A new coding mutation in the tnf-α leader sequence in tuberculosis-sensitive i/st mice causes higher secretion levels of soluble tnf-α

ABSTRACT I/St and A/Sn mice are polar extremes in terms of several parameters defining sensitivity to _Mycobacterium tuberculosis_. TNF-α, mainly produced by activated macrophages, can

mediate both physiological and pathophysiological processes. Adequate TNF-α levels are essential for a forceful protective response to _M. tuberculosis_. We have functionally characterized a

nonsynonymous substitution, Arg8His, in the highly conserved cytoplasmic domain of the pro-TNF-α leader peptide from extremely _M. tuberculosis-_sensitive I/St mice. This was compared to

the common pro-TNF-α variant found in A/Sn mice. Using cDNA constructs, both variants were constitutively expressed in HEK293A cells. A significantly higher secretion level of Arg8His TNF-α

was shown using flow cytometry and ELISA analysis (_P_=0.0063), while intracellular levels were similar for both protein variants. An even TNF-α distribution throughout the cells was seen

using confocal microscopy. This suggests that the Arg8His substitution affects pro-TNF-α processing. The I/St mouse may serve as a model to further explore the function of the well-conserved

cytoplasmic region of TNF-α. However, other identified substitutions in the I/St promoter, introns and 3′UTR of _Tnf-_α, as well as the cellular environment _in vivo_ may affect the balance

between soluble and intracellular Arg8His TNF-α before and during _M. tuberculosis_ infection. Access through your institution Buy or subscribe This is a preview of subscription content,

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HOST-PROTECTIVE IMMUNE RESPONSES AGAINST TUBERCULOSIS Article Open access 06 August 2024 ACCESSION CODES ACCESSIONS GENBANK/EMBL/DDBJ * Y00467 REFERENCES * Nikonenko BV, Apt AS, Moroz AM,

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and Informatics Laboratory School of Medicine, University of Pennsylvania, Pennsylvania, 1997. Download references ACKNOWLEDGEMENTS This study was supported by grants from Swedish Medical

Research Council, Karolinska Institutet Foundation, Magnus Bergwall Foundation, and Swedish Heart and Lung Foundation. We thank Ricardo Giscombe for skilful assistance with the flow

cytometry experiments. AUTHOR INFORMATION Author notes * H Källström Present address: Department of Medical Biochemistry and Microbiology, The Biomedical Center, Uppsala University, Box 582,

751 23, Uppsala, Sweden AUTHORS AND AFFILIATIONS * Department of Molecular Medicine, Karolinska Institutet, CMM, Karolinska Hospital, Stockholm, Sweden A K Kähler, A-S Persson, F Sánchez 

& C Lavebratt * Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm, Sweden H Källström * Laboratory for Immunogenetics, Central Institute for Tuberculosis of the

Russian Academy of Medical Sciences, Moscow, Russia A S Apt * McGill Center for the Study of Host Resistance, Montreal General Hospital, Montreal, Quebec, Canada E Schurr Authors * A K

Kähler View author publications You can also search for this author inPubMed Google Scholar * A-S Persson View author publications You can also search for this author inPubMed Google Scholar

* F Sánchez View author publications You can also search for this author inPubMed Google Scholar * H Källström View author publications You can also search for this author inPubMed Google

Scholar * A S Apt View author publications You can also search for this author inPubMed Google Scholar * E Schurr View author publications You can also search for this author inPubMed Google

Scholar * C Lavebratt View author publications You can also search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to C Lavebratt. RIGHTS AND PERMISSIONS

Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Kähler, A., Persson, AS., Sánchez, F. _et al._ A new coding mutation in the _Tnf_-α leader sequence in tuberculosis-sensitive

I/St mice causes higher secretion levels of soluble TNF-α. _Genes Immun_ 6, 620–627 (2005). https://doi.org/10.1038/sj.gene.6364249 Download citation * Received: 01 March 2005 * Revised: 13

June 2005 * Accepted: 13 June 2005 * Published: 14 July 2005 * Issue Date: 01 October 2005 * DOI: https://doi.org/10.1038/sj.gene.6364249 SHARE THIS ARTICLE Anyone you share the following

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SharedIt content-sharing initiative KEYWORDS * TNF-α * tuberculosis * leader peptide * pro-TNF-α