Biomedical Chemistry: Research and Methods 2025, 8(4), e00309

COMPARISON OF THE EFFICIENCY OF DENDRITIC CELL MATURATION UPON STIMULATION WITH BACTERIAL LIPOPOLYSACCHARIDE OR TUMOR NECROSIS FACTOR ALFA

E.A. Titov1,2, V.K. Plisova1,2, I.A. Pokusaeva1, R.Yu. Saryglar1, I.V. Kholodenko1, O.A. Bystrykh3, A.V. Kuprin3, K.N. Yarygin1, A.Yu. Lupatov*1

1Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia; *e-mail: biocell@inbox.ru
2Moscow Gymnasium in the Southwest No. 1543 named after Yu.V. Zavelsky,
3 build. 5, 26 Baku Komissarov, Moscow, 119571 Russia
3A.V. Vishnevsky Institute of Surgery, 27 Bolshaya Serpukhovskaya, Moscow, 117997 Russia

Keywords:dendritic cells; induced maturation; tumor necrosis factor; lipopolysaccharide; cell-based vaccines

DOI:10.18097/BMCRM00309

The whole version of this paper is available in Russian.

Dendritic cells (DCs) are the professional antigen-presenting cells capable of presenting antigens to T-lymphocytes, thereby initiating the primary immune response. The unique immunological properties of this cell population make their use as a cellular vaccine relevant for the treatment of oncological and chronic infectious diseases. A critical stage in obtaining DCs with immunostimulatory properties is their maturation. To compare the effectiveness of alternative methods for stimulating maturation, DCs were obtained in vitro from peripheral blood monocytes through their induced differentiation in the presence of the cytokines GM-CSF and IL-4. As biochemical stimuli for inducing maturation, bacterial lipopolysaccharide (LPS) or tumor necrosis factor alpha combined with prostaglandin E2 (TNFα+PGE2) were used. No significant differences were found in the ability of these factors to stimulate the expression of HLA-DR and costimulatory molecules (CD80, CD83, CD86). The use of alternative maturations did not lead to differences in the ability of DCs to stimulate the proliferation of allogeneic lymphocytes. At the same time, there were morphological signs indicating the ability of LPS to stimulate differentiation into macrophages.
Figure 1. Cell cultures derived from peripheral blood mononuclear cells. A – Monocytes after attachment to the surface of the culture vessel. B – Immature dendritic cells (DCs) on the third day of differentiation. Phase-contrast light microscopy.

Figure 2. Expression of the monocyte/macrophage marker CD14 and the immature dendritic cell marker CD1a on cells after 5 days of cultivation. A – unstained control. B – cells cultured without GM-CSF and IL-4. C – cells cultured in the presence of GM-CSF and IL-4 in the medium. Fluorescence intensity is shown on both axes. The percentage of cells included in each quadrant is indicated.

Figure 3. Dendritic cell cultures after induction of their maturation using LPS (A) or TNFα + PGE2 (B). Phase-contrast light microscopy.

Figure 4. Distribution of lymphocytes across cell cycle phases during co-cultivation with immature (iDC) or mature (LPS; TNFα+PGE2) dendritic cells. The x-axis represents fluorescence intensity; the y-axis represents the number of recorded events (cells).

Figure 5. Proliferative activity of allogeneic lymphocytes during their co-cultivation with immature (iDC) or mature (LPS; TNFα+PGE2) dendritic cells. The x-axis represents fluorescence intensity; the y-axis represents the number of recorded events (cells).

Figure 6. Expression of HLA-DR and costimulatory molecules on dendritic cells during their maturation. iDC – immature dendritic cells; LPS, TNFα+PGE2 – mature dendritic cells stimulated to mature by the corresponding method; gray peak – unstained control (immature dendritic cells). The x-axis represents fluorescence intensity; the y-axis represents the normalized number of recorded events (cells).

FUNDING

This work was performed within the framework of the Program of Fundamental Scientific Research in the Russian Federation for the long-term period (2021-2030) No. 122022800499-5.

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