Comparison of flagellin and an oil-emulsion adjuvant in inactivated Newcastle disease vaccine in stimulation of immunogenic parameters

The present study was designed to investigate the potential application of native (N) and recombinant (truncated modified [tmFliC] and full-length [flFliC]) flagellin proteins along with inactivated Newcastle disease virus (NDV). Fifty six SPF chickens were immunized twice with PBS (control), inactivated NDV (Ag), inactivated NDV/flFliC (AgF), inactivated NDV/tmFliC (AgT), inactivated NDV/N (AgN), commercial vaccine containing Montanide (Vac) and Vac/N (VacN), with a two-week interval. Blood was collected weekly and spleens were harvested after chickens were sacrificed. Interleukin-6 (IL-6) and tumor necrotic factor-α (TNF-α) gene expression in peripheral blood mononuclear cells were analyzed by Real-Time PCR. Antibody response was assessed by haemagglutination inhibition (HI). Cellular activity was quantified by MTT assay. Results showed that the most IL-6 and TNF-α gene expression was observed in AgF group (P < 0.01). The lowest gene expression among vaccinated groups was observed in Ag group for IL-6 and Ag and Vac group for TNF-α. The highest HI titer was observed in Vac, VacN, AgF and AgT groups. The AgF group showed the highest cellular activity (P < 0.01). In conclusion, flagellin-adjuvanted groups showed a pro-inflammatory effect and acted similarly to or better than the Vac group. Hence, flagellin can be proposed as a potential adjuvant for ND vaccine.     

New approaches in vaccine development

In the last century, vaccines have been one of the most powerful tools for preventing infectious diseases. Smallpox has been eradicated and other diseases such as poliomyelitis or measles have been reduced to very low levels in many regions of the world. However, infectious diseases remain the leading cause of death worldwide. Thus, the development of vaccines to prevent diseases for which no vaccine currently exists such as AIDS or malaria as well as the improvement of efficacy and safety of existing vaccines remains a high priority. Achieving such ambitious goals in a near future will certainly require a strong modification of the methods that have been used so far to identify vaccine candidates. In particular, modern vaccinology could strongly benefit of the latest developments of molecular biology and immunology. Here, we will discuss some potential applications of the increasing knowledge of pathogen genomes as well as the immune system for the discovery of new antigenic targets and the development of new strategies of vaccination.     

Protective efficacy of a live attenuated Mycoplasma hyopneumoniae vaccine with an ISCOM-matrix adjuvant in pigs

An attenuated Mycoplasma hyopneumoniae vaccine that requires intrathoracic administration is commercially available for use against mycoplasmal pneumonia in China. Given the limitations of such a route of administration, this study was undertaken to assess the capacity of an ISCOM-matrix adjuvant to enhance immunogenicity following intramuscular use. Immune responses in pigs following vaccination and subsequent intra-tracheal bacterial inoculation were examined using lymphocyte proliferation, serology and mucosal IgA in both nasal and saliva swabs.Vaccination induced clear lymphocyte proliferation, but only slight serum antibody responses although these were significantly increased following experimental infection. Mucosal IgA was not detected in either nasal or salivary secretions. Following bacterial challenge, animals vaccinated with the adjuvant-containing live vaccine exhibited less severe pulmonary lesions (median score 3.67) than unvaccinated pigs (median score 13.58). The degree of ciliary loss on the respiratory tract surface was reduced in vaccinated pigs compared with experimentally infected controls. The findings indicated that the adjuvant vaccine administered IM provided protection against experimentally induced mycoplasmal pneumonia and could have commercial potential.     

助剂对高效氯氰菊酯在甘蓝叶片表皮渗透性的影响

 研究了3类助剂（矿物油、高级脂肪酸、非离子表面活性剂）对非内吸性杀虫剂高效氯氰菊酯在甘蓝叶片表皮渗透性的影响,证明药液的渗透性在一定范围内与药剂中助剂含量和给药后的时间成正相关。助剂间一定比例的配合对该渗透性具有协同作用。通过添加助剂后药液的各种物理性状进行测试及助剂溶液改变甘蓝表面蜡质结构的电镜观察,初步讨论了各类助剂影响渗透性的作用机理。     

Immune response of BALB/c mouse immunized with recombinant MSPs proteins of Anaplasma marginale binding to immunostimulant complex (ISCOM)

Anaplasmosis, caused by Anaplasma marginale, results in significant economic losses of cattle in tropical and subtropical regions worldwide. Six major surface proteins (MSPs) were well characterized and designated as MSP1, MSP2, MSP3, MSP4, and MSP5. The objective of this study was to evaluate the humoral immune response of BALB/c mice against the recombinant MSPs, incorporated into immunostimulating complex (ISCOM). The recombinant proteins purified by Ni-NTA columns were incorporated into ISCOM and ISCOMATRIX by the lipid film hydration method. BALB/c mice immunized with ISCOM/rMSPs and ISCOMATRIX/rMSPs vaccines produced whole IgG, IgG1, and IgG2a, in contrast to the negative groups (PBS and ISCOMATRIX adjuvant). All groups that received antigen responded specifically against the rMSPs by Western blotting, showing the rMSP1a (60-105kDa), rMSP1b (100kDa), rMSP4 (47kDa), and rMSP5 (29kDa). Additional studies will have to be performed in cattle to evaluate the humoral and cellular mechanisms of this subunit vaccine and their possible use as protective vaccines against homologous and heterologous strains of A. marginale.     

淫羊藿—蜂胶佐剂对雏鸡免疫器官早期发育和形态结构的影响

３日龄雏鸡按两个剂量组分别于皮下注射淫羊藿－蜂胶佐剂（简称淫蜂佐剂）０．２ｍＬ和０．４ｍＬ,２４日龄时重复注射１次,于７,１４,２１,２８,３５,４２日龄时随机抽样扑杀,采取免疫器官称重并进行光镜和电镜检查,比较不同剂量的淫蜂佐剂对雏鸡免疫器官的增重以及形态结构的影响。结果表明,淫蜂佐剂能显著促进雏鸡免疫器官的早期发育,并能刺激Ｔ,Ｂ淋巴细胞活化。重复注射后,可再次引起脾脏增重,且与剂量有相关性,     

Immunoadjuvant effects of bacterial genomic DNA and CpG oligodeoxynucleotides on avian influenza virus subtype H5N1 inactivated oil emulsion vaccine in chicken

This study investigated the immunoadjuvant effects of three types of bacterial genomic DNA and CpG oligonucleotides (CpG ODN) on the avian influenza virus (AIV) subtype H5N1 inactivated oil emulsion vaccine under two immunization strategies. The genomic DNA extracted from Escherichia coli O₂, Staphylococcus aureus, Streptococcus faecalis FQ68, and synthetic CpG ODN were used as adjuvants, and their effects on the AIV oil emulsion vaccine were examined in chickens. The results indicated that when administered separately from the vaccine, adjuvants induced lower haemagglutination inhibition (HI) titres and serum IgG titres but resulted in higher concentrations of IFN-γ and IL-10. In contrast, when combined with the oil emulsion vaccine prior to inoculation, CpG ODN induced higher HI, IgG titres and IFN-γ concentration but resulted in lower IL-10 concentration. These data suggest that, depending on the immunization approaches, adjuvants may exert distinct immune effects in chickens receiving AIV H5N1 oil emulsion vaccine: the prior incorporation of CpG ODN into the vaccine may augment both the humoral and Th1 type immune responses, while separate inoculation of adjuvants has not shown better adjuvanticity.     

Effects of adjuvants on deposition efficiency of fenhexamid sprays applied to Chardonnay grapevine foliage

Adequate spray deposition on susceptible grapevine tissue is an essential requirement for effective chemical control of economically important diseases, such as grey mould, powdery mildew and downy mildew. The objective of this study was to evaluate the potential of some agricultural adjuvants to improve foliar spray deposition. Deposition quantity and quality was assessed by means of a spray assessment protocol using fluorometry, photomicrography and digital image analyses. The visual droplet rating technique developed by G Furness, Australia, was also included in initial assessments. Both assessment protocols showed that spray deposition quantity increased with increasing spray volume applications of 27 l ha⁻¹ to 581 l ha⁻¹ with a motorised backpack mistblower, but decreased at 698 l ha⁻¹, possibly due to run-off. Addition of selected spray adjuvants at 526 l ha⁻¹ volume demonstrated improved deposition quantity and quality. Agral 90 (ethoxylated alkylphenol), BB5 (acidifier), Nu-film-P (terpene oil), and Solitaire (silicone/plant oil) significantly improved deposition on upper and lower leaf surfaces compared with the fenhexamid-only and water sprayed control. Break-thru S240 (organosilicone) and Villa 51 (alkylpolyethylene glycol ether) did not improve deposition quantity, although remarkably better deposition quality was obtained. An adjuvant concentration effect (within the registered concentration range) was evident at spray volumes of 502 l ha⁻¹, especially those retained on the upper leaf surfaces. Agral 90 and Nu-film-P effected significant improvement of spray deposition at the higher concentrations (18 ml and 50 ml hl⁻¹, respectively), but not at the lower concentrations (6 ml and 20 ml hl⁻¹, respectively). Solitaire improved deposition at the lower concentration tested (50 ml hl⁻¹), whereas reduced deposition at the higher concentration (100 ml per hl⁻¹) was attributed to excessive spray run-off. No significant improvement of spray deposition was observed for both concentrations tested with Villa 51 (50 and 100 ml hl⁻¹). Spray mixtures with adjuvants Agral 90 and Solitaire yielded similar deposition values at 500 l ha⁻¹ compared with the fenhexamid-only control at 720 l ha⁻¹, but reduced deposition at the higher spray volume, possibly due to spray run-off. This study clearly demonstrated the potential of adjuvants to improve deposition quantity and quality, but highlights the necessity to match adjuvant concentrations and application volumes on the spray target to achieve maximum spray deposition.     

副猪嗜血杆菌灭活疫苗佐剂的筛选

为了评价铝胶、蜂胶及白油佐剂对副猪嗜血杆菌的免疫增强效果,我们将铝胶、蜂胶和白油3种佐剂分别与副猪嗜血杆菌血清5型菌株联合制备灭活疫苗,然后接种雄性成年家兔,采用ELSIA方法分别检测3种灭活苗在免疫期间副猪嗜血杆菌P5蛋白的抗体效价。结果显示:3次免疫后,白油佐剂免疫组的平均效价为1∶4000,铝胶佐剂免疫组的平均效价为1∶700,蜂胶佐剂免疫组的平均效价为1∶100,无佐剂对照组的平均效价为1∶100。说明这3种佐剂中免疫增强效果最好的为白油,铝胶次之。     

Evaluation of Cisplatin as an electrochemotherapy agent for the treatment of incompletely excised mast cell tumors in dogs

Background: Electrochemotherapy (ECT) couples the administration of anticancer drugs with the delivery of electric pulses that increase the drug uptake through the cell membranes, resulting in an improved efficacy. Hypothesis: To evaluate the tolerability and efficacy of cisplatin (CDDP) as an ECT agent to prevent recurrence of incompletely resected mast cell tumors (MCTs). Animals: Thirty‐seven dogs. Methods: Prospective study recruiting dogs with incompletely excised MCTs as confirmed by surgeon and pathology reports. After debulking, the tumor bed and margins were infiltrated with CDDP, and then exposed to trains of biphasic electrical pulses under sedation. Five minutes after the injection of the chemotherapy agent, sequences of 8 biphasic pulses lasting 50 + 50 μs each, were delivered in bursts of 1,300 V/cm for sclerosed and of 800 V/cm for exposed lesions, with caliper or needle array electrodes, respectively. A second session was performed 1 or 2 weeks later based on clinical considerations. Results: The treatment was well tolerated with minimal adverse effects. Twenty‐nine dogs had no evidence of recurrence over the 6‐year study period, 6 had tumor recurrence, 1 died of multiple cutaneous MCTs, and 1 died of unrelated causes. The estimated median time to recurrence was 1,200 days. Recurrence was not observed among the long‐term (>1 year) treated dogs. Conclusions and Clinical Importance: ECT with CDDP appears effective in the treatment of incompletely resected MCT in dogs and could be a useful addition to the current options based on its low cost, limited toxicity, and ease of administration.