Molecular Markers of EBV- and HHV6-Associated Mononucleosis

Epstein–Barr virus (EBV) and human herpesvirus type 6 (HHV6) are causative agents of infectious mononucleosis and can lead to the development of lymphoproliferative diseases. Means of radical therapy for this disease are yet to be found. Key transcripts involved in the pathogenesis can be used as molecular markers and also as potential therapeutic targets.

The aim of the study was to identify molecular markers associated with infection caused by EBV and HHV6; specifically, we looked into the markers localized in blood leukocytes of patients with infectious mononucleosis.

Materials and Methods. We studied the transcriptome of peripheral blood leukocytes in children and adolescents with infectious mononucleosis caused by Epstein–Barr virus (EBV-IM) and human herpesvirus type 6 (HHV6-IM), as well as healthy subjects matched by gender and age. Using our original DNA biochips, we determined the expression of 403 genes (total representation level of all mRNA of one gene) and 712 transcripts (individual spliced mRNA of one gene) essential for the proliferation and apoptosis of immunocompetent cells. Data analysis was performed using a combination of machine learning and traditional statistics. The genes and transcripts which are highly important for paired classification and have the statistically significant differences in the expression between patients and healthy subjects were selected to serve molecular markers of the infection.

Results. Unique groups of candidate markers for EBV-IM and HHV6-IM were identified. EBV-IM was characterized by a decreased expression of the AR transcript 5 and ASCC1 transcript 4 and also of the CAD gene and FADD mRNA; an increased expression of the HLA-DPA1 transcript 2 and RIPK1 transcript 4 were found. In patients with HHV6-IM, an increase in the expression of AVEN mRNA, CHUK transcript 2, CIRBP transcript 2, and TRAF3 transcript 2, as well as a decrease in the expression of IRAK4 transcript 10 was observed. In the post-infection period, the expression levels of most of the markers returned to normal.

Conclusion. The sets of identified markers are uniquely characteristic of the two infections (EBV-IM and HHV6-IM) and can be used as targets for new therapies.

1. Filatova E.N., Solntsev L.A., Presnyakova N.B., Kulova E.A., Utkin O.V. Determination of some immunological features of hhv-6-mediated infectious mononucleosis in children by the method of discriminatory analysis. Infektsiya i immunitet 2018; 8(2): 223–229, https://doi.org/10.15789/2220-7619-2018-2-223-229.
2. Dojcinov S., Fend F., Quintanilla-Martinez L. EBV-positive lymphoproliferations of B- T- and NK-cell derivation in non-immunocompromised hosts. Pathogens 2018; 7(1): 28, https://doi.org/10.3390/pathogens7010028.
3. Nakayama-Ichiyama S., Yokote T., Oka S., Iwaki K., Kobayashi K., Hirata Y., Hiraoka N., Takayama A., Akioka T., Miyoshi T., Takubo T., Tsuji M., Hanafusa T. Diffuse large B-cell lymphoma, not otherwise specified, associated with coinfection of human herpesvirus 6 and 8. J Clin Oncol 2011; 29(21): e636–e637, https://doi.org/10.1200/jco.2011.35.1254.
4. Razzaque A. Oncogenic potential of human herpesvirus-6 DNA. Oncogene 1990; 5(9): 1365–1370.
5. Li B., Zeng Q. Personalized identification of differentially expressed pathways in pediatric sepsis. Mol Med Rep 2017; 16(4): 5085–5090, https://doi.org/10.3892/mmr.2017.7217.
6. Omar M., Klawonn F., Brand S., Stiesch M., Krettek C., Eberhard J. Transcriptome-wide high-density microarray analysis reveals differential gene transcription in periprosthetic tissue from hips with chronic periprosthetic joint infection vs aseptic loosening. J Arthroplasty 2017; 32(1): 2342–2340, https://doi.org/10.1016/j.arth.2016.06.036.
7. Sano D., Tazawa M., Inaba M., Kadoya S., Watanabe R., Miura T., Kitajima M., Okabe S. Selection of cellular genetic markers for the detection of infectious poliovirus. J Appl Microbiol 2018; 124(4): 1001–107, https://doi.org/10.1111/jam.13621.
8. Scicluna B.P., van Vught L.A., Zwinderman A.H., Wiewel M.A., Davenport E.E., Burnham K.L., Nürnberg P., Schultz M.J., Horn J., Cremer O.L., Bonten M.J., Hinds C.J., Wong H.R., Knight J.C., van der Poll T.; MARS consortium. Classification of patients with sepsis according to blood genomic endotype: a prospective cohort study. Lancet Respir Med 2017; 5(10): 816–826, https://doi.org/10.1016/s2213-2600(17)30294-1.
9. Knyazev D.I., Starikova V.D., Utkin О.V., Solntsev L.А., Sakharnov N.А., Efimov E.I. Splicing-sensitive DNA-microarrays: peculiarities and applicationin biomedical research (review). Sovremennye tehnologii v medicine 2015; 7(4): 162–173, https://doi.org/10.17691/stm2015.7.4.23.
10. Pirooznia M., Yang J.Y., Yang M.Q., Deng Y. A comparative study of different machine learning methods on microarray gene expression data. BMC Genomics 2008; 9(Suppl 1): S13, https://doi.org/10.1186/1471-2164-9-s1-s13.
11. Solntsev L.A., Starikova V.D., Sakharnov N.A., Knyazev D.I., Utkin O.V. Strategy of probe selection for studying mRNAs that participate in receptor-mediated apoptosis signaling. Mol Biol 2015; 49(3): 457–465, https://doi.org/10.1134/s0026893315030164.
12. Wu Z., Aryee M.J. Subset quantile normalization using negative control features. J Comput Biol 2010; 17(10): 1385–1395, https://doi.org/10.1089/cmb.2010.0049.
13. Torices S., Alvarez-Rodríguez L., Grande L., Varela I., Muñoz P., Pascual D., Balsa A., López-Hoyos M., Martinez-Taboada V., Fernández-Luna J.L. A truncated variant of ASCC1, a novel inhibitor of NF-B, is associated with disease severity in patients with rheumatoid arthritis. J Immunol 2015; 195(11): 5415–5420, https://doi.org/10.4049/jimmunol.1501532.
14. Michel M., Wilhelmi I., Schultz A.-S., Preussner M., Heyd F. Activation-induced tumor necrosis factor receptor-associated factor 3 (Traf3) alternative splicing controls the noncanonical nuclear factor κB pathway and chemokine expression in human T cells. J Biol Chem 2014; 289(19): 13651–13660, https://doi.org/10.1074/jbc.m113.526269.
15. Filatova E.N., Utkin O.V. The role of noncoding MRNA isoforms in the regulation of gene expression. Russ J Genet 2018; 54(8): 879–887, https://doi.org/10.1134/s1022795418080057.
16. Feoktistova M., Leverkus M. Programmed necrosis and necroptosis signalling. FEBS J 2014; 282(1): 19–31, https://doi.org/10.1111/febs.13120.
17. Haan K.M., Kwok W.W., Longnecker R., Speck P. Epstein-Barr virus entry utilizing HLA-DP or HLA-DQ as a coreceptor. J Virol 2000; 74(5): 2451–2454, https://doi.org/10.1128/jvi.74.5.2451-2454.2000.

Filatova E.N., Sakharnov N.A., Knyazev D.I., Tsybusova T.N., Utkin O.V. Molecular Markers of EBV- and HHV6-Associated Mononucleosis. Sovremennye tehnologii v medicine 2019; 11(3): 7, https://doi.org/10.17691/stm2019.11.3.01