TY  - JOUR
AU  - Božić, Dragana
AU  - Ćirković, Ivana
AU  - Milovanović, Jovica
AU  - Bufan, Biljana
AU  - Folić, Miljan
AU  - Savić Vujović, Katarina
AU  - Pavlović, Bojan
AU  - Jotić, Ana
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5341
AB  - Bacterial biofilms play an important role in the pathogenesis of chronic upper respiratory tract infections. In addition to conventional antimicrobial therapy, N-acetyl-L-cysteine (NAC) and propolis are dietary supplements that are often recommended as supportive therapy for upper respiratory tract infections. However, no data on the beneficial effect of their combination against bacterial biofilms can be found in the scientific literature. Therefore, the aim of our study was to investigate the in vitro effect of N-acetyl-L-cysteine (NAC) and dry propolis extract in fixed combinations (NAC/dry propolis extract fixed combination) on biofilm formation by bacterial species isolated from patients with chronic rhinosinusitis, chronic otitis media, and chronic adenoiditis. The prospective study included 48 adults with chronic rhinosinusitis, 29 adults with chronic otitis media, and 33 children with chronic adenoiditis. Bacteria were isolated from tissue samples obtained intraoperatively and identified using the MALDI-TOF Vitek MS System. The antimicrobial activity, synergism, and antibiofilm effect of NAC/dry propolis extract fixed combination were studied in vitro. A total of 116 different strains were isolated from the tissue samples, with staphylococci being the most frequently isolated in all patients (57.8%). MICs of the NAC/dry propolis extract fixed combination ranged from 1.25/0.125 to 20/2 mg NAC/mg propolis. A synergistic effect (FICI ≤ 0.5) was observed in 51.7% of strains. The majority of isolates from patients with chronic otitis media were moderate biofilm producers and in chronic adenoiditis they were weak biofilm producers, while the same number of isolates in patients with chronic rhinosinusitis were weak and moderate biofilm producers. Subinhibitory concentrations of the NAC/propolis combination ranging from 0.625–0.156 mg/mL to 10–2.5 mg/mL of NAC combined with 0.062–0.016 mg/mL to 1–0.25 mg/mL of propolis inhibited biofilm formation in all bacterial strains. Suprainhibitory concentrations ranging from 2.5–10 mg/mL to 40–160 mg/mL of NAC in combination with 0.25–1 mg/mL to 4–16 mg/mL of propolis completely eradicated the biofilm. In conclusion, the fixed combination of NAC and dry propolis extract has a synergistic effect on all stages of biofilm formation and eradication of the formed biofilm in bacteria isolated from upper respiratory tract infections.
PB  - MDPI
T2  - Pharmaceuticals
T1  - In Vitro Antibiofilm Effect of N-Acetyl-L-cysteine/Dry Propolis Extract Combination on Bacterial Pathogens Isolated from Upper Respiratory Tract Infections
VL  - 16
IS  - 11
DO  - 10.3390/ph16111604
ER  - 

@article{
author = "Božić, Dragana and Ćirković, Ivana and Milovanović, Jovica and Bufan, Biljana and Folić, Miljan and Savić Vujović, Katarina and Pavlović, Bojan and Jotić, Ana",
year = "2023",
abstract = "Bacterial biofilms play an important role in the pathogenesis of chronic upper respiratory tract infections. In addition to conventional antimicrobial therapy, N-acetyl-L-cysteine (NAC) and propolis are dietary supplements that are often recommended as supportive therapy for upper respiratory tract infections. However, no data on the beneficial effect of their combination against bacterial biofilms can be found in the scientific literature. Therefore, the aim of our study was to investigate the in vitro effect of N-acetyl-L-cysteine (NAC) and dry propolis extract in fixed combinations (NAC/dry propolis extract fixed combination) on biofilm formation by bacterial species isolated from patients with chronic rhinosinusitis, chronic otitis media, and chronic adenoiditis. The prospective study included 48 adults with chronic rhinosinusitis, 29 adults with chronic otitis media, and 33 children with chronic adenoiditis. Bacteria were isolated from tissue samples obtained intraoperatively and identified using the MALDI-TOF Vitek MS System. The antimicrobial activity, synergism, and antibiofilm effect of NAC/dry propolis extract fixed combination were studied in vitro. A total of 116 different strains were isolated from the tissue samples, with staphylococci being the most frequently isolated in all patients (57.8%). MICs of the NAC/dry propolis extract fixed combination ranged from 1.25/0.125 to 20/2 mg NAC/mg propolis. A synergistic effect (FICI ≤ 0.5) was observed in 51.7% of strains. The majority of isolates from patients with chronic otitis media were moderate biofilm producers and in chronic adenoiditis they were weak biofilm producers, while the same number of isolates in patients with chronic rhinosinusitis were weak and moderate biofilm producers. Subinhibitory concentrations of the NAC/propolis combination ranging from 0.625–0.156 mg/mL to 10–2.5 mg/mL of NAC combined with 0.062–0.016 mg/mL to 1–0.25 mg/mL of propolis inhibited biofilm formation in all bacterial strains. Suprainhibitory concentrations ranging from 2.5–10 mg/mL to 40–160 mg/mL of NAC in combination with 0.25–1 mg/mL to 4–16 mg/mL of propolis completely eradicated the biofilm. In conclusion, the fixed combination of NAC and dry propolis extract has a synergistic effect on all stages of biofilm formation and eradication of the formed biofilm in bacteria isolated from upper respiratory tract infections.",
publisher = "MDPI",
journal = "Pharmaceuticals",
title = "In Vitro Antibiofilm Effect of N-Acetyl-L-cysteine/Dry Propolis Extract Combination on Bacterial Pathogens Isolated from Upper Respiratory Tract Infections",
volume = "16",
number = "11",
doi = "10.3390/ph16111604"
}

Božić, D., Ćirković, I., Milovanović, J., Bufan, B., Folić, M., Savić Vujović, K., Pavlović, B.,& Jotić, A.. (2023). In Vitro Antibiofilm Effect of N-Acetyl-L-cysteine/Dry Propolis Extract Combination on Bacterial Pathogens Isolated from Upper Respiratory Tract Infections. in Pharmaceuticals
MDPI., 16(11).
https://doi.org/10.3390/ph16111604

Božić D, Ćirković I, Milovanović J, Bufan B, Folić M, Savić Vujović K, Pavlović B, Jotić A. In Vitro Antibiofilm Effect of N-Acetyl-L-cysteine/Dry Propolis Extract Combination on Bacterial Pathogens Isolated from Upper Respiratory Tract Infections. in Pharmaceuticals. 2023;16(11).
doi:10.3390/ph16111604 .

Božić, Dragana, Ćirković, Ivana, Milovanović, Jovica, Bufan, Biljana, Folić, Miljan, Savić Vujović, Katarina, Pavlović, Bojan, Jotić, Ana, "In Vitro Antibiofilm Effect of N-Acetyl-L-cysteine/Dry Propolis Extract Combination on Bacterial Pathogens Isolated from Upper Respiratory Tract Infections" in Pharmaceuticals, 16, no. 11 (2023),
https://doi.org/10.3390/ph16111604 . .

TY  - JOUR
AU  - Božić, Dragana
AU  - Pavlović, Bojan
AU  - Milovanović, Jovica
AU  - Jotić, Ana
AU  - Colović, Jelena
AU  - Cirković, Ivana
PY  - 2018
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/3212
AB  - Microbial biofilms have been implicated in the pathogenesis of chronic rhinosinusitis with nasal polyposis (CRSwNP). The aim of our study was to evaluate in vitro effects of amoxicillin-clavulanic acid and levofloxacin on biofilm formation by bacterial species isolated from sinus tissue in patients with CRSwNP. The sinus mucosal specimens were harvested from the upper parts and roof of ethmoid cavity of 48 patients with CRSwNP. Each sample was washed thoroughly in three separate beakers of sterile saline to remove any planktonic bacteria and further subjected to microbiology analysis. The biofilm-forming capacity of isolated strains was detected by microtiter-plate method and the effects of subinhibitory (1/2x to 1/16x MIC) and suprainhibitory concentrations (4, 8, 16, 32, and 64 A mu g/ml) of amoxicillin-clavulanic acid and levofloxacin on biofilm production were investigated. Bacterial strains were isolated in 42 (87.5%) patients: one microorganism in 80.9% and two microorganisms in 19.1% of patients. The most prevalent bacteria in CRSwNP biofilms were Staphylococcus epidermidis (34%) and S. aureus (28%) followed by S. haemolyticus (12%), Pseudomonas aeruginosa (8%), Moraxella catarrhalis (6%), Streptococcus pneumoniae (6%), and other staphylococci (6%). Subinhibitory concentrations of amoxicillin-clavulanic acid and levofloxacin significantly reduced biofilm formation (p  lt  0.01 and p  lt  0.05, respectively), with better efficacy of amoxicillin-clavulanic acid (1/2-1/8x MIC) on staphylococci and levofloxacin (1/2- 1/4x MIC) on M. catarrhalis and P. aeruginosa biofilm formation. Suprainhibitory concentrations of both tested antibiotics (4-64 A mu g/ml) significantly eradicated mature biofilms of staphylococci (p  lt  0.01). The effect of levofloxacin on eradication of staphylococcal biofilms was more noticeable, compared to the effect of amoxicillin-clavulanic acid (p  lt  0.01). Suprainhibitory concentrations of both tested antibiotics had no effect on eradication of previously formed M. catarrhalis and P. aeruginosa biofilms (p > 0.05). The amoxicillin-clavulanic acid and levofloxacin are shown to be potent antibiofilm agents in patients with CRSwNP. The effects of tested compounds depend on bacterial species and the volume of formed biofilm.
PB  - Springer, New York
T2  - European Archives of Oto-Rhino-Laryngology
T1  - Antibiofilm effects of amoxicillin-clavulanic acid and levofloxacin in patients with chronic rhinosinusitis with nasal polyposis
VL  - 275
IS  - 8
SP  - 2051
EP  - 2059
DO  - 10.1007/s00405-018-5049-6
ER  - 

@article{
author = "Božić, Dragana and Pavlović, Bojan and Milovanović, Jovica and Jotić, Ana and Colović, Jelena and Cirković, Ivana",
year = "2018",
abstract = "Microbial biofilms have been implicated in the pathogenesis of chronic rhinosinusitis with nasal polyposis (CRSwNP). The aim of our study was to evaluate in vitro effects of amoxicillin-clavulanic acid and levofloxacin on biofilm formation by bacterial species isolated from sinus tissue in patients with CRSwNP. The sinus mucosal specimens were harvested from the upper parts and roof of ethmoid cavity of 48 patients with CRSwNP. Each sample was washed thoroughly in three separate beakers of sterile saline to remove any planktonic bacteria and further subjected to microbiology analysis. The biofilm-forming capacity of isolated strains was detected by microtiter-plate method and the effects of subinhibitory (1/2x to 1/16x MIC) and suprainhibitory concentrations (4, 8, 16, 32, and 64 A mu g/ml) of amoxicillin-clavulanic acid and levofloxacin on biofilm production were investigated. Bacterial strains were isolated in 42 (87.5%) patients: one microorganism in 80.9% and two microorganisms in 19.1% of patients. The most prevalent bacteria in CRSwNP biofilms were Staphylococcus epidermidis (34%) and S. aureus (28%) followed by S. haemolyticus (12%), Pseudomonas aeruginosa (8%), Moraxella catarrhalis (6%), Streptococcus pneumoniae (6%), and other staphylococci (6%). Subinhibitory concentrations of amoxicillin-clavulanic acid and levofloxacin significantly reduced biofilm formation (p  lt  0.01 and p  lt  0.05, respectively), with better efficacy of amoxicillin-clavulanic acid (1/2-1/8x MIC) on staphylococci and levofloxacin (1/2- 1/4x MIC) on M. catarrhalis and P. aeruginosa biofilm formation. Suprainhibitory concentrations of both tested antibiotics (4-64 A mu g/ml) significantly eradicated mature biofilms of staphylococci (p  lt  0.01). The effect of levofloxacin on eradication of staphylococcal biofilms was more noticeable, compared to the effect of amoxicillin-clavulanic acid (p  lt  0.01). Suprainhibitory concentrations of both tested antibiotics had no effect on eradication of previously formed M. catarrhalis and P. aeruginosa biofilms (p > 0.05). The amoxicillin-clavulanic acid and levofloxacin are shown to be potent antibiofilm agents in patients with CRSwNP. The effects of tested compounds depend on bacterial species and the volume of formed biofilm.",
publisher = "Springer, New York",
journal = "European Archives of Oto-Rhino-Laryngology",
title = "Antibiofilm effects of amoxicillin-clavulanic acid and levofloxacin in patients with chronic rhinosinusitis with nasal polyposis",
volume = "275",
number = "8",
pages = "2051-2059",
doi = "10.1007/s00405-018-5049-6"
}

Božić, D., Pavlović, B., Milovanović, J., Jotić, A., Colović, J.,& Cirković, I.. (2018). Antibiofilm effects of amoxicillin-clavulanic acid and levofloxacin in patients with chronic rhinosinusitis with nasal polyposis. in European Archives of Oto-Rhino-Laryngology
Springer, New York., 275(8), 2051-2059.
https://doi.org/10.1007/s00405-018-5049-6

Božić D, Pavlović B, Milovanović J, Jotić A, Colović J, Cirković I. Antibiofilm effects of amoxicillin-clavulanic acid and levofloxacin in patients with chronic rhinosinusitis with nasal polyposis. in European Archives of Oto-Rhino-Laryngology. 2018;275(8):2051-2059.
doi:10.1007/s00405-018-5049-6 .

Božić, Dragana, Pavlović, Bojan, Milovanović, Jovica, Jotić, Ana, Colović, Jelena, Cirković, Ivana, "Antibiofilm effects of amoxicillin-clavulanic acid and levofloxacin in patients with chronic rhinosinusitis with nasal polyposis" in European Archives of Oto-Rhino-Laryngology, 275, no. 8 (2018):2051-2059,
https://doi.org/10.1007/s00405-018-5049-6 . .

TY  - THES
AU  - Pavlović, Bojan
PY  - 2018
UR  - http://eteze.bg.ac.rs/application/showtheses?thesesId=6204
UR  - http://nardus.mpn.gov.rs/handle/123456789/10196
UR  - https://fedorabg.bg.ac.rs/fedora/get/o:18799/bdef:Content/download
UR  - http://vbs.rs/scripts/cobiss?command=DISPLAY&base=70036&RID=2048300642
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/3676
AB  - Otkriće dendritskih ćelija (DC) predstavlja jedno od najznačajnijih dostignuća u imunologiji, s obzirom da su ove ćelije ključne za započinjanje i regulaciju imunskog odgovora protiv patogena i malignih ćelija, kao i protiv sopstvenih ćelija tokom autoimunskog odgovora. Ove ćelije predstavljaju heterogenu grupu čija je zajednička osobina sposobnost indukcije antigen-specifičnog adaptivnog imuniteta. Tokom recirkulacije između perifernih tkiva i limfnih organa, DC preuzimaju antigene iz mikrosredine i prezentuju ih naivnim T limfocitima. U uslovima homeostaze prezentacija antigena od strane nezrelih DC dovodi do uspostavljanja tolerancije na antigen, koja uključuje ispoljavanje koinhibitornih molekula (PD-L1, ILT3, ILT4, i dr.) na DC i indukciju regulatornih T ćelija (Treg). Sa druge strane, aktivacija DC, posredstvom receptora za prepoznavanje molekulskih obrazaca i proinflamacijskih citokina, dovodi do maturacije DC što se ogleda u ispoljavanju kostimulatornih molekula i prezentacije antigena u imunogenoj formi. Zrele DC aktiviraju i pomoćničke T (engl. T helper, Th) limfocite i citotoksične T limfocite i indukuju njihovu proliferaciju i diferencijaciju u efektorske i memorijske ćelije. Citokini koje produkuju DC imaju najznačajniju ulogu u regulaciji diferencijacije Th limfocita ka različitim subpopulacijama (Th1, Th17, Th2 ili Treg). Nastale efektorske T ćelije dalje regulišu ćelijski i humoralni imunski odgovor. Imajući u vidu sve navedeno, jasno je da bi imunogene DC, koje su sposobne da indukuju tumor-specifične Th1 ćelije i citotoksične CD8+ T limfocite, bile izuzetno aktraktivne za primenu u imunoterapiji kancera. Predloženi su brojni protokoli za dobijanje imunogenih DC od monocita in vitro, uključujući i postupak ubrzane diferencijacije, koji je usled skraćenja vremena diferencijacije znatno jeftiniji i fiziološki relevantniji u odnosu na konvencionalni postupak dobijanja DC. Međutim, za sada nema dovoljno podataka koji bi potvrdili prednost DC dobijenih protokolom ubrzane diferencijacije (engl. fast DC, fDC) u odnosu na konvencionalne DC (engl. conventional, cDC), u pogledu njihove sposobnosti da indukuju efikasan anti-tumorski imunski odgovor. Takođe, nije poznato u kojoj meri razlike u fenotipu i funkciji monocita iz različitih donora utiču na njihov potencijal da se diferenciraju u DC. Pokazano je da je protokolom ubrzane diferencijacije u prisustvu proinflamacijskog koktela moguće dobiti imunogene DC, isto kao i konvencionalnim protokolom, u pogledu fenotipskog sazrevanja, produkcije IL-12 i indukcije Th1 ćelija, ali samo kod dela donora koji su označeni kao dobri responderi. Suprotno, fDC dobijene iz monocita loših respondera, stimulisanih na isti način, bile su fenotipski manje zrele (ispoljavale su niže nivoe CD1a, MHC II, CD83, CD86, CCR7),iiprodukovale su manje IL-12 i slabije su stimulisale proliferaciju aloreaktivnih T ćelija i usmeravale diferencijaciju ka Th1 ćelijama...
AB  - The discovery of dendritic cells (DC) is one of the most important findings in immunology, especially when the role of these cells in regulation of the immune response against pathogens, and cancer cells is considered. DC are heterogeneous cells, all of which are able to induce antigen-specific adaptive immunity. DC recirculate between peripheral and lymph organs during which they sample the microenvironment for antigens and present them to naïve T lymphocytes. Immature DC present antigen during homeostasis which leads to induction of antigen tolerance, and the response is triggered actively via expression of co-inhibitory molecules (PD-L1, ILT3, ILT4, etc.) and induction of regulatory T cells (Treg). In contrast, the activation of DC via pattern recognition receptors and proinflammatory cytokines induces up-regulation of co-stimulatory molecules and immunogenic antigen presentation. Thereby, DC activate the proliferation and differentiation of T lymphocytes towards effector and memory T helper (Th) cells and cytotoxic T cells. The cytokines produced by DC thereby, are critically involved in the regulation of T cell differentiation (Th polarization) and development of specific Th1, Th17, Th2 or Treg-mediated immune response. Subsequently, effector T cells regulate cellular and humoral immune response. These facts make DC extremely attractive for the application in immunotherapy, such as cancer immunotherapy which is focused on induction of immunogenic DC that are capable of inducing tumor-specific Th1 and cytotoxic CD8+ T cells. Various protocols for the induction of monocyte-derived DC in vitro have been proposed, including fast DC protocols which is claimed to be much cheaper and more relevant physiologically than the conventional protocol. However, a strong evidence which supports that fast DC (fDC) are better than conventional DC (cDC) in inducing an efficient anti-tumor response, is still missing. Moreover, it is still unknown how donor-related differences in the phenotypic and functional properties of precursor monocytes relate to the capacity of these cells to differentiate into DC. Here we showed that fast DC protocol which included proinflammatory cytokines cocktail is as good as the conventional DC protocol in inducing phenotypic maturation, IL-12 production and Th1 polarization by certain donors only (responders). fDC from non-responders expressed lower levels of CD1a, MHC class II, CD83, CD86, CCR7 and IL-12, displayed lower allostimulatory capacity and induction of Th1 cells. These properties correlated with a higher surface expression of CD16 by non-responder monocytes, and their higher capacity to produce IL-6 and IL-1β early in culture, compared to responder monocytes. Since proinflammatory cocktail may exhibit inhibitory effect on DC differentiation,vwe tested whether Poly (I:C), a strong Th1 polarizing agent, can improve the efficacy of obtaining immunogenic fDC...
PB  - Универзитет у Београду, Фармацеутски факултет
T2  - Универзитет у Београду
T1  - Sazrevanje i funkcija humanih dendritskih ćelija dobijenih od monocita skraćenjem vremena diferencijacije
UR  - https://hdl.handle.net/21.15107/rcub_nardus_10196
ER  - 

@phdthesis{
author = "Pavlović, Bojan",
year = "2018",
abstract = "Otkriće dendritskih ćelija (DC) predstavlja jedno od najznačajnijih dostignuća u imunologiji, s obzirom da su ove ćelije ključne za započinjanje i regulaciju imunskog odgovora protiv patogena i malignih ćelija, kao i protiv sopstvenih ćelija tokom autoimunskog odgovora. Ove ćelije predstavljaju heterogenu grupu čija je zajednička osobina sposobnost indukcije antigen-specifičnog adaptivnog imuniteta. Tokom recirkulacije između perifernih tkiva i limfnih organa, DC preuzimaju antigene iz mikrosredine i prezentuju ih naivnim T limfocitima. U uslovima homeostaze prezentacija antigena od strane nezrelih DC dovodi do uspostavljanja tolerancije na antigen, koja uključuje ispoljavanje koinhibitornih molekula (PD-L1, ILT3, ILT4, i dr.) na DC i indukciju regulatornih T ćelija (Treg). Sa druge strane, aktivacija DC, posredstvom receptora za prepoznavanje molekulskih obrazaca i proinflamacijskih citokina, dovodi do maturacije DC što se ogleda u ispoljavanju kostimulatornih molekula i prezentacije antigena u imunogenoj formi. Zrele DC aktiviraju i pomoćničke T (engl. T helper, Th) limfocite i citotoksične T limfocite i indukuju njihovu proliferaciju i diferencijaciju u efektorske i memorijske ćelije. Citokini koje produkuju DC imaju najznačajniju ulogu u regulaciji diferencijacije Th limfocita ka različitim subpopulacijama (Th1, Th17, Th2 ili Treg). Nastale efektorske T ćelije dalje regulišu ćelijski i humoralni imunski odgovor. Imajući u vidu sve navedeno, jasno je da bi imunogene DC, koje su sposobne da indukuju tumor-specifične Th1 ćelije i citotoksične CD8+ T limfocite, bile izuzetno aktraktivne za primenu u imunoterapiji kancera. Predloženi su brojni protokoli za dobijanje imunogenih DC od monocita in vitro, uključujući i postupak ubrzane diferencijacije, koji je usled skraćenja vremena diferencijacije znatno jeftiniji i fiziološki relevantniji u odnosu na konvencionalni postupak dobijanja DC. Međutim, za sada nema dovoljno podataka koji bi potvrdili prednost DC dobijenih protokolom ubrzane diferencijacije (engl. fast DC, fDC) u odnosu na konvencionalne DC (engl. conventional, cDC), u pogledu njihove sposobnosti da indukuju efikasan anti-tumorski imunski odgovor. Takođe, nije poznato u kojoj meri razlike u fenotipu i funkciji monocita iz različitih donora utiču na njihov potencijal da se diferenciraju u DC. Pokazano je da je protokolom ubrzane diferencijacije u prisustvu proinflamacijskog koktela moguće dobiti imunogene DC, isto kao i konvencionalnim protokolom, u pogledu fenotipskog sazrevanja, produkcije IL-12 i indukcije Th1 ćelija, ali samo kod dela donora koji su označeni kao dobri responderi. Suprotno, fDC dobijene iz monocita loših respondera, stimulisanih na isti način, bile su fenotipski manje zrele (ispoljavale su niže nivoe CD1a, MHC II, CD83, CD86, CCR7),iiprodukovale su manje IL-12 i slabije su stimulisale proliferaciju aloreaktivnih T ćelija i usmeravale diferencijaciju ka Th1 ćelijama..., The discovery of dendritic cells (DC) is one of the most important findings in immunology, especially when the role of these cells in regulation of the immune response against pathogens, and cancer cells is considered. DC are heterogeneous cells, all of which are able to induce antigen-specific adaptive immunity. DC recirculate between peripheral and lymph organs during which they sample the microenvironment for antigens and present them to naïve T lymphocytes. Immature DC present antigen during homeostasis which leads to induction of antigen tolerance, and the response is triggered actively via expression of co-inhibitory molecules (PD-L1, ILT3, ILT4, etc.) and induction of regulatory T cells (Treg). In contrast, the activation of DC via pattern recognition receptors and proinflammatory cytokines induces up-regulation of co-stimulatory molecules and immunogenic antigen presentation. Thereby, DC activate the proliferation and differentiation of T lymphocytes towards effector and memory T helper (Th) cells and cytotoxic T cells. The cytokines produced by DC thereby, are critically involved in the regulation of T cell differentiation (Th polarization) and development of specific Th1, Th17, Th2 or Treg-mediated immune response. Subsequently, effector T cells regulate cellular and humoral immune response. These facts make DC extremely attractive for the application in immunotherapy, such as cancer immunotherapy which is focused on induction of immunogenic DC that are capable of inducing tumor-specific Th1 and cytotoxic CD8+ T cells. Various protocols for the induction of monocyte-derived DC in vitro have been proposed, including fast DC protocols which is claimed to be much cheaper and more relevant physiologically than the conventional protocol. However, a strong evidence which supports that fast DC (fDC) are better than conventional DC (cDC) in inducing an efficient anti-tumor response, is still missing. Moreover, it is still unknown how donor-related differences in the phenotypic and functional properties of precursor monocytes relate to the capacity of these cells to differentiate into DC. Here we showed that fast DC protocol which included proinflammatory cytokines cocktail is as good as the conventional DC protocol in inducing phenotypic maturation, IL-12 production and Th1 polarization by certain donors only (responders). fDC from non-responders expressed lower levels of CD1a, MHC class II, CD83, CD86, CCR7 and IL-12, displayed lower allostimulatory capacity and induction of Th1 cells. These properties correlated with a higher surface expression of CD16 by non-responder monocytes, and their higher capacity to produce IL-6 and IL-1β early in culture, compared to responder monocytes. Since proinflammatory cocktail may exhibit inhibitory effect on DC differentiation,vwe tested whether Poly (I:C), a strong Th1 polarizing agent, can improve the efficacy of obtaining immunogenic fDC...",
publisher = "Универзитет у Београду, Фармацеутски факултет",
journal = "Универзитет у Београду",
title = "Sazrevanje i funkcija humanih dendritskih ćelija dobijenih od monocita skraćenjem vremena diferencijacije",
url = "https://hdl.handle.net/21.15107/rcub_nardus_10196"
}

Pavlović, B.. (2018). Sazrevanje i funkcija humanih dendritskih ćelija dobijenih od monocita skraćenjem vremena diferencijacije. in Универзитет у Београду
Универзитет у Београду, Фармацеутски факултет..
https://hdl.handle.net/21.15107/rcub_nardus_10196

Pavlović B. Sazrevanje i funkcija humanih dendritskih ćelija dobijenih od monocita skraćenjem vremena diferencijacije. in Универзитет у Београду. 2018;.
https://hdl.handle.net/21.15107/rcub_nardus_10196 .

Pavlović, Bojan, "Sazrevanje i funkcija humanih dendritskih ćelija dobijenih od monocita skraćenjem vremena diferencijacije" in Универзитет у Београду (2018),
https://hdl.handle.net/21.15107/rcub_nardus_10196 .

TY  - JOUR
AU  - Cirković, Ivana
AU  - Pavlović, Bojan
AU  - Božić, Dragana
AU  - Jotić, Ana
AU  - Bakić, Ljubica
AU  - Milovanović, Jovica
PY  - 2017
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/3005
AB  - Microbial biofilms have been implicated in the pathogenesis of chronic rhinosinusitis with nasal polyposis (CRSwNP). Intranasal application of corticosteroids and saline is a reliable option for their management. The aim of our study was to evaluate in vitro antibiofilm effects of corticosteroids and isotonic and hypertonic nasal saline in CRSwNP patients. The sinus mucosal specimens were harvested from the ethmoid cavity of 48 patients with CRSwNP and further subjected to hematoxylin-eosin staining and microbiology analysis. The biofilm-forming capacity of isolated bacterial strains was detected by microtiter-plate method and the effects of therapeutic doses of mometasone, fluticasone, isotonic and hypertonic saline on biofilm production were investigated. Bacterial strains were isolated in 42 (87.5%) patients: one organism in 34 (80.9%) and two organisms in 8 (19.1%). Staphylococcus epidermidis (34%) and Staphylococcus aureus (28%) were the most prevalent bacteria in biofilms of CRSwNP patients. Corticosteroids and saline solutions significantly reduced biofilm formation (p  lt  0.01 and p  lt  0.05, respectively) with better efficacy of fluticasone and isotonic nasal saline. Treatment with fluticasone, mometasone, isotonic and hypertonic nasal saline completely prevented biofilm production in 66, 50, 84 and 38% of bacterial strains, respectively. The most significant density reduction was observed in biofilm formed by Staphylococcus aureus, Pseudomonas aeruginosa and Streptococcus pneumoniae compared to other bacterial species (p  lt  0.01, p  lt  0.05, p  lt  0.05, respectively). The antibiofilm effects of corticosteroids and saline solutions also greatly depended on bacterial biomass (p  lt  0.05), with the most significant effect on high compared to small amount of formed biofilm. The topical steroids and nasal saline are shown to be potent antibiofilm agents in patients with CRSwNP. The effects of tested compounds depend on bacterial species and volume of formed biofilm.
PB  - Springer, New York
T2  - European Archives of Oto-Rhino-Laryngology
T1  - Antibiofilm effects of topical corticosteroids and intranasal saline in patients with chronic rhinosinusitis with nasal polyps depend on bacterial species and their biofilm-forming capacity
VL  - 274
IS  - 4
SP  - 1897
EP  - 1903
DO  - 10.1007/s00405-017-4454-6
ER  - 

@article{
author = "Cirković, Ivana and Pavlović, Bojan and Božić, Dragana and Jotić, Ana and Bakić, Ljubica and Milovanović, Jovica",
year = "2017",
abstract = "Microbial biofilms have been implicated in the pathogenesis of chronic rhinosinusitis with nasal polyposis (CRSwNP). Intranasal application of corticosteroids and saline is a reliable option for their management. The aim of our study was to evaluate in vitro antibiofilm effects of corticosteroids and isotonic and hypertonic nasal saline in CRSwNP patients. The sinus mucosal specimens were harvested from the ethmoid cavity of 48 patients with CRSwNP and further subjected to hematoxylin-eosin staining and microbiology analysis. The biofilm-forming capacity of isolated bacterial strains was detected by microtiter-plate method and the effects of therapeutic doses of mometasone, fluticasone, isotonic and hypertonic saline on biofilm production were investigated. Bacterial strains were isolated in 42 (87.5%) patients: one organism in 34 (80.9%) and two organisms in 8 (19.1%). Staphylococcus epidermidis (34%) and Staphylococcus aureus (28%) were the most prevalent bacteria in biofilms of CRSwNP patients. Corticosteroids and saline solutions significantly reduced biofilm formation (p  lt  0.01 and p  lt  0.05, respectively) with better efficacy of fluticasone and isotonic nasal saline. Treatment with fluticasone, mometasone, isotonic and hypertonic nasal saline completely prevented biofilm production in 66, 50, 84 and 38% of bacterial strains, respectively. The most significant density reduction was observed in biofilm formed by Staphylococcus aureus, Pseudomonas aeruginosa and Streptococcus pneumoniae compared to other bacterial species (p  lt  0.01, p  lt  0.05, p  lt  0.05, respectively). The antibiofilm effects of corticosteroids and saline solutions also greatly depended on bacterial biomass (p  lt  0.05), with the most significant effect on high compared to small amount of formed biofilm. The topical steroids and nasal saline are shown to be potent antibiofilm agents in patients with CRSwNP. The effects of tested compounds depend on bacterial species and volume of formed biofilm.",
publisher = "Springer, New York",
journal = "European Archives of Oto-Rhino-Laryngology",
title = "Antibiofilm effects of topical corticosteroids and intranasal saline in patients with chronic rhinosinusitis with nasal polyps depend on bacterial species and their biofilm-forming capacity",
volume = "274",
number = "4",
pages = "1897-1903",
doi = "10.1007/s00405-017-4454-6"
}

Cirković, I., Pavlović, B., Božić, D., Jotić, A., Bakić, L.,& Milovanović, J.. (2017). Antibiofilm effects of topical corticosteroids and intranasal saline in patients with chronic rhinosinusitis with nasal polyps depend on bacterial species and their biofilm-forming capacity. in European Archives of Oto-Rhino-Laryngology
Springer, New York., 274(4), 1897-1903.
https://doi.org/10.1007/s00405-017-4454-6

Cirković I, Pavlović B, Božić D, Jotić A, Bakić L, Milovanović J. Antibiofilm effects of topical corticosteroids and intranasal saline in patients with chronic rhinosinusitis with nasal polyps depend on bacterial species and their biofilm-forming capacity. in European Archives of Oto-Rhino-Laryngology. 2017;274(4):1897-1903.
doi:10.1007/s00405-017-4454-6 .

Cirković, Ivana, Pavlović, Bojan, Božić, Dragana, Jotić, Ana, Bakić, Ljubica, Milovanović, Jovica, "Antibiofilm effects of topical corticosteroids and intranasal saline in patients with chronic rhinosinusitis with nasal polyps depend on bacterial species and their biofilm-forming capacity" in European Archives of Oto-Rhino-Laryngology, 274, no. 4 (2017):1897-1903,
https://doi.org/10.1007/s00405-017-4454-6 . .

TY  - JOUR
AU  - Pavlović, Bojan
AU  - Božić, Dragana
AU  - Milovanović, Jovica
AU  - Jotić, Ana
AU  - Đukić, Vojko
AU  - Đukić, Slobodanka
AU  - Konstantinović, Neda
AU  - Cirković, Ivana
PY  - 2016
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/2695
AB  - Objectives: Biofilms are associated with persistent infections and resistant to conventional therapeutic strategies. The aim of this study was to investigate the quantity of biofilm produced on silicone intranasal splints. Methods: Quantity of biofilm formation on silicone splints (SS) was tested on 15 strains of Staphylococcus aureus and Moraxella catarrhalis, respectively. Antimicrobial susceptibility testing was performed in accordance with European Committee on Antimicrobial Susceptibility Testing recommendations. Results: All tested strains formed different amounts of biofilm on SS: 66.7% S. aureus and 93.3% M. catarrhalis were weak biofilm producers and 33.3% S. aureus and 6.7% M. catarrhalis were moderate biofilm producers. S. aureus formed significantly higher quantity of biofilm compared with M. catarrhalis (p  lt  0.05). Multidrug resistant S. aureus produced significantly higher amount of biofilm compared with non-multidrug resistant strains (p  lt  0.05). Conclusion: Quantity of biofilm on SS is highly dependent on bacterial species and their resistance patterns. Future studies are needed to ascertain another therapeutic option for prophylaxis prior to SS placement.
PB  - Akademiai Kiado Rt, Budapest
T2  - Acta Microbiologica et Immunologica Hungarica
T1  - Quantification of biofilm formation on silicone intranasal splints: An in vitro study
VL  - 63
IS  - 3
SP  - 301
EP  - 311
DO  - 10.1556/030.63.2016.006
ER  - 

@article{
author = "Pavlović, Bojan and Božić, Dragana and Milovanović, Jovica and Jotić, Ana and Đukić, Vojko and Đukić, Slobodanka and Konstantinović, Neda and Cirković, Ivana",
year = "2016",
abstract = "Objectives: Biofilms are associated with persistent infections and resistant to conventional therapeutic strategies. The aim of this study was to investigate the quantity of biofilm produced on silicone intranasal splints. Methods: Quantity of biofilm formation on silicone splints (SS) was tested on 15 strains of Staphylococcus aureus and Moraxella catarrhalis, respectively. Antimicrobial susceptibility testing was performed in accordance with European Committee on Antimicrobial Susceptibility Testing recommendations. Results: All tested strains formed different amounts of biofilm on SS: 66.7% S. aureus and 93.3% M. catarrhalis were weak biofilm producers and 33.3% S. aureus and 6.7% M. catarrhalis were moderate biofilm producers. S. aureus formed significantly higher quantity of biofilm compared with M. catarrhalis (p  lt  0.05). Multidrug resistant S. aureus produced significantly higher amount of biofilm compared with non-multidrug resistant strains (p  lt  0.05). Conclusion: Quantity of biofilm on SS is highly dependent on bacterial species and their resistance patterns. Future studies are needed to ascertain another therapeutic option for prophylaxis prior to SS placement.",
publisher = "Akademiai Kiado Rt, Budapest",
journal = "Acta Microbiologica et Immunologica Hungarica",
title = "Quantification of biofilm formation on silicone intranasal splints: An in vitro study",
volume = "63",
number = "3",
pages = "301-311",
doi = "10.1556/030.63.2016.006"
}

Pavlović, B., Božić, D., Milovanović, J., Jotić, A., Đukić, V., Đukić, S., Konstantinović, N.,& Cirković, I.. (2016). Quantification of biofilm formation on silicone intranasal splints: An in vitro study. in Acta Microbiologica et Immunologica Hungarica
Akademiai Kiado Rt, Budapest., 63(3), 301-311.
https://doi.org/10.1556/030.63.2016.006

Pavlović B, Božić D, Milovanović J, Jotić A, Đukić V, Đukić S, Konstantinović N, Cirković I. Quantification of biofilm formation on silicone intranasal splints: An in vitro study. in Acta Microbiologica et Immunologica Hungarica. 2016;63(3):301-311.
doi:10.1556/030.63.2016.006 .

Pavlović, Bojan, Božić, Dragana, Milovanović, Jovica, Jotić, Ana, Đukić, Vojko, Đukić, Slobodanka, Konstantinović, Neda, Cirković, Ivana, "Quantification of biofilm formation on silicone intranasal splints: An in vitro study" in Acta Microbiologica et Immunologica Hungarica, 63, no. 3 (2016):301-311,
https://doi.org/10.1556/030.63.2016.006 . .

TY  - JOUR
AU  - Jotić, Ana
AU  - Božić, Dragana
AU  - Milovanović, Jovica
AU  - Pavlović, Bojan
AU  - Jesić, Snežana
AU  - Pelemis, Mijomir
AU  - Novaković, Marko
AU  - Cirković, Ivana
PY  - 2016
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/2561
AB  - Bacterial biofilm formation has been implicated in the high incidence of persistent otorrhoea after tympanostomy tube insertion. The aim of the study was to investigate whether biofilm formation on tympanostomy tubes depends on the genetic profile of methicillin-resistant Staphylococcus aureus (MRSA) strains. Capacity of biofilm formation on fluoroplastic tympanostomy tubes (TTs) was tested on 30 MRSA strains. Identification and methicillin resistance were confirmed by PCR for nuc and mecA genes. Strains were genotypically characterised (SCCmec, agr and spa typing). Biofilm formation was tested in microtiter plate and on TTs. Tested MRSA strains were classified into SCCmec type I (36.7 %), III (23.3 %), IV (26.7 %) and V (13.3 %), agr type I (50 %), II (36.7 %) and III (13.3 %), and 5 clonal complexes (CCs). All tested MRSA strains showed ability to form biofilm on microtiter plate. Capacity of biofilm formation on TTs was as following: 13.3 % of strains belonged to the category of no biofilm producers, 50 % to the category of weak biofilm producers and 36.7 % to moderate biofilm producers. There was a statistically significant difference between CC, SCCmec and agr types and the category of biofilm production on TTs tubes (p  lt  0.001): CC5, SCCmecI type and agrII type with a moderate amount of biofilm, and CC8 and agrI type with a low amount of biofilm. Biofilm formation by MRSA on TTs is highly dependent on genetic characteristics of the strains. Therefore, MRSA genotyping may aid the determination of the possibility of biofilm-related post-tympanostomy tube otorrhea.
PB  - Springer, New York
T2  - European Archives of Oto-Rhino-Laryngology
T1  - Biofilm formation on tympanostomy tubes depends on methicillin-resistant Staphylococcus aureus genetic lineage
VL  - 273
IS  - 3
SP  - 615
EP  - 620
DO  - 10.1007/s00405-015-3607-8
ER  - 

@article{
author = "Jotić, Ana and Božić, Dragana and Milovanović, Jovica and Pavlović, Bojan and Jesić, Snežana and Pelemis, Mijomir and Novaković, Marko and Cirković, Ivana",
year = "2016",
abstract = "Bacterial biofilm formation has been implicated in the high incidence of persistent otorrhoea after tympanostomy tube insertion. The aim of the study was to investigate whether biofilm formation on tympanostomy tubes depends on the genetic profile of methicillin-resistant Staphylococcus aureus (MRSA) strains. Capacity of biofilm formation on fluoroplastic tympanostomy tubes (TTs) was tested on 30 MRSA strains. Identification and methicillin resistance were confirmed by PCR for nuc and mecA genes. Strains were genotypically characterised (SCCmec, agr and spa typing). Biofilm formation was tested in microtiter plate and on TTs. Tested MRSA strains were classified into SCCmec type I (36.7 %), III (23.3 %), IV (26.7 %) and V (13.3 %), agr type I (50 %), II (36.7 %) and III (13.3 %), and 5 clonal complexes (CCs). All tested MRSA strains showed ability to form biofilm on microtiter plate. Capacity of biofilm formation on TTs was as following: 13.3 % of strains belonged to the category of no biofilm producers, 50 % to the category of weak biofilm producers and 36.7 % to moderate biofilm producers. There was a statistically significant difference between CC, SCCmec and agr types and the category of biofilm production on TTs tubes (p  lt  0.001): CC5, SCCmecI type and agrII type with a moderate amount of biofilm, and CC8 and agrI type with a low amount of biofilm. Biofilm formation by MRSA on TTs is highly dependent on genetic characteristics of the strains. Therefore, MRSA genotyping may aid the determination of the possibility of biofilm-related post-tympanostomy tube otorrhea.",
publisher = "Springer, New York",
journal = "European Archives of Oto-Rhino-Laryngology",
title = "Biofilm formation on tympanostomy tubes depends on methicillin-resistant Staphylococcus aureus genetic lineage",
volume = "273",
number = "3",
pages = "615-620",
doi = "10.1007/s00405-015-3607-8"
}

Jotić, A., Božić, D., Milovanović, J., Pavlović, B., Jesić, S., Pelemis, M., Novaković, M.,& Cirković, I.. (2016). Biofilm formation on tympanostomy tubes depends on methicillin-resistant Staphylococcus aureus genetic lineage. in European Archives of Oto-Rhino-Laryngology
Springer, New York., 273(3), 615-620.
https://doi.org/10.1007/s00405-015-3607-8

Jotić A, Božić D, Milovanović J, Pavlović B, Jesić S, Pelemis M, Novaković M, Cirković I. Biofilm formation on tympanostomy tubes depends on methicillin-resistant Staphylococcus aureus genetic lineage. in European Archives of Oto-Rhino-Laryngology. 2016;273(3):615-620.
doi:10.1007/s00405-015-3607-8 .

Jotić, Ana, Božić, Dragana, Milovanović, Jovica, Pavlović, Bojan, Jesić, Snežana, Pelemis, Mijomir, Novaković, Marko, Cirković, Ivana, "Biofilm formation on tympanostomy tubes depends on methicillin-resistant Staphylococcus aureus genetic lineage" in European Archives of Oto-Rhino-Laryngology, 273, no. 3 (2016):615-620,
https://doi.org/10.1007/s00405-015-3607-8 . .

TY  - JOUR
AU  - Pavlović, Bojana
AU  - Cvijetić, Nataša
AU  - Dragacević, Luka
AU  - Ivković, Branka
AU  - Vujić, Zorica
AU  - Kuntić, Vesna
PY  - 2016
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/2597
AB  - One of the most commonly used surfactants in the production of split virus influenza vaccine is nonionic surfactant Triton X-100. After splitting of the virus is accomplished, Triton X-100 is removed from the vaccine by subsequent production steps. Because of toxicity of Triton X-100, which remains in the vaccine in residual amounts, a sufficiently sensitive method for its detection and quantification needs to be defined. Two methods for determination of Triton X-100 residuals were developed: the UV-spectrophotometry and HPLC methods. For both methods, preparation of vaccine samples and removal of proteins and virus particles were crucial: samples were treated with methanol (1:1) and then centrifuged at 25000 x g for 30 min. After such treatment, the majority of vaccine components that interfered in the UV region were removed, and diluted samples could be directly measured. The chromatographic system included C18 column, step methanol gradient, and detection at 225 nm with a single peak of Triton X-100 at 12.6 min. Both methods were validated and gave satisfactory results for accuracy, precision, specificity, linearity, and robustness. LOQ was slightly lower for the HPLC method. Hence, it was shown that both methods are suitable for analysis of residual amounts of Triton X-100, with the advantages of the UV method being its simplicity and availability in most laboratories.
PB  - AOAC Int, Gaithersburg
T2  - Journal of AOAC International
T1  - Direct UV Spectrophotometry and HPLC Determination of Triton X-100 in Split Virus Influenza Vaccine
VL  - 99
IS  - 2
SP  - 396
EP  - 400
DO  - 10.5740/jaoacint.15-0201
ER  - 

@article{
author = "Pavlović, Bojana and Cvijetić, Nataša and Dragacević, Luka and Ivković, Branka and Vujić, Zorica and Kuntić, Vesna",
year = "2016",
abstract = "One of the most commonly used surfactants in the production of split virus influenza vaccine is nonionic surfactant Triton X-100. After splitting of the virus is accomplished, Triton X-100 is removed from the vaccine by subsequent production steps. Because of toxicity of Triton X-100, which remains in the vaccine in residual amounts, a sufficiently sensitive method for its detection and quantification needs to be defined. Two methods for determination of Triton X-100 residuals were developed: the UV-spectrophotometry and HPLC methods. For both methods, preparation of vaccine samples and removal of proteins and virus particles were crucial: samples were treated with methanol (1:1) and then centrifuged at 25000 x g for 30 min. After such treatment, the majority of vaccine components that interfered in the UV region were removed, and diluted samples could be directly measured. The chromatographic system included C18 column, step methanol gradient, and detection at 225 nm with a single peak of Triton X-100 at 12.6 min. Both methods were validated and gave satisfactory results for accuracy, precision, specificity, linearity, and robustness. LOQ was slightly lower for the HPLC method. Hence, it was shown that both methods are suitable for analysis of residual amounts of Triton X-100, with the advantages of the UV method being its simplicity and availability in most laboratories.",
publisher = "AOAC Int, Gaithersburg",
journal = "Journal of AOAC International",
title = "Direct UV Spectrophotometry and HPLC Determination of Triton X-100 in Split Virus Influenza Vaccine",
volume = "99",
number = "2",
pages = "396-400",
doi = "10.5740/jaoacint.15-0201"
}

Pavlović, B., Cvijetić, N., Dragacević, L., Ivković, B., Vujić, Z.,& Kuntić, V.. (2016). Direct UV Spectrophotometry and HPLC Determination of Triton X-100 in Split Virus Influenza Vaccine. in Journal of AOAC International
AOAC Int, Gaithersburg., 99(2), 396-400.
https://doi.org/10.5740/jaoacint.15-0201

Pavlović B, Cvijetić N, Dragacević L, Ivković B, Vujić Z, Kuntić V. Direct UV Spectrophotometry and HPLC Determination of Triton X-100 in Split Virus Influenza Vaccine. in Journal of AOAC International. 2016;99(2):396-400.
doi:10.5740/jaoacint.15-0201 .

Pavlović, Bojana, Cvijetić, Nataša, Dragacević, Luka, Ivković, Branka, Vujić, Zorica, Kuntić, Vesna, "Direct UV Spectrophotometry and HPLC Determination of Triton X-100 in Split Virus Influenza Vaccine" in Journal of AOAC International, 99, no. 2 (2016):396-400,
https://doi.org/10.5740/jaoacint.15-0201 . .

TY  - JOUR
AU  - Čolić, Miodrag
AU  - Mojsilović, Slavko
AU  - Pavlović, Bojan
AU  - Vučićević, Dragana
AU  - Majstorović, Ivana
AU  - Bufan, Biljana
AU  - Stojić-Vukanić, Zorica
AU  - Vasilijić, Saša
AU  - Vučević, Dragana
AU  - Gašić, Sonja
AU  - Balint, Bela
PY  - 2004
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/547
AB  - Background. Dendritic cells (DC) have been used for immunotherapy of malignant tumors, different kinds of infections, and other clinical conditions. For that purpose, optimal conditions for the generation of functionally mature DC in vitro are required. Two different protocols for the induction of maturation of monocyte-derived DC (MDDC) were compared in this study. Methods. MDDC were generated in vitro by cultivating adherent monocytes of healthy volunteers with granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin 4 (IL-4) during 6-days period. The immature DC thus prepared were induced to mature using two protocols. DC were stimulated for 2 days with lipopolysaccharide (LPS), or with a cocktail of proinflammatory mediators (PM) containing IL-1b, IL-6, tumor necrosis factor α (TNFα), and prostaglandin E2 (PGE2), respectively. Phenotypic characteristics of MDDC and their endocytic activity were studied by flow cytometry. Allostimulatory activity of these cells was tested in the mixed leukocyte reaction (MLR), whereas the production of cytokines was determined by ELISA kits. Results. MDDC matured with PM (PM-DC) were predominantly non-adherent cells, while about 30% of LPS-matured DC were adherent cells. In comparison with LPS-DC, PM-DC expressed higher levels of CD86 and CD83, had lower endocytic activity, produced higher levels of IL-10 and lower levels of IL-12, and more strongly stimulated proliferation of allogeneic lymphocytes. Conclusion The protocol based on the combination of proinflammatory cytokines and PGE2 is better for the induction of maturation of human MDDC in vitro than the protocol using LPS alone.
AB  - Uvod. Dendritične ćelije (DC) se koriste u imunoterapiji malignih tumora, različitih vrsta infekcija i drugih oboljenja. U tom cilju neophodni su optimalni uslovi za dobijanje funkcionalno zrelih DC in vitro. U ovom radu smo poredili dva različita protokola za indukciju maturacije DC monocitnog porekla (MDDC). Metode. MDDC su dobijene in vitro kultivisanjem adherentnih monocita zdravih dobrovoljnih davalaca krvi pomoću faktora stimulacije granulocitno-makrofagnih kolonija (GM-CSF) i interleukina-4 (IL-4) u toku 6 dana. Tako pripremljene nezrele DC su indukovane na sazrevanje pomoću dva protokola. DC su stimulisane u toku 2 dana lipopolisaharidom (LPS) ili koktelom proinflamatornih medijatora (PM) koji je sadržavao IL-1b, IL-6, faktor nekroze tumora α (TNFα) i prostaglandin E2 (PGE2). Fenotipske karakteristike MDDC i njihova endocitozna aktivnost su ispitivani pomoću protočne citometrije. Alostimulatorna aktivnost ovih ćelija je ispitivana u testu mešane leukocitne reakcije (MLR), dok je stvaranje citokina određivano ELISA kompletima. Rezultati. MDDC koje su sazrevale u prisustvu PM (PM-DC) su bile predominantno neadherentne ćelije, dok su oko 30% DC koje su sazrevale pod uticajem LPS (LPS-DC) bile adherentne. U poređenju sa LPS-DC, PM-DC su ispoljavale više nivoe CD86 i CD83 molekula, imale slabiju endocitoznu aktivnost, produkovale više IL-10, a manje IL-12 i snažnije stimulisale proliferaciju alogenih limfocita. Zaključak. Protokol koji se bazira na primeni kombinacije proinflamatornih citokina i PGE2 je bolji za indukciju maturacije humanih MDDC in vitro nego LPS protokol.
PB  - Vojnomedicinska akademija - Institut za naučne informacije, Beograd
T2  - Vojnosanitetski pregled
T1  - Comparison of two different protocols for the induction of maturation of human dendritic cells in vitro
T1  - Poređenje dva različita protokola za indukciju maturacije humanih dendritičnih ćelija in vitro
VL  - 61
IS  - 5
SP  - 471
EP  - 478
DO  - 10.2298/VSP0405471C
ER  - 

@article{
author = "Čolić, Miodrag and Mojsilović, Slavko and Pavlović, Bojan and Vučićević, Dragana and Majstorović, Ivana and Bufan, Biljana and Stojić-Vukanić, Zorica and Vasilijić, Saša and Vučević, Dragana and Gašić, Sonja and Balint, Bela",
year = "2004",
abstract = "Background. Dendritic cells (DC) have been used for immunotherapy of malignant tumors, different kinds of infections, and other clinical conditions. For that purpose, optimal conditions for the generation of functionally mature DC in vitro are required. Two different protocols for the induction of maturation of monocyte-derived DC (MDDC) were compared in this study. Methods. MDDC were generated in vitro by cultivating adherent monocytes of healthy volunteers with granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin 4 (IL-4) during 6-days period. The immature DC thus prepared were induced to mature using two protocols. DC were stimulated for 2 days with lipopolysaccharide (LPS), or with a cocktail of proinflammatory mediators (PM) containing IL-1b, IL-6, tumor necrosis factor α (TNFα), and prostaglandin E2 (PGE2), respectively. Phenotypic characteristics of MDDC and their endocytic activity were studied by flow cytometry. Allostimulatory activity of these cells was tested in the mixed leukocyte reaction (MLR), whereas the production of cytokines was determined by ELISA kits. Results. MDDC matured with PM (PM-DC) were predominantly non-adherent cells, while about 30% of LPS-matured DC were adherent cells. In comparison with LPS-DC, PM-DC expressed higher levels of CD86 and CD83, had lower endocytic activity, produced higher levels of IL-10 and lower levels of IL-12, and more strongly stimulated proliferation of allogeneic lymphocytes. Conclusion The protocol based on the combination of proinflammatory cytokines and PGE2 is better for the induction of maturation of human MDDC in vitro than the protocol using LPS alone., Uvod. Dendritične ćelije (DC) se koriste u imunoterapiji malignih tumora, različitih vrsta infekcija i drugih oboljenja. U tom cilju neophodni su optimalni uslovi za dobijanje funkcionalno zrelih DC in vitro. U ovom radu smo poredili dva različita protokola za indukciju maturacije DC monocitnog porekla (MDDC). Metode. MDDC su dobijene in vitro kultivisanjem adherentnih monocita zdravih dobrovoljnih davalaca krvi pomoću faktora stimulacije granulocitno-makrofagnih kolonija (GM-CSF) i interleukina-4 (IL-4) u toku 6 dana. Tako pripremljene nezrele DC su indukovane na sazrevanje pomoću dva protokola. DC su stimulisane u toku 2 dana lipopolisaharidom (LPS) ili koktelom proinflamatornih medijatora (PM) koji je sadržavao IL-1b, IL-6, faktor nekroze tumora α (TNFα) i prostaglandin E2 (PGE2). Fenotipske karakteristike MDDC i njihova endocitozna aktivnost su ispitivani pomoću protočne citometrije. Alostimulatorna aktivnost ovih ćelija je ispitivana u testu mešane leukocitne reakcije (MLR), dok je stvaranje citokina određivano ELISA kompletima. Rezultati. MDDC koje su sazrevale u prisustvu PM (PM-DC) su bile predominantno neadherentne ćelije, dok su oko 30% DC koje su sazrevale pod uticajem LPS (LPS-DC) bile adherentne. U poređenju sa LPS-DC, PM-DC su ispoljavale više nivoe CD86 i CD83 molekula, imale slabiju endocitoznu aktivnost, produkovale više IL-10, a manje IL-12 i snažnije stimulisale proliferaciju alogenih limfocita. Zaključak. Protokol koji se bazira na primeni kombinacije proinflamatornih citokina i PGE2 je bolji za indukciju maturacije humanih MDDC in vitro nego LPS protokol.",
publisher = "Vojnomedicinska akademija - Institut za naučne informacije, Beograd",
journal = "Vojnosanitetski pregled",
title = "Comparison of two different protocols for the induction of maturation of human dendritic cells in vitro, Poređenje dva različita protokola za indukciju maturacije humanih dendritičnih ćelija in vitro",
volume = "61",
number = "5",
pages = "471-478",
doi = "10.2298/VSP0405471C"
}

Čolić, M., Mojsilović, S., Pavlović, B., Vučićević, D., Majstorović, I., Bufan, B., Stojić-Vukanić, Z., Vasilijić, S., Vučević, D., Gašić, S.,& Balint, B.. (2004). Comparison of two different protocols for the induction of maturation of human dendritic cells in vitro. in Vojnosanitetski pregled
Vojnomedicinska akademija - Institut za naučne informacije, Beograd., 61(5), 471-478.
https://doi.org/10.2298/VSP0405471C

Čolić M, Mojsilović S, Pavlović B, Vučićević D, Majstorović I, Bufan B, Stojić-Vukanić Z, Vasilijić S, Vučević D, Gašić S, Balint B. Comparison of two different protocols for the induction of maturation of human dendritic cells in vitro. in Vojnosanitetski pregled. 2004;61(5):471-478.
doi:10.2298/VSP0405471C .

Čolić, Miodrag, Mojsilović, Slavko, Pavlović, Bojan, Vučićević, Dragana, Majstorović, Ivana, Bufan, Biljana, Stojić-Vukanić, Zorica, Vasilijić, Saša, Vučević, Dragana, Gašić, Sonja, Balint, Bela, "Comparison of two different protocols for the induction of maturation of human dendritic cells in vitro" in Vojnosanitetski pregled, 61, no. 5 (2004):471-478,
https://doi.org/10.2298/VSP0405471C . .