Cytokine Profile and Expression of FYN, ZAP-70 and LAT During Concanavalin A Stimulation in Patients with Resistant Bronchial Asthma

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Abstract


Background: Bronchial asthma (BA) is one of the most spreading chronic lung pathology in the world. The disease is characterized by high heterogeneity of clinical phenotypes including resistant forms which provoke significant clinical problem. Immune shift from Th2 to alternative immunological response is considered to be a mechanism of drug-resistance in BA treatment but this issue is not considerably studied yet. Aims: Detection of distinctive patterns in cytokine secretion and genetic expression (ZAP-70, FYN and LAT) of naïve and concanavalin A stimulated lymphocytes in patients with resistant BA. Materials and methods: The study enrolled ten patients in each group: subjects with treatment resistant BA, severe BA, and controls (30 in total). During the experiment, all patients with BA received treatment according to the condition. For each participant lymphocytes isolation from venous blood was performed. Cells were cultured with concanavalin A and without stimulation. Concentrations of cytokines IL-2, IL-12, TNF-α, IL-4, IL-5, and IL-6 in supernatants were measured with ELISA. Reverse transcription polymerase chain reaction was used to detect the mRNA expression of LAT, ZAP-70, and FYN genes. Results: Significant disease contribution to the lymphocyte secretion profile was established without concanavalin A stimulation: increased levels of IL-2 and IL-4 was observed in lymphocytes of patients with resistant BA if compared to the results of gorup with severe BA. Patients with resistant BA were characterized by weak cytokine response to the stimulation: only TNF-α and IL-5 levels were significantly increased whereas in group with severe BA all cytokines concentrations increased except IL-12, in controls — except IL-12 and IL-2. Significant FYN upregulation was identified in resistant BA group if compared with other groups, and in severe BA patients if compared with controls. The concanavalin A-stimulated cells showed increased expression of ZAP-70 in cells of patients with resistant BA compared to control group. Conclusions: Lymphocytes from patients with resistant BA are characterized by lack of cytokine response to concanavalin A stimulation, alteration of cytokine secretion, and genetic expression profile similar to cells with low sensitivity to apoptosis. The FYN gene is a perspective target for finding approaches to overcome resistance to steroid drugs in bronchial asthma.


Vyacheslav A. Petrov

Siberian State Medical University

Author for correspondence.
Email: vyacheslav.a.petrov@mail.ru
ORCID iD: 0000-0002-5205-9739

Russian Federation

2g bld.18, Moskovskiy trakt, 634001 Tomsk.

SPIN-код: 9635-2243

Irina V. Saltykova

Siberian State Medical University

Email: ira.saltikova@mail.ru
ORCID iD: 0000-0002-0457-5392

Russian Federation

Tomsk.

SPIN-код: 9432-3873

Ksenia V. Nevskaya

Siberian State Medical University

Email: nevskayaksenia@gmail.com
ORCID iD: 0000-0003-1659-8812

Russian Federation

Tomsk.

SPIN-код: 1405-0472

Julia B. Dorofeeva

Siberian State Medical University

Email: julia.dorofeeva25@gmail.com
ORCID iD: 0000-0002-9301-4465

Russian Federation

Tomsk.

SPIN-код: 9890-8870

Sofiya P. Lezhava

Siberian State Medical University

Email: lezhavasofiya@gmail.com
ORCID iD: 0000-0002-8806-8231

Russian Federation

Tomsk.

SPIN-код: 9629-7318

Natalya A. Kirillova

Siberian State Medical University

Email: kirillova.natalya@gmail.com
ORCID iD: 0000-0001-9549-9614

Russian Federation

Tomsk.

SPIN-код: 8308-5833

Evgeny S. Kulikov

Siberian State Medical University

Email: evgeny.s.kulikov@gmail.com
ORCID iD: 0000-0002-0088-9204

Russian Federation

MD, PhD, Professor.

Tomsk.

SPIN-код: 9934-1476

Alexey E. Sazonov

Siberian State Medical University

Email: sazonov_al@mail.ru
ORCID iD: 0000-0001-8611-5770

Russian Federation

Tomsk.

SPIN-код: 6177-6729

Ludmila M. Ogorodova

Siberian State Medical University

Email: lm-ogorodova@mail.ru
ORCID iD: 0000-0002-2962-1076

Russian Federation

MD, PhD, Professor.

Tomsk.

SPIN-код: 4362-8431

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