, sheep red blood cells (SRBCs) and the T-cell-independent antigen, trinitrophenyl-lipopoccharide (TNP-LPS), were determined in genetically inbred mice. In addition, the induction of hepatic microsomal ethoxyresorufin O-deethylase (EROD) activity was also measured. The highly chlorinated biphenyls suppressed the splenic PFC response to SRBCs in C57BL/6 and DBA/2 mice and were relatively more active in the former strain. The C57BL/6 mice are more responsive to aryl hydrocarbon (Ah) receptor agonists than DBA/2 mice and these data support a possible role for the Ah receptor in mediating this response. However, previous studies with polychlorinated biphenyls (PCBs) indicate that congeners with 3 or 4 ortho-chloro substituents are inactive as Ah receptor agonists and this was consistent with the minimal induction of hepatic microsomal EROD activity by the highly chlorinated biphenyls in both strains of mice. Thus, the results suggest that the inhibition of the splenic PFC response to SRBCs observed in this study was primarily an Ah receptor-independent response. Some of the highly chlorinated diphyenyl ethers namely decachlorodiphenyl ether and 2,2′,3,3′,4,4′,5,6,6′-nonachlorodiphenyl ether, inhibited the antigenic response to TNP-LPS in C57 BL/6 mice. The results indicate that the suppression of the TNP-LPS-mediated immune response may be a more reliable indicator of the Ah receptor-dependent immunotoxicity of halogenated hydrocarbons.

Synergistic activity of polynuclear aromatic hydrocarbon mixtures as aryl hydrocarbon (Ah) receptor agonists

The relative potencies of benzo[ a]pyrene and a complex mixture of polynuclear aromatic hydrocarbons (PAHs) produced as by-products of manufactured gas plant (MGP) residues as inducers of hepatic microsomal ethoxyresorufin O-deethylase (EROD) activity were determined in the B6C3F1 mouse. The ED 50 values for the induction response were 78 and 65 mg/kg for benzo[ a]pyrene and the MGP-PAH mixture, respectively. Analysis of the MGP-PAH mixture indicated that benzo[ a]pyrene and other compounds containing four or more rings and which are known to induce EROD activity were only present as trace components of this mixture. A comparison of the EROD induction potencies of benzo[ a]pyrene and the MGP-PAH mixture showed that the mixture was approximately 706 times more potent than expected based on its benzo[ a]pyrene content (0.17%). This induced P-450 activity could significantly increase the metabolism of the carcinogenic PAHs and thereby modulate the overall carcinogenicity of the mixture. The apparent synergistic activity of the MGP-PAH mixture was further investigated by comparing the activities of this mixture and benzo[ a]pyrene for several other aryl hydrocarbon (Ah) receptor-mediated responses including (i) induction of hepatic CYP1A1 mRNA levels, (ii) transformation of the rat cytosolic Ah receptor to a complex which binds to a dioxin responsive element, (iii) induction of EROD activity and (iv) antiestrogenicity in MCF-7 human breast cancer cells, and (v) inhibition of the splenic plaque-forming cell (PFC) response to both T cell-dependent and independent antigens in B6C3F1 mice. For the EROD and CYP1A1 mRNA induction and cytosolic transformation activities and immunosuppressive effects, the MGP-PAH mixture was approximately 100–900 times more potent as an Ah receptor agonist than expected based on its benzo[ a]pyrene content. The synergistic activity was lower (19-fold) for the antiestrogenic response in MCF-7 cells. The reason for the synergistic effects of the MGP-PAH mixture were not due to contamination of the mixture by 2,3,7,8-tetrachlorodibenzo- p-dioxin and related compounds and the results suggest that the enhanced potency of the mixture is due to unknown interactions between the individual PAHs present in the mixture.

Immunotoxic potencies of polychlorinated biphenyl (PCB), dibenzofuran (PCDF) and dibenzo- p-dioxin (PCDD) congeners in C57BL/6 and DBA/2 mice

The dose-dependent effects of a single acute exposure of 2,3,7,8-tetrachlorodibenzo- p-dioxin (TCDD), 2,3,4,7,8-pentachlorodibenzofuran (PeCDF), 1,2,3,7,9-PeCDF, 1,3,6,8-tetrachlorodibenzofuran (TCDF), 3,3′,4,4′,5-pentachlorobiphenyl (pentaCB), and 3,3′,4,4′,5,5′-hexaCB on the suppression of the splenic plaque-forming cell (PFC) response to the T-cell-independent antigen trinitrophenyl-lipopolysaccharide were determined in C57BL/6 and DBA/2 mice. In addition, the induction of hepatic microsomal ethoxyresorufin O-deethylase (EROD) activity was also measured in these animals. 2,3,7,8-TCDD and 2,3,4,7,8-PeCDF were the most immunotoxic congeners in both strains of mice and with the exception of the latter congener, the ED 50 values for each compound were lower in the C57BL/6 than the DBA/2 mice. 2,3,7,8-TCDD induced hepatic microsomal EROD activity in both strains of mice whereas the other congeners were considerably less active or inactive as inducers. The results of this study demonstrated that for the halogenated aromatic hydrocarbons the immunotoxic response was a more sensitive indicator of exposure than the induction of CYP1A1 activity. The rank order for the immunotoxic potencies of the chlorinated aromatic compounds used in this study was 2,3,7,8-TCDD ≈ 2,3,4,7,8-PeCDF > 3,3′,4,4′,5-pentaCB ≈ 3,3′,4,4′,5,5′-hexaCB > 1,2,3,7,9-PeCDF > 1,3,6,8-TCDF. The order of activity for these congeners was similar for other Ah receptor-mediated responses and these results coupled with the differential responsiveness of the C57BL/6 and DBA/2 mice confirms the role of aryl hydrocarbon (Ah) receptor in mediating the suppression of this T-cell-independent response.

Immunosuppressive and Monooxygenase Induction Activities of Highly Chlorinated Diphenyl Ether Congeners in C57BL/6 and DBA/2 Mice

Immunosuppressive and Monooxygenase Induction Activities of Highly Chlorinated Diphenyl Ether Congeners in C57BL/6 and DBA/2 Mice. Harper, N., Howie, L, Connor, K., Arellano, L., Craig, A., Dickerson, R., and Safe, S. (1993). Fundam. Appl. Toxicol. 20, 496-502.The dose-response effects of 2.2′,3,3′,4,5,5′,6,6′-,2,2′,3,3′,4, 4′,5,6,6′- and 2,2′,3,3′,4,4′,5,5′6-nonachlodiphenyl ether (nonaCDE) and decachlorodiphenyl ether (decaCDE) on the splenic plaque-forming cell (PFC) response to sheep red blood cells (SRBCs) and the induction of hepatic microsomal ethoxyresorufin O-deethylase (EROD) activity was determined in aryl hydrocarbon (Ah)-responsive C57B1,/6 and less All responsive DBA/2 mice. All the congeners exhibited immunotoxicity at doses between 2.5 and 10 μmol/kg in C57BL/6 mice whereas in DBA/2 mice doses ⩾ 25 μmol/kg were required to cause inhibition of the PFC response to SRBCs. The results also showed that the nonaCDE isomers and decaCDE were more active as inducers of hepatic EROD activity in C57BL/6 than DBA/2 mice; However, there was not a correlation between the induced EROD activity and the CVP1A1 and CYP1A2 mRNA levels in the C57BL/6 mice. These data suggested that the immunotoxicity of these compounds was mediated through the Ah receptor. However, the results showed that the immunotoxicity of the nonaCDE isomers and decaCDE was unexpectedly high compared to that of lower chlorinated diphenyl ethers and there were no apparent structure-activity relationships among the higher chlorinated congeners. This suggests that some of the immunosuppressive effects observed for the nonaCDE isomers and decaCDE may be Ah receptor-independent.

T-cell-dependent and T-cell-independent mechanisms of tolerance to glucuronoxylomannan of Cryptococcus neoformans serotype A

Glucuronoxylomannan (GXM), a type 2 T-independent antigen, is the major component of the capsular polysaccharide (CnCAP) of Cryptococcus neoformans. Previous studies have described the tolerogenic effects of high doses of CnCAP on the specific humoral response. In this investigation, evidence for both high-dose and low-dose tolerance to GXM is presented. BALB/cBy female mice, primed with either 5 ng or 50 micrograms of GXM, then coimmunized 3 days later with immunogenic doses of both GXM and type 3 pneumococcal polysaccharide (SSS-III), showed an antigen-specific inhibition in their splenic plaque-forming cell (PFC) responses to GXM compared with control groups primed with normal saline. SSS-III PFCs remained unchanged between GXM-primed and normal saline-primed groups. Low-dose tolerance appeared to be T dependent, whereas high-dose tolerance appeared to be T independent. Low-dose tolerance to GXM could not be induced in athymic BALB/c nu/nu mice, whereas high-dose tolerance in the same mice could be induced. Furthermore, low-dose tolerance was adoptively transferred with B-cell-depleted splenocytes to naive BALB/c mice, while high-dose tolerance was not. Complement-mediated depletion of CD4+ but not CD8+ splenocytes from low-dose-primed mice abrogated the transfer of low-dose tolerance. These findings indicate T-dependent and T-independent mechanisms of antigen-specific B-cell tolerance to GXM in BALB/c mice at low and high antigen doses, respectively.

Teratogenicity and immunotoxicity of 3,3′,4,4′,5-pentachlorobiphenyl in C57BL/6 mice

Administration of 3,3′,4,4′,5-pentachlorobiphenyl (pentaCB) to female C57BL/6 mice at doses from 130.5 to 522 μg/kg body weight resulted in the dose-dependent formation of fetal cleft palate and hydronephrosis. The estimated relative potency of 3,3′,4,4′,5-pentaCB compared to 2,3,7,8-tetrachlorodibenzo- p-dioxin (TCDD) was in the range of <0.07-0.04. The immunotoxicity of 3,3′4,4′,5-pentaCB and two structurally-related congeners, 3,3,′,4,4′-tetraCB and 3,3′4,4t́,5,5′-hexaCB, was investigated in male C57BL/6 mice by determining their suppression of the splenic plaque-forming cell response to sheep red blood cells. The potencies of these compounds relative to TCDD were determined from the ratios of their corresponding ED 50 values and were 0.77-0.55 (3,3′,4,4′,5-pentaCB), 1,1-0.29 (3,3′,4,4′,5,5′-hexaCB) and 0.14-0.03 (3,3′,4,4′-tetraCB). These results demonstrate that the immunosuppressive activities of the PCB congeners relative to TCDD were much higher than observed for many other TCDD-like responses in mice and other laboratory animals.