Supplementary data for the article: Spasojević, D.; Zmejkoski, D.; Glamočlija, J.; Nikolić, M.; Soković, M.; Milošević, V.; Jarić, I.; Stojanović, M.; Marinković, E.; Barisani-Asenbauer, T.; et al. Lignin Model Compound in Alginate Hydrogel: A Strong Antimicrobial Agent with High Potential in Wound Treatment. International Journal of Antimicrobial Agents 2016, 48 (6), 732–735. https://doi.org/10.1016/j.ijantimicag.2016.08.014

Supplementary material for: [https://doi.org/10.1016/j.ijantimicag.2016.08.014]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2364

Modulation of functional characteristics of murine peritoneal macrophages by dehydrogenate polymer from coniferyl alcohol and alginate

The dehydrogenate polymer from coniferyl alcohol (DHP; a lignin model compound) in alginate hydrogel (ALG) has been shown to exert a strong antibacterial activity. To broadens a spectrum of potential DHP/ALG application, we aimed this study to evaluate the immunomodulatory activity of DHP/ALG. DHP and ALG were tested separately and in mixture (1:2 w/w) for their impact on in vitro production of cytokines (IL-6, IL-12, and IL-10) and reactive oxygen (ROS) and nitrogen (RNS) species by resident (RMs) and thioglycolate-elicited (TGMs) peritoneal macrophages of BALB/c mice. RMs and TGMs were stimulated (48h) with ALG and DHP in concentrations previously shown to be non-cytotoxic (up to 50 and 25 μg/ml, respectively). DHP/ALG promotes simultaneous production of inflammatory (IL-6, IL-12) and regulatory cytokines by RMs in a positive dose-dependent manner. Production of inflammatory cytokines was stimulated by ALG, while an increase in IL-10 production positively correlated to the concentration of DHP. ALG also stimulated the production of IL-12 by TGMs, which was mirrored in the outcome of ALG/DHP stimulation. The significant increase in the activity of myeloperoxidase (MPO) due to DHP and/or ALG stimulation was recorded in TGMs, while a slight increase in MPO activity in RMs was recorded only upon stimulation with the higher amount of ALG. ALG in a positive dose-dependent manner stimulated the production of ROS and RNS by both RMS and TGMs. In all cases, except ROS production by RMs, the impact of ALG stimulation was mirrored in the outcome of ALG/DHP stimulation. Our results suggest that DHP/ALG exerts an immunomodulatory activity that could complement already reported antimicrobial activity and warrants further investigation on the use of DHP/ALG in the treatment of infectious diseases

Recombinant banana lectin as mucosal immunostimulator

To explore the potential usage of recombinant banana lectin as an immunostimulator it was of interest to analyze the effect it exerted in vivo in mice, upon two different routes of exposure. By Western blotting analysis of the gastric and intestinal luminal content it was found that recombinant banana lectin is stable in vivo in the mouse digestive tract, where it specifically interacts with the mucosal surfaces which can be inhibited by the addition of glucose. Its attachment to mice jejunum has been confirmed by immunofluorescence. In addition, it was able to induce systemic immunity, evidenced by specific antibody production, by oral application, a property that might be exploited for the induction of systemic immune responses. (c) 2012 Elsevier Ltd. All rights reserved

JSCS–3827 Original scientific paper

The monoclonal antibody 26 raised against tetanus toxoid also recognizes tetanus toxin and β2-glycoprotein I – its binding properties in vitro and potential application

Supplementary data for article: Marinkovic, E.; Djokic, R.; Lukic, I.; Filipovic, A.; Inic-Kanada, A.; Kosanovic, D.; Gavrovic-Jankulovic, M.; Stojanovic, M. Modulation of Functional Characteristics of Resident and Thioglycollate-Elicited Peritoneal Murine Macrophages by a Recombinant Banana Lectin. PLoS ONE 2017, 12 (2). https://doi.org/10.1371/journal.pone.017246

Supporting information for: [https://doi.org/10.1371/journal.pone.017246]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2417

Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion

Allergen-specific immunotherapy (AIT) is a desensitizing treatment for allergic diseases that corrects the underlined pathological immune response to innocuous protein antigens, called allergens. Recombinant allergens employed in the AIT allowed the production of well-defined formulations that possessed consistent quality but were often less efficient than natural allergen extracts. Combining recombinant allergens with an adjuvant or immunomodulatory agent could improve AIT efficacy. This study aimed to perform structural and functional characterization of newly designed recombinant chimera composed of the Bet v 1, the major birch pollen allergen, and Banana Lectin (BanLec), TLR2, and CD14 binding protein, for the application in AIT. rBet v 1-BanLec chimera was designed in silico and expressed as a soluble fraction in Escherichia coli. Purified rBet v 1-BanLec (33.4 kDa) retained BanLec-associated biological activity of carbohydrate-binding and preserved IgE reactive epitopes of Bet v 1. The chimera revealed secondary structures with predominant β sheets. The immunomodulatory capacity of rBet v 1-BanLec tested on macrophages showed changes in myeloperoxidase activity, reduced NO production, and significant alterations in the production of cytokines when compared to both rBanLec and rBet v 1. Comparing to rBet v 1, rBet v 1-BanLec was demonstrated to be more efficient promoter of IL-10 production as well as weaker inducer of NO production and secretion of pro-inflammatory cytokines TNFα, and IL-6. The ability of rBet v 1-BanLec to promote IL-10 in together with the preserved 3D structure of Bet v 1 part implies that the construct might exert a beneficial effect in the allergen-specific immunotherapy

Supplementary data for article: Marinkovic, E.; Djokic, R.; Lukic, I.; Filipovic, A.; Inic-Kanada, A.; Kosanovic, D.; Gavrovic-Jankulovic, M.; Stojanovic, M. Modulation of Functional Characteristics of Resident and Thioglycollate-Elicited Peritoneal Murine Macrophages by a Recombinant Banana Lectin. PLoS ONE 2017, 12 (2). https://doi.org/10.1371/journal.pone.017246

Supporting information for: [https://doi.org/10.1371/journal.pone.017246]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2417