TLR4 is constitutively expressed in chick thymic epithelial cells

Huang, Haibo; Xiang, Quanhang; Wu, Hui; Ansari, Abdur Rahman; Wen, Le; Ge, Xin; Wang, Jixiang; Peng, Kaiman; Liu, Huazhen

doi:10.1016/j.vetimm.2014.01.005

Cited by 11 publications

(11 citation statements)

References 40 publications

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“…For immunochemistry, the antibodies of TLR4 (1:50), LC3A/B (1:50), PCNA (1:100), PCK (1:50) and S100 (1:50) were used, which either target the amino acid sequences sharing high identity (>70%) with those found in ostriches, or their applicability for birds are declared by each company ( S1 Table ). The immunochemistry staining was performed as described [ 16 ]. For apoptotic staining, a mouse IgM anti-ssDNA monoclonal antibody (1:10; EMD Millipore, Billerica, USA) was used.…”

Section: Methodsmentioning

confidence: 99%

“…The expressions of TLR4, LC3A/B and PCNA were measured using integrated optical density (IOD) ratio as described [ 16 ], and the total numbers of ssDNA positive cell per 1 × 10 5 μm 2 in thymic cortex were quantified.…”

Section: Methodsmentioning

confidence: 99%

“…Toll like receptors (TLRs) are major components of innate immune system in that they can recognize multiple endogenous and exogenous stress signals and induce an immune response [ 14 ]. TLRs are expressed in the thymus of chickens and are suggested to participate in thymus development by inducing downstream factors [ 15 , 16 ]. Furthermore, abnormal thymic development is associated with increased expression of TLR4 [ 17 ], aberrant activation of transcription regulators (NF-κB, MAPK) [ 18 , 19 ], and overexpression of proinflammatory cytokines (IL-6 and IFN) [ 20 , 21 ].…”

Section: Introductionmentioning

confidence: 99%

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Increased Thymic Cell Turnover under Boron Stress May Bypass TLR3/4 Pathway in African Ostrich

Huang

Xiao

et al. 2015

PLoS ONE

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Previous studies revealed that thymus is a targeted immune organ in malnutrition, and high-boron stress is harmful for immune organs. African ostrich is the living fossil of ancient birds and the food animals in modern life. There is no report about the effect of boron intake on thymus of ostrich. The purpose of present study was to evaluate the effect of excessive boron stress on ostrich thymus and the potential role of TLR3/4 signals in this process. Histological analysis demonstrated that long-term boron stress (640 mg/L for 90 days) did not disrupt ostrich thymic structure during postnatal development. However, the numbers of apoptotic cells showed an increased tendency, and the expression of autophagy and proliferation markers increased significantly in ostrich thymus after boron treatment. Next, we examined the expression of TLR3 and TLR4 with their downstream molecular in thymus under boron stress. Since ostrich genome was not available when we started the research, we first cloned ostrich TLR3 TLR4 cDNA from thymus. Ostrich TLR4 was close to white-throated Tinamou. Whole avian TLR4 codons were under purify selection during evolution, whereas 80 codons were under positive selection. TLR3 and TLR4 were expressed in ostrich thymus and bursa of fabricius as was revealed by quantitative real-time PCR (qRT-PCR). TLR4 expression increased with age but significantly decreased after boron treatment, whereas TLR3 expression showed the similar tendency. Their downstream molecular factors (IRF1, JNK, ERK, p38, IL-6 and IFN) did not change significantly in thymus, except that p100 was significantly increased under boron stress when analyzed by qRT-PCR or western blot. Taken together, these results suggest that ostrich thymus developed resistance against long-term excessive boron stress, possibly by accelerating intrathymic cell death and proliferation, which may bypass the TLR3/4 pathway. In addition, attenuated TLRs activity may explain the reduced inflammatory response to pathogens under boron stress.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Increased Thymic Cell Turnover under Boron Stress May Bypass TLR3/4 Pathway in African Ostrich

Huang

Xiao

et al. 2015

PLoS ONE

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“…For example, TLR4 signaling plays a key role in immune cell recruitment in the MG thymus (Cordiglieri et al 2014), and TLR7 and TLR9 are also involved in the pathogenesis of the MG thymus (Cavalcante et al 2016). TLR4 is constitutively expressed on TECs (Huang et al 2014), and the expression of TLR4 was enhanced on the TECs of the involuted thymuses of myasthenic patients (Bernasconi et al 2005). The indirect effect of endotoxin stimulation on thymic involution caused alterations in the gene expression downstream of TLR3 and TLR4 (Fig.…”

Section: Mechanisms Of Ati Role Of Tlrs and Atimentioning

confidence: 99%

Acute Thymic Involution and Mechanisms for Recovery

Ansari

Liu

2017

Arch. Immunol. Ther. Exp.

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“…Sections were then reacted with 5% normal goat serum (Thermo Scientific, Rockford, USA) for 10 min at 37 • C to prevent any non-specific reactions, and incubated with a rabbit polyclonal antibody to human TLR4 (1:100 dilution; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) at 4 • C overnight. This antibody to TLR4 was raised against a peptide near the C-terminus of human TLR4, and has previously been used to identify TLR4 in chickens (Huang et al, 2014). After incubation with the primary antibody, sections were washed in PBS three times, and incubated for 20 min with goat anti-rabbit IgG antibody (Thermo Scientific, Rockford, USA) labeled with peroxidase.…”

Section: Immunohistochemical Stainingmentioning

confidence: 99%