2026-10-13T02:44:52Z https://api.figshare.com/v2/oai

oai:figshare.com:article/34026925 2026-09-29T17:38:44Z category_4 category_7 category_8 category_12 category_46 category_64 category_132 portal_5 item_type_1 month_year_09_2026

<p>Pools of <i>Francisella</i> MHC-II peptides elicit time-dependent, specific and CD4⁺ T cell-mediated IFN-γ response in immune splenocytes.</p> Pavlina Laskova (25138447) Stanislava Porkertova (25138450) Ondrej Ballek (11568544) Lucie Balonova (1392496) Jana Pavloskova (25138453) Ales Neuwirth (13913055) Ivona Pavkova (2339947) Ivo Fabrik (1849519) Pavol Lukac (25138456) Luca Vannucci (14358999) Dominik Filipp (1849615) Jiri Stulik (151536) Marek Link (151527) Biochemistry Medicine Microbiology Cell Biology Immunology Cancer Infectious Diseases sup >+</ sup severe pneumonic form induced immune responses identified 57 mhc div >< p immunopeptidomics reveals conserved protective immune response peptide vaccine containing fully virulent strains ensure complete protection cells following immunization francisella tularensis </ tularensis </ using immunopeptidomics based immunopeptidomics vaccination studies subsequent screening significant role potential role least 80 ii peptides human tularemia highly expressed fsc200 strain five epitopes epitopes proved early control dominant epitopes characterizing antigen cells playing cell epitopes bacterial proteome 23 different <p>C57Bl/6 mice were subcutaneously injected with 3 × 10⁴ CFU of <i>F. tularensis</i> LVS and immune lymphocytes, pooled from groups of 3-4 mice, were stimulated with bacterial MHC-II peptides assembled in pools for immunogenicity assessment. The numbers of IFN-γ producing cells (SFCs) were determined at 3, 4 and 5 weeks p.i. following stimulation of (A) 5 × 10⁵ whole splenocytes or (B) cocultures of 3 × 10⁵ spleen CD4⁺ T cells and 3 × 10⁴ BMDCs with peptide pools (PP). Data of individual assayed wells per condition and mean values are shown. PPs with significant response against control without peptides (unstim) were detected with one-way ANOVA followed by Sidak’s post hoc test. The specificity with which the peptide-induced IFN-γ production depended on immune status of lymphocytes was verified by stimulation of (C) 5 × 10⁵ whole splenocytes (3 weeks p.i.) or (D) cocultures of 3 × 10⁵ spleen CD4⁺ T cells (4 weeks p.i.) and 3 × 10⁴ BMDCs with PP-3. Lymphocytes were isolated from LVS immunized (immune) or age-matched control (control) mice. Data of individual assayed wells per condition and mean values are shown. Responses were assessed for significant difference using a two-tailed t-test. (E) IFN-γ and (F) TNF-α concentrations were determined by ELISA in cell culture supernatants of 8 × 10⁵ LVS-immune splenocytes (3 weeks p.i.) after 48 h of PPs stimulation. Data of 3 biological replicates and mean values are shown. PPs with significant response against control without peptides were detected with one-way ANOVA followed by Dunnett’s post hoc test. PPs representing a subset of 15 peptides (Table B in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1014597#ppat.1014597.s002" target="_blank">S1 Appendix</a>) were used for all assays with 5 µM final concentration of each peptide. Ova II peptide (5 µM) was used as a negative control in some experiments. Asterisks indicate p-values (**p < 0.01, ***p < 0.001, ****p < 0.0001, ns for p ≥ 0.05). Data are results of one experiment (A, B, C) or representative results of two biological replicates (D).</p> 2026-09-29T17:54:09Z Image Figure 10.1371/journal.ppat.1014597.g003 https://figshare.com/articles/figure/_p_Pools_of_i_Francisella_i_MHC-II_peptides_elicit_time-dependent_specific_and_CD4_sup_sup_T_cell-mediated_IFN-_response_in_immune_splenocytes_p_/34026925 CC BY 4.0