Document details Prepared by Roger Drew, PhD, DABT Tarah Hagen, MSc ToxConsult Pty Ltd. ABN: 55 158 303 167 PO Box 316 Darling South, VIC 3145 Tel: 03 9569 3918/ 03 9572 1448 Fax: 03 9563 5330 Prepared for Food Standards Australia New Zealand ToxConsult document ToxCR300816-RF 6th December 2016 Roger Drew, PhD, DABT (Diplomate American Board of Toxicology) Tarah Hagen, MSc (Environmental Toxicology) Disclaimer This report was prepared by ToxConsult Pty Ltd as an account of work for FSANZ (the ‘Client’). This report should be read, and used in its entirety. The material in it reflects ToxConsult’s best judgement in the light of the information available to it at the time of preparation. However, as ToxConsult cannot control the conditions under which this report may be used, ToxConsult will not be responsible for damages of any nature resulting from use of or reliance upon this report. ToxConsult’s responsibility for the information herein is subject to the terms of engagement with the client. Information provided by the client has been used in good faith; ToxConsult has not, and was not required to, verify its veracity. Copyright and any other Intellectual Property associated with this report belongs to ToxConsult Pty Ltd and may not be reproduced in any form without the written consent of ToxConsult. The Client, and only the client, is granted an exclusive licence for the use of the report for the purposes described in the report.
~ 5 > Calibrat’n 12.47 ± 0.61 7.13 ± 1.0 10.75 ± 0.82 8.21 ± 1.15 8.21 ± 1.15 11.6 ± 0.2 0.95 ± 0.13 1.5 ± 0.23 25 ND ~ 9 21.64 ± 4.4 22.6 ± 2.29 24.5 ± 5.56 50.8 ± 2.5 ~40 110 ± 6.18 97.3 ± 7.6 13.45 ±1.48 15.4 ±1.56 50 65.43 ± 11.7 51.71 ± 3.8 59.7 ± 12.2 59.7 ± 12.2 ~100 ND ~ 50 96.7± 5.2 125.8 ± 3.9 20.93 ± 2.36 31.93 ± 3.59 125 120.7 ± 21.8 114.2 ± 23.7 ~ 140 281 ± 16.3 250 ± 20.1 ~ 200 29.88 ± 3.53 43.2 ± 3.95
Appendix B: Summary of immunomodulation animal studies Abbreviations and symbols used in this Appendix are: Symbols ↓ Decrease relative to control ↓↓ Marked decrease relative to control ↑ Increase relative to control Sl↑ Slight increase relative to control ↔ No change relative to controls ♂ Male ♀ Female Units Kg Kilogram L Litre mL Millilitre µg Microgram Ng Nanogram Abreviations APFO Ammonium perfluorooctanoate PFOA is the anion to this substance Bw Body weight B B-cell Conc Concentration D Day DST Delayed hypersensitivity test GD Gestation day Ig Immunoglobulin IL Interleukin lPFOS Linear perfluorooctane sulphonate lAPFO Linear ammonium perfluorooctanoate KO Knock out ND Not determined NK Natural killer cell PFC Plaque forming assay PND Postnatal day Prolif Proliferation
Gavage 60d lPFOS K+ Water 0.02% Tween 80 83.3 5 8.21 ± 1.15 ↔ ↔ ↔ ↑Apopt’s ↑ liver wt ↓ MMP Spleen 833.3 50 59.74 ± 12.2 ↓ ↓ ↓ ↓ # ↑Apopt’s ↑ liver wt ↓ MMP Spleen In Dong et al. (2012a) a variety of end points have been measured (cell viability, cell cycle, cell apoptosis/necrosis, mitochondrial membrane potential [MMP], gene expression e.g. p53]). Increased apoptosis (decreased cell viability) in spleen and thymus cell suspensions at ≥ 5 mg/kg TAD and MMP splenocytes decreased. Immuno-NOEL (↑ splenocyte apoptosis, ↓ MMP) = 4.35 mg/L. (↑ liver wt is likely adaptive & PPARα mediated). NOEL = 4.35 mg/L based on increased apoptosis and decreased MMP in splenocytes. LOEL = 8.21 mg/L. Dong et al. (2012b) Mouse C57BL/6♂ Gavage 60d lPFOS K+ Water 0.02% Tween 80 0 0 0.04 ± 0.01 8.3 0.5 0.58 ± 0.19 ↔ ↔ ↔ ↔ ↔ ↔ 16.7 1 4.35 ± 0.63 ↔ ↔ ↔ ↔ ↔ ↔ 83.3 5 8.21 ± 1.15 ↔ ↔ ↔ ↑ IL-1β ↔ Others Macrophages not splenocytes ↑ Liver wt 416.7 25 24.53 ± 5.56 ↓ ↓ ↓ ↔ IL-β ↑ Liver wt 833.3 50 59.74 ± 12.2 ↓ ↓ ↓ ↑ IL-1β ↑ IL-6 Cytokines ↑ ex-vivo with in vivo exposure with & without iv LPS ↑ Liver wt 2,083 125 114.19 ± 23.72 ↓ ↓ ↓ ↑ IL-1β ↑ IL-6 ↑ Liver wt
(2010a). Mouse C57BL/6♂ Diet 10d (w/w) mg/kg 0 0.005 ~ 100 125.8 ± 3.9 ↔ ↔ ↔ ↔ Spleen ↓ TNF-α, IFN-γ, IL-4 in liver ↔ in IgM & IFN-γ response to LPS or Con A ↑ liver wt (centrilobular hypertrophy) ↓ serum TGs & cholesterol ↑ serum ALP This study investigated the effect of PFOS & PFOA on hepatic immune status. Numbers of hepatic cells that appear immunophenotypically to be erythrocyte progenitors were increased, other intrahepatic immune cells were unaffected. There were decreased cytokines in the liver, but ex vivo production of IgM by hepatic B cells and IFN-γ by hepatic T cells with specific stimulators (LPS and Con A, respectively) was not statistically different from controls, but appeared to be larger in PFOS treated animals. Data for splenocytes not included. Study not suitable for NOEL determination. Qazi et al. (2010b). Mouse B6C3F1♂ % diet (w/w) µg/kg bw/d TAD mg/kg 0 0 0.04 ± 0.002 0.000156 250 5.55 11.6 ± 0.2 ↓ bw gain ↔ ↔ ↔ ↔ (also for ↔ serum IgM & IgG ↔ with SRBC ↑ liver wt
immunocompetent as control animals. Ex-vivo splenocyte responses to antigen stimulation were not altered. In considering the above it is suggested lymphocyte apoptosis in thymus of males (LOEL 20.9 mg/L) may be a more appropriate endpoint for a NOEL in this study, i.e. NOEL of 13.5 mg/L. In females the NOEL was 43.2 mg/L. The authors suggest immune responses in the rat may, in part, be secondary to hepatic changes. Keil et al. 2008 Mouse Gavage 1-17 lPFOS in water plus 0.5% Tween 80 mg/kg/d TAD mg/kg calculated 0 0.1 1.7 ~ 1 ↔ ↔ ↔ ↔ NK at 4 wks old ↔ ↓ liver wt ♀ at 4 wks but ↔ at 8 wks ↔ ♂. 1.0 17 ~9 ↔ ↔ ↔ ↔ NK at 4 wks old ↔ ↔ liver wt ♂♀ at 4 & 8 wks old ↓ NK ♂, ↔♀, at 8 wks old 5.0 85 ~ 50 ↔ ↔ ↔ ↓ B cell ♀ ↔ ♂ 4 wks old. ↔ ♂♀ at 8 wks old ↔ CD3+ & CD4+ ♂♀ at 4 wks old. ↓ CD3+ CD4+ ♂ at 8 wks. ↔ NK at 4 wks old. ↓ NK ♂ ♀ at 8 wks old Not done at 4 wks ↓ ♂ at 8 wk old ↔ ♀ ↑ liver wt ♂ ↓kid wt ♀ at 4 wks old. ↔ ♂♀ at 8 wks old.