The B and T Cell Responses to Orf Virus Infection of Ovine Skin

Abstract— Local scarification and infection with orf virus induced an influx into the dermis of a significantly greater number of T cells than scarification alone. These cells accumulated underneath the early lesion caused by the scarification and later within epidermal pustules which formed as a result of infection. Cells of the γδ, T₄ and T₈ subsets were all implicated in this response. Dermal B cell numbers also rose in response to infection but relatively fewer B cells were found within pustules. Dendritic cells reactive for IgM were prominent under the initial lesion caused by scarification and dense aggregations occurred under the degenerating epidermis about 72 h after infection. Résumé— L'infection par scarification locale de virus de l'ecthyma contagieux du mouton a provoqué un influx de lymphocytes T dans le derme significativement plus important que lors de scarification seule. Ces cellules s'accumulaient sous les lésions précoces provoquées par la scarification et plus tard au niveau des pustules resultant de l'infection. Les lymphocytes T3, T4 et γδ, étaient impliqués dans cette reaction. Le nombre de lymphocytes B dermiques augmentait aussi en réponse à l'infection, mais peu d'entre eux étaient retrouvér dans les pustules. Les cellules répondant à l'lgM étaient majoritaires sous les seules lésions provoquées par scarification et des agrégations denses apparaissaient sous l'épiderme dégénéré 72 heures aprés l'infection. Zusammenfassung— Die lokale Skarifikation und Infektion mit dem Orf-Virus führte zum Einwandern von einer signifikant größeren Anzahl von T-Zellen in die Dermis, als die Skarifikation allein. Die Zellen akkumulierten sich unter der frühen Skarifikationsläsion und später innerhalb der epidermalen Pusteln, die sich durch die Infektion ergaben. Zellen der Gamma-delta-, T4-und T8-SubpopuIationen waren an dieser Reaktion beteiligt. Auch die Anzahl dermaler B-Zellen stieg als Antwort auf die Infektion, aber innerhalb der Pusteln wurden, relativ gesehen, weniger B-Zellen gefunden. Die auf IgM reaktiven Denritenzellen hoben sich unter der initialen Läsion durch die Skarifikation hervor, dichte Aggregationen fanden sich unter der degenerierenden Epidermis ungefähr 72 Stunden nach der Infektion. Resumen Infección e introducción por medio de multiples punturas cutáneas del virus Orf, produjo un mayor aflujo de linfocitos T en la dermis que punturas de manera aislada. Éstas células se acumularon debajo de la lesión primaria causada por las punturas y de forma tardía en las pústulas epidérmicas, las cuales se formaron como resultado de la infección. Las células del tipo yó de los subgrupos T4 y T8, se encontraron todas implicadas en esta respuesta. Los números de linficitos B en la dermis se vieron aumentados como reacción a la infección, pero menos en las pústulas, relativamente. Células dentríticas reactivas a la IgM fueron prevalentes en la lesión inicial por las multiples punturas, y 72 horas despues de la infección, se detectaron agregaciones densas que ocurrieron bajo la epidermis en degeneración.     

血红素铁对仔猪生长性能和血液指标的影响

选取120头28日龄断奶仔猪,随机分成4个处理组,即对照组和3个试验组,每组3个重复,每个重复10头。对照组饲喂基础日粮,试验1组、试验2组、试验3组分别饲喂基础日粮中血红素铁净添加量为27.5mg/kg、55.0mg/kg、85.0mg/kg的3种试验日粮,研究日粮中添加血红素铁对断奶仔猪的生长性能、血液指标(血红蛋白浓度、免疫球蛋白浓度)的影响。试验正试期14d。结果表明,血红素铁各添加量组均能够提高仔猪的生长性能,日粮中血红素铁添加量为55mg/kg时,仔猪平均日增重、饲料转化率较对照组分别提高12.02%和10.27%(P<0.05)。血红素铁对断奶仔猪有良好的促生长作用,也在一定程度上提高了仔猪血液中血红蛋白和IgG浓度。     

外燥对小鼠呼吸道分泌功能与抗体的影响

目的:观察外燥对小鼠呼吸道分泌功能(RS)与呼吸道IgG(IgG-R)的影响.方法:SPF级昆明小鼠72只随机分为4组:常温常湿组(A组)、常温燥组(B组)、温燥组(C组)和凉燥组(D组),每组18只.第7天与第14天检测各组RS(μg/mL)与IgG-R.结果:B组与D组第7天与第14天IgG-R高于A组(P＜0.05);C组第7天RS增高、第14天下降(P＜0.01),但IgG-R均低于其它3组(P＜0.05);D组RS第7天减少、第14天升高(P＜0.01).结论:温燥致病与致肺津生成障碍及气道御邪相关的IgG明显减少相关.温燥致气道IgG长期低水平现象值得研究.     

微波处理条件对蛋清结构和IgG结合能力的影响

为探究降低蛋清致敏性的新方法,在不同的微波条件下处理蛋清蛋白,通过SDS-PAGE(十二烷基磺酸钠-聚丙烯酰氨凝胶电泳)分析分子量变化、圆二色光谱分析二级结构变化、紫外光谱分析三级结构变化、外源性荧光光谱分析疏水性变化、间接ELISA(酶联免疫吸附试验)分析IgG(免疫球蛋白G)结合能力的变化,研究微波对蛋清的影响。研究证实,微波处理降低蛋清蛋白的潜在致敏性,其中400 W/30 s处理后的蛋白显示出最低的IgG结合能力,所有条件下处理后的蛋清蛋白的二、三级结构均发生显著改变。80 W的微波功率下,蛋白质的结构先折叠,该条件下处理30 s后,蛋白结构又展开;当微波功率达到640 W时,处理时间较长的蛋白分子产生堆叠。蛋白质分子的结构变化导致了其抗原性的变化。     

Evaluation of Brix Refractometry for the Estimation of Colostrum Quality in Jennies

Donkey placenta does not allow the passage of immunoglobulins; thus, foals are born hypogammaglobulinemic and an adequate intake of high-quality colostrum in the first 24 hours of life is crucial for the surviving. The study aims to assess the relation between colostrum immunoglobulin G (IgG) concentration evaluated by the single radial immunodiffusion (SRID) test and the Brix refractometer in donkeys to establish a cutoff value for high quality of colostrum based on Brix refractometry. Colostrum was collected at foaling, and at 6, 12, and 24 hours after foaling from the left and the right half of nine Amiata jennies. A total of 72 colostrum samples were analyzed. A Friedman test with a Dunn’s test for multiple comparisons was used for assessing the differences between the left and right half at each sampling time. No differences were found between the left and right halves; the average value was used to analyze the effect of sampling time on the IgG concentrations and Brix values. The relationship between colostrum IgG concentrations (SRID test) versus Brix value and Brix value versus time was analyzed using two different mixed linear models. A strong statistically significant relation has been found between IgG concentrations and Brix values (R² = 0.84). The relation between IgG concentrations and Brix refractometer showed a cutoff point of 17% Brix score for the identification of high-quality colostrum. The Brix value (%) decreased continuously from 16.29 by 0.29 × hour. Jennies’ and donkey foals’ management may be greatly improved using this simple and cheap device.     

猪血浆IgG提取物对小白鼠免疫功能的影响

研究灌胃猪血浆IgG提取物对小白鼠免疫功能的影响。采用腹腔注射环磷酰胺制造试验小白鼠的免疫功能抑制模型,选用齐墩果酸为阳性对照组,猪血浆IgG提取物为试验组,预饲5d后,然后将猪血浆IgG提取物按每千克体重45、90和180mg3种剂量给小白鼠连续灌胃10d后测定小白鼠免疫水平的变化。小白鼠灌胃猪血浆IgG提取物后,对抗由环磷酰胺引起的小白鼠免疫器官萎缩有一定的保护作用,并对环磷酰胺所致动物DTH反应低下有一定的对抗作用;能显著增加小白鼠血清中IgG的含量和免疫球蛋白的含量(P<0.05),能显著增加小白鼠血清中球蛋白的含量(P<0.05),灌胃猪血浆IgG提取物能显著提高小白鼠的免疫功能。     

Assessment of the IgG index in dogs by indirect immunoenzimatic assays as diagnostic tool for inflammatory diseases of central nervous system

The IgG index measures the intrathecal immunoglobulin production and it is a useful tool for diagnosis of inflammatory diseases involving the central nervous system. This index is based on the precise quantification of albumin and IgG in canine cerebrospinal fluid and serum. Here, we report the development of an indirect competitive ELISAs for the detection of both antigens. Thirty-two dogs were included in this study, divided into three experimental groups. Group A was composed of 22 healthy animals, as determined by standard clinical examination. In group B, six animals, presented neurological pathologies associated with endogenous IgG production and, in group C four animals presented neurological diseases or symptoms not associated with intrathecal IgG production. Cerebrospinal fluid and serum samples were obtained from these animals. As expected, by using the indirect ELISAs proposed here, the IgG indexes obtained in healthy animals (A) were 0.371+/-0.252 (SD). In B and C, the values (3.002+/-1.897; 0.36+/-0.306, respectively), were in agreement with the pathologic conditions of the individuals in each group. Thus, the immunometric competition ELISA methods proposed here allow the discrimination of abnormal intrathecal IgG production, in a variety of inflammatory pathologic conditions of the central nervous system.     

Use of albumin quotient and IgG index to differentiate blood- vs brain-derived proteins in the cerebrospinal fluid of cats with feline infectious peritonitis

BACKGROUND: Inflammation of the central nervous system (CNS) is a frequent condition in cats but etiology often remains unsolved. Routine cerebrospinal fluid (CSF) analysis can be extended through the calculation of the albumin quotient (Q(alb)), a marker of the integrity of the blood-brain barrier (BBB), and IgG index, an estimate of intrathecal IgG synthesis. OBJECTIVES: The purpose of this study was to validate nephelometric methods for CSF protein analysis, and to use the Q(alb) and IgG index to discriminate blood- and brain-derived immunoglobulin fractions in cats with feline infectious peritonitis (FIP). METHODS: Cats presented to our clinic between 2001 and 2005 were included in the study based on clinical and laboratory data and histopathologic findings at necropsy. Cats were grouped as having nonneurologic disease (controls; n=37), brain tumors (n=8), FIP involving the CNS (n=12), and extraneural FIP (n=12). CSF-total protein (TP) was measured and albumin and IgG concentrations were measured in paired CSF/serum samples; Q(alb) and IgG index were calculated. Intraassay and interassay precision of the nephelometric assays were determined using pooled samples. RESULTS: Coefficients of variation for the nephelometric assays ranged from 2.7% to 7.2%. In control cats, CSF-TP concentration ranged from 0.06 to 0.36 g/L, Q(alb) ranged from 0.6 to 5.7 x 10(-3), and IgG index ranged from 0.3 to 0.6. Q(alb) and IgG index were significantly higher in cats with brain tumors and cats with CNS-FIP compared with other groups. Compared with control cats, pleocytosis was evident in 8 of 12 (67%) cats and CSF-TP was increased in 3 of 12 (25%) cats with CNS-FIP. CONCLUSION: Nephelometry is a reliable method for measurement of CSF protein, albumin, and IgG in cats. The Q(alb) and IgG index did not identify a CSF protein pattern specific for BBB dysfunction or intrathecal IgG synthesis in cats with CNS-FIP.     

抗猪IgG单克隆抗体的制备与应用

采用辛酸一硫酸铵盐析和SuperdexTM-200 凝胶层析分离纯化猪血清IgG,免疫BALB/c小鼠,利用细胞融合技术获得8株稳定分泌抗猪IgG 单克隆抗体的杂交瘤细胞.Ig 亚类分析表明,所制备单克隆抗体的轻链亚类均为K,重链除6H3为IgG2b外,其余7株均属于IgG1.Western blot分析显示,5株单克隆抗体识别猪IgG轻链,3株单克隆抗体识别猪IgG重链.以蛋白G亲和层析纯化单克隆抗体7H1腹水,用改良过碘酸钠法进行辣根过氧化物酶(HRP)标记,结果显示,HRP 标记抗猪IgG单克隆抗体对纯化猪IgG的工作浓度达1:12 800.将HRP标记鼠抗猪IgG单克隆抗体与HRP标记羊抗猪IgG多克隆抗体应用于猪血清圆环病毒抗体检测,二者符合率为95.12%,其ELISA和Westem blot的工作浓度为1:2 000和1:10000,表明HRP标记抗猪IgG单克隆抗体具有高度的敏感性.本研究制备的酶标抗猪IgG单克隆抗体为猪病的免疫诊断试剂研究和猪细胞生物学研究提供了一种基础工具.     

Immunoglobulin G4‐related disease in a dog

Immunoglobulin G4‐related disease (IgG4‐RD), which affects many organ systems, has been recognized as a distinct clinical entity in human medicine for just over a decade but has not been previously identified in dogs. In humans, IgG4‐RD is characterized by diffuse IgG4‐positive lymphoplasmacytic infiltrates that commonly lead to increased serum concentrations of IgG4 and IgE, peripheral eosinophilia, tumorous swellings that often include the parotid salivary glands, obliterative phlebitis, and extensive fibrosis. Herein we describe the diagnosis, clinical progression, and successful treatment of IgG4‐RD in an 8‐year‐old female spayed Husky mixed breed dog. Immunoglobulin G4‐related disease should be considered as a differential diagnosis for dogs with vague clinical signs, lymphoplasmacytic swellings, restricted polyclonal gammopathy, eosinophilia or some combination of these findings.