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Diesel exhaust particles, the local inflammatory response, and the systemic IgE response to ovalbumin
Workshop 2. Air Pollution and Allergy OZONE-INDUCED IMPAIRMENT OF PULMONARY TYPE IV HYPERSENSITIVITY AND AIRWAY HYPERRESPONSIVENESSIN MICE Henk Van Loveren * ], Leendert Van Bree 2, Helen Van Der Vliet t, Johan Garssen t. t Laboratory of Pathology and Immunobiology,
2 Laboratory of Health Effects Research, National Institute of Public Health and Environment (RIVM), P.O. Box 1, 3720 BA Bilthoven, The Netherlands
15
IDENTIFICATION OF CHEMICAL SENSITIZERS ACCORDING TO CYTOKINE EXPRESSION INDUCED IN DRAINING LYMPH NODES Rebecca J. Dearman *, Ian Kimber. Zeneca Central Toxicology Laboratory, Alderley Park, Macclesfield, Cheshire, SKIO 4TJ, UK
Individuals suffering from COPD or asthma are groups at risk with respect to the effects of ozone. The majority of studies dealing with ozone effects on pulmonary allergy is restricted to IgE mediated allergy (type I allergy) in which Th-2 cells are important regulatory cells. In animal models for type I allergy it has been demonstrated that ozone can potentiate the induction as well as the effector phase of this type of hyperimmune reaction (e.g. allergic asthma). Recently it has been demonstrated in a murine model that in non-IgE mediated "asthma" T cells may also play a crucial role. It was demonstrated that low molecular weight compounds can induce delayed type hypersensitivity-like reactions in the lung. Since in delayed type hypersensitivity reactions Th-1 cells play a crucial role it may be suggested that in non-IgE mediated pulmonary diseases Th-1 cells are involved. We have demonstrated that acute ozone exposure can inhibit pulmonary delayed type hypersensitivity induced by small molecular weight compounds in the mouse. The inflammatory component as well as the hyperresponsiveness of the trachea that is induced by this type of hypersensitivity reaction are significantly inhibited by ozone. These results further underline the effect of ozone on the Th-1/Th2 balance in the respiratory tract.
We have shown previously that characteristic cytokine secretion profiles consistent with the selective activation of T helper 1 (Thl)-type and Th2-type cells are induced following topical exposure of mice to chemical contact allergens such as 2,4-dinitrochlorobenzene (DNCB) or respiratory allergens such as trimellitic anhydride (TMA), respectively. We have now examined cytokine expression provoked by hexyl cinnamic aldehyde (HCA), a contact allergen recommended as a positive control for skin sensitization predictive tests by (OECD) Organization for Economic Cooperation and Development. BALB/c mice were exposed topically to HCA (50, 25 or 10%) in 4:1 acetone: olive oil (AOO). Control animals were treated concurrently with 1% DNCB or 10% TMA in AOO. Thirteen days after the initiation of exposure, draining auricular lymph nodes were excised, lymph node cells (LNC) cultured for 12-120 hr and cytokine content of supernatants analyzed by enzyme-linked immunosorbent assays. TMA-stimulated LNC produced high levels of the Th2-type cytokines interleukins 4 and 10, but little of the Thl-type product interferon y. Exposure to DNCB resulted in the converse pattern. At all concentrations of HCA tested a profile of cytokine expression similar to the contact allergen DNCB was provoked. These data demonstrate that, in common with DNCB, Thl-type cytokine production is stimulated by topical exposure to HCA, suggesting that it may be possible to identify and classify chemical allergens according to induced cytokine expression.
Keywords: ozone; respiratory allergy; Th-I/Th-2 balance
Keywords: chemical allergens; Thl- and Th2-type cells; cytokines
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DIESEL EXHAUST PARTICLES, THE LOCAL INFLAMMATORY RESPONSE, AND THE SYSTEMIC IgE RESPONSE TO OVALBUMIN
SYSTEMIC ANAPHYLAXIS IN THE GUINEA-PIG: EFFECT OF SEVEN DIFFERENT HUMAN VACCINES
Martinus L~vik *, Per Ivar Guarder, Ann-Kristin Hcgseth, Randi Hagemann I, Ingvar Eide i National Institute of Public
Bertrand Brutzkus l, Bernard Coquet 2, Bernard Danve 2, Jacques Descotes * 2. t Department of Pharmacology and Medical
Health, Oslo, Trondheim, Norway; I Statoil Research Centre, Trondheim, Norway
Toxicology, INSERM U80, Lyon, France; 2 Pasteur-M~rieux S~rums & Vaccins, Marcy-l'Etoile, France
The mouse (BALB/c) popliteal lymph node assay (PLNA) was used to study the possible adjuvant effect of diesel exhaust particles (DEP), carbon black (CB) and AI(OH)3 on the immune response to the model allergen ovalbumin (OA). Three different lymph node parameters were determined as a measure of the local inflammatory response: increase in weight, increase in cell numbers, and increase in cell proliferation (thymidine in-corporation). In serum, total IgE and specific anti-OA IgE were measured. OA with DEP gave a significant increase in all three lymph node parameters as compared to either DEP or OA alone. Also, the duration of the local inflammatory response was increased when OA was injected together with DEP. OA with DEP also gave a marked increase in IgE anti-OA antibodies. No effect was seen on total serum IgE levels. DEE CB and AI(OH)3 when injected with OA all had grossly similar effects on the lymph node parameters as well as the specific IgE response, indicating that the non-extractable particle core may be responsible for a substantial part of the adjuvant activity of DEE When similar experiments were performed in T-cell deficient nu/nu (nude) mice, no increase was seen in the lymph node parameters, indicating that the local lymph node reaction observed in immunologically normal BALB/c mice is mediated by a specific immune response, rather than being a mere irritative reaction.
The use of guinea-pigs for predicting the human risk of anaphylaxis due to heterologous macromolecules is debated. During further validation of a standardized protocol [1], 7 human vaccines have been tested: 2 purified proteins (tetanus toxoid: TT, hepatitis B), 1 polyoside conjugated to TT (haemophilus), 2 inactivated viruses (influenza, mumps), 1 viral combination (mumps-measles-rubella: MMR), and 1 multiple combination (diphteria-tetanus-pertussispolio). Groups of 5 adult young male Dunkin-Hartley guinea-pigs received 0.08 to 2 human vaccine doses subcutaneously at day 1 and 8, and 0.5 human dose intravenously at day 22. A positive control group received ovalbumin as described [1]. MMR caused anaphylaxis, but no significant reactions were noted with the 6 other vaccines. Reactions caused by MMR were shown to be due to the human albumin of the formulation. Although anaphylaxis is a typical, but very rare, adverse event of vaccines in man, the lack of clinical responses in guinea-pigs support the view that guinea-pigs should not be discarded a priori as predictors of the anaphylactic risk due to the administration of heterologous macromolecules to man.
Keywords: diesel exhaust particles; adjuvant; IgE; popliteal lymph node assay; carbon black; nude mice
[1] Brutzkus et al., The Toxicologist. 15, 99, 1995.
2 Laboratory of Health Effects Research, National Institute of Public Health and Environment (RIVM), P.O. Box 1, 3720 BA Bilthoven, The Netherlands
15
IDENTIFICATION OF CHEMICAL SENSITIZERS ACCORDING TO CYTOKINE EXPRESSION INDUCED IN DRAINING LYMPH NODES Rebecca J. Dearman *, Ian Kimber. Zeneca Central Toxicology Laboratory, Alderley Park, Macclesfield, Cheshire, SKIO 4TJ, UK
Individuals suffering from COPD or asthma are groups at risk with respect to the effects of ozone. The majority of studies dealing with ozone effects on pulmonary allergy is restricted to IgE mediated allergy (type I allergy) in which Th-2 cells are important regulatory cells. In animal models for type I allergy it has been demonstrated that ozone can potentiate the induction as well as the effector phase of this type of hyperimmune reaction (e.g. allergic asthma). Recently it has been demonstrated in a murine model that in non-IgE mediated "asthma" T cells may also play a crucial role. It was demonstrated that low molecular weight compounds can induce delayed type hypersensitivity-like reactions in the lung. Since in delayed type hypersensitivity reactions Th-1 cells play a crucial role it may be suggested that in non-IgE mediated pulmonary diseases Th-1 cells are involved. We have demonstrated that acute ozone exposure can inhibit pulmonary delayed type hypersensitivity induced by small molecular weight compounds in the mouse. The inflammatory component as well as the hyperresponsiveness of the trachea that is induced by this type of hypersensitivity reaction are significantly inhibited by ozone. These results further underline the effect of ozone on the Th-1/Th2 balance in the respiratory tract.
We have shown previously that characteristic cytokine secretion profiles consistent with the selective activation of T helper 1 (Thl)-type and Th2-type cells are induced following topical exposure of mice to chemical contact allergens such as 2,4-dinitrochlorobenzene (DNCB) or respiratory allergens such as trimellitic anhydride (TMA), respectively. We have now examined cytokine expression provoked by hexyl cinnamic aldehyde (HCA), a contact allergen recommended as a positive control for skin sensitization predictive tests by (OECD) Organization for Economic Cooperation and Development. BALB/c mice were exposed topically to HCA (50, 25 or 10%) in 4:1 acetone: olive oil (AOO). Control animals were treated concurrently with 1% DNCB or 10% TMA in AOO. Thirteen days after the initiation of exposure, draining auricular lymph nodes were excised, lymph node cells (LNC) cultured for 12-120 hr and cytokine content of supernatants analyzed by enzyme-linked immunosorbent assays. TMA-stimulated LNC produced high levels of the Th2-type cytokines interleukins 4 and 10, but little of the Thl-type product interferon y. Exposure to DNCB resulted in the converse pattern. At all concentrations of HCA tested a profile of cytokine expression similar to the contact allergen DNCB was provoked. These data demonstrate that, in common with DNCB, Thl-type cytokine production is stimulated by topical exposure to HCA, suggesting that it may be possible to identify and classify chemical allergens according to induced cytokine expression.
Keywords: ozone; respiratory allergy; Th-I/Th-2 balance
Keywords: chemical allergens; Thl- and Th2-type cells; cytokines
~'~-~
DIESEL EXHAUST PARTICLES, THE LOCAL INFLAMMATORY RESPONSE, AND THE SYSTEMIC IgE RESPONSE TO OVALBUMIN
SYSTEMIC ANAPHYLAXIS IN THE GUINEA-PIG: EFFECT OF SEVEN DIFFERENT HUMAN VACCINES
Martinus L~vik *, Per Ivar Guarder, Ann-Kristin Hcgseth, Randi Hagemann I, Ingvar Eide i National Institute of Public
Bertrand Brutzkus l, Bernard Coquet 2, Bernard Danve 2, Jacques Descotes * 2. t Department of Pharmacology and Medical
Health, Oslo, Trondheim, Norway; I Statoil Research Centre, Trondheim, Norway
Toxicology, INSERM U80, Lyon, France; 2 Pasteur-M~rieux S~rums & Vaccins, Marcy-l'Etoile, France
The mouse (BALB/c) popliteal lymph node assay (PLNA) was used to study the possible adjuvant effect of diesel exhaust particles (DEP), carbon black (CB) and AI(OH)3 on the immune response to the model allergen ovalbumin (OA). Three different lymph node parameters were determined as a measure of the local inflammatory response: increase in weight, increase in cell numbers, and increase in cell proliferation (thymidine in-corporation). In serum, total IgE and specific anti-OA IgE were measured. OA with DEP gave a significant increase in all three lymph node parameters as compared to either DEP or OA alone. Also, the duration of the local inflammatory response was increased when OA was injected together with DEP. OA with DEP also gave a marked increase in IgE anti-OA antibodies. No effect was seen on total serum IgE levels. DEE CB and AI(OH)3 when injected with OA all had grossly similar effects on the lymph node parameters as well as the specific IgE response, indicating that the non-extractable particle core may be responsible for a substantial part of the adjuvant activity of DEE When similar experiments were performed in T-cell deficient nu/nu (nude) mice, no increase was seen in the lymph node parameters, indicating that the local lymph node reaction observed in immunologically normal BALB/c mice is mediated by a specific immune response, rather than being a mere irritative reaction.
The use of guinea-pigs for predicting the human risk of anaphylaxis due to heterologous macromolecules is debated. During further validation of a standardized protocol [1], 7 human vaccines have been tested: 2 purified proteins (tetanus toxoid: TT, hepatitis B), 1 polyoside conjugated to TT (haemophilus), 2 inactivated viruses (influenza, mumps), 1 viral combination (mumps-measles-rubella: MMR), and 1 multiple combination (diphteria-tetanus-pertussispolio). Groups of 5 adult young male Dunkin-Hartley guinea-pigs received 0.08 to 2 human vaccine doses subcutaneously at day 1 and 8, and 0.5 human dose intravenously at day 22. A positive control group received ovalbumin as described [1]. MMR caused anaphylaxis, but no significant reactions were noted with the 6 other vaccines. Reactions caused by MMR were shown to be due to the human albumin of the formulation. Although anaphylaxis is a typical, but very rare, adverse event of vaccines in man, the lack of clinical responses in guinea-pigs support the view that guinea-pigs should not be discarded a priori as predictors of the anaphylactic risk due to the administration of heterologous macromolecules to man.
Keywords: diesel exhaust particles; adjuvant; IgE; popliteal lymph node assay; carbon black; nude mice
[1] Brutzkus et al., The Toxicologist. 15, 99, 1995.