Variability in non-core vaccination rates of dogs and cats in veterinary clinics across the United States

Vaccination guidelines for dogs and cats indicate that core vaccines (for dogs, rabies, distemper, adenovirus, parvovirus; for cats, feline parvovirus, herpes virus-1, calicivirus) are essential to maintain health, and that non-core vaccines be administered according to a clinician’s assessment of a pet’s risk of exposure and susceptibility to infection. A reliance on individual risk assessment introduces the potential for between-practice inconsistencies in non-core vaccine recommendations. A study was initiated to determine non-core vaccination rates of dogs (Leptospira, Borrelia burgdorferi, Bordetella bronchiseptica, canine influenza virus) and cats (feline leukemia virus) in patients current for core vaccines in veterinary practices across the United States. Transactional data for 5,531,866 dogs (1,670 practices) and 1,914,373 cats (1,661 practices) were retrieved from practice management systems for the period November 1, 2016 through January 1, 2020, deidentified and normalized. Non-core vaccination status was evaluated in 2,798,875 dogs and 788,772 cats that were core-vaccine current. Nationally, median clinic vaccination rates for dogs were highest for leptospirosis (70.5%) and B. bronchiseptica (68.7%), and much lower for canine influenza (4.8%). In Lyme-endemic states, the median clinic borreliosis vaccination rate was 51.8%. Feline leukemia median clinic vaccination rates were low for adult cats (34.6%) and for kittens and 1-year old cats (36.8%). Individual clinic vaccination rates ranged from 0 to 100% for leptospirosis, B. bronchiseptica and feline leukemia, 0–96% for canine influenza, and 0–94% for borreliosis. Wide variation in non-core vaccination rates between clinics in similar geographies indicates that factors other than disease risk are driving the use of non-core vaccines in pet dogs and cats, highlighting a need for veterinary practices to address gaps in patient protection. Failure to implement effective non-core vaccination strategies leaves susceptible dogs and cats unprotected against vaccine-preventable diseases.     

Vaccination inhibits the human adenoviral transduction in a mouse keratoconjunctivitis model

Adenovirus infections are a major cause of epidemic keratoconjunctivitis (EKC), which can lead to corneal subepithelial infiltrates and multifocal corneal opacity. In the current study, we investigated the use of an E1/E3-deleted adenovirus serotype 5 (Ad5) vector as a vaccine administered intramuscularly (IM) or intranasally (IN) against subsequent challenges with a luciferase-expressing Ad5 (Ad5-Luci) vector via eyedrop. We evaluated the adaptive immune response to Ad5 vector vaccination and confirmed a robust polyfunctional CD8 T cell response in splenic cells. Neutralizing Ad5 antibodies were also measured in the sera of vaccinated mice as well as Ad5 antibody in the eye wash solutions. Upon challenge with Ad5-Luci vector 8 weeks post the primary immunization, transduction was significantly reduced by > 70% in the vaccinated mice, which was slightly better in IM- vs. that in IN-vaccinated animals. Resistance to subsequent challenge was observed 10 months post primary IM vaccination, with sustained reduction up to 60% in the Ad5-Luci vector transduction. Passive immunization of naive mice with antisera from IM to vaccinated mice subsequently challenged with the Ad5-Luci vector resulted in approximately 40% loss in transduction efficiency. Furthermore, the mice that received IM immunization with or without CD8 T cell depletion showed > 40% and 70% reductions, respectively, in Ad8 genomic copies after Ad8 topical challenge. We conclude that Ad-vector vaccination successfully induced an adaptive immune response that prevented subsequent Ad transduction in the cornea and conjunctiva-associated tissues in a mouse model of adenovirus keratoconjunctivitis, and that both cellular and humoral immunity play an important role in preventing Ad transduction.     

Ad-CD40L mobilizes CD4 T cells for the treatment of brainstem tumors

BackgroundDiffuse midline glioma, formerly DIPG (diffuse intrinsic pontine glioma), is the deadliest pediatric brainstem tumor with median survival of less than one year. Here, we investigated (i) whether direct delivery of adenovirus-expressing cluster of differentiation (CD)40 ligand (Ad-CD40L) to brainstem tumors would induce immune-mediated tumor clearance and (ii) if so, whether therapy would be associated with a manageable toxicity due to immune-mediated inflammation in the brainstem.MethodsSyngeneic gliomas in the brainstems of immunocompetent mice were treated with Ad-CD40L and survival, toxicity, and immune profiles determined. A clinically translatable vector, whose replication would be tightly restricted to tumor cells, rAd-Δ24-CD40L, was tested in human patient–derived diffuse midline gliomas and immunocompetent models.ResultsExpression of Ad-CD40L restricted to brainstem gliomas by pre-infection induced complete rejection, associated with immune cell infiltration, of which CD4+ T cells were critical for therapy. Direct intratumoral injection of Ad-CD40L into established brainstem tumors improved survival and induced some complete cures but with some acute toxicity. RNA-sequencing analysis showed that Ad-CD40L therapy induced neuroinflammatory immune responses associated with interleukin (IL)-6, IL-1β, and tumor necrosis factor α. Therefore, to generate a vector whose replication, and transgene expression, would be tightly restricted to tumor cells, we constructed rAd-Δ24-CD40L, the backbone of which has already entered clinical trials for diffuse midline gliomas. Direct intratumoral injection of rAd-Δ24-CD40L, with systemic blockade of IL-6 and IL-1β, generated significant numbers of cures with readily manageable toxicity.ConclusionsVirus-mediated delivery of CD40L has the potential to be effective in treating diffuse midline gliomas without obligatory neuroinflammation-associated toxicity.     

白细胞介素-18基因修饰的胎肝细胞经脾移植对小鼠免疫功能的影响

目的：观察表达mIL-18的重组腺病毒基因修饰的胎肝细胞（AdmIL-18/BNL.CL2)经脾移植对正常小鼠免疫功能的影响。方法：实验组小鼠经脾移植AdmIL-18/BNL.CL2，同时设LacZ病毒对照组（Ad-LacZ/BNL.CL2),BNL.CL2细胞对照组及空白对照组。2周后处死，留取血清，制备腹腔巨噬细胞、脾淋巴细胞、肝组织匀浆液，提取肝组织总RNA。采用ELISA法检测各组小鼠血清、腹腔Mφ和脾细胞培养上清、肝匀浆中细胞因子的含量；采用半定量RT－PCR法，检测肝组织细胞因子mRNA相对表达量；以LDH释放法测定腹腔Mφ杀伤活性和脾NK细胞活性，用MTT还原比色法测定脾淋巴细胞的增殖活性。结果：实验组小鼠血清、细胞培养上清及肝匀浆中，IL－18、IL－2、IFN－γ、TNF-α稔均高于其它对照组，而IL－4、IL－10水平则低于对照组；半定量RT－PCR结果与ELISA检测结果一致；同时，实验组腹腔Mφ的杀伤活性和脾NK细胞活性，及脾淋巴细胞增殖活性也明显高于对照组。结论：AdmIL-18能有效转染至胎肝细胞并稳定表达mIL-18;AdmIL-18基因修饰的胎肝细胞经脾移植后，可显著提高肝脏、脾脏免疫细胞活性，活化腹腔Mφ，促进Th1类细胞因子表达，抑制Th2类细胞因子的分泌。     

大鼠MMP-13重组腺病毒载体的构建

目的　构建大鼠MMP1 3重组腺病毒载体。方法　将MMP1 3cDNA目的片段插入腺病毒柯氏质粒中 ,再将此质粒与腺病毒DNA末端蛋白复合物共转染 2 93细胞 ,通过同源重组构建MMP1 3 重组腺病毒载体。结果　MMP1 3 重组腺病毒载体经PCR鉴定正确 ,包装纯化后 ,检测病毒滴度为 4× 1 0 9PFU/ml。结论　成功地构建了RatMMP1 3 重组腺病毒载体 ,为下一步行MMP 1 3转基因治疗低顺应性膀胱的研究提供了条件     

BMP7腺病毒基因转染对骨髓基质干细胞生物学的影响

目的 探讨BMP7腺病毒基因转染对骨髓基质干细胞(BMSCs)的体外、体内的生物学功能影响.方法 抽取羊的骨髓,分离、培养骨髓基质干细胞.用重组骨形成蛋白7腺病毒(adenovirus bone morphogenetic protein 7;Adeno-BMP7)转染(M.O.I=100)70%～80%融合时的第二代细胞,三天后分别进行透射电镜观察、流式细胞仪检测、Western-blot、钙结节染色以及珊瑚和细胞复合物皮下回植.结果 在体外:细胞超微结构观察显示Adeno-BMP7基因转染后的BMSCs的物质合成代谢功能活跃;流式细胞仪检测表明Adeno-BMP7基因转染对BMSCs的细胞周期无明显影响;Western-blot检测证明转染Adeno-BMP7后的BMSCs有BMP7在蛋白水平上的表达;染色表明转染Adeno-BMP7后的BMSCs可形成较大的钙结节.在体内:转染Adeno-BMP7的BMSCs可明显促进新骨的形成,与没转染Adeno-BMP7的BMSCs有差异.结论 Adeno-BMP7转染可促进BMSCs的体外成骨定向分化,Adeno-BMP7转染后的BMSCs具有更强的体内成骨能力.     

Regulators of Cardiac Cell Growth, Differentiation, and Apoptosis

The pathogenesis of heart hypertrophy and failure have been the focus of intense clinical and basic science investigation, yet the signal transduction pathways and molecular process that underlie the compensatory growth process that ultimately leads to heart failure remain enigmatic. Since ventricular myocytes have exited the cell cycle, growth of the myocardium in response to hemodynamic load occurs by cellular hypertrophy and not by hyperplasia. In this article, we document the potential involvement of tumor suppressor pocket proteins and cell cycle regulators that may impinge on the growth, differentiation, and apoptosis of cardiac muscle. This revised version was published online in August 2006 with corrections to the Cover Date.     

人IL-10基因重组复制缺陷型腺病毒DNA的构建

目的 构建人IL-10基因的重组腺病毒并进行体外表达和检测。方法从质粒pcDNA3IL-10的CMV启动子下游完整切下IL-10 cDNA的片段,将其定向插入Gateway载体,在克隆酶的作用下将阳性克隆质粒转入目的腺病毒载体。PacI酶线性化IL-10腺病毒DNA后阳离子质脂体转染293A细胞,获得人IL-10的复制缺陷型重组腺病毒。体外感染人胰腺癌细胞株Bxpc-3和大鼠胰腺细胞株AR-42J;Western blot检测人IL-10的表达。结果 成功地构建人IL-10重组腺病毒,病毒滴度达2×109PFU/mL。体外感染的细胞株均检测到IL-10的表达。结论 构建复制缺陷型重组腺病毒能够介导IL-10的基因表达,为细胞因子的抗炎冶疗奠定实验基础。