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Immune Responses Induced by chlamydial Polymorphic Membrane Protein C formulated with probiotic Lactobacillus rhamnosus as an Adjuvant in Conjunctival Immunization
No full textPurpose : Our strategy in designing an efficient vaccine against Chlamydia trachomatis (Ct) relies on the assumptions that the subunit vaccine is based on proteins involved in the very first contact of Ct and host-cell, the chosen route via the ocular mucosa mimics the natural infection, and its application is needle-free and safe. We evaluated the immunogenicity of N-terminal portion of Ct polymorphic membrane protein C, involved in Ct-host cell interaction, when applied via the ocular mucosa; and examined the adjuvantic influence of Lactobacillus rahmnosus (LB) on the characteristics of the promoted immune response in a BALB/c mouse model system.
Methods : Mice were immunized via the conjunctiva with N-PmpC alone (N-PmpC/PBS), and N-PmpC mixed LB (N-PmpC/1x106 CFU/ml/PBS). Concentration of N-PmpC in all vaccines prepared for the conjunctival administration was 1.5 mg/ml and each mouse was immunized with 15 mg N-PmpC in 10 ml (5 ml per eye administered). Detection of PmpC-specific and CtB-specific immunoglobulins in mouse sera and tears was done by the ELISA. Phenotyping of lymphocytes was done by flow cytometry. The statistical significance of the observed differences was evaluated using 1-Way Repeated Measures ANOVA. A probability (P) value of 0.05 was set as a limit of significance (software: ORIGIN 8.0).
Results : Ocular immunization with N-PmpC alone and in combination with LB was well tolerated in all animals. No signs of toxicity and pathology were observed. N-PmpC applied via the conjunctiva in combination with LB stimulated the production of specific IgA (p<0.05) and IgG (p<0.005) at both local and systemic level that was significantly enhanced in comparison with N-PmpC alone. In addition, N-PmpC mixed with LB initiated a T cell response characterized by the Th1 skewing and a rise of the effector CD8+ T cells and Tregs (p<0.05).
Conclusions : Conjunctival application of a chlamydial specific subunit antigen combined with a particulated carrier induced significant humoral and cellular immune responses. Further studies are needed to optimize relevant antigen and adjuvant combinations
Burkholderia cepacia complex in Serbian patients with cystic fibrosis: prevalence and molecular epidemiology
No full textThe Burkholderia cepacia complex (Bcc) organisms remain significant pathogens in patients with cystic fibrosis (CF). This study was performed to evaluate the prevalence, epidemiological characteristics, and presence of molecular markers associated with virulence and transmissibility of the Bcc strains in the National CF Centre in Belgrade, Serbia. The Bcc isolates collected during the four-year study period (2010-2013) were further examined by 16 s rRNA gene, pulsed-field gel electrophoresis of genomic DNA, multilocus sequence typing analysis, and phylogenetic analysis based on concatenated sequence of seven alleles. Fifty out of 184 patients (27.2 %) were colonized with two Bcc species, B. cenocepacia (n = 49) and B. stabilis (n = 1). Thirty-four patients (18.5 %) had chronic colonization. Typing methods revealed a high level of similarity among Bcc isolates, indicating a person-to-person transmission or acquisition from a common source. New sequence types (STs) were identified, and none of the STs with an international distribution were found. One centre-specific ST, B. cenocepacia ST856, was highly dominant and shared by 48/50 (96 %) patients colonized by Bcc. This clone was characterized by PCR positivity for both the B. cepacia epidemic strain marker and cable pilin, and showed close genetic relatedness to the epidemic strain CZ1 (ST32). These results indicate that the impact of Bcc on airway colonization in the Serbian CF population is high and virtually exclusively limited to a single clone of B. cenocepacia. The presence of a highly transmissible clone and probable patient-to-patient spread was observed
Smallpox as an actual biothreat: lessons learned from its outbreak in ex-Yugoslavia in 1972
Get PDFVariola (smallpox) virus is classified as class A of potential biological weapons, due to its microbiological, genetic, antigenic and epidemiological characteristics. The potential danger is more real because vaccination against smallpox has stopped since disease eradication in 1979. That is why we want to share our unique, rich experience and acquired knowledge in the fight against this highly contagious and deadly disease during the smallpox outbreak in ex-Yugoslavia in 1972. It was the largest postwar outbreak in Europe when there were officially registered 175 ill patients, 35 of them with lethal outcome. This outbreak was specific by the time of its occurrence, the affected territory, dimensions and some epidemiological characteristics, but also by the well-organized, synchronized and efficient reaction of the competent state services in the fight against it
Starenje utiče na M1/M2 polarizaciju rezidentnih peritonsalnih makrofaga pacova stimulisanih In vitro
No full textCilj rada bio je da se ispita uticaj starenja na aktivnost makrofaga aktivisanih M1/M2
polarizujućim faktorima. Peritonealne rezidentne makrofage (rMf) i tioglikolatom“
indukovane makrofage (pMf) mladih (3 mesoca) im starih (18-19 meseci) pocova su
kultivisane u prisustvu stimulatora klasične (M1) - lipopolisaharida (LPS) i faktora
stimulacije kolonija granulocita i makrofaga (GM-CSF), i alternativis (M2) aktivacije
makrofaga - ingerleukina-4 (IL-4), ili u odsustvu poznatih stimulatora. Ispitivana je
sposobnost fagocitoze zimozana i sekrecije inflamatornih medijlora. Starsnjem se
povećavala učestalost makrofaga monoci:nog porekla (SSK7'S068' ćelije) u okviru
populacije rMf, dok je u okviru populacije tMf nađena povećana učestalost makrofaga
najzrelijeg fenotipa (CD163'°CD68" ćelije). Nijedan od ispitivanih stimulatora nije uticao
na fagocitnu sposobnost rMf i ztimf Povećana sekrecija IL-1D, 1:-6 i #10 i smanjena
sekrecija IGF-D u odgovoru na stimulaciju LPS-om je nađena kod rMf i Md pacova obe
starosti. GM-CSF je povećao sekreciju IL-1B i IL-6 kod rMf starih pacova i tMf mladih
pacova. Paradoksalno, IL-4 je povećao sekreciju pro-inflamatornih citokina, IL-16 i IL-6, kod
rMf starih pacova. Starenjem se metabolizam arginina i u rMf i u tgMf usmeravao ka
sintezi ypee LPS je, za razliku od GM-CSF, favorizovao sintezu azot monoksida u rMf i pMf
pacova obe starosti. Nasuprot tome, kod rRMf i pMf mladih pacova IL-4 je usmerio
metabolizam arginina prema sintezi uree. Rezultati su pokazali da sa starenjem nM gube
sposobnost polarizacije pod uticajem GM-CSF, i da rMf pod uticajom IL-4 polarizuju prema
pro-inflamatornom Mi sekretornom fenotipu. Gubitak kontrole sekrecije inflamatornih
medijatora iz makrofaga u odgovoru na M1/M2 aktivatore mogao bi da doprinese povećanom
riziku od oboljevanja od infektivnih i inflamatornih bolesti u starosti
Direct UV Spectrophotometry and HPLC Determination of Triton X-100 in Split Virus Influenza Vaccine
No full textOne 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
Ontogeny of pileus shape in Natrix natrix and N. tessellata
No full textWe examined the divergence in pileus shape and ontogenetic allometric changes in pileus shape in two closely related species of snakes - Natrix natrix and N. tessellata. These two species have similar ecology, but different microhabitat and diet preferences. We analysed the ontogenetic series from hatchlings to adults. Hatchlings of two species significantly diverged in pileus size and shape. At the adult stage, significant sexual dimorphism in pileus size and shape was found within both species. Adults of N. natrix and N. tessellata (females and males) diverged in pileus shape, but not in size. Allometric shape changes accounted for significant amount of variance in pileus shape among different age classes, and appear to be species- and sex-specific. Our results suggested that allometries of pileus shape are highly evolvable traits, which may result from natural selection for functional optimisation of head shape in Natricine species
Lactobacillus Helveticus L10 Supplementation Modulates Immunity Parameters in Elite Athletes
No full textAging affects the responsiveness of rat peritoneal macrophages to GM-CSF and IL-4
No full textMacrophages undergo significant functional alterations during aging. The aim of the present study was to investigate changes of rat macrophage functions and response to M1/M2 polarization signals with age. Therefore, resident and thioglycollate-elicited peritoneal macrophages from young (3-month-old) and aged (18-19-month-old) rats were tested for phagocytic capacity and ability to secrete inflammatory mediators following in vitro stimulation with LPS and GM-CSF, and IL-4, prototypic stimulators for classically (M1) and alternatively activated (M2) macrophages, respectively. Aging increased the frequency of monocyte-derived (CCR7+ CD68+) and the most mature (CD163+ CD68+) macrophages within resident and thioglycollate-elicited peritoneal macrophages, respectively. The ability to phagocyte zymosan of none of these two cell subsets was affected by either LPS and GM-CSF or IL-4. The upregulated production of IL-1 beta, IL-6 and IL-10 and downregulated that of TGF-beta was observed in response to LPS in resident and thioglycollate-elicited macrophages from rats of both ages. GM-CSF elevated production of IL-1 beta and IL-6 in resident macrophages from aged rats and in thioglycollate-elicited macrophages from young rats. Unexpectedly, IL-4 augmented production of proinflammatory mediators, IL-1 beta and IL-6, in resident macrophages from aged rats. In both resident and thioglycollate-elicited macrophages aging decreased NO/urea ratio, whereas LPS but not GM-SCF, shifted this ratio toward NO in the macrophages from animals of both ages. Conversely, IL-4 reduced NO/urea ratio in resident and thioglycollate-elicited macrophages from young rats only. In conclusion, our study showed that aging diminished GM-CSF-triggered polarization of elicited macrophages and caused paradoxical IL-4-driven polarization of resident macrophages toward proinflammatory M1 phenotype. This age-related deregulation of macrophage inflammatory mediator secretion and phagocytosis in response to M1/M2 activators may lead to the deficient control of infectious and/or inflammatory diseases in advanced age
Lignin model compound in alginate hydrogel: a strong antimicrobial agent with high potential in wound treatment
No full textNowadays bacterial resistance to known antibiotics is a serious health problem. In order to achieve more efficient treatment, lately there is an effort to find new substances, such as certain biomaterials, that are non-toxic to humans with antibiotic potential. Lignins and lignin-derived compounds have been proposed to be good candidates for use in medicine and health maintenance. In this study, the antibacterial activity of the lignin model polymer dehydrogenate polymer (DHP) in alginate hydrogel (Alg) was studied. The obtained results show that DHP-Alg has strong antimicrobial activity against several bacterial strains and biofilms and does not have a toxic effect on human epithelial cells. These results strongly suggest its application as a wound healing agent or as an adjunct substance for wound treatments. (C) 2016 Elsevier B.V. and International Society of Chemotherapy. All rights reserved
Peripubertal ovariectomy influences thymic adrenergic network plasticity in adult rats
No full textThe study investigated the influence of peripubertal ovariectomy on the thymic noradrenaline (NA) concentration, and the thymocyte NA content and beta(2)- and alpha(1)-adrenoceptor (AR) expression in adult 2- and 11-month-old rats. In control rats, the thymic NA concentration increased with age. This increase reflected rise in the density of catecholamine (CA)-containing fluorescent nerve fibers and cells and their CA content. Additionally, the average beta(2)- and alpha(1)-AR thymocyte surface density changed in the opposite direction with age; the density of beta(2)-AR decreased, whereas that of alpha(1)-AR increased. Ovariectomy diminished the thymic NA concentration in 2-month-old rats. This reflected the decrease in the density of fluorescent nerve fibers, and CA content in fluorescent nerve fibers and non-lymphoid cells, since the thymocyte NA content was increased in ovariectomized (Ox) rats. Estrogen supplementation prevented the ovariectomy-induced changes. In Ox rats, the density of CA-synthesizing nerve fibers and non-lymphoid cells diminished with age. To the contrary, NA content in thymocytes increased with age, but it did not exceed that in 11-month-old controls. Additionally, ovariectomy diminished the average thymocyte surface density of beta(2)-ARs, but it increased that of alpha(1)-ARs in 2-month-old-rats (due to estrogen, and estrogen and progesterone deficiency, respectively). These changes, despite of the rise in circulating estrogen level post-ovariectomy, remained stable with age. This most likely reflected a decreased sensitivity to estrogen action, as a consequence of the hormone misprinting in peripubertal age. The analysis of thymocyte proliferation in culture suggested that age-and ovariectomy-induced alterations in thymocyte NA synthesis and AR expression altered NA autocrine/paracrine action on thymocytes. In conclusion, the study indicates that the ovarian hormone deficiency in peripubertal age affects ovarian steroid-dependent remodeling of thymic adrenergic regulatory network in adult rats. (C) 2016 Elsevier B.V. All rights reserved