Effects of oil-based adjuvants on the immune response of pigs after dermal administration of antigen and evaluation of the immunization level after a subsequent Actinobacillus pleuropneumoniae challenge in pigs.

Route of vaccine delivery can greatly impact the immunogenicity, efficacy and safety of the vaccine. Four groups of piglets were immunised transdermally (t.d.), intradermally (i.d.) or intramuscularly (i.m.) with the same doses of antigen in combination with a water-in-oil-in-water emulsion adjuvant Montanide™ ISA 201 VG or with a microemulsion adjuvant Montanide™ IMS 1313 VG N ST (Seppic, France). The last group was left without vaccination as a control group. All animals were subsequently exposed to the infection induced by Actinobacillus pleuropneumoniae (App). The immune response was evaluated with respect to the intensity of systemic and mucosal antibody formation, their isotype characterisation and rate of cell-mediated immunity. These findings were compared with the intensity of adverse local reactions and level of protection in experimental challenge. Monitoring of the local reaction at the injection site after each administration showed that microemulsion adjuvant IMS 1313 was less reactogenic than the water-in-oil-in-water emulsion ISA 201. In terms of efficacy, both dermal administrations were less immunogenic than the i.m route. The i.m. injection induced higher anti-App9 IgG and IgM titres. Nevertheless, IgG1 and IgG2 isotypes analysis revealed a close immunological profile between i.m. and i.d. routes. The concentration of IFN-γ from peripheral blood after in vitro restimulation with the specific antigen was only increased in the i.m. group at the day of challenge (D35) and two weeks after (D49). Interestingly, the smallest gross pulmonary lesions were observed in the i.d. vaccinated group (3.4%) compared to the control group (39.4%) and to groups with other routes of administration. Taken together, these results suggest that i.d. administration of vaccines is a promising approach. Even the i.d. vaccine was more reactogenic and slightly less immunogenic than the i.m. vaccine, its protection effectiveness seemed to be superior.

Effects of IFNγ and IL4 rich microenvironment on porcine monocyte-derived dendritic cell activation in vitro.

Dendritic cells (DCs) represent a heterogeneous group of major antigen-presenting cells, responding to different stimuli in their microenvironment. They are able to activate naïve T-cells and drive their polarization towards effector types. However, the effect of different Th-polarizing cytokine microenvironment on porcine DCs remains poorly described. Therefore, the effect of IFNγ or IL4 rich microenvironment on porcine monocyte-derived dendritic cells and their activation, antigen recognition and cytokine secretion towards T-cell polarization were studied in vitro. IFNγ-rich microenvironment induced a higher proinflammatory response and release of Th1/Th17-polarizing cytokines (IL1ß, IL12p35, IL23p19), while anti-inflammatory properties (IL10, TGFß) was not affected by a cytokine microenvironment. From the achieved results, we propose that different cytokine microenvironment has the potential to modulate dendritic cells and their ability to activate T-cells.

Aspects of intradermal immunization with different adjuvants: The role of dendritic cells and Th1/Th2 response.

Intradermal (i.d.) application of vaccine is promising way how to induce specific immune response against particular pathogens. Adjuvants, substances added into vaccination dose with the aim to increase immunogenicity, play important role in activation of dendritic cells with subsequent activation of lymphocytes. They can, however, induce unwanted local reactions. The aim of the study was to determine the effect of i.d. administration of model antigen keyhole limped hemocyanine alone or with different adjuvants-aluminium hydroxide and oil-based adjuvants-on local histopathological reaction as well as dendritic cell activation at the site of administration and local cytokine and chemokine response. This was assessed at 4 and 24 hours after application. Selection of the adjuvants was based on the fact, that they differently enhance antibody or cell-mediated immunity. The results showed activation of dendritic cells and both Th1 and Th2 response stimulated by oil-based adjuvants. It was associated with higher expression of set of genes, incl. chemokine receptor CCR7 or Th1-associated chemokine CXCL10 and cytokine IFNγ. Application of the antigen with aluminium hydroxide induced higher expression of Th2-associated IL4 or IL13. On the other hand, both complete and incomplete Freund´s adjuvants provoked strong local reaction associated with influx of neutrophils. This was accompanied with high expression of proinflammatory IL1 or neutrophil chemoattractant CXCL8. Surprisingly, similarly strong local reaction was detected also after application of aluminium hydroxide-based adjuvant. The best balanced local reaction with sufficient activation of immune cells was detected after application of oil-based adjuvants Montanide and Emulsigen.

Oil-based adjuvants delivered intradermally induce high primary IgG2 immune response in swine.

The effects of intradermal application of antigen with or without different adjuvants and activation of immune response are presented in this study. Six groups of six piglets each were immunized with keyhole limpet haemocyanin (KLH) antigen in combination with aluminium hydroxide or oil-based adjuvants (complete and incomplete Freund's adjuvants, Montanide ISA 206 and Emulsigen). IgG1 and IgG2 levels in sera were measured by KLH-specific ELISA. Interestingly, oil-based adjuvants induced high primary IgG2 response, suggesting the Th1 lymphocyte polarization. Also, considering the similarities between human and porcine organism, we suggest that intradermal application could be considered as an effective vaccine delivery route in both veterinary and human medicine.

Treatment of postoperative pain after total hip arthroplasty: comparison between metamizol and paracetamol as adjunctive to opioid analgesics-prospective, double-blind, randomised study.

INTRODUCTION: Metamizole use has been limited because of its risk of agranulocytosis. However, more recent literature seems to support its safety. This prospective, randomised, double-blind study was conducted to compare the analgesic effects of intravenous metamizole or intravenous paracetamol in combination with morphine PCA during the first 24 h following total hip arthroplasty. MATERIALS AND METHODS: One hundred ten consecutive patients were selected for study. The two study groups were (A) metamizole, (B) paracetamol. Postoperative pain therapy was provided by Morphine PCA pump. In the first treatment group (A group), all patients received intravenous metamizole 1.5 g every 8 h during the first 24 postoperative hours. In the second treatment group (B group), all patients received intravenous paracetamol 1 g every 8 h during the first 24 postoperative hours. Postoperative pain intensity was measured 1, 2, 3, 4, 6, 8, 10, 14, 18, 22 h after the end of surgery by a VAS. RESULTS: Statistically significant differences in VAS pain values favoring metamizole were reported at 6-h (p = 0.038), 8-h (p = 0.036), 14-h (p = 0.011), 18-h (p < 0.001) and 22-h (p = 0.025) post-baseline. Mean cumulative pain values were 17.9 for metamizole and 30.6 for paracetamol. CONCLUSIONS: In this study, we have also shown excellent efficacy of paracetamol and metamizole combined with opioids, but metamizole proved to be a better analgesic than paracetamol. It is also necessary to mention the financial aspect considering that intravenous paracetamol is about ten times more expensive than an equivalent analgesic doses of intravenous metamizole.

[Characteristics of anesthesia in patients with osteogenesis imperfecta undergoing orthopedic surgical procedures]. / Osobitosti anestezije kod ortopedskih zahvata u bolesnika s osteogenesis imperfect.

The aim is to show our experience in anesthesia of patients with osteogenesis imperfecta (OI) who have undergone orthopedic surgical procedures. This is a retrospective analysis of OI patients treated at our Department from 1980 to 2012. We analyzed demographics, comorbidities, preoperative characteristics, anesthesia types, anesthetics and intraoperative and postoperative complications. In the given period, 26 OI patients were treated, using 103 surgeries, and 103 anesthesia procedures. Most procedures, a total of 68, were used in children aged 0-10 years. According to the diagnosis, OI type III was mostly encountered. The rating of the American Society of Anesthesiologist (ASA) physical status was II in most cases, a total of 99. General anesthesia was used in 89 cases, and regional anesthesia in 14. Fourteen intraoperative complications were seen, mostly difficult intubation, and six postoperative cardiovascular instability cases. With careful preparation, and knowledge of pitfalls, anesthesia in these patients should be a safe procedure.