Skip to main content

Table 4 Experimental studies observing modulation of NF-κB by PFAS (see also Modulation of NF-κB regulated gene transactivation section, Results)

From: Consideration of pathways for immunotoxicity of per- and polyfluoroalkyl substances (PFAS)

PFAS

Study / Method

Effect

Reference

PFNA

In vivo: p.o. 14 day treatment of mice (2008) and rats (2009)

No effect on NF-κB observed measured with RT PCR of thymus cells;

[93, 94]

PFOS, PFOA

In vitro: THP-1 cells, treated with 10-100 μg/ml PFOA and 1-100 μg/ml PFOS in the presence of LPS for 30 min or 3 hrs (NF-κB promoter activity);

↓ of NF-κB by PFOS and PFOA in a dose dependent manner

PFOS: ↓ of LPS-induced I-κB activation, NF-κB binding to DNA, p65 phosphorylation, and transcription according (independent of PPARα);

PFOA: ↓ p65 phosphorylation and NF-κB mediated transcription (through PPARα)

[205, 206]

PFOS

In vivo: BALB/c mice, regular (RD) or high-fat diet (HFD); then exposed to PFOS (0, 5, and 20 mg/kg/day) for 14 days.

(to investigate interference with lipid metabolism)

RD group: atrophy of immune organs, bw ↓ (highest dose), histopathological alterations of thymus and spleen, ↑ apoptosis of thymocytes

HFD group: More serious atrophy was seen in the immune organs; PFOS exposure did not suppress the NF-κB signalling pathway (↑IL-1β)

[207]

PFBS, PFOS, PFOA, PFDA, fluorotelomer

In vitro promyelotic cell line THP-1 and PHA-stimulated human PBLs;

Cells treated with 0.1 – 10 μg/ml PFAS and LPS for 30 min or 3 hrs (NF-κB promoter activity);

↓of LPS-induced phosphorylation of p65 (RELA) and NF-κB driven gene transcription (only PFOA activated PPARα)

PFBS and PFDA prevented LPS-induced I-κB degradation

All PFCs ↓ TNFα, while effects on other cytokines where unequal

[206]

PFOA

In vitro: HMC-1 cells, 50-400 μM PFOA for 12 hrs; proteins measured by Western blot.

[in vivo mouse allergy model; 10 and 50 mg/kg bw PFOA for 4 days on dorsal surface of each ear]

In vitro: ↑ of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, and IL-8) was NF-κB dependent;

↑ of: phosphorylation of p38, translocation of p65 NF- κB to the nucleus and degradation of IκB)

[In vivo: ↑IgE-dependent allergic local reaction in mice]

[128]

PFOA

In vivo: zebrafish (0.05, 0.1, 0.5, and 1 mg/L) for 21 d, expression analysis of spleen

↑pro-inflammatory cytokine (IL-1β and IL-21) at a low exposure concentration (0.05 mg/L) and ↓at higher exposure concentrations (≥ 0.1 mg/L) via Myd88/NF-κB pathway

↓TLR2 expression (at 1 mg/L) to 56% compared to control

[112]

PFDA

In vitro: human AGS cell line treated with 5-50 μM PFDA for 24 hrs, protein and mRNA level

↑ NF-κB activity at 5 μM, also c-Rel and p52 were ↑; (↑IL-13, IL-18 and NLRP3)

[208]

PFDA (C = 10) and PFUnDA (C = 11), no effect: PFHpA C = 7, PFNA C = 9;

In vitro: IgE stimulated mast cells, RBL -2H3, transfected with NF-κB luciferase transporter construct

Treated with respective PFAS 100 μM, 30 min

[In vivo: ovalbumin induced system of anaphylaxis]

In vitro: IgE stimulated mast cells: ↑ pro-inflammatory cytokines (PFDA and PFUnDA only) ↑intracellular Ca, ↑ histamine

RBL cells: ↑ NF-κB activity (PFDA and PFUnDA only)

[In vivo: ↑allergic reactions only with PFDA, PFNA and PFUnDA]

[111]

PFOS

In vitro: IgE stimulated RBL-2H3 cells; treated with 100 and 500 μM PFOS for 30 min; assayed with Western blot and luciferase assay

[In vivo: OVA induced anaphylaxis ICR mouse model (50-150 mg/kg PFOS p.o. 3x on day 9, 11 and 13)]

In vitro: ↑ NF-κB (luciferase activity); ↑ degradation of I-κB and nuclear translocation of NF-κB;

↑ gene expression of pro-inflammatory cytokines; pre-treatment with a NF-κB inhibitor: ↓TNF-α production and luciferase activity;

[In vivo: allergic symptoms were ↑ by PFOS]

[160]

PFOS

In vitro: rat primary KCs and hepatocytes treated with 100 μM PFOS for 48 hrs

In vivo: male SD rats 1 or 10 mg/kg bw PFOS per gavage for 20 days;

In vitro: KC cells - ↑ NF-κB activation and p65 translocation. ↑ I-κB and JNK phosphorylation in hepatocytes and KCs;

↑ production of TNF-α and IL-6 in KCs, and was ↓ by NF-kB inhibitor

↑ hepatocyte proliferation by altering regulatory proteins (↑ PCNA, c-Jun, c-MYC and CyD1 in vitro & in vivo)

In vivo: hepatocellular damage and inflammation, ↑serum TNFα and IL-6 level

[209]

PFOA and PFOS

In vitro: macrophages treated with six EDCs via sirtuin (SIRT) regulation using the murine macrophage RAW 264.7 cell line

PFOS and PFOA did not alter NF-κB expression (only Mono(2-ethylhexyl) phthalate did)

[210]

PFOS

In vivo (hepatotoxicity and immunotoxicity) in zebrafish: 0, 0.02, 0.04 and 0.08 mg/L of PFOS for 7, 14, and 21 days

Immune-regulatory function in the liver was disturbed by affecting liver structure, enzyme activities (↓ACP, AKP, lysozyme, ↑MPO) via ↑ NF-κB signalling,↑ ROS

[211]

PFOA

In vivo zebrafish

↑ TLR2/Myd88/p65 pathway →

(↓ IFN and BAFF mRNA expression →

↓ of Ig secretion)

lipid metabolism disorder enhances the immune toxicity level in the spleen

[212]

PFOS

In vitro: mural BMDMs; human cells: THP-1 cells (10-200 nM PFOS according to the authors corresponding to PFOS-serum levels in most human subjects)

[In vivo: wild type (WT) C57BL/6 J mice were injected i.p., acute: 5, 15, or 25 mg/kg/d for 5 days & chronic: 0.066 mg/kg/d for 30 days; OVA induced asthmatic exacerbation model]

THP-1 cells: ↑ NF-κB signalling; Ca2+–PKC-dependent pathway; ↑mRNA levels and release of TNF-α and IL-6

BMDMs: ↑ phosphorylation of NF-κB p65 and degradation of IκBα

[In vivo: PFOS induced inflammation, ↑ IL-6, TNF-α, IL-1ß) tissue damage (lungs, liver, kidneys) via activation of the AIM2 inflammasome; asthmatic exacerbation, ↑IL-4, IL-1ß]

[158]

PFOA

In vivo: zebrafish were exposed to 0.05, 0.1, 0.5, and 1 mg/L for 21 days

↑TLR2 /Myd88/NF-κB (p65) pathway ↑ proinflammatory cytokines (IFN and IL-1β) → regulation of antibody expression and regulation of expression of other cytokines (↓IL-4) → immune disorders

[213]

Modulation of NF-kB in different cell types (less immune relevant)

 PFOS

In vivo: mice fed a regular (RD) or high-fat diet (HFD) and then exposed to PFOS (0, 5, and 20 mg/kg/day) for 14 days.

More serious atrophy of immune organs seen in HFD group; HFD might ↑ apoptosis caused by PFOS; no effect on NF-κB signalling pathway

[207]

 PFNA

In vivo: Rats treated with PFNA or PFNA & gadolinium chloride, an inhibitor of KCs, for 14 days.

In vitro: primary rat hepatocytes

↑ NF-κB, ↑ TNFα and IL-1β were involved in ↓of PPARα promoter activity.

[214]

 PFOA

In vitro (cancer research): breast cancer cells MDA-MB-231

↑ NF-κB translocation into the nucleus

↑mRNA and protein levels of MMP-2/−9 → invasiveness increased

[215]

 PFOS

In vitro (neurotoxicity, in this case: immunotoxicity in central nervous system): murine BV2 microglial cells

↑ NF-κB, ↑ TNF-alpha and IL-6 expression.

In part via c-Jun N-terminal protein kinase, ERK and NF-κB signalling; related to neurodegenerative diseases;

[216]

 PFOA

In vitro (cancer research): human colorectal cancer cell DLD-1

↑ NF-κB activity by stimulating translocation into nucleus; (↑ MMP2/9 expression → invasiveness increased)

[217]

 PFOS

In vitro (neurotoxicity): HAPI rat microglia (i.e. innate immune system of the CNS)

↑ NF-κB p65 and PKC were activated, ↑TNF-α secretion via Ca2+-dependent PKC-NF-кB signalling

[218]

 PFOA

(cardiotoxicity) fertile chicken eggs, +/− l-carnitine co-treatment

↑ p65 translocation in ED19 embryo hearts and hatchling hearts, alleviated by l-carnithine (antioxidant, NO modulatory); ↑ ROS levels

[219]

 PFOA

In vitro (cancer research): A2780 human ovarian cancer cell line

↑ NF-κB signalling through ERK1/2 phosphorylation (↑of MMP-2/−9 expression associated with tumor invasion)

[220]

 PFOS

In vivo (hepatotoxicity): in male mice, 10 mg/kg/day i.g. alone or with narginin for 3 weeks

↑ NF-κB activity and ↑inflammatory cytokines TNF-α and IL-6 in the liver (also Bax and caspase 3), suppressed by narginin (anti-oxidative, anti-inflammatory and anti-apoptotic)

[175]

 PFOS

In vitro (neurotoxicity): astrocytes, C6 glioma cells (rat)

↑ phosphorylation and degradation of IκBα, and translocation of NF-κB p65 to the nucleus; ↑pro-inflammatory cytokines (IL-1β) via AKT-dependent NF-κB signalling pathway.

[221]

 PFOA

In vitro (cancer research): in human follicular thyroid carcinoma cells (FTC133)

↑ phosphorylation of NF-κB p65 and nuclear translocation. Reversed by NF-κB inhibitor; ↑ MMP-2 and tumor invasion

[222]

 PFOA

In vivo rat model of PFOA-induced patellar instability

↑ of the NF-κB signalling pathway; associated with early patellofemoral articular cartilage degeneration

[223]

  1. For discussion of results see Results, Modulation of NF-κB regulated gene transactivation section
  2. ↑ induction, ↓ reduction, AGS Human gastric adenocarcinoma cell line, BAFF B cell-activating factor, BMDMs Bone marrow-derived macrophages, CNS Central nervous system, EDCs Endocrine disruptors, HAPI Highly aggressive proliferating immortalized rat microglia cells, HMC-1 Cells human mastoid cell line, IFN Interferon, i.g Intragastrically, Ig Immunoglobulin, IL Interleukin, IκBα NF-κB inhibitor alpha, KCs Kupffer cells, LPS Lipopolysaccharide, MMP-2/−9 Matrix metalloproteinases, Myd88 Myeloid differentiation factor 88, NF-κB Nuclear factor kappa B, OVA Ovalbumin, p65 NF-kappa-B p65 subunit (see also RelA), RBL-2H3 Rat basophilic leukaemia cells, RelA (= p65, i.e. NF-κB subunit) REL-associated protein, ROS Reactive oxygen species, SD Sprague-Dawley, THP-1 Human monocytic leukaemia cells, TLR 2 Toll-like receptor 2