Recombinant Pneumolysin of Pneumococci Induces TLR4 Expression and Maturation of Dendritic Cells In Vitro.

Pneumolysin (Ply) is a target for the development of serotype-independent pneumococcal vaccines, an important condition for the efficacy of which is their ability to activate innate immunity with the subsequent formation of adaptive immunity. In this study, the ability of recombinant full-length Ply (rPly) of pneumococci to induce TLR expression and maturation of dendritic cells generated from mouse bone marrow was evaluated. It was shown that rPly in vitro increased the number of dendritic cells expressing Toll-like receptor 4 (TLR4) on the membrane. rPly caused maturation of dendritic cells generated from mouse bone marrow, which manifested in a decrease in the number of progenitor cells (CD34), an increase in the number of cells expressing the adhesion molecule CD38, costimulatory molecules CD80 and CD86, molecules of terminal differentiation of dendritic cells CD83, as well as molecules of antigenic presentation of the major histocompatibility complex class II.

[BALANCE OF Thl/Th2/Th9/Thl7/Th22 CYTOKINES IN POST-OPERATION PERIOD IN PATIENTS WITH MALIGNANT TUMOR OF LIVER].

AIM: Evaluate cytokine status in patients with malignant liver cells after surgery. MATERIALS AND METHODS: 33 patients aged 35 to 76 years were included into the study. Blood was obtained before the operation and in the post-operation period: after 6 and 24 hours and at day 7 Cytokine profile (IL-Ib, IL-2, TNF-α, IFN-γ, IL-12p70, IL-4, IL-5,IL-6, IL-10, IL-13, IL-9, I-17a, IL-22) was evaluated using Multiplex- 13 system (Bender MedSystems, Austria). RESULTS: In patients levels of all the studied cytokines (Thl/Th2/Th9/Thl7/Th22) were already increased before the operations, that gives evidence of the presence of an inflammatory proc- ess connected with activation of immune system effectors. CONCLUSION: Disbalance of cytokine system helper cells resulting in functional and organic alterations through induction of the "cytokine storm" may aggravate the state of these patients. Further studies on the correction of cytokine system in these patients are thus needed.

[EFFECT OF INFLUENZA VACCINES ON SUBPOPULATIONS OF BLOOD DENDRITIC CELLS].

AIM: Study the effect of Vaxigrip split, Influvac subunit and Grippol plus immune-adjuvanted vaccines on the content of myeloid (mDC) and plasmacytoid (pDC) dendritic cells (DC) in blood of vaccinated healthy women. Materials andmethods. Blood of 30 healthy women aged 18-50 years was studied at days 7 and 30 after the vaccination. pDC (CD14+CD16-/CD85k(ILT3)-PE/ CD123-PC5) and mDC (CD14+CD-16-/CD85k(ILT3)-PE/CD33 -PC5) immune phenotyping was carried out using mAbs (Beckman Coulter, France) and flow cytometer Cytomix FC-500 (Beckman Coulter, USA). RESULTS: Use of unadjuvanted vaccines Vaxigrip and Influvac resulted in an increase of the numbers of mDC and pDC (p<0.05) in blood of the vaccinated only at day 7 of the observation. Grippol resulted in a more significant (2.2 - 3.6 times, p<0.05) increase of DC subpopulations (compared with unadjuvanted vaccines) at both day 7 and a month after the vaccination. CONCLUSION: Influenza vaccination activated innate effectors - the first component on the way of infection penetration - dendritic cells of both myeloid and lymphoid origin. Wherein, a more pronounced and prolonged effect of such activation is observed when immune-adjuvanted vaccine is used compared with subunit and split vaccines.

[SYNTHETIC CONJUGATED ANALOGUES OF CAPSULE POLYSACCHARIDES OF PNEUMOCOCCUS - AN INSTRUMENT FOR DETECTION OF POST-VACCINATION ANTIBODIES].

AIM: Evaluation ofthe ability of capsule polysaccharides (CP) of Streptococcus pneumoniae serotype 3 and 14 and their synthetic structure analogues, conjugated with bovine serum albumin (BSA), to detect antibodies in post-vaccination sera of mice. Materials andmethods. Oligosaccharides correspond- ing to one, one and a half and two repeating links of serotype 3 and 14 S. pneumoniae CP were synthe- sized, their conjugates with BSA were produced by squarate method as well. Ligand content-per BSA molecule was controlled by MALDI-TOF spectrometry. Immune sera were obtained after 2 intraperi- toneal administrations to mice of glucoconjugates adsorbed on aluminum hydroxide or 13-valent pneumococcal conjugated vaccine. Determination of levels of post-vaccination class G antibodies and their sub-isotypes was carried out in EIA. RESULTS: Immunization of mice with neoglucoconjugates resulted in formation of predominantly IgGl recognizing serotype 3 and 14 S. pneumoniae C. IgG1 in mice immunized with a 13-valent conjugated vaccine recognized serotype 3 S. pneumoniae CP, but detected serotype 14 S. pneumoniae CP weakly. All the conjugated synthetic oligosaccharides were characterized by a high ability to bind antibodies in blood of mice immunized with the polysaccharide conjugated vaccine. BSA-tetrasaccharide of serotype 3 S. pneumoniae and BSA-tetrasaccharide of serotype 14 S.pneumoniae were characterized by the highest ability to detect IgG1 against C. CONCLUSION: Synthetic oligosaccharides, conjugated with BSA protein-carrier, may be used to develop diagnostic test-systems for determination of antibodies in post-vaccination sera.

[COMPARATIVE ACTIVITY OF INFLUENZA VACCINES: EFFECT ON LYMPHO- CYTE SUBPOPULATION STRUCTURE].

AIM: Study subpopulation structure of lymphocytes in healthy individuals under the effect of various influenza vaccines in an in vitro system. MATERIALS AND METHODS: Evaluation of immune- phenotype features of PBMCs, activated in vitro by immune-adjuvanted and unadjuvanted vaccines against influenza in healthy individuals, was carried out by using flow cytometry method. RESULTS: Grippol plus vaccine caused a more pronounced stimulating effect compared with subunit and split-vaccines on NK-cells, cells with markers of early activation CD45/CD25, induced the quantity of natural regulatory cells (CD4/CD25/Foxp3), increase of the number of B-cells and reduced the amount of cell types with apoptosis marker CD45/CD95. CONCLUSION: Immune-adjuvanted vaccine Grippol plus induced formation of effectors of both innate and adaptive immunity and possessed the most powerful potential regarding activation of various types of immune-competent cells compared with unadjuvanted vaccines.