Cytokine expression provides clues to the pathophysiology of Gulf War illness and myalgic encephalomyelitis

Authors: Svetlana F. Khaiboullina, Kenny L. DeMeirleir, Shanti Rawat, Grady S. Berk, Rory S. Gaynor-Berk, Tatjana Mijatovic, Natalia Blatt, Albert A. Rizvanov, Sheila G. Young, Vincent C. Lombardi
Publication: Cytokine Volume 72, Issue 1, March 2015, Pages 1–8

Gulf War illness (GWI) is a chronic disease of unknown etiology characterized by persistent symptoms such as cognitive impairment, unexplained fatigue, pervasive pain, headaches, and gastrointestinal abnormalities. Current reports suggest that as many as 200,000 veterans who served in the 1990–1991 Persian Gulf War were afflicted. Several potential triggers of GWI have been proposed including chemical exposure, toxins, vaccines, and unknown infectious agents. However, a definitive cause of GWI has not been identified and a specific biological marker that can consistently delineate the disease has not been defined. Myalgic encephalomyelitis (ME) is a disease with similar and overlapping symptomology, and subjects diagnosed with GWI typically fit the diagnostic criteria for ME. For these reasons, GWI is often considered a subgroup of ME. To explore this possibility and identify immune parameters that may help to understand GWI pathophysiology, we measured 77 serum cytokines in subjects with GWI and compared these data to that of subjects with ME as well as healthy controls. Our analysis identified a group of cytokines that identified ME and GWI cases with sensitivities of 92.5% and 64.9%, respectively. The five most significant cytokines in decreasing order of importance were IL-7, IL-4, TNF-α, IL-13, and IL-17F. When delineating GWI and ME cases from healthy controls, the observed specificity was only 33.3%, suggesting that with respect to cytokine expression, GWI cases resemble control subjects to a greater extent than ME cases across a number of parameters. These results imply that serum cytokines are representative of ME pathology to a greater extent than GWI and further suggest that the two diseases have distinct immune profiles despite their overlapping symptomology.

Death-domain associated protein-6 (DAXX) mediated apoptosis in hantavirus infection is counter-balanced by activation of interferon-stimulated nuclear transcription factors.

Authors: Khaiboullina SF, Morzunov SP, Boichuk SV, Palotás A, St Jeor S, Lombardi VC, Rizvanov AA.
Publication: Virology. 2013 Sep 1;443(2):338-48. doi: 10.1016/j.virol.2013.05.024. Epub 2013 Jul 3.

Hantaviruses are negative strand RNA species that replicate predominantly in the cytoplasm. They also activate numerous cellular responses, but their involvement in nuclear processes is yet to be established. Using human umbilical vein endothelial cells (HUVECs), this study investigates the molecular finger-print of nuclear transcription factors during hantavirus infection. The viral-replication-dependent activation of pro-myelocytic leukemia protein (PML) was followed by subsequent localization in nuclear bodies (NBs). PML was also found in close proximity to activated Sp100 nuclear antigen and interferon-stimulated gene 20 kDa protein (ISG-20), but co-localization with death-domain associated protein-6 (DAXX) was not observed. These data demonstrate that hantavirus triggers PML activation and localization in NBs in the absence of DAXX-PLM-NB co-localization. The results suggest that viral infection interferes with DAXX-mediated apoptosis, and expression of interferon-activated Sp100 and ISG-20 proteins may indicate intracellular intrinsic antiviral attempts.

Biological insight, high-throughput datasets and the nature of neuro-degenerative disorders.

Authors: Valente AX, Oliveira PJ, Khaiboullina SF, Palotás A, Rizvanov AA.
Publication: Curr Drug Metab. 2013 Sep;14(7):814-8.

Life sciences are experiencing a historical shift towards a quantitative, data-rich regime. This transition has been associated with the advent of bio-informatics: mathematicians, physicists, computer scientists and statisticians are now commonplace in the field, working on the analysis of ever larger data-sets. An open question regarding what should drive scientific progress in this new era remains: will biological insight become increasingly irrelevant in a world of hypothesis-free, unbiased data analysis? This piece offers a different perspective, pin-pointing that biological thought is more-than-ever relevant in a data-rich setting. Some of the novel highthroughput information being acquired in the field of neuro-degenerative disorders is highlighted here. As but one example of how theory and experiment can interact in this new reality, our efforts in developing an idiopathic neuro-degenerative disease hematopoietic stemcell ageing theory are described.

Human dendritic cells transfected with a human papilloma virus-18 construct display decreased mobility and upregulated cytokine production.

Authors: Khaiboullina SF, Morzunov SP, Hall MR, De Meirleir KL, Rizvanov AA, Lombardi VC.
Publication: Int J Oncol. 2013 Nov;43(5):1701-9. doi: 10.3892/ijo.2013.2074. Epub 2013 Aug 21.

The marked depletion of dendritic cells (DCs) in skin cancers, as well as preneoplastic and neoplastic cervical epithelium, suggests a central role for DCs in productive human papillomavirus (HPV) infection and cancer promotion. It has been suggested that HPV may facilitate tumor development by reducing DC density, contributing to a decrease in local immune surveillance. In this study, we have examined the response of human DCs transfected with a construct containing the HPV18 genome and their subsequent expression of papilloma virus proteins. Transfected cells expressed the L1 major capsid protein and upregulated E6 and E7 oncoprotein transcripts as detected by RT-PCR. Transfection of DCs also resulted in a significant increase in cytokine production. Finally, we observed that HPV18 transfection decreased the migratory activity of DCs. Our data indicate that HPV transfection of DCs leads to changes in migratory activity and cytokine production, which potentially can suppress or delay immune responses to viral antigens. Additionally, changes in cytokine production by HPV-transformed human fibroblasts and human cervical epithelial cells revealed that the migratory and antigen-presenting functions of DCs may be impaired by the suppressive effects of cytokines produced by HPV-infected epithelial and stromal cells.

Andes-virus-induced cytokine storm is partially suppressed by ribavirin.

Authors: Khaiboullina SF, Rizvanov AA, Lombardi VC, Morzunov SP, Reis HJ, Palotás A, St Jeor S.
Publication: Antivir Ther. 2013;18(4):575-84. doi: 10.3851/IMP2524. Epub 2013 Jan 8.

BACKGROUND: Microbe-induced over-activation of cytokines, especially tumour necrosis factor (TNF)-α, is key to the pathogenesis of hantavirus infection leading to severe inflammation with high mortality rate. Although ribavirin showed promise in inhibiting viral replication in vitro, its clinical efficacy remains controversial.

METHODS: Various concentrations of ribavirin were used to determine its effect on cytokine activation in our infectious model system.
RESULTS: Ribavirin decreased the virus load and dose-dependently inhibited the accumulation of RANTES messenger RNA in Andes-virus (ANDV)-infected human endothelial cells, but failed to suppress TNF-α-induced activation of RANTES and interleukin-6 in ANDV-inoculated cultures. This report also shows, for the first time, that the deleterious over-stimulation by TNF-α is mediated by nuclear factor-κB, and describes the effect of ribavirin on cytokine production following ANDV infection.

CONCLUSIONS: Although highly effective in preventing ANDV replication and suppressing activation of select inflammatory mediators, the therapeutic efficacy of ribavirin is limited due to its inability to fully inhibit cytokine outburst triggered by hantavirus infection.

Screening Models of Anticancer Drugs

Authors: Natalia L. Blatt, Rimma N. Mingaleeva, Svetlana F. Khaiboullina, Alexander Kotlyar, Vincent C. Lombardi,
Albert A. Rizvanov*
Publication: Life Science Journal 2013;10(4)

Abstract: Animal models have been indispensable when conducting research to further the understanding of cancer biology and when developing anticancer drugs. This article presents an overview of the most commonly utilized animal models for preclinical screening of anticancer agents. These models can be roughly divided into two groups:

models in which tumors are transplanted into mice, and models in which tumors develop in situ, either spontaneously or induced. Special attention is paid to the widely used subcutaneous xenotransplant and the orthotopic tumor models. We will also highlight the development and use of genetically modified mice.

[Blatt N.L., Mingaleeva R.N., Khaiboullina S.F., Kotlyar A., Lombardi V.C., Rizvanov A.A. IN VIVO Screening
Models of Anticancer Drugs. Life Sci J 2013;10(4):1892-1900] (ISSN:1097-8135).