Enhanced nasal cytokine production in human beings after in vivo challenge with diesel exhaust particles David Diaz-Sanchez, PhD, Albert Tsien, MD, Adrian.

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Presentation transcript:

Enhanced nasal cytokine production in human beings after in vivo challenge with diesel exhaust particles David Diaz-Sanchez, PhD, Albert Tsien, MD, Adrian Casillas, MD, Anton Robert Dotson, MD, Andrew Saxon, MD Journal of Allergy and Clinical Immunology Volume 98, Issue 1, Pages 114-123 (July 1996) DOI: 10.1016/S0091-6749(96)70233-6 Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 1 An IgE response induced by in vivo DEP challenge. Subjects were challenged intranasally with saline solution or a total of 0.3 mg of DEPs. Each bar represents the mean IgE level of 14 subjects, determined immediately before or 4 days after challenge. IgE levels were determined by ELISA. Error bars represent 1 standard deviation. **p < 0.01, paired t test between pre- and postchallenge levels. Journal of Allergy and Clinical Immunology 1996 98, 114-123DOI: (10.1016/S0091-6749(96)70233-6) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 2 In vivo challenge with DEPs increases cytokine mRNA expression. Cells from nasal lavages performed before and 18 hours after in vivo exposure to 0.3 mg of DEPs were used to obtain cDNA and amplified with β-actin mRNA-specific primers. Complementary DNA samples were adjusted to produce similar intensity bands on an ethidium bromide–stained agarose gel for β-actin after PCR amplification, and then these samples were used in PCR amplification with mRNA-specific primers for: IL-2, IL-4, and IL-5 (A) and for IL-6, IL-10, IL-13, IFN-γ, and CD3δ (B). The amplified cDNA was separated on an agarose gel. Lane 1 shows the results from PBMCs stimulated with phorbol myristate acetate and ionomycin, a regimen known to express all cytokine mRNA, which served as a positive control. Amplified cDNAs from cells recovered after challenge from two representative subjects (lanes 2 and 3) were compared with those obtained before challenge (lanes 4 and 5). Although results for only two average subjects are shown, similar results were observed in all 14 subjects. Journal of Allergy and Clinical Immunology 1996 98, 114-123DOI: (10.1016/S0091-6749(96)70233-6) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 2 In vivo challenge with DEPs increases cytokine mRNA expression. Cells from nasal lavages performed before and 18 hours after in vivo exposure to 0.3 mg of DEPs were used to obtain cDNA and amplified with β-actin mRNA-specific primers. Complementary DNA samples were adjusted to produce similar intensity bands on an ethidium bromide–stained agarose gel for β-actin after PCR amplification, and then these samples were used in PCR amplification with mRNA-specific primers for: IL-2, IL-4, and IL-5 (A) and for IL-6, IL-10, IL-13, IFN-γ, and CD3δ (B). The amplified cDNA was separated on an agarose gel. Lane 1 shows the results from PBMCs stimulated with phorbol myristate acetate and ionomycin, a regimen known to express all cytokine mRNA, which served as a positive control. Amplified cDNAs from cells recovered after challenge from two representative subjects (lanes 2 and 3) were compared with those obtained before challenge (lanes 4 and 5). Although results for only two average subjects are shown, similar results were observed in all 14 subjects. Journal of Allergy and Clinical Immunology 1996 98, 114-123DOI: (10.1016/S0091-6749(96)70233-6) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 2 In vivo challenge with DEPs increases cytokine mRNA expression. Cells from nasal lavages performed before and 18 hours after in vivo exposure to 0.3 mg of DEPs were used to obtain cDNA and amplified with β-actin mRNA-specific primers. Complementary DNA samples were adjusted to produce similar intensity bands on an ethidium bromide–stained agarose gel for β-actin after PCR amplification, and then these samples were used in PCR amplification with mRNA-specific primers for: IL-2, IL-4, and IL-5 (A) and for IL-6, IL-10, IL-13, IFN-γ, and CD3δ (B). The amplified cDNA was separated on an agarose gel. Lane 1 shows the results from PBMCs stimulated with phorbol myristate acetate and ionomycin, a regimen known to express all cytokine mRNA, which served as a positive control. Amplified cDNAs from cells recovered after challenge from two representative subjects (lanes 2 and 3) were compared with those obtained before challenge (lanes 4 and 5). Although results for only two average subjects are shown, similar results were observed in all 14 subjects. Journal of Allergy and Clinical Immunology 1996 98, 114-123DOI: (10.1016/S0091-6749(96)70233-6) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 2 In vivo challenge with DEPs increases cytokine mRNA expression. Cells from nasal lavages performed before and 18 hours after in vivo exposure to 0.3 mg of DEPs were used to obtain cDNA and amplified with β-actin mRNA-specific primers. Complementary DNA samples were adjusted to produce similar intensity bands on an ethidium bromide–stained agarose gel for β-actin after PCR amplification, and then these samples were used in PCR amplification with mRNA-specific primers for: IL-2, IL-4, and IL-5 (A) and for IL-6, IL-10, IL-13, IFN-γ, and CD3δ (B). The amplified cDNA was separated on an agarose gel. Lane 1 shows the results from PBMCs stimulated with phorbol myristate acetate and ionomycin, a regimen known to express all cytokine mRNA, which served as a positive control. Amplified cDNAs from cells recovered after challenge from two representative subjects (lanes 2 and 3) were compared with those obtained before challenge (lanes 4 and 5). Although results for only two average subjects are shown, similar results were observed in all 14 subjects. Journal of Allergy and Clinical Immunology 1996 98, 114-123DOI: (10.1016/S0091-6749(96)70233-6) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 2 In vivo challenge with DEPs increases cytokine mRNA expression. Cells from nasal lavages performed before and 18 hours after in vivo exposure to 0.3 mg of DEPs were used to obtain cDNA and amplified with β-actin mRNA-specific primers. Complementary DNA samples were adjusted to produce similar intensity bands on an ethidium bromide–stained agarose gel for β-actin after PCR amplification, and then these samples were used in PCR amplification with mRNA-specific primers for: IL-2, IL-4, and IL-5 (A) and for IL-6, IL-10, IL-13, IFN-γ, and CD3δ (B). The amplified cDNA was separated on an agarose gel. Lane 1 shows the results from PBMCs stimulated with phorbol myristate acetate and ionomycin, a regimen known to express all cytokine mRNA, which served as a positive control. Amplified cDNAs from cells recovered after challenge from two representative subjects (lanes 2 and 3) were compared with those obtained before challenge (lanes 4 and 5). Although results for only two average subjects are shown, similar results were observed in all 14 subjects. Journal of Allergy and Clinical Immunology 1996 98, 114-123DOI: (10.1016/S0091-6749(96)70233-6) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 2 In vivo challenge with DEPs increases cytokine mRNA expression. Cells from nasal lavages performed before and 18 hours after in vivo exposure to 0.3 mg of DEPs were used to obtain cDNA and amplified with β-actin mRNA-specific primers. Complementary DNA samples were adjusted to produce similar intensity bands on an ethidium bromide–stained agarose gel for β-actin after PCR amplification, and then these samples were used in PCR amplification with mRNA-specific primers for: IL-2, IL-4, and IL-5 (A) and for IL-6, IL-10, IL-13, IFN-γ, and CD3δ (B). The amplified cDNA was separated on an agarose gel. Lane 1 shows the results from PBMCs stimulated with phorbol myristate acetate and ionomycin, a regimen known to express all cytokine mRNA, which served as a positive control. Amplified cDNAs from cells recovered after challenge from two representative subjects (lanes 2 and 3) were compared with those obtained before challenge (lanes 4 and 5). Although results for only two average subjects are shown, similar results were observed in all 14 subjects. Journal of Allergy and Clinical Immunology 1996 98, 114-123DOI: (10.1016/S0091-6749(96)70233-6) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 2 In vivo challenge with DEPs increases cytokine mRNA expression. Cells from nasal lavages performed before and 18 hours after in vivo exposure to 0.3 mg of DEPs were used to obtain cDNA and amplified with β-actin mRNA-specific primers. Complementary DNA samples were adjusted to produce similar intensity bands on an ethidium bromide–stained agarose gel for β-actin after PCR amplification, and then these samples were used in PCR amplification with mRNA-specific primers for: IL-2, IL-4, and IL-5 (A) and for IL-6, IL-10, IL-13, IFN-γ, and CD3δ (B). The amplified cDNA was separated on an agarose gel. Lane 1 shows the results from PBMCs stimulated with phorbol myristate acetate and ionomycin, a regimen known to express all cytokine mRNA, which served as a positive control. Amplified cDNAs from cells recovered after challenge from two representative subjects (lanes 2 and 3) were compared with those obtained before challenge (lanes 4 and 5). Although results for only two average subjects are shown, similar results were observed in all 14 subjects. Journal of Allergy and Clinical Immunology 1996 98, 114-123DOI: (10.1016/S0091-6749(96)70233-6) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 2 In vivo challenge with DEPs increases cytokine mRNA expression. Cells from nasal lavages performed before and 18 hours after in vivo exposure to 0.3 mg of DEPs were used to obtain cDNA and amplified with β-actin mRNA-specific primers. Complementary DNA samples were adjusted to produce similar intensity bands on an ethidium bromide–stained agarose gel for β-actin after PCR amplification, and then these samples were used in PCR amplification with mRNA-specific primers for: IL-2, IL-4, and IL-5 (A) and for IL-6, IL-10, IL-13, IFN-γ, and CD3δ (B). The amplified cDNA was separated on an agarose gel. Lane 1 shows the results from PBMCs stimulated with phorbol myristate acetate and ionomycin, a regimen known to express all cytokine mRNA, which served as a positive control. Amplified cDNAs from cells recovered after challenge from two representative subjects (lanes 2 and 3) were compared with those obtained before challenge (lanes 4 and 5). Although results for only two average subjects are shown, similar results were observed in all 14 subjects. Journal of Allergy and Clinical Immunology 1996 98, 114-123DOI: (10.1016/S0091-6749(96)70233-6) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 2 In vivo challenge with DEPs increases cytokine mRNA expression. Cells from nasal lavages performed before and 18 hours after in vivo exposure to 0.3 mg of DEPs were used to obtain cDNA and amplified with β-actin mRNA-specific primers. Complementary DNA samples were adjusted to produce similar intensity bands on an ethidium bromide–stained agarose gel for β-actin after PCR amplification, and then these samples were used in PCR amplification with mRNA-specific primers for: IL-2, IL-4, and IL-5 (A) and for IL-6, IL-10, IL-13, IFN-γ, and CD3δ (B). The amplified cDNA was separated on an agarose gel. Lane 1 shows the results from PBMCs stimulated with phorbol myristate acetate and ionomycin, a regimen known to express all cytokine mRNA, which served as a positive control. Amplified cDNAs from cells recovered after challenge from two representative subjects (lanes 2 and 3) were compared with those obtained before challenge (lanes 4 and 5). Although results for only two average subjects are shown, similar results were observed in all 14 subjects. Journal of Allergy and Clinical Immunology 1996 98, 114-123DOI: (10.1016/S0091-6749(96)70233-6) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 Increased cytokine mRNA expression by DEPs is not due to increased T cell numbers alone. Cells from nasal lavages performed before and 18 hours after in vivo exposure to 0.3 mg of DEPs were used to obtain cDNA and amplified with CD3δ mRNA-specific primers. Complementary DNA samples were adjusted to produce similar intensity bands on an ethidium bromide–stained agarose gel for CD3δ after PCR amplification, and then these samples were used in PCR amplification with mRNA-specific primers for IL-2, IL-4, and IL-5 (A) and for IL-6, IL-10, IL-13, and IFN-γ (B). The amplified cDNA was separated on an agarose gel. Lane 1 shows the results from PBMCs stimulated with phorbol myristate acetate and ionomycin, a regimen known to express all cytokine mRNA, which served as a positive control. Amplified cDNAs from cells recovered after challenge from two representative subjects (lanes 2 and 3) were compared with those obtained before challenge (lanes 4 and 5). Although results for only two subjects are shown, similar results were observed in all 14 subjects. Journal of Allergy and Clinical Immunology 1996 98, 114-123DOI: (10.1016/S0091-6749(96)70233-6) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 Increased cytokine mRNA expression by DEPs is not due to increased T cell numbers alone. Cells from nasal lavages performed before and 18 hours after in vivo exposure to 0.3 mg of DEPs were used to obtain cDNA and amplified with CD3δ mRNA-specific primers. Complementary DNA samples were adjusted to produce similar intensity bands on an ethidium bromide–stained agarose gel for CD3δ after PCR amplification, and then these samples were used in PCR amplification with mRNA-specific primers for IL-2, IL-4, and IL-5 (A) and for IL-6, IL-10, IL-13, and IFN-γ (B). The amplified cDNA was separated on an agarose gel. Lane 1 shows the results from PBMCs stimulated with phorbol myristate acetate and ionomycin, a regimen known to express all cytokine mRNA, which served as a positive control. Amplified cDNAs from cells recovered after challenge from two representative subjects (lanes 2 and 3) were compared with those obtained before challenge (lanes 4 and 5). Although results for only two subjects are shown, similar results were observed in all 14 subjects. Journal of Allergy and Clinical Immunology 1996 98, 114-123DOI: (10.1016/S0091-6749(96)70233-6) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 Increased cytokine mRNA expression by DEPs is not due to increased T cell numbers alone. Cells from nasal lavages performed before and 18 hours after in vivo exposure to 0.3 mg of DEPs were used to obtain cDNA and amplified with CD3δ mRNA-specific primers. Complementary DNA samples were adjusted to produce similar intensity bands on an ethidium bromide–stained agarose gel for CD3δ after PCR amplification, and then these samples were used in PCR amplification with mRNA-specific primers for IL-2, IL-4, and IL-5 (A) and for IL-6, IL-10, IL-13, and IFN-γ (B). The amplified cDNA was separated on an agarose gel. Lane 1 shows the results from PBMCs stimulated with phorbol myristate acetate and ionomycin, a regimen known to express all cytokine mRNA, which served as a positive control. Amplified cDNAs from cells recovered after challenge from two representative subjects (lanes 2 and 3) were compared with those obtained before challenge (lanes 4 and 5). Although results for only two subjects are shown, similar results were observed in all 14 subjects. Journal of Allergy and Clinical Immunology 1996 98, 114-123DOI: (10.1016/S0091-6749(96)70233-6) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 Increased cytokine mRNA expression by DEPs is not due to increased T cell numbers alone. Cells from nasal lavages performed before and 18 hours after in vivo exposure to 0.3 mg of DEPs were used to obtain cDNA and amplified with CD3δ mRNA-specific primers. Complementary DNA samples were adjusted to produce similar intensity bands on an ethidium bromide–stained agarose gel for CD3δ after PCR amplification, and then these samples were used in PCR amplification with mRNA-specific primers for IL-2, IL-4, and IL-5 (A) and for IL-6, IL-10, IL-13, and IFN-γ (B). The amplified cDNA was separated on an agarose gel. Lane 1 shows the results from PBMCs stimulated with phorbol myristate acetate and ionomycin, a regimen known to express all cytokine mRNA, which served as a positive control. Amplified cDNAs from cells recovered after challenge from two representative subjects (lanes 2 and 3) were compared with those obtained before challenge (lanes 4 and 5). Although results for only two subjects are shown, similar results were observed in all 14 subjects. Journal of Allergy and Clinical Immunology 1996 98, 114-123DOI: (10.1016/S0091-6749(96)70233-6) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 Increased cytokine mRNA expression by DEPs is not due to increased T cell numbers alone. Cells from nasal lavages performed before and 18 hours after in vivo exposure to 0.3 mg of DEPs were used to obtain cDNA and amplified with CD3δ mRNA-specific primers. Complementary DNA samples were adjusted to produce similar intensity bands on an ethidium bromide–stained agarose gel for CD3δ after PCR amplification, and then these samples were used in PCR amplification with mRNA-specific primers for IL-2, IL-4, and IL-5 (A) and for IL-6, IL-10, IL-13, and IFN-γ (B). The amplified cDNA was separated on an agarose gel. Lane 1 shows the results from PBMCs stimulated with phorbol myristate acetate and ionomycin, a regimen known to express all cytokine mRNA, which served as a positive control. Amplified cDNAs from cells recovered after challenge from two representative subjects (lanes 2 and 3) were compared with those obtained before challenge (lanes 4 and 5). Although results for only two subjects are shown, similar results were observed in all 14 subjects. Journal of Allergy and Clinical Immunology 1996 98, 114-123DOI: (10.1016/S0091-6749(96)70233-6) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 Increased cytokine mRNA expression by DEPs is not due to increased T cell numbers alone. Cells from nasal lavages performed before and 18 hours after in vivo exposure to 0.3 mg of DEPs were used to obtain cDNA and amplified with CD3δ mRNA-specific primers. Complementary DNA samples were adjusted to produce similar intensity bands on an ethidium bromide–stained agarose gel for CD3δ after PCR amplification, and then these samples were used in PCR amplification with mRNA-specific primers for IL-2, IL-4, and IL-5 (A) and for IL-6, IL-10, IL-13, and IFN-γ (B). The amplified cDNA was separated on an agarose gel. Lane 1 shows the results from PBMCs stimulated with phorbol myristate acetate and ionomycin, a regimen known to express all cytokine mRNA, which served as a positive control. Amplified cDNAs from cells recovered after challenge from two representative subjects (lanes 2 and 3) were compared with those obtained before challenge (lanes 4 and 5). Although results for only two subjects are shown, similar results were observed in all 14 subjects. Journal of Allergy and Clinical Immunology 1996 98, 114-123DOI: (10.1016/S0091-6749(96)70233-6) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 Increased cytokine mRNA expression by DEPs is not due to increased T cell numbers alone. Cells from nasal lavages performed before and 18 hours after in vivo exposure to 0.3 mg of DEPs were used to obtain cDNA and amplified with CD3δ mRNA-specific primers. Complementary DNA samples were adjusted to produce similar intensity bands on an ethidium bromide–stained agarose gel for CD3δ after PCR amplification, and then these samples were used in PCR amplification with mRNA-specific primers for IL-2, IL-4, and IL-5 (A) and for IL-6, IL-10, IL-13, and IFN-γ (B). The amplified cDNA was separated on an agarose gel. Lane 1 shows the results from PBMCs stimulated with phorbol myristate acetate and ionomycin, a regimen known to express all cytokine mRNA, which served as a positive control. Amplified cDNAs from cells recovered after challenge from two representative subjects (lanes 2 and 3) were compared with those obtained before challenge (lanes 4 and 5). Although results for only two subjects are shown, similar results were observed in all 14 subjects. Journal of Allergy and Clinical Immunology 1996 98, 114-123DOI: (10.1016/S0091-6749(96)70233-6) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 Increased cytokine mRNA expression by DEPs is not due to increased T cell numbers alone. Cells from nasal lavages performed before and 18 hours after in vivo exposure to 0.3 mg of DEPs were used to obtain cDNA and amplified with CD3δ mRNA-specific primers. Complementary DNA samples were adjusted to produce similar intensity bands on an ethidium bromide–stained agarose gel for CD3δ after PCR amplification, and then these samples were used in PCR amplification with mRNA-specific primers for IL-2, IL-4, and IL-5 (A) and for IL-6, IL-10, IL-13, and IFN-γ (B). The amplified cDNA was separated on an agarose gel. Lane 1 shows the results from PBMCs stimulated with phorbol myristate acetate and ionomycin, a regimen known to express all cytokine mRNA, which served as a positive control. Amplified cDNAs from cells recovered after challenge from two representative subjects (lanes 2 and 3) were compared with those obtained before challenge (lanes 4 and 5). Although results for only two subjects are shown, similar results were observed in all 14 subjects. Journal of Allergy and Clinical Immunology 1996 98, 114-123DOI: (10.1016/S0091-6749(96)70233-6) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 4 In vivo challenge with DEPs induces IL-4 protein production. IL-4 protein was measured in nasal washes performed before and 18 hours after in vivo challenge with 0.3 mg of DEPs. Results from six subjects are shown. Journal of Allergy and Clinical Immunology 1996 98, 114-123DOI: (10.1016/S0091-6749(96)70233-6) Copyright © 1996 Mosby, Inc. Terms and Conditions