鼠疫F1-V重组蛋白疫苗滴鼻免疫应答效果的研究

目的 以重组霍乱毒素B亚单位（rCT-B）为鼠疫F1-V重组蛋白的佐剂制备黏膜疫苗,观察小鼠诱导的黏膜免疫和系统免疫应答效果。方法以制备的鼠疫黏膜疫苗滴鼻免疫小鼠4次免疫后,采用间接ELISA检测血清特异性抗F1-V的IgG和IgA抗体及抗体亚型分类,检测鼻咽喉、肺、小肠及阴道灌洗液中特异性抗F1-V的黏膜分泌型IgA;采用流式细胞术检测鼻相关淋巴组织淋巴细胞、脾淋巴细胞、肠系膜淋巴结及小肠PP结T淋巴细胞表型的变化。结果以rCT-B为佐剂的鼠疫F1-V重组蛋白黏膜疫苗滴鼻免疫后,能够诱导血清中IgG、IgA抗体比正常对照组显著升高（P〈0．01）,同时诱导鼻咽、肺、小肠和阴道内特异性黏膜抗体升高,尤其是肺和生殖道冲冼液内抗体升高极为显著（P〈0．01）。与单纯的F1-V组相比,不同剂量比例疫苗组都能诱导较高、较快的血清IgG、IgA和黏膜sIgA,其中1：2疫苗组能诱导更强的系统免疫和黏膜免疫,但是相比之下,5：1疫苗组是最合适的免疫剂量。结论rCT-B佐剂不仅能提高鼠疫F1-V黏膜疫苗的系统全身免疫应答,还能促进诱导呼吸道、消化道和生殖道等局部黏膜sIgA抗体,增强局部免疫应答,提示rCT-B佐剂能显著提高鼠疫感染的免疫应答作用,这为下一步疫苗的免疫保护评价奠定了基础。     

重组幽门螺杆菌尿素酶B亚单位疫苗鼻腔免疫的实验研究

目的:探讨基因工程疫苗Hp重组尿素酶B亚单位(rUreB)鼻腔接种的免疫效果。方法:以rUreB不同剂量或加不同佐剂滴鼻免疫BALB/c小鼠。末次免疫7 d后,收集血清及胃黏膜、小肠黏膜、鼻黏膜及气管黏膜冲洗液,用ELISA法检测抗rUreB特异性抗体。结果:rUreB鼻腔免疫后各实验组血清特异性IgG及各黏膜冲洗液中特异性IgA的水平均明显增高,与对照组相比较差异显著(P<0.01)。20μg剂量组与10μg剂量组相比较,仅血清特异性IgG水平增高,其它黏膜特异性IgA的水平未见增高。大肠杆菌不耐热肠毒素B亚单位(LTB)的佐剂效果较霍乱毒素B亚单位(CTB)强,卡泊波可增强鼻腔接种疫苗在胃黏膜洗液中的抗体应答水平。结论:CTB、LTB、卡泊波均可作为rUreB鼻腔黏膜接种的佐剂。HprUreB鼻黏膜接种,不仅可诱导血清特异性抗体反应,而且能引起多个黏膜部位的免疫应答,是一种方便、有效、廉价的免疫途径。     

蛋白体佐剂及用于鼠疫F1-Ⅴ抗原的免疫效果

目的以蛋白体(proteosomes)佐剂,非共价结合鼠疫F1-V重组蛋白为免疫原,探讨滴鼻免疫BALB/c小鼠后诱导的免疫应答和免疫保护效果。方法佐剂与鼠疫F1-V重组蛋白为免疫原非共价结合,滴鼻免疫BALB/c小鼠4次后,采用间接ELISA检测血清特异性抗F1-V的IgG和IgA抗体及抗体亚型分类,并检测鼻咽、肺、小肠及阴道灌洗液中特异性抗F1-V的黏液分泌型IgA;并用流式细胞术检测鼻相关淋巴组织淋巴细胞、脾淋巴细胞、肠系膜淋巴结及小肠PP结T淋巴细胞表型的变化。第4次免疫后7d,用100 LD_(50)的鼠疫141强毒株进行腹腔攻毒。结果(1)以蛋白体为佐剂的鼠疫F1-V抗原与单纯的F1-V组相比,蛋白体疫苗组诱导血清IgG、IgA抗体显著升高(P<0.01),同时蛋白体疫苗组能诱导鼻咽、肺、小肠和阴道内多个黏膜部位特异性IgA抗体的产生,尤其是肺和生殖道冲冼液内抗体升高极为显著(P<0.01);(2)蛋白体疫苗组主要引起IgG1型抗体,主要诱导T_H2型免疫反应;(3)蛋白体疫苗组NALT和SP中CD4~ /CD8~ 比值比PBS对照有显著增高(P<0.01),MLN和PP中CD4~ /CD8~ 比值与PBS对照差异无统计学意义(P>0.05)。(4)小鼠在100 LD_(50)的鼠疫141强毒株腹腔攻毒后鼠疫F1-V重组蛋白组小鼠免疫保护率为0,而蛋白体佐剂疫苗组小鼠免疫保护率为67%。结论以自制的蛋白体为鼠疫F1-V抗原的佐剂滴鼻免疫小鼠,蛋白体不仅提高鼠疫F1-V抗原的系统免疫应答,而且能诱导小鼠呼吸道、消化道和生殖道局部黏膜免疫应答。蛋白体佐剂鼠疫疫苗对100 LD_(50)的鼠疫141强毒株腹腔攻毒具有一定的免疫保护作用,这为鼠疫黏膜疫苗的研制提供候选材料,也为鼠疫黏膜疫苗深入研究奠定了基础。     

不同免疫途径对chitosan-DNA疫苗诱导CVB3特异性免疫应答的影响及其意义

目的：确定新型chitosan-DNA疫苗的有效免疫途径。方法：将chitosan-pcDN3-VPI疫分别苗以肌注、口服、滴鼻3种免疫方式免疫Balb/c小鼠；以ELISA检测免疫小鼠血清中IgG、IgM、、IgA，评估其特异性体液免疫应答；以特异性淋巴细胞增殖反应和CTL活性反映其诱导细胞免疫；以5LD50致死剂量CVB3攻击免疫小鼠，评价不同免疫途径的免疫保护效果。结果：①在诱导CVB3特异性体液免疫方面：chitosan-pcDNA3-VPI疫苗肌注组诱生了高水平IgM和IgG，但未能诱生黏膜IgA；口服免疫组仅诱生低水平的黏膜IgA，未能诱生特异性IgM和IgG；滴鼻组可诱生低水平的I埘及高水平的IgG和黏膜IgA。②在诱导CVB3特异性细胞免疫方面：仅滴鼻组诱导了较高水平的淋巴细胞特异性增殖反应和CTL活性；口服组的淋巴细胞增殖活性和CTL活性稍弱；肌注组几乎不能诱导特异性细胞免疫应答。③免疫保护作用：滴鼻组可保护33．3％小鼠长期存活；口服组仅达到16．7％的保护率；肌注组无保护作用。结论：滴鼻免疫途径可能是chitosan-pcDNA3-VPI基因疫苗最合适的诱导全面免疫应答的免疫途径。     

汉坦病毒经不同途径黏膜免疫效果研究

目的 以大肠埃希菌不耐热肠毒素8亚单位为佐剂研究汉坦病毒经不同黏膜途径免疫的效果.方法 以乳糖为诱导剂,在大肠埃希菌表达不耐热肠毒素B亚单位(LTB),Ni2+亲和层析进行纯化.以灭活汉坦病毒84Fli株为疫苗,LTB为佐剂,分别采用滴鼻、口服、经阴道3种黏膜途径接种C57BL/6小鼠.ELISA检测血清中特异IgG和阴道冲洗液中特异IgA.结果 免疫印迹和神经节苷脂结合活性研究,证实了LTB的表达.经滴鼻、口服、阴道免疫,均可诱导分泌型IgA抗体和血清IgG抗体反应,不同途径接种组间差异无统计学意义.结论 以LTB为佐剂的灭活汉坦病毒通过3种黏膜途径均能产生抗汉坦病毒黏膜免疫和系统免疫应答.     

HIV-1CN54 DNA疫苗滴鼻免疫小鼠诱发系统和黏膜免疫应答

目的：探讨重组痘苗病毒rVVsyngp120或rVVmCN54gp120候选疫苗是否增强HIV-1CN54合成gp120基因(syngp120)DNA疫苗的免疫原性。方法：第0、7、14、21天用DNA疫苗滴鼻免疫小鼠,第28、35、42天再滴鼻接种rVVsyngp120或rVVmCN54gp120。体外测脾和肠系膜淋巴结(MLN)淋巴细胞增殖应答与CD8^ CTL应答。测血清和黏膜洗液特异的IgG和IgA,并测其是否中和实验室适应株HIV-1SF33。结果：单纯DNA免疫后,脾和MLN淋巴细胞在体外发生增殖应答和CTL应答,且测出血清特异的IgG和黏膜洗液特异的IgA。重组痘苗病毒末次免疫后第2周(第56天),发现rVVmCN54gp120增强MLN淋巴细胞增殖应答和CTL应答,脾CTL应答也增强。rVVsyngp120则增强MLN CTL应答。同时发现：2组重组痘苗病毒免疫的动物其血清中特异IgG抗体滴度均有所增高,但黏膜(粪便和阴道)洗液特异IgA抗体滴度却未增高,未测出血清特异IgA和黏膜洗液特异IgG。免疫血清可中和HIV-1SF33,而阴道洗液却不能。结论：单纯DNA疫苗滴鼻免疫可诱发较弱的系统和黏膜体液免疫与细胞免疫,但维持时间短。重组痘苗病毒主要增强局部黏膜的细胞免疫应答,且稍增强系统体液免疫应答,未增强黏膜的IgA应答。免疫血清有中和作用。     

不同佐剂与鼠疫F1-V融合重组蛋白抗原滴鼻免疫效果的研究

目的 研究不同佐剂与鼠疫F1-V融合重组蛋白抗原滴鼻免疫Balb/c小鼠,观察机体产生体液免疫和局部粘膜免疫反应的效果,为发展黏膜疫苗提供理论基础.方法 鼠疫F1-V融合重组蛋白抗原按比例分别与PorB(2类外膜蛋白)重组蛋白、蛋白体佐剂制备黏膜疫苗,滴鼻免疫Balb/c小鼠3次,取尾静脉血,采用ELISA检测血清IgG及抗体亚型分类,并检测鼻咽、肺、小肠及阴道灌洗液sIsA;采用FAC检测脾淋巴细胞表型的变化.结果 PorB重组蛋白佐剂疫苗组和蛋白体佐剂疫苗组较无佐剂组体液免疫抗体水平高、蛋白体佐剂疫苗组好于PorB重组蛋白佐剂疫苗组,但无显著性差异.结论 PorB重组蛋白佐剂疫苗和蛋白体佐剂疫苗均能诱导较强的系统免疫和黏膜免疫应答,且PorB重组蛋白佐剂疫苗免疫效果可与蛋白体佐剂疫苗相媲美,可进一步论证是否可用PorB重组蛋白佐剂替代蛋白体佐剂,这为鼠疫粘膜疫苗的研制奠定了基础.     

EHF疫苗不同途径免疫后血清及黏膜抗体应答情况

目的探讨EHF疫苗经不同剂量和途径免疫后血清IgG及黏膜IgA产生情况,以探讨合适的免疫剂量和接种途径。方法以不同剂量EHF双价灭活疫苗分别经皮下和灌胃免疫小鼠,共3次（第0、5、10天）,末次接种后5d收集血清和小肠冲洗液,用间接免疫荧光法（IFA）检测血清EHF IgG抗体和小肠冲洗液IgA抗体。结果皮下注射能诱导血清特异性IgG和黏膜IgA的产生,灌胃免疫未见抗体产生,1．4 TCID50的EHF疫苗剂量在皮下注射组血清IgG和小肠冲洗液IgA有100％阳性率。结论EHF疫苗皮下注射能诱导血清特异性IgG和黏膜IgA的产生,1．4 TCID50的剂量为较佳剂量。     

高致病性人禽流感H5N1透皮疫苗促渗剂的初步筛选

目的 初步筛选用于高致病性人禽流感H5N1透皮疫苗的较有效的促渗剂.方法 选用乙醇、丙二醇、二甲基亚砜、维甲酸、油酸作为促渗剂,将其应用于BALB/c小鼠,再用灭活的高致病性人禽流感H5N1透皮疫苗免疫小鼠,通过评价透皮免疫应答的效果对促渗剂进行初步筛选.结果 二甲基亚砜、维甲酸组和油酸组血清IgG抗体效价明显高于其他促渗剂组(P＜0.05).结论 在高致病性人禽流感H5N1透皮疫苗的小鼠模型中,二甲基亚砜、维甲酸和油酸是较好的促渗剂.     

二价轮状病毒灭活疫苗的免疫效果评价

目的 评价二价轮状病毒灭活疫苗免疫效果,探讨其开发的可行性.方法 首先将G1型和G3型轮状病毒灭活疫苗等量混合制备二价疫苗,设立抗原量相等的G1型、G3型单价疫苗对照及PBS阴性对照,然后肌内注射免疫小鼠,通过ELISA检测血清和肠道中病毒特异性IgG和IgA、微量中和实验检测血清中病毒特异性中和抗体,ELISPOT分析免疫小鼠脾细胞中病毒特异性IFN-γ分泌细胞和IL-4分泌细胞的数量,评价二价疫苗的免疫效果.结果 与阴性对照组相比,二价疫苗显著诱发了G1和G3特异性血清IgG、IgA、中和抗体和肠道IgG、IgA.与单价疫苗组相比,二价疫苗诱发的G1特异性血清IgG和IgA、肠道IgG和IgA及血清中和抗体与G1单价疫苗组诱发的相应抗体间差异无统计学意义;但二价疫苗组诱发的G3特异性血清IgG(t=2.691,P＜0.05)及中和抗体(t=2.561,P＜0.05)却显著高于G3单价疫苗组诱发的相应抗体水平,其余G3特异性抗体水平两者无差异.同时,与阴性对照组相比,二价疫苗免疫小鼠脾细胞中IFN-γ和IL-4分泌细胞的数量均显著增加.与单价疫苗组相比,G1刺激产生的IFN-γ和IL-4分泌细胞的数量,二价疫苗组与G1单价疫苗组无显著差异;但在二价疫苗组,G3刺激产生的IL-4分泌细胞的数量显著高于G3单价疫苗组(t=2.327,P＜0.05),而IFN-γ分泌细胞的数量两组无显著性差异.结论 二价疫苗可以有效的诱发免疫应答,疫苗不同成分诱发的免疫应答未发生干扰抑制,这些结果为临床二价轮状病毒灭活疫苗的开发提供了实验依据.     

Vaccines with the MF59 adjuvant do not stimulate antibody responses against squalene

Squalene is a naturally occurring oil which has been used in the development of vaccine adjuvants, such as the oil-in-water emulsion MF59. In past years, by use of noncontrolled and nonvalidated assays, a claim was made that antisqualene antibodies were detectable in the sera of individuals with the so-called Gulf War syndrome. Using a validated enzyme-linked immunosorbent assay for the quantitation of immunoglobulin G (IgG) and IgM antibodies against squalene, we demonstrated that antisqualene antibodies are frequently detectable at very low titers in the sera of subjects who were never immunized with vaccines containing squalene. More importantly, vaccination with a subunit influenza vaccine with the MF59 adjuvant neither induced antisqualene antibodies nor enhanced preexisting antisqualene antibody titers. In conclusion, antisqualene antibodies are not increased by immunization with vaccines with the MF59 adjuvant. These data extend the safety profile of the MF59 emulsion adjuvant.     

MF59 adjuvant enhances diversity and affinity of antibody-mediated immune response to pandemic influenza vaccines

Oil-in-water adjuvants have been shown to improve immune responses against pandemic influenza vaccines as well as reduce the effective vaccine dose, increasing the number of doses available to meet global vaccine demand. Here, we use genome fragment phage display libraries and surface plasmon resonance to elucidate the effects of MF59 on the quantity, diversity, specificity, and affinity maturation of human antibody responses to the swine-origin H1N1 vaccine in different age groups. In adults and children, MF59 selectively enhanced antibody responses to the hemagglutinin 1 (HA1) globular head relative to the more conserved HA2 domain in terms of increased antibody titers as well as a more diverse antibody epitope repertoire. Antibody affinity, as inferred by greatly diminished (≥10-fold) off-rate constants, was significantly increased in toddlers and children who received the MF59-adjuvanted vaccine. Moreover, MF59 also improved antibody affinity maturation after each sequential vaccination against avian H5N1 in adults. For both pandemic influenza vaccines, there was a close correlation between serum antibody affinity and virus-neutralizing capacity. Thus, MF59 quantitatively and qualitatively enhances functional antibody responses to HA-based vaccines by improving both epitope breadth and binding affinity, demonstrating the added value of such adjuvants for influenza vaccines.     

Comparison of the Immunogenicity and Safety of the Conventional Subunit,MF59-Adjuvanted,and Intradermal Influenza Vaccines in the Elderly

The influenza vaccination is known as the most effective method for preventing influenza infection and its complications in the elderly. Conventional subunit (Agrippal S1; Novartis), MF59-adjuvanted (Fluad; Novartis), and intradermal (IDflu15; Sanofi Pasteur) influenza vaccines are widely used throughout South Korea. However, few comparative studies evaluating the safety and immunogenicity of these vaccines are available. Prior to the beginning of the 2011-2012 influenza season, 335 healthy elderly volunteers randomly received one of three seasonal trivalent influenza vaccines, the conventional subunit, MF59-adjuvanted, or intradermal influenza vaccine. Serum hemagglutination-inhibiting antibody levels were measured at the time of vaccination and at 1 and 6 months after vaccination. Adverse events were recorded prospectively. A total of 113 conventional subunit, 111 MF59-adjuvanted, and 111 intradermal influenza vaccine volunteers were followed up during a 6-month postvaccination period. One month after vaccination, all three vaccines satisfied Committee for Medical Products for Human Use (CHMP) immunogenicity criteria for the A/H1N1 and A/H3N2 strains but not for the B strain. Compared with the subunit vaccine, the intradermal vaccine exhibited noninferiority, while the MF59-adjuvanted vaccine exhibited superiority. Furthermore, the MF59-adjuvanted vaccine was more immunogenic against the A/H3N2 strain than was the subunit vaccine up to 6 months postvaccination. The most common local and systemic reactions to the conventional subunit, MF59-adjuvanted, and intradermal influenza vaccines were pain at the injection site (7.1%, 10.8%, and 6.3%, respectively) and generalized myalgia (0.9%, 8.1%, and 5.4%, respectively). Local and systemic reactions were similar among the three vaccine groups. MF59-adjuvanted vaccine exhibited superior immunogenicity compared with a conventional subunit vaccine and had a comparable safety profile. For older adults, the MF59-adjuvanted vaccine is preferable for providing superior immunogenicity.     

Effective influenza vaccines for children: A critical unmet medical need and a public health priority

Seasonal influenza causes clinical illness and hospitalization in all age groups; however, conventional inactivated vaccines have only limited efficacy in young children. MF59 (?) , an oil-in-water emulsion adjuvant, has been used since the 1990s to enhance the immunogenicity of influenza vaccines in the elderly, a population with waning immune function due to immunosenescence. Clinical trials now provide information to support a favorable immunogenicity and safety profile of MF59-adjuvanted influenza vaccine in young children. Published data indicate that Fluad (?) , a trivalent seasonal influenza vaccine with MF59, was immunogenic and well tolerated in young children, with a benefit/risk ratio that supports routine clinical use. A recent clinical trial also shows that Fluad provides high efficacy against PCR-confirmed influenza. Based on the results of clinical studies in children, the use of MF59-adjuvanted vaccine offers the potential to enhance efficacy and make vaccination a viable prevention and control strategy in this population.     

MF59 is a safe and potent vaccine adjuvant that enhances protection against influenza virus infection

In preclinical studies, MF59 adjuvant offered improved protection against influenza virus challenge and significantly reduced the viral load in the lungs of challenged mice. In humans, MF59 is a safe and potent vaccine adjuvant that has been licensed in more than 20 countries (Fluad [Novartis Vaccines and Diagnostics Inc., MA, USA]). The safety profile of an MF59-adjuvanted vaccine is well established through a large safety database. MF59 adjuvant has had a significant impact on the immunogenicity of influenza vaccines in the elderly and in adults who are chronically ill. MF59 has also been shown to have a significant impact on the immunogenicity of pandemic influenza vaccines. MF59 allows for broader cross-reactivity against viral strains not included in the vaccine. MF59 has been shown to be more potent for both antibody and T-cell responses than aluminum-based adjuvants. MF59 has broad potential to be used as a safe and effective vaccine adjuvant for a wide range of vaccine types.     

Detection of anti-H5 responses in human sera by HI using horse erythrocytes following MF59-adjuvanted influenza A/Duck/Singapore/97 vaccine

Haemagglutination-inhibition (HI) tests are a simple method used to assess immune responses to influenza haemagglutinin. However, HI tests are insensitive at detection of antibody responses to avian haemagglutinin after vaccination or natural infection, even in the presence of high titres of neutralising antibody or virus isolation. Avian influenza viruses preferentially bind to sialic acid receptors that contain N-acetylneuraminic acid alpha2,3-galactose (alpha2,3Gal) linkages while human viruses preferentially bind to those containing N-acetylneuraminic acid alpha2,6-galactose (alpha2,6Gal) linkages. By using horse erythrocytes in the HI test and thereby increasing the proportion of alpha2,3Gal linkages available for binding, we are able to demonstrate improved detection of antibody to avian H5 in human sera following vaccination with MF59-adjuvanted A/Duck/Singapore/97 surface antigen vaccine. This modified HI test was more sensitive in detection of anti-H5 antibody evoked by revaccination of primed subjects and may be useful in assessing potential avian HA vaccine candidates.     

MF59&#x2122; as a vaccine adjuvant: a review of safety and immunogenicity

Approximately 70 years passed between the licensing of alum salts as vaccine adjuvants and that of MF59? MF59, an oil-in-water emulsion, is currently licensed for use in the elderly as an adjuvant in seasonal influenza vaccines. Its mechanism of action is not fully understood, but enhancement of the interaction between the antigen and the dendritic cell seems to be involved. When used with seasonal influenza vaccines, an increase occurs in the hemagglutination inhibition antibody titers against some, but not all, seasonal vaccine influenza strains. The adjuvant effect is more pronounced when MF59 is combined with novel influenza antigens such as H9 and H5. The use of the adjuvant is associated with an increase in the frequency of local and systemic early post-vaccine adverse events (3-7 days), but no increase in adverse events was observed thereafter. Currently, MF59 is under evaluation as an adjuvant with other antigens such as pandemic influenza antigens and cytomegalovirus antigens.     

Comparison of the Long-Term Immunogenicity of Two Pandemic Influenza A/H1N1 2009 Vaccines,the MF59-Adjuvanted and Unadjuvanted Vaccines,in Adults

Since the first reports of the A/H1N1 virus in April 2009, the pandemic influenza virus spread globally and circulated for a long time. The primary method for the control of influenza is vaccination, but levels of influenza vaccine-induced antibody are known to decline rapidly during a 6-month period. In adults aged 18 to 64 years, we compared the long-term immunogenicity of two of the influenza A/H1N1 2009 monovalent vaccines, 3.75-μg MF59-adjuvanted vaccine and 15-μg unadjuvanted vaccine. The serum hemagglutinin inhibition (HI) titers were determined prevaccination and at 1, 6, and 10 months after vaccination. One hundred six (88.3%) of the 120 subjects were monitored for the entire 10-month period after receiving the influenza A/H1N1 2009 monovalent vaccine. There were 60 patients who received the unadjuvanted vaccine and 46 patients who received the MF59-adjuvanted vaccine. The seroprotection rates, seroconversion rates, and the geometric mean titer (GMT) folds fulfilled the criteria of the European Medicines Agency (EMA) for influenza A/California/7/2009 (H1N1) at 1 month after vaccination irrespective of the vaccine composition. Although the GMTs at 1 month postvaccination were somewhat higher in the unadjuvanted vaccine recipients than in the MF59-adjuvanted vaccine recipients, the difference was not significant (P = 0.29). The seroprotection rates at 6 and 10 months postvaccination were preserved above 70% but only in the MF59-adjuvanted vaccine recipients. In conclusion, low-dose MF59-adjuvanted influenza vaccine, even with 3.75 μg hemagglutinin antigen, might induce excellent long-term immunity that is comparable to the conventional dose of unadjuvanted vaccine among healthy adults aged 18 to 64 years.     

In vitro assessment of the allergenicity of novel MF59-adjuvanted pandemic H1N1 influenza vaccine produced in dog kidney cells

A licensed inactivated MF59-adjuvanted seasonal influenza vaccine (Optaflu) produced in canine kidney cells (MDCK 33016-PF) contained no egg proteins and did not trigger degranulation in rat basophilic leukemia (RBL) cells passively sensitized with human anti-dog IgE, supporting its safe use in dog-allergic individuals. The cell-derived pandemic H1N1 influenza vaccine was also adjuvanted with the emulsion adjuvant MF59, and support for its similar safe use was sought. We sought to evaluate in vitro allergenicity of the MF59-adjuvanted cell-derived pandemic H1N1 influenza vaccine in subjects with dog allergy, with a mediator release assay. RBL-2H3 cells transfected with human Fcε receptor type 1 were sensitized with sera from adult dog-allergic subjects and stimulated with serial dilutions of pandemic H1N1 influenza vaccine and dog dander extract. β-N-hexosaminidase release (NHR) was used as a marker of RBL degranulation.. Median dog dander-specific IgE in 30 dog-allergic subjects was 27.7 kUA/L (range 10.1; > 100); and in 5 dog non-allergic subjects was < 0.35 kUA/L (UniCAP system). Median (range) maximum NHR in dog-allergic subjects was: pandemic H1N1 influenza vaccine 1.1% (0; 4.4) and dog dander 6.9% (0.7; 37.3), P < 0.001. In conclusion, MF59-adjuvanted pandemic H1N1 influenza vaccine produced in continuous canine kidney cells did not trigger degranulation in RBL cells passively sensitized with human anti-dog IgE, supporting its safe use in dog-allergic individuals.