B Cell Profiling in Patients with Pemphigus Vulgaris.

Pemphigus vulgaris is a severe, socially significant autoimmune disease associated with autoantibodies to the desmoglein 3 antigen. The disease affects all age groups, beginning at 18 years of age; the mortality rate of pemphigus can reach as high as 50%, depending on a patient's age and a number of other factors. There is no highly selective or personalized therapy for pemphigus vulgaris at the moment. One of the well-known therapeutic approaches to the disease is to use rituximab, an anti-CD20 antibody that can help achieve B cell depletion in peripheral blood. To solve the problem of nonspecific elimination of B cells in patients with pemphigus vulgaris, it is reasonable to use specific immunoligands, their choice being based on an assessment of the level of autoantibodies specific to each of the fragments of desmoglein. In this work, the proportion of autoreactive B cells in patients diagnosed with pemphigus vulgaris is found to be 0.09-0.16%; a positive correlation was revealed between the antibody level and the number of autoreactive B cells to various fragments of desmoglein.

[Narrow-band medium-wave ultraviolet therapy in patients with atopic dermatitis: efficacy and safety]. / Uzkopolosnaya srednevolnovaya ul'trafioletovaya terapiya bol'nykh atopicheskim dermatitom: effektivnost' i bezopasnost'.

For the treatment of patients with atopic dermatitis of moderate and heavy severity level, narrow-band medium-wave ultraviolet therapy (narrow-band phototherapy) can be used. An analysis of the results of studies of the efficacy and safety of narrow-band medium-wavelength ultraviolet therapy in patients with atopic dermatitis is presented, and a characteristic of the regimens of the phototherapy carried out is given. It has been shown that narrow-band phototherapy is an effective and safe method of treating patients with atopic dermatitis, but its effectiveness varies widely. Data were obtained on the absence of an increase in the effect during therapy with higher doses of radiation, about the higher efficiency of narrow-band phototherapy with concurrent medication, with an increase in the number of irradiation procedures, as well as in patients with a higher minimum erythemal dose, which indicates the possible existence of factors characterizing the individual characteristics of the response of patients to narrow-band phototherapy.

Analysis of the Specificity of Auto-Reactive Antibodies to Individual Fragments of the Extracellular Domain of Desmoglein 3 in Patients with Pemphigus Vulgaris.

A method for the analysis of the epitope specificity of auto-reactive antibodies to desmoglein 3 (Dsg3) using competitive ELISA has been developed. It is based on a two-stage solid-phase ELISA with initial "depletion" of auto-reactive antibodies against the studied epitope and subsequent quantitative assessment of antibodies against full-length extracellular domain Dsg3. The proposed approach for assessing the specificity of the autoimmune response in patients with pemphigus vulgaris can provide in the future the possibility to personalize the therapy using plasmapheresis by preliminary selection of the antigenic composition of the extracorporeal immunosorbent.

The Effect of Janus Kinase Inhibitors and Phosphodiesterase-4 Inhibitors on Skin and Plasma Cytokine Levels in Patients with Psoriasis.

In patients with moderate-to-severe and severe psoriasis and high efficacy of therapy (PASI≥75) with signaling pathway inhibitors (apremilast, tofacitinib), cytokine spectra in the skin and blood plasma were studied using xMAP technology at baseline and on weeks 14 and 26 of treatment. Comparison of cytokine levels in psoriatic lesional skin and plasma samples of patients treated with apremilast or tofacitinib revealed statistical difference only for IFNγ level (р<0.05) at week 26.

Recombinant Fragment of the Extracellular Domain of Human Desmoglein 3 Fused with the Fc-Fragment of Human IgG1 Selectively Adsorbs Autoreactive Antibodies from the Sera of Pemphigus Patients.

Using the recombinant second fragment of the extracellular domain (EC2) of human desmoglein type 3 (Dsg3) as an affinity ligand, an immunosorbent was obtained that selectively binds autoreactive antibodies to this domain from the immune sera of patients with pemphigus. The EC2 protein was obtained in the form of a fusion protein with the Fc-fragment of human IgG1. The production was carried out in CHO cells using the method of transient expression.

Experimental Modeling of Leprosy in BALB/c, BALB/c Nude, CBA, and C57BL/6ТNF-/- Mice.

Leprosy was modeled in an experiment on BALB/c, BALB/cNude, CBA, and C57BL/6ТNF-/- mice using three Mycobacterium leprae strains obtained from patients with a diagnosis of A30 according to ICD-10 from different regions of the Russian Federation. Proliferation of M. leprae of the used strains showed a temporal-quantitative dependence on the used mouse line. CBA and BALB/cNude mice were optimal for strain R and BALB/c and BALB/cNude lines were optimal for strain I. BALB/cNude mice infected with strain I had low lifespan. M. leprae strain M showed low proliferation activity in BALB/cNude and C57BL/6ТNF-/- mice.

[Role of IL-17A and neutrophilic granulocytes in the pathogenesis of psoriasis]. / Rol' IL-17A i neitrofil'nykh granulotsitov v patogeneze psoriaza.

OBJECTIVE: To study the expression of IL-17A in the inflammatory infiltrating cells in the plaques as one of the key points in the pathological process in moderate to severe psoriasis. MATERIAL AND METHODS: The material obtained from 50 patients with moderate and severe psoriasis was examined by an indirect immunofluorescence assay to determine the composition of cell infiltrate and the type of IL-17A-producing cells in the foci of lesion. The markers of lymphocytes (CD3), dendritic cells (CD11c), neutrophilic granulocytes (Mpo), and mast cells (Trp) were used. The proliferative activity of basal keratinocytes (Ki-67), the expression of IL-17A, and the co-expression of IL-17A, Mpo, and Trp were also studied. RESULTS: A positive correlation was established between the count of neutrophilic granulocytes in the infiltrate, the expression of IL-17A, and the number of neutrophils expressing IL-17A with the duration of the disease and the severity of the patient's condition. CONCLUSION: Neutrophilic granulocytes and their expression of IL-17A play one of the key roles in the pathogenesis of psoriasis.

[The refinement of leprosy PCR diagnostics by the amplification of specie-specific repeated fragment of the Mycobacterium leprae genome.]

Certain level of new registered cases of leprosy in a number of endemic countries in the world, as well as growing rate of transboundary migratory flows, raise the issue of effective diagnosis of this disease in countries with sporadic incidence of leprosy, including the Russian Federation. The purpose of the study was to develop a highly sensitive PCR test for detecting the genetic material of Mycobacterium leprae and to compare the test robustness and sensitivity with the commercially available Leprosy Genesig Standard Kit (Primerdesign Ltd., UK). The proposed approach uses real time PCR of non-coding repeating element RLEP, unique for the M. leprae genome, using TaqMan probe. The high test specificity was shown using the reference DNA samples of pathogenic and conditionally pathogenic mycobacterium, as well and its comparison with single-copy genes of M. leprae (rrs, fbp, MntH) PCR detection. The use of a commercially available test system based on the single-copy rpoB gene detection provided 59.4% sensitivity to the detection of M. leprae in the clinical material, while the application of the developed approach increased this index to 96.8%. The developed PCR diagnostics test of leprosy is submitted for state clinical approval process, whereupon the practical use of the test diagnostics allows solving a wide range of tasks to identify and confirm new cases of leprosy, and monitoring both the effectiveness of leprosy treatment, and epidemiological (including transboundary) the spread of the disease.

[Current approaches to the morphologic diagnosis of different types of congenital epidermolysis bullosa]. / Sovremennye podkhody k morfologicheskoi diagnostike razlichnykh tipov vrozhdennogo bulleznogo epidermoliza.

Congenital epidermolysis bullosa (CEB) is an extensive group of hereditary skin diseases, the differential diagnosis of which is a challenge due to the rarity of this pathology and the diversity of its clinical manifestations. The determination of the type of CEB makes it possible to estimate its prognosis and to facilitate a prenatal diagnosis. AIM: to optimize the morphological diagnosis of different types of CEB. MATERIAL AND METHODS: 28 skin biopsies from 14 patients with different types of CEB were investigated. The investigators performed routine histological examination of skin fragments taken from a bullous area and immunofluorescence antigen mapping using the indirect immunofluorescence test (IIFT) with antibodies against structural proteins of the dermal-epidermal junction (laminin α3, ß3, and γ2 chains, keratins 5 and 14, types VII and XVII collagen, α6 and ß4 integrin subunits, desmoplakin, plectin, kindlin-1, and plakophillin) of the apparently unaffected skin. The intact skin of healthy individuals, which had been obtained during cosmetic operations, was used as controls in IIFT. RESULTS: Immunofluorescence antigen mapping could determine the type of CEB in all cases and in 86% of cases identify the protein, the impaired production of which was responsible for the development of the disease. CONCLUSION: Immunofluorescence antigen mapping is an integral part of the comprehensive morphological diagnosis of CEB, acting as an intermediate between the morphological verification of CEB diagnosis and the targeted search for mutations by a molecular genetic method.

[Role of the skin expression of neuropeptides, neurotrophins and their receptors in the pathogenesis of dermatoses].

OBJECTIVE: to define the role of neurotransmitters and their receptors in the development of itch and in the maintenance of a skin inflammatory response in patients with psoriasis and atopic dermatitis. MATERIAL AND METHODS: Skin biopsy specimens from 30 patients with psoriasis and 30 patients with atopic dermatitis were investigated by histological, immunoperoxidase, and indirect immunofluorescence assays. The investigators determined the expression of protein gene product 9.5 (PGP9.5), amphiregulin, semaphorin 3A, calcitonin gene-related peptide (CGRP) and its receptor (CGRP-R), nerve growth factor (NGF) and its receptor TrkA, and substance P (SP) and its receptor SP-R. The indirect immunofluorescence assay was used for quantitative analysis. The findings were statistically analyzed using a Statistica 10 program. RESULTS: Immunoperoxidase examination of the skin biopsy specimens from patients with atopic dermatitis and psoriasis revealed enhanced expression of amphiregulin, NGF, and PGP9.5, appearance of positively stained epidermal nerve fibers, and decreased expression of the nerve reduction factor semaphorin 3A in all cases. Some patients with atopic dermatitis and psoriasis showed increased expression of CGRP and CGRP-R, SP, SP-R, and TrkA. A pronounced inflammatory response was generally observed in these cases. CONCLUSION: The investigation performed suggests that atopic dermatitis and psoriasis are characterized by a larger number of epidermal nerve fibers and by a direct correlation between this indicator, disease severity, and itch intensity. The production of neuropeptides and neurotrophins is closely related to the development of a skin inflammatory response irrespective of its cause and dysregulation of these processes is likely to favor the body's sensitization and the chronic pattern the course of diseases.

[Future Therapeutic Targets in Management of Autoimmune Skin Diseases].

Pemphigus is a severe, potentially fatal bullous skin disease, caused by desmoglein autoantibody production and immune-mediated regulation of T-cells subsets. Conventional therapy including systemic corticosteroids with or without other immunosupressants causes numerous adverse effects and becomes inefficient in refractory patients. In this work, the authors showed a modern view on the pathogenesis ofpemphigus. This article describes the detailed action mechanism of rituximab, a chimeric monoclonal antibody directed against CD20 antigen of B-cells. The authors conduct the results of meta-analyses of rituximab's efficiency in pemphigus patients. Moreover, in this article, the authors consider new promising treatment tions and potential targets for biological therapy of pemphigus diseases.

Detection of mutant Mycoplasma hominis strains resistant to 16-membered macrolide antibiotic josamycin in clinical samples.

The incidence of point mutations responsible for josamycin resistance was studied by PCR in 48 strains of M. hominis isolated from patients with bacterial vaginosis. Mutant M. hominis strains were detected in 48% cases.