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2. AU Content in the MicroRNA Sequence Influences its Stability after Heat Treatment

Author

López, Agnès Garcias, Brogaard, Louise, Heegaard, Peter Mikail Helwag, Cirera, Susanna, Skovgaard, Kerstin, López, Agnès Garcias, Brogaard, Louise, Heegaard, Peter Mikail Helwag, Cirera, Susanna, and Skovgaard, Kerstin

Abstract

BACKGROUND: MicroRNAs (miRNAs) are short non-coding RNA molecules which regulate gene expression post-transcriptionally and are involved in a multitude of cellular processes. MiRNAs are known to be very stable compared to messenger RNAs (mRNAs), making them excellent candidates as biomarkers for disease. Recently, studies have suggested that miRNA stability in formalin fixed samples might depend on their nucleotide composition.OBJECTIVE: To explore the stability of a panel of miRNAs isolated from porcine blood and lung tissue after heat and enzyme treatment.METHOD: Porcine RNA isolated from lung tissue and blood leukocytes was used for this study. RNA samples were exposed to heat treatment and RNAse A digestion. The levels of selected miRNAs were measured by means of qPCR before and after heat and enzyme treatment.RESULTS: Fourteen miRNAs were successfully analysed, and they were found to degrade differently after exposure to heat or RNAse A. MiRNAs with <60% of adenine (A) and uracil (U) in their sequence were found to be more stable.CONCLUSION: This is the first study showing that different miRNAs isolated from lung tissue display unequal stability after heat treatment, probably based on their nucleotide composition, highlighting the importance of considering the miRNA sequence when investigating their value as biomarkers.

Published
2019

3. Migration of intestinal dendritic cell subsets upon intrinsic and extrinsic TLR3 stimulation

Author

López, Agnès Garcias, primary, Bekiaris, Vasileios, additional, Luda, Katarzyna Müller, additional, Hütter, Julia, additional, Muleta, Konjit Getachew, additional, Nakawesi, Joy, additional, Ulmert, Isabel, additional, Kotarsky, Knut, additional, Malissen, Bernard, additional, O’Keeffe, Meredith, additional, Holzmann, Bernhard, additional, Agace, William, additional, and Lahl, Katharina, additional

Published
2019
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5. avß8 integrin-expression by BATF3-dependent dendritic cells facilitates early IgA responses to Rotavirus

Author

Nakawesi, J., This, S., Hütter, J., Boucard-Jourdin, M., Barateau, V., Muleta, K. Getachew, Gooday, L. J., Thomsen, K. Fog, López, A. Garcias, Ulmert, I., Poncet, D., Malissen, B., Greenberg, H., Thaunat, O., Defrance, T., Paidassi, H., and Lahl, K.

Abstract

Secretory intestinal IgA can protect from re-infection with rotavirus (RV), but very little is known about the mechanisms that induce IgA production during intestinal virus infections. Classical dendritic cells (cDCs) in the intestine can facilitate both T cell-dependent and -independent secretory IgA. Here, we show that BATF3-dependent cDC1, but not cDC2, are critical for the optimal induction of RV-specific IgA responses in the mesenteric lymph nodes. This depends on the selective expression of the TGFß-activating integrin avß8 by cDC1. In contrast, avß8 on cDC1 is dispensible for steady state immune homeostasis. Given that cDC2 are crucial in driving IgA during steady state but are dispensable for RV-specific IgA responses, we propose that the capacity of DC subsets to induce intestinal IgA responses reflects the context, as opposed to an intrinsic property of individual DC subsets.

Published
2021
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