Diesel exhaust particles induce a Th2 phenotype in mouse naïve mononuclear cells in vitro

  • Authors:
    • Ken-ichiro Inoue
    • Eiko Koike
    • Akiko Endoh
    • Daigo Sumi
    • Yoshito Kumagai
    • Kazuichi Hayakawa
    • Masako Kiyono
    • Michitaka Tanaka
    • Hirohisa Takano
  • View Affiliations

  • Published online on: July 21, 2010     https://doi.org/10.3892/etm.2010.129
  • Pages: 761-767
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

It has been shown that pulmonary exposure to diesel exhaust particles (DEP) exacerbates allergic manifestations (i.e., allergic asthma) in vivo. However, the underlying mechanisms remain enigmatic. Furthermore, DEP components responsible for the exacerbation remain unidentified. Hypothesizing that DEP induce a helper T (Th)2-biased condition in immune mononuclear cells even in the absence of an allergen, the present study elucidated the effects of DEP and their components on the characterization of primary splenocytes and T cells in vitro. ICR mouse-derived splenic mononuclear and T cells isolated from the splenocytes were co-cultured with organic chemicals in DEP extracted with dichloromethane (DEP-OC 0.5-50 µg/ml), residual carbonaceous nuclei of DEP (washed DEP 0.5-50 µg/ml) or ‘whole’ DEP (1-100 µg/ml); thereafter, interleukin (IL)-4 and IgE production by these cells and the surface expression of CD19, IL-4R, CD69 and the CD40 Ligand (L) were evaluated by means of ELISA and flow cytometry. DEP and their components increased IL-4 and total IgE levels in a concentration-dependent manner 24 h after culture, and the effects were greater with washed DEP than with DEP-OC. On the other hand, each DEP component increased the surface expression of CD19 on splenocytes and that of IL-4R, CD69 or CD40L on both splenocytes and isolated T cells; however, the impact was greater with DEP-OC. These results suggest that DEP cause naïve mononuclear cells, including T cells to enter a Th2-biased state and that each DEP component plays a differential role in the efficacy.

Introduction

It is generally accepted that diesel exhaust particles (DEP), derived from diesel engine-powered automobiles and major constituents in atmospheric particulate matter, are associated with several allergic disorders, which perhaps can be referred to as adjuvanticity (1). We and others have experimentally demonstrated that pulmonary exposure to DEP aggravates allergic airway inflammation and amplifies the lung expression of helper T (Th)2 cytokines in vivo (25). However, detailed mechanisms underlying the exacerbating effects of DEP on the allergic pathology remain unclear, particularly when analyzed only in in vivo samples.

DEP reportedly affect/disrupt several cell populations such as epithelial cells (6,7), endothelial cells (8), macrophages (9), eosinophils (10) and mast cells/basophils (11,12), which play roles in the pathogenesis of allergic inflammation, mainly in vitro. Furthermore, we (13) and others (14) have shown that DEP activate dendritic cells, one of the pivotal cell types involved in adaptive immunity, leading to maladaptive immune responses in vitro. However, the cellular contribution of DEP-mediated exacerbation of allergy has not been fully clarified.

Peripheral lymphoid organs and their resident mononuclear cells, including lymphocytes, are key players in the pathogenesis of allergic pathology; thus, the effects of DEP on these compartments might generate much information regarding their adjuvant effects on allergy. Previous studies have shown that DEP and their components directly/indirectly activate B cells to produce IgE in vitro (15,16), implicating B/plasma cells in the mechanism of the adjuvant effects of DEP. However, less has been studied regarding the impact of DEP on T cells, in particular, in the context of Th lymphocyte differentiation/activation.

Another important issue is the fact that DEP have carbonaceous nuclei, which absorb organic chemicals, including polycyclic aromatic hydrocarbons (17). Previous studies have demonstrated that organic chemicals extracted from DEP have adjuvant potential for the production of allergen-specific immunogloblins in vivo (18) and in vitro (15). We previously reported that extracted organic chemicals from DEP with benzene-ethanol (referred to as ‘DEP-OC’), rather than residual carbonaceous nuclei of DEP after extraction (referred to as ‘washed DEP’), predominantly intensified allergen-related airway inflammation in mice, although ‘whole’ DEP (before extraction) exhibited much greater adjuvant effects than DEP-OC or washed DEP (19). Nonetheless, the distinct effect/role of DEP components in naïve immune cells in vitro has never been reported.

Therefore, the aim of the present study was i) to examine the effects of DEP on naïve splenic mononuclear cells and T cells isolated from splenocytes and ii) to compare the effects of organic chemical components (DEP-OC) and residual particles of DEP (washed DEP).

Materials and methods

Animals

Male ICR mice at 6 weeks of age and weighing 29–33 g (Japan Clea Co., Tokyo, Japan) were used in all experiments. They were fed a commercial diet (Japan Clea Co.) and given water ad libitum. Mice were housed in an animal facility maintained at 24–26°C with 55–75% humidity and a 12-h light/dark cycle. The studies adhered to the National Institutes of Health guidelines for the experimental use of animals. All animal studies were approved by the Institutional Review Board of the National Institute for Environmental Studies.

Preparation of DEP

A 4JB1-type, light-duty, four-cylinder, 2.74 L Isuzu diesel engine (Isuzu Automobile Co., Tokyo, Japan) under computer control was connected to a dynamometer (Meiden-sha, Tokyo, Japan). DEP were collected in the dilution tunnel (stainless steel tubing) of a diesel inhalation facility by scraping the inside surface of the tubing as previously described (20). The collected DEP were referred to as ‘whole’ DEP and stored at −80°C until use. Whole DEP were suspended in phosphate-buffered saline (PBS; Sigma, St. Louis, MO, USA) at pH 7.4 containing 0.05% Tween-80 (Nakarai Tesque, Kyoto, Japan) and 0.25% dimethyl sulfoxide (DMSO) (Nakalai Tesque). The whole DEP suspension was sonicated for 3 min with an ultrasonic disrupter (UD-201; Tomy Seiko, Tokyo, Japan), as previously described (21,22).

Preparation of DEP-OC and washed DEP

DEP were extracted with benzene-ethanol as previously described (19). Briefly, 10 g of DEP was suspended in 800 ml of benzene-ethanol (3:1, v/v) and was ultrasonicated for 30 min. The suspension was centrifuged at 600 × g for 20 min. The supernatants were transferred to another tube and then were filtered with a membrane filter (pore size 0.45 μm). This procedure was repeated twice. The residual (solid) particles of DEP were prepared as washed DEP. The benzene-ethanol solution was evaporated to dryness in a rotary evaporator and then a vacuum pump, and the residue was dissolved in 100% DMSO and prepared as DEP-OC. They were also stored at −80°C until use.

Splenocyte preparation and DEP exposure

Spleens were removed from the ICR mice and placed in a Petri dish with PBS. The spleens were pushed through a 200-mesh stainless steel sheet, and the resulting cells were suspended in PBS. Cells were collected by centrifugation at 400 × g for 10 min at 20°C. Thereafter, red blood cells were removed by incubating with hypotonic lysis buffer for 3 min. Cells were washed twice with PBS and resuspended in R10 [Gibco RPMI-1640 medium (Invitrogen, Grand Island, NY, USA), supplemented with 10% heat-inactivated fetal bovine serum (FBS; MP Biomedicals Inc., Eschwege, Germany), 100 U/ml penicillin, 100 μg/ml streptomycin (Sigma) and 50 μM 2-mercaptoethanol (Invitrogen)]. The number of viable cells was determined using the trypan blue dye exclusion method. Splenocytes (1×106) were cultured in 1 ml of R10 containing DEP-OC (0.5, 5 or 50 μg/ml), washed DEP (0.5, 5 or 50 μg/ml), whole DEP (1, 10 or 100 μg/ml) or the control (0.1% DMSO, 0.05% Tween-80) in 12-well plates at 37°C in a 5% CO2/95% air atmosphere. After a 24-h incubation, cells were analyzed for FACS, and the culture supernatants were collected and frozen for subsequent interleukin (IL)-4 and total IgE analyses. Since the organic fraction of DEP constitutes ∼50% of total particle mass (16,23), we set the concentration of each component.

T-cell isolation and DEP exposure

T lymphocytes were isolated by passing splenocytes (1×108) over a nylon fiber column (Wako Pure Chemical Industries Ltd., Osaka, Japan) based on a T-cell purification method. This method utilizes the principle of nylon fiber's affinity to B cells; thus, T-cell preparation of sufficient purity and without severe damages was achieved through the preferential adsorption of B cells to the fiber in the column (24). The column was incubated for 45 min at 37°C in a 5% CO2/95% air atmosphere; thereafter, it was washed with R10 and the fraction of non-adherent cells (T cells) was collected. The T cells were then resuspended in R10. The DEP exposure protocol and examination points and parameters (except for IgE) in the experiments were the same as for the splenocytes.

ELISA for interleukin (IL)-4

ELISA for IL-4 (Amersham, Buckinghamshire, UK) was conducted according to the manufacturer's instructions. The secondary antibodies were conjugated to horseradish peroxidase. Values generated by subtracting readings obtained at 450 nm from those at 550 nm were converted to pg/ml using values obtained from standard curves generated with the limits of detection at 5 pg/ml.

Total IgE determination

Total IgE antibody level was measured by IgE-capture ELISA (2,25). In brief, microplate wells were coated with a rat anti-mouse IgE monoclonal antibody (BD Biosciences Pharmingen, San Diego, CA, USA) and incubated overnight at 4°C. After washing with PBS containing 0.05% Tween-20 (PBST; Nakalai Tesque), microplate wells were incubated with 1% BSA-PBS and 0.01% thimerosal at room temperature for 1 h. After washing with PBS containing PBST, diluted samples were added to the microplate and incubated overnight at 4°C. After washing with PBST, biotinylated rat anti-mouse IgE was added to each well and incubated for 1 h at room temperature with β-D-galactosidase-conjugated streptavidin (Zymed Laboratories, San Francisco, CA, USA). After the final washing, the wells were incubated with 4-methylumbelliferyl-β-galactoside (Sigma) as the enzyme substrate at 37°C for 2 h. The enzyme reaction was stopped with 0.1 M glycine-NaOH (pH 10.3). The fluorescene intensity was read using a microplate fluorescene reader (Fluoroskan Flow Laboratories, Costa Mesa, CA, USA). A450 readings of the samples were converted to nanograms per milliliter, using a standard curve generated with double dilutions of the mouse IgE κ isotype standard (BD Biosciences Pharmingen).

FACS analysis

For FACS analysis, the following monoclonal antibodies were used: IL-4R (CD124: mIL4R-M1, PE-conjugated), CD69 (H1. 2F3), CD40 Ligand (L) (CD154: gp39: MR1, PE-conjugated) and CD19 (FITC-conjugated) (all from BD Biosciences Pharmingen). Cells (1–3×105) were resuspended in 100 μl PBS with 0.3% BSA and 0.05% sodium azide (Wako Pure Chemical Industries Ltd.) and stained with antibodies at 1 μg for 30 min on ice. After incubation, the cells were washed, and fluorescence was measured using a FACSCalibur (Becton Dickinson and Company, NJ, USA). For each sample, fluorescence data from 10,000 cells were collected, and positive cells were expressed as the percentage of total events.

Statistical analysis

Data are expressed as the mean ± SEM of four animals from one experiment, representative of three experiments. Differences among groups were analyzed by ANOVA (Stat View version 4.0; Abacus Concepts, Inc., Berkeley, CA, USA). When significant differences were detected, post hoc comparisons within each group were evaluated with the Fisher's LSD test. Significance was assigned to P-values <0.05.

Results

Effects of DEP on IL-4 production from naïve splenocytes and their T cells

To determine the effects of DEP exposure on naïve splenocytes and their T cells in the context of the Th2 milieu, we compared the protein levels of IL-4 in the supernatants 24 h after co-culture (Fig. 1). Exposure to whole DEP and their components elevated IL-4 levels in the supernatants from splenocytes and isolated T cells in a concentration-dependent manner as compared to IL-4 levels in the cells exposed to the control medium [P<0.01 vs. whole DEP (100 μg/ml), DEP-OC (50 μg/ml) or washed DEP (50 μg/ml), except for T-cells exposed to DEP-OC (50 μg/ml) (P<0.05)].

Effects of DEP on total IgE production from naïve splenocytes

To further determine whether DEP exposure biases these mononuclear cells towards a Th2 response, we compared protein levels of total IgE in the supernatants from the splenocytes 24 h after co-culture (Fig. 2). Whole DEP and their components increased the total IgE level in a concentration-dependent manner (data not shown). As compared to exposure to medium alone, exposure to whole DEP and their components at each maximal concentration elevated the value in the supernatants (P<0.01). However, in this context, whole DEP did not reveal synergistic effects of DEP-OC plus washed DEP, as observed in the previous in vivo study (19).

Effects of DEP on CD19 expression on naïve splenocytes and IL-4R, CD69 and CD40L expression on naïve splenocytes and their T cells

To determine the effects of exposure to DEP on the surface expression of molecules related to lymphocyte activation 24 h after the culture of naïve splenocytes and their T cells, we analyzed the expression patterns of CD19 (on splenocytes; Fig. 2), IL-4R, CD69 and CD40L (on splenocytes and T cells) by FACS (Fig. 3). Exposure to whole DEP, washed DEP or DEP-OC significantly increased CD19 expression on splenocytes as compared to the level of CD19 expression on splenocytes exposed to the control medium (P<0.01; Fig. 2). The expression levels of IL-4R, CD69 or CD40L were significantly higher on both splenocytes and isolated T cells treated with whole DEP (100 μg/ml) or DEP-OC (50 μg/ml) than the expression levels on cells treated with medium alone (P<0.01). Also, the level was higher on cells treated with washed DEP (P<0.01 for IL-4R on both cells, CD69 or CD40L on splenocytes; P<0.05 for CD40L on T cells) compared to the level on cells treated with medium; however, the level was lower on cells treated with DEP-OC (P<0.01 for IL-4R on splenocytes, CD69 or CD40L; data not shown).

Discussion

The present study demonstrated that DEP exposure induces IL-4 and IgE production and several surface markers related to lymphocyte activation by mouse naïve mononuclear cells and/or isolated T cells in vitro. Studies on their constituents show that both DEP components induce IL-4 production by splenocytes and isolated T cells and consequent IgE synthesis in splenocytes; however, the effect is generally greater for washed DEP than for DEP-OC. In contrast, DEP-OC only increased the surface expression of CD19 on splenocytes. Furthermore, although both components increased the surface expression of IL-4R, CD69 and CD40L on both splenocytes and isolated T cells, the impact was significantly greater for DEP-OC than for washed DEP with an overall trend.

With respect to the adjuvant activity of DEP on atopic pathophysiology, there are two major aspects, i.e., impacts on allergens/antigens and on host immunity. DEP reportedly absorb several allergens, such as Lol p1, Bet v 1, Der p 1, Fel d 1 and Can f 1 (26,27). Furthermore, D'Amato et al demonstrated that, by adhering to the surface of airborne allergens, such as pollen grains and/or plant-derived paucimicronic components, air pollutants including DEP, modify their antigenic properties (28), although the underlying mechanisms remain to be determined. On the other hand, DEP and/or DE were reported to directly/indirectly influence several host cell populations, particularly immune cells, leading to the progression/development/exacerbation of allergic disorders in vitro. For example, DEP and their components directly activated B cells to enhance IgE production (29,16), chemical constituents of DEP induced IL-4 production/release by human-derived basophils (12) and DE exposure decreased IL-12 (Th1 cytokine) production from alveolar macrophages or a macrophage cell line (RAW264.7 cell) in vitro (30) as well as that in lung homogenates in vivo (2). Furthermore, DEP was found to suppress T-bet expression and interferon-γ production by isolated T cells from healthy humans, implicating DEP in promoting Th2-skewed responses in lymphocytes (31,32). In the present study, whole DEP and their components (washed DEP and DEP-OC) induced IL-4 production and IgE synthesis from splenocytes. Our present study was, accordingly, consistent with these previous studies (31,32) with an overall trend; however, in the present study, we first demonstrated that DEP components differentially contributed to the adjuvant effects on allergic reaction, i.e., washed DEP and DEP-OC induced a Th2 response (IL-4 production with IgE de novo synthesis) in splenocytes, including T cells, whereas DEP-OC had a marked potential to up-regulate IL-4R (with CD69, CD40L and CD19) on these cells, which conceivably culminates in the promotion of Th2-skewed immune reactions and consequent atopic immunopathology (Fig. 4). In addition, we and another group found that DEP activated dendritic cells, professional antigen-presenting cells, to favor allergen-specific Th2 immunogenicity in vitro (33,13). Thus, the present study also expanded the previous ones in that DEP and their components favor a Th2 milieu in naïve splenic mononuclear cells and T cells isolated from splenocytes, even in the absence of antigen-presenting cells (plus allergen), indicating that these toxicants have the potential to promote an allergic pathophysiology irrespective of allergen sensitization and/or DC/T cell interaction.

We previously demonstrated that DEP components differentially affect allergic lung inflammation in vivo, i.e., DEP-OC predominantly exacerbated allergic inflammation, whereas washed DEP did not aggravate the pathology (19). Also, in the present study, DEP-OC induced IL-4/IgE production/synthesis, implying a role in the Th2 milieu even in the absence of allergen-priming. Furthermore, DEP-OC markedly induced the surface expression of CD19, suggesting a role in B cell-plasma cell activation. These in vitro effects of DEP-OC support the adjuvant property against Th2-mediated pathobiology in vivo (19) and also might be one of the mechanisms explaining the phenomenon (19). Notably, however, washed DEP also significantly elevated IL-4 production with IgE synthesis in the present study. Conversely, our previous in vivo study demonstrated that pulmonary exposure to washed DEP in asthmatic mice did not increase the lung expression of Th2-type cytokines, such as IL-4, IL-5 and IL-13, but did exacerbate Th1-type cytokines, such as interferon-γ (19). These inconsistent results may be explained by a difference between in vitro and in vivo studies. As compared to an in vivo condition (19), washed DEP easily come into contact with immune cells and stimulate/activate them in vitro. In support of this, we confirmed that carbon (black) nanoparticles, which are ultrafine particles less than 100 nm in mass median aerodynamic diameter compared to washed DEP (200 nm), exacerbate allergic airway inflammation even in vivo (25). Furthermore, the enhancement was greater with smaller (14 nm) than with larger (56 nm) nanoparticles (25), suggesting that more mobile particles have a higher adjuvant potential. On the contrary, a soluble component, DEP-OC, may influence systemic allergic response (broadly) likely reaching the spleen, thereby yielding profound adverse effects on allergic pathophysiology in vivo. Alternatively, DEP-derived carbonaceous compounds (washed DEP) may have antiallergic properties against alveolar macrophages and/or lung resident cells, including endothelial cells, epithelial cells and fibroblasts rather than mononuclear cells, which can hinder the enhancing effects in vivo. Further studies are required to address this issue.

CD69 is an early activation marker of hematopoietic cells. The molecule is reportedly linked to the activation of Th1 lineage in vitro (34) and Th1-dominant immunopathology in vivo (35). CD40-CD40L (CD154) interaction is crucial in the regulation of crosstalk between both dendritic (DC)-T and DC-B cells (36). In the present study, DEP-OC exposure dramatically up-regulated CD69 and CD40L on both splenocytes and T cells. It is attractive to speculate that the amplifying effect on CD69 expression partially contributes to relatively weaker IL-4 synthesis by DEP-OC causally via altering Th1/Th2 homeostasis than that by washed DEP. Alternatively, DEP-OC may fully show their adjuvant effects on allergic reactions in cases of (allergen)/antigen-presenting cells/lymphocytes rather than naïve (physiological) conditions/cells. Future investigations are necessary employing the same protocol involving Th2 cells with or without allergen-priming.

There are some limitations regarding this study. We used splenocytes and isolated T cells from these cells, not completely matching the real situation in terms of the exposure pattern, as humans are exposed to particulate matter (PM), including DEP mainly through the airway route. However, our recent ex vivo studies showed that pulmonary exposure to DEP potentiates CD4 polarization (37) with an allergenspecific Th2 response in splenocytes of asthmatic mice (13), suggesting that these extrathoracic lymphoid cells are a target. In the future, nevertheless, the same protocol as in the present study should be employed involving draining lymph node cells around respiratory systems, such as cervical and mediastinal nodes. In any case, the present study may provide a clue for future novel screening tests to detect the pro-allergic activity of other conceivable pollutants, such as airborne PM, nanomaterials and environmental chemicals to elicit allergic reactions. Nonetheless, investigation of intracellular signaling regarding the differential effects of DEP components in depth to identify molecular targets is warranted. We are currently examining this using a T-cell line (Jurkat cells). In the cell line, whole DEP and washed DEP significantly and DEP-OC modestly activated ERK1/2 of MAP kinase (unpublished data) consistent with a previous report (32).

In summary, whole DEP and their components induced IL-4 production with IgE synthesis from naïve splenocytes and/or splenocyte-derived T cells in vitro. The effects were stronger with residual carbonaceous nuclei in DEP than with organic chemical components, whereas, organic chemical components markedly increased the surface expression of molecules related to B-cell activation, such as CD19, and T-cell activation, such as IL-4R, CD69 and CD40L, on these cells. These data indicate the pro-allergic effects of DEP on lymphoid-lineage cells, even in the absence of allergen-priming in vitro, and raise the possibility that each component in DEP differentially affects immune cells, which culminates in an increase in the DEP-mediated exacerbation of allergic inflammatory conditions with maladaptive Th2 immunity.

Acknowledgements

This study was partly funded by grants from the Grant-in-Aid (#20120014 to H. Takano) for Scientific Research on Innovative Areas. The authors thank Rie Yanagisawa, Naoko Ueki, Satomi Abe and Rieko Shibahara for the assistance throughout the study.

References

1. 

Ichinose T, Furuyama A and Sagai M: Biological effects of diesel exhaust particles (DEP). II. Acute toxicity of DEP introduced into lung by intratracheal instillation. Toxicology. 99:153–167. 1995. View Article : Google Scholar : PubMed/NCBI

2. 

Takano H, Yoshikawa T, Ichinose T, Miyabara Y, Imaoka K and Sagai M: Diesel exhaust particles enhance antigen-induced airway inflammation and local cytokine expression in mice. Am J Respir Crit Care Med. 156:36–42. 1997. View Article : Google Scholar : PubMed/NCBI

3. 

Ichinose T, Takano H, Miyabara Y and Sagai M: Long-term exposure to diesel exhaust enhances antigen-induced eosinophilic inflammation and epithelial damage in the murine airway. Toxicol Sci. 44:70–79. 1998. View Article : Google Scholar

4. 

Miyabara Y, Takano H, Ichinose T, Lim HB and Sagai M: Diesel exhaust enhances allergic airway inflammation and hyperresponsiveness in mice. Am J Respir Crit Care Med. 157:1138–1144. 1998. View Article : Google Scholar : PubMed/NCBI

5. 

Takano H, Ichinose T, Miyabara Y, Yoshikawa T and Sagai M: Diesel exhaust particles enhance airway responsiveness following allergen exposure in mice. Immunopharmacol Immunotoxicol. 20:329–336. 1998. View Article : Google Scholar : PubMed/NCBI

6. 

Terada N, Maesako K, Hiruma K, Hamano N, Houki G, Konno A, Ikeda T and Sai M: Diesel exhaust particulates enhance eosinophil adhesion to nasal epithelial cells and cause degranulation. Int Arch Allergy Immunol. 114:167–174. 1997. View Article : Google Scholar

7. 

Takizawa H, Abe S, Okazaki H, Kohyama T, Sugawara I, Saito Y, Ohtoshi T, Kawasaki S, Desaki M, Nakahara K, Yamamoto K, Matsushima K, Tanaka M, Sagai M and Kudoh S: Diesel exhaust particles upregulate eotaxin gene expression in human bronchial epithelial cells via nuclear factor-kappa B-dependent pathway. Am J Physiol Lung Cell Mol Physiol. 284:L1055–L1062. 2003. View Article : Google Scholar

8. 

Terada N, Hamano N, Maesako KI, Hiruma K, Hohki G, Suzuki K, Ishikawa K and Konno A: Diesel exhaust particulates upregulate histamine receptor mRNA and increase histamine-induced IL-8 and GM-CSF production in nasal epithelial cells and endothelial cells. Clin Exp Allergy. 29:52–59. 1999. View Article : Google Scholar

9. 

Beck-Speier I, Dayal N, Karg E, Maier KL, Schumann G, Schulz H, Semmler M, Takenaka S, Stettmaier K, Bors W, Ghio A, Samet JM and Heyder J: Oxidative stress and lipid mediators induced in alveolar macrophages by ultrafine particles. Free Radic Biol Med. 38:1080–1092. 2005. View Article : Google Scholar : PubMed/NCBI

10. 

Hirota R, Akimaru K and Nakamura H: In vitro toxicity evaluation of diesel exhaust particles on human eosinophilic cell. Toxicol In Vitro. 22:988–994. 2008. View Article : Google Scholar : PubMed/NCBI

11. 

Saneyoshi K, Nohara O, Imai T, Shiraishi F, Moriyama H and Fujimaki H: IL-4 and IL-6 production of bone marrow-derived mast cells is enhanced by treatment with environmental pollutants. Int Arch Allergy Immunol. 114:237–245. 1997. View Article : Google Scholar : PubMed/NCBI

12. 

Devouassoux G, Saxon A, Metcalfe DD, Prussin C, Colomb MG, Brambilla C and Diaz-Sanchez D: Chemical constituents of diesel exhaust particles induce IL-4 production and histamine release by human basophils. J Allergy Clin Immunol. 109:847–853. 2002. View Article : Google Scholar : PubMed/NCBI

13. 

Inoue K, Koike E, Takano H, Yanagisawa R, Ichinose T and Yoshikawa T: Effects of diesel exhaust particles on antigen-presenting cells and antigen-specific Th immunity in mice. Exp Biol Med. 234:200–209. 2009. View Article : Google Scholar : PubMed/NCBI

14. 

Porter M, Karp M, Killedar S, Bauer SM, Guo J, Williams D, Breysse P, Georas SN and Williams MA: Diesel-enriched particulate matter functionally activates human dendritic cells. Am J Respir Cell Mol Biol. 37:706–719. 2007. View Article : Google Scholar : PubMed/NCBI

15. 

Takenaka H, Zhang K, Diaz-Sanchez D, Tsien A and Saxon A: Enhanced human IgE production results from exposure to the aromatic hydrocarbons from diesel exhaust: direct effects on B-cell IgE production. J Allergy Clin Immunol. 95:103–115. 1995. View Article : Google Scholar : PubMed/NCBI

16. 

Tsien A, Diaz-Sanchez D, Ma J and Saxon A: The organic component of diesel exhaust particles and phenanthrene, a major polyaromatic hydrocarbon constituent, enhances IgE production by IgE-secreting EBV-transformed human B cells in vitro. Toxicol Appl Pharmacol. 142:256–263. 1997. View Article : Google Scholar

17. 

IARC: Monographs on the Evaluation of Carcinogenic Risks to Humans: Diesel and Gasoline Exhaust and Some Nitoarenes. Lyon, France: pp. 1989

18. 

Heo Y, Saxon A and Hankinson O: Effect of diesel exhaust particles and their components on the allergen-specific IgE and IgG1 response in mice. Toxicology. 159:143–158. 2001. View Article : Google Scholar : PubMed/NCBI

19. 

Yanagisawa R, Takano H, Inoue K, Ichinose T, Sadakane K, Yoshino S, Yamaki K, Yoshikawa T and Hayakawa K: Components of diesel exhaust particles differentially affect Th1/Th2 response in a murine model of allergic airway inflammation. Clin Exp Allergy. 36:386–395. 2006. View Article : Google Scholar : PubMed/NCBI

20. 

Sagai M, Furuyama A and Ichinose T: Biological effects of diesel exhaust particles (DEP). III. Pathogenesis of asthma like symptoms in mice. Free Radic Biol Med. 21:199–209. 1996. View Article : Google Scholar : PubMed/NCBI

21. 

Takano H, Yanagisawa R, Ichinose T, Sadakane K, Yoshino S, Yoshikawa T and Morita M: Diesel exhaust particles enhance lung injury related to bacterial endotoxin through expression of proinflammatory cytokines, chemokines, and intercellular adhesion molecule-1. Am J Respir Crit Care Med. 165:1329–1335. 2002. View Article : Google Scholar

22. 

Yanagisawa R, Takano H, Inoue K, Ichinose T, Sadakane K, Yoshino S, Yamaki K, Kumagai Y, Uchiyama K, Yoshikawa T and Morita M: Enhancement of acute lung injury related to bacterial endotoxin by components of diesel exhaust particles. Thorax. 58:605–612. 2003. View Article : Google Scholar : PubMed/NCBI

23. 

Kawasaki S, Takizawa H, Takami K, Desaki M, Okazaki H, Kasama T, Kobayashi K, Yamamoto K, Nakahara K, Tanaka M, Sagai M and Ohtoshi T: Benzene-extracted components are important for the major activity of diesel exhaust particles: effect on interleukin-8 gene expression in human bronchial epithelial cells. Am J Respir Cell Mol Biol. 24:419–426. 2001. View Article : Google Scholar

24. 

Julius MH, Simpson E and Herzenberg LA: A rapid method for the isolation of functional thymus-derived murine lymphocytes. Eur J Immunol. 3:645–649. 1973. View Article : Google Scholar : PubMed/NCBI

25. 

Inoue K, Takano H, Yanagisawa R, Sakurai M, Ichinose T, Sadakane K and Yoshikawa T: Effects of nano particles on antigen-related airway inflammation in mice. Respir Res. 6:1062005. View Article : Google Scholar : PubMed/NCBI

26. 

Knox RB, Suphioglu C, Taylor P, Desai R, Watson HC, Peng JL and Bursill LA: Major grass pollen allergen Lol p 1 binds to diesel exhaust particles: implications for asthma and air pollution. Clin Exp Allergy. 27:246–251. 1997. View Article : Google Scholar : PubMed/NCBI

27. 

Ormstad H, Johansen BV and Gaarder PI: Airborne house dust particles and diesel exhaust particles as allergen carriers. Clin Exp Allergy. 28:702–708. 1998. View Article : Google Scholar : PubMed/NCBI

28. 

D'Amato G, Liccardi G, D'Amato M and Cazzola M: Outdoor air pollution, climatic changes and allergic bronchial asthma. Eur Respir J. 20:763–776. 2002.PubMed/NCBI

29. 

Zhang K, Clark EA and Saxon A: CD40 stimulation provides an IFN-gamma-independent and IL-4-dependent differentiation signal directly to human B cells for IgE production. J Immunol. 146:1836–1842. 1991.

30. 

Saito Y, Azuma A, Kudo S, Takizawa H and Sugawara I: Effects of diesel exhaust on murine alveolar macrophages and a macrophage cell line. Exp Lung Res. 28:201–217. 2002. View Article : Google Scholar : PubMed/NCBI

31. 

Ohtani T, Nakagawa S, Kurosawa M, Mizuashi M, Ozawa M and Aiba S: Cellular basis of the role of diesel exhaust particles in inducing Th2-dominant response. J Immunol. 174:2412–2419. 2005. View Article : Google Scholar : PubMed/NCBI

32. 

Sasaki Y, Ohtani T, Ito Y, Mizuashi M, Nakagawa S, Furukawa T, Horii A and Aiba S: Molecular events in human T cells treated with diesel exhaust particles or formaldehyde that underlie their diminished interferon-gamma and interleukin-10 production. Int Arch Allergy Immunol. 148:239–250. 2009. View Article : Google Scholar

33. 

Chan RC, Wang M, Li N, Yanagawa Y, Onoe K, Lee JJ and Nel AE: Pro-oxidative diesel exhaust particle chemicals inhibit LPS-induced dendritic cell responses involved in T-helper differentiation. J Allergy Clin Immunol. 118:455–465. 2006. View Article : Google Scholar : PubMed/NCBI

34. 

Harimaya A, Himi T, Fujii N, Tarkkanen J, Carlson P, Ylikoski J and Mattila P: Induction of CD69 expression and Th1 cytokines release from human peripheral blood lymphocytes after in vitro stimulation with Alloiococcus otitidis and three middle ear pathogens. FEMS Immunol Med Microbiol. 43:385–392. 2005. View Article : Google Scholar : PubMed/NCBI

35. 

Hernandez-Garcia C, Fernandez-Gutierrez B, Morado IC, Banares AA and Jover JA: The CD69 activation pathway in rheumatoid arthritis synovial fluid T cells. Arthritis Rheum. 39:1277–1286. 1996. View Article : Google Scholar : PubMed/NCBI

36. 

Ma DY and Clark EA: The role of CD40 and CD154/CD40L in dendritic cells. Semin Immunol. 21:265–272. 2009. View Article : Google Scholar : PubMed/NCBI

37. 

Inoue K, Koike E, Yanagisawa R and Takano H: Effects of pulmonary exposure to diesel exhaust particles on extrathoracic CD4 polarization in asthmatic mice. Immunopharmacol Immunotoxicol. 31:71–74. 2009. View Article : Google Scholar : PubMed/NCBI

Related Articles

Journal Cover

September 2010
Volume 1 Issue 5

Print ISSN: 1792-0981
Online ISSN:1792-1015

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
APA
Inoue, K., Koike, E., Endoh, A., Sumi, D., Kumagai, Y., Hayakawa, K. ... Takano, H. (2010). Diesel exhaust particles induce a Th2 phenotype in mouse naïve mononuclear cells in vitro. Experimental and Therapeutic Medicine, 1, 761-767. https://doi.org/10.3892/etm.2010.129
MLA
Inoue, K., Koike, E., Endoh, A., Sumi, D., Kumagai, Y., Hayakawa, K., Kiyono, M., Tanaka, M., Takano, H."Diesel exhaust particles induce a Th2 phenotype in mouse naïve mononuclear cells in vitro". Experimental and Therapeutic Medicine 1.5 (2010): 761-767.
Chicago
Inoue, K., Koike, E., Endoh, A., Sumi, D., Kumagai, Y., Hayakawa, K., Kiyono, M., Tanaka, M., Takano, H."Diesel exhaust particles induce a Th2 phenotype in mouse naïve mononuclear cells in vitro". Experimental and Therapeutic Medicine 1, no. 5 (2010): 761-767. https://doi.org/10.3892/etm.2010.129