Evaluation of the effects in the in vitro system of synthetic thymic hexapeptide on the expression levels of NF-κB, IFNα/βR and CD119 neutrophilic granulocytes in patients with chronic herpes viral co-infections

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Abstract

Background. Strategies used by herpes viruses with human cells are complex and multifaceted. On one hand, inborn defects in antiviral immune defense have been unveiled, which also affect interferon (IFN) system underlying development of chronic recalcitrant relapsing viral infections such as remittent respiratory viral infections, herpesvirus infections, and papillomavirus infections. On the other hand, numerous viruses are able to damage both immune system and IFN network. During inborn and acquired defects in IFN network, inborn or induced mutation in gene products involved in signaling cascades aimed at upregulating gene expression responsible for IFN production are observed. One of the strategies used by diverse viruses is altering some signaling pathways resulting in activated transcription factors including nuclear factor NF-kB. However, antiviral mechanisms executed by neutrophilic granulocytes (NGs), particularly affecting NF-kB expression have not been elucidated. Aim of the study: to study in vitro features of NF-kB expression and number of neutrophilic granulocytes (NG) expressing membrane IFNα/βR and IFNγR in patients with atypical chronic active herpes virus infections (AChA-HVI), followed by assessing an effect of arginyl-alpha-aspartyl-lysyl-valyl-tyrosyl-arginine — hexapeptide (HP), a synthetic analogue of the active center of the thymopoietin (active substance of drug “Imunofan”, Russia), on the expression of NG NF-kB and IFNα/βR and IFNγR. Materials and methods. We observed 25 patients of both sexes aged 23 to 64 years with AChA-HVI, manifested by chronic fatigue syndrome and cognitive disorders. Study design: stage 1 — clinical, ELISA, PCR methods, FC was used. Stage 2 — the in vitro experiment: 32 blood samples from 8 healthy adults and 375 blood samples from 25 patients with AChA-HVI were analyzed: % NG expressing NF-kB, IFNα/βR, IFNγR and the relevant MFI levels by using FC before and after incubation with HP. Results. Our study demonstrated low level (MFI) of NF-kB expression in 100% NG associated with decreased % of NG expressing IFNα/βR and IFNγR in all patients with AChA-HVI and low serum level for IFNα and IFNγ in comparison with healthy individuals. In the in vitro experiment there was shown that 100% of NG expressed NF-kB after exposure to HP. However, only 48% patients (SG 2) restored NF-kB expression level (MFI) to normal range and 52% of cases (SG 1) had no response. HP increased % of NG expressing IFNα/βR in SG 2 and increased % of NG expressing IFNγR in SG 1. Conclusions. It was shown, that influence of HP “in vitro” has ambiguous effects on the expression of NF-kB, % of NG expressing IFNα/βR and IFNγR in patients with AChA-HVI. We assume that different NF-kB response to HP is associated with inborn or secondary NF-kB deficiency.

About the authors

I. V. Nesterova

The Peoples’ Friendship University of Russia; Kuban State Medical University

Email: inesterova1@yandex.ru

PhD, MD (Medicine), Professor, Department of Allergology and Immunology

Russian Federation, Moscow; Krasnodar

E. O. Khalturina

I.M. Sechenov First Moscow State Medical University (Sechenov University)

Author for correspondence.
Email: jane_k@inbox.ru

PhD (Medicine), Associate Professor, Department of Microbiology, Virology and Immunology

Russian Federation, Moscow

V. N. Nelubin

Moscow State University of Medicine and Dentistry

Email: vlnelyubin@mail.ru

PhD, MD (Medicine), Professor

Russian Federation, Moscow

S. V. Khaidukov

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry

Email: hsv@mail.ibch.ru

PhD, MD (Biology), Senior Researcher, Carbohydrate Laboratory

Russian Federation, Moscow

G. A. Chudilova

Kuban State Medical University

Email: chudilova2015@yandex.ru

PhD, MD (Biology), Associate Professor, Department of Clinical Immunology, Allergology and Laboratory Diagnostics

Russian Federation, Krasnodar

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Supplementary files

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2. Figure 1. Expression levels of nuclear factor NF-B in neutrophilic granulocytes of patients suffering from AChA-HVI and in control group (conditionally healthy individuals) according to MFI distribution

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3. Figure 2 Comparison of the expression levels (MFI) for NF-B in neutrophilic granulocytes from patients with AChA-HVI before and after exposure to HP in in vitro experimental system

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4. Figure 3. Count of NG expressing membrane receptors IFNα/βR and IFNγ (CD119) before and after HP exposure in patients suffering from AChA-HVI

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Copyright (c) 2022 Nesterova I.V., Khalturina E.O., Nelubin V.N., Khaidukov S.V., Chudilova G.A.

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