Supplementary Material, Supplemental_Figure_S1 – International Journal of Immunopathology and Pharmacology

<p>Supplementary Material, Supplemental_Figure_S1 for International Journal of Immunopathology and Pharmacology by Eiko Koike, Rie Yanagisawa, Tin-Tin Win-Shwe and Hirohisa Takano in International Journal of Immunopathology and Pharmacology</p

Effects of lactational exposure to low-dose BaP on allergic and non-allergic immune responses in mice offspring

<p>Benzo[a]pyrene (BaP) can induce developmental and reproductive toxicity; however, the full scope of its immunotoxic effects remains unknown. This study aimed to assess effects of lactational exposure to low-dose BaP (comparable to human exposure) on potential allergic\non-allergic immune responses in murine offspring. Lactating C3H/HeJ dams were orally dosed with BaP at 0, 0.25, 5.0, or 100 pmol/animal/week) at post-natal days [PND] 1, 8, and 15. Five-weeks-old pups then received intratracheally ovalbumin (OVA) every 2 weeks for 6 weeks. Following the final exposure, mice were processed to permit analyses of bronchoalveolar lavage (BAL) fluid cell profiles as well as levels of lung inflammatory cytokines and chemokines, serum OVA-specific immunoglobulin, and mediastinal lymph node (MLN) cell activation/proliferation. In OVA-sensitized male offspring, lactational low-dose BaP exposure led to enhanced (albeit not significantly) macrophage, neutrophil, and eosinophil infiltration to, and increased T-helper (T_H)-2 cytokine production in, the lungs. In females, BaP exposure, regardless of dose, led to slightly enhanced lung levels of macrophages and eosinophils, and of inflammatory molecules. Protein levels of interleukin (IL)-33 in the OVA + BaP (middle dose) group, and interferon (IFN)-γ in the OVA + BaP (low dose) group, were higher than that of the OVA (no BaP) group. <i>Ex vivo</i> studies showed lactational exposure to BaP partially induced activation of T-cells and antigen-presenting cells (APCs) in the MLN cells of both male and female offspring, with or without OVA sensitization. Further, IL-4 and IFNγ levels in MLN culture supernatants were elevated even without OVA-re-stimulation in OVA + BaP groups. In conclusion, lactational exposure to low-dose BaP appeared to exert slight effects on later allergic and non-allergic immune responses in offspring by facilitating development of modest T_H2 responses and activating MLN cells. In addition, lactational exposures to BaP might give rise to gender differences in allergic/non-allergic immune responses of offspring.</p

Results of the <i>t</i>-test in terms of average values for all channels (1 to 42) comparing z scores for oxyHb between MCS patients and controls.

<p>Values are expressed as means (± standard deviations).</p><p>*Significant at <i>p</i><0.05.</p><p>Abbreviations: MO, mandarin orange; Pf, perfume; NO, non-odorant; JC, Japanese cypress; Mt, menthol. Numbers in parentheses in column 1 indicate the order of the 10 repetitions (1 to 10).</p

Ratings of hedonic (A) and irritating (B) odours by MCS patients (<i>n</i> = 12) and controls (<i>n</i> = 11) after the olfactory stimulation.

<p>Abbreviations: MO: mandarin orange, Pf: perfume, NO: non-odorant, JC: Japanese cypress, Mt: menthol. Numbers in parentheses indicate orders of ten repetitions (1 to 10). Statistically significant differences between groups are indicated. ^*<i>p</i><0.05, ^**<i>p</i><0.01.</p

Results of the <i>t</i>-test for the physical and psychological scales.

<p>Values are expressed as means (± standard deviations).</p><p>*Significant at <i>p</i><0.05.</p>a<p>Because of missing values, <i>t</i>-test results included the following numbers of patients. MCS and control: CSS-SHR, MUSS, APQ, TMAS, MCSD, TAS, TAS-20, TAS-20 DIF, TAS-20 DDF and TAS-20 EOT, <i>n</i> = 11 and <i>n</i> = 10; SSAS, CHS, CNSS and CSAS, <i>n</i> = 11 and <i>n</i> = 11; NAS, <i>n</i> = 10 and <i>n</i> = 11.</p><p>Abbreviations: CI, chemical intolerance; OI, other intolerance; SS, symptom severity.</p

Average <i>t</i> value of each channel comparing z scores for oxyHb between MCS patients (<i>n</i> = 12) and controls (<i>n</i> = 11).

<p>Statistically significant differences between groups are indicated as underlined values. ^*<i>p</i><0.05, ^**<i>p</i><0.01. Significant tendencies are indicated: ⁺<i>p</i><0.10.</p

Topographical maps of average z scores for oxyHb in MCS patients (<i>n</i> = 12) and controls (<i>n</i> = 11).

<p>Abbreviations: MO: mandarin orange, Pf: perfume, NO: non-odorant, JC: Japanese cypress, Mt: menthol. Numbers in parentheses indicate the order of the 10 repetitions (1 to 10).</p

Correlation coefficient (r) between rCBF after the first and second exposures to the odor in terms of z scores for all channels (1 to 42).

<p>Values are expressed as Pearson product-moment correlation coefficients.</p><p>*Significant at <i>p</i><0.05.</p><p>Abbreviations: MO, mandarin orange; Pf, perfume; NO, non-odorant; JC, Japanese cypress; Mt, menthol.</p

Experimental setting and NIRS channel orientation.

<p>Detectors and illuminators are shown as gray circles (1 to 14). Channels are shown as white squares (1 to 42). The international 10–20 standard positions and other positional information are indicated. One holder with 42 NIRS channels was set on the PFC of each patient so that the midpoint of channels 38 and 39 corresponded to the intersection point of the F7, F8 and Fz of the international 10–20 system and channels 35 to 38 and 39 to 42 aligned with F8 and F7, respectively.</p

Demographic characteristics of the study population.

<p>Demographic characteristics of the study population.</p