Effects of Aflatoxin B1 on T-Cell Subsets and mRNA Expression of Cytokines in the Intestine of Broilers
Abstract
:1. Introduction
2. Results
2.1. T-Cell Subsets in the Small Intestinal Intraepithelial Lymphocytes (IELs)
2.2. T-Cell Subsets in the Small Intestinal Lamina Propria Lymphocytes (LPLs)
2.3. The Expression Levels of the IL-2, IL-4, IL-6, IL-10, IL-17, IFN-γ and TNF-α (LITAF) mRNA in the Small Intestinal Mucosa
3. Discussion
4. Experimental Section
4.1. Animals and Diets
Composition | Content (%) | Nutrient | Content (%) |
---|---|---|---|
Corn | 51.95 | Crude protein (CP) | 21.50 |
Soybean | 39.50 | Methionine (Met) | 0.50 |
Rapeseed oil | 4.10 | Calcium (Ca) | 1.00 |
dl-Metionine | 0.20 | All phosphorus (P) | 0.70 |
Calcium hydrogen phosphate | 1.85 | Methionine + cysteine (Met + Cys) | 0.84 |
Calcium carbonate | 1.30 | Lysine (Lys) | 1.15 |
Sodium chloride | 0.40 | Threonine (Thr) | 0.83 |
Trace element premix a | 0.50 | Metabolizable energy (ME) (MJ/Kg) | 29.90 |
Choline | 0.17 | ||
Multivitamins b | 0.03 | ||
Total | 100 |
4.2. Determination of the Proportion of T-Cell Subsets in the Intestine by FCM
4.2.1. Isolation of the Intra-Epithelial Lymphocytes (IELs)
4.2.2. Isolation of Lamina Propria Lymphocytes (LPLs)
4.2.3. Flow Cytometry (FCM) Method
4.3. Determination of Cytokines mRNA Expression Levels in the Intestinal Mucosa by qRT-PCR
Gene | Primer | Sequences (5'–3') | Product Size (bp) | Accession Number |
---|---|---|---|---|
IL-2 | F | CCCGTGGCTAACTAATCTGC | 114 | AF000631 |
R | TTGAGCCCGTAGGTTACAGAA | |||
IL-4 | F | TCTTCCTCAACATGCGTCAG | 108 | NM_1007079.1 |
R | GGTCTGCTAGGAACTTCTCCAT | |||
IL-6 | F | CAAGGTGACGGAGGAGGAC | 254 | AJ309540 |
R | TGGCGAGGAGGGATTTCT | |||
IL-10 | F | CGGGAGCTGAGGTGAA | 272 | AJ621614 |
R | GTGAAGAAGCGGTGACAGC | |||
IL-17 | F | CAGATGCTGGATGCCTAACC | 123 | AJ493595 |
R | CCAGTGAGCGTTTGCTGATA | |||
INF-γ | F | AGCTGACGGTGGACCTATTATT | 259 | Y07922 |
R | GGCTTTGCGCTGGATTC | |||
TNF-α (LITAF) | F | TGTGTATGTGCAGCAACCCGTAGT | 229 | AY765397 |
R | GGCATTGCAATTTGGACAGAAGT | |||
β-Actin | F | TGCTGTGTTCCCATCTATCG | 150 | L08165 |
R | TTGGTGACAATACCGTGTTCA |
4.4. Statistics
5. Conclusions
Acknowledgments
Conflicts of Interest
References
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Jiang, M.; Peng, X.; Fang, J.; Cui, H.; Yu, Z.; Chen, Z. Effects of Aflatoxin B1 on T-Cell Subsets and mRNA Expression of Cytokines in the Intestine of Broilers. Int. J. Mol. Sci. 2015, 16, 6945-6959. https://doi.org/10.3390/ijms16046945
Jiang M, Peng X, Fang J, Cui H, Yu Z, Chen Z. Effects of Aflatoxin B1 on T-Cell Subsets and mRNA Expression of Cytokines in the Intestine of Broilers. International Journal of Molecular Sciences. 2015; 16(4):6945-6959. https://doi.org/10.3390/ijms16046945
Chicago/Turabian StyleJiang, Min, Xi Peng, Jing Fang, Hengmin Cui, Zhengqiang Yu, and Zhengli Chen. 2015. "Effects of Aflatoxin B1 on T-Cell Subsets and mRNA Expression of Cytokines in the Intestine of Broilers" International Journal of Molecular Sciences 16, no. 4: 6945-6959. https://doi.org/10.3390/ijms16046945