猪生物医学模型的基因组学研究进展

本综述对猪生物医学模型的多样性和基因组学在这些模型持续开发中的重要性作了一个简要的更新。由于猪在体型大小、生理条件、器官发育和疾病发展等方面与人类相似,因而已作为一种重要的哺乳动物医学模型为人类所用。将猪作为人类医学模型,可以使研究人员从容不迫地安排研究时间,并可利用普通的人类医学技术对机体内部的血管和器官进行成像处理,且可以采集重复的外周样本,并且在扑杀后可采集复杂的粘膜组织样本。以猪为医学模型,能够利用同窝的猪或克隆猪和转基因猪,因而使比较分析和遗传作图更为容易。以猪作为人类医学的动物模型可获得大量代表各种组织的分化方向明确的细胞系,从而可更加有利于基因表达和药物敏感性的检测。因此,对人而言,猪是一种出色的生物医学模型。由于猪在基因序列和染色体结构上与人类高度同源,因而对基因组应用而言,其不失为一个宝贵的资源。由于目前猪基因组序列已被人类充分掌握,因而有改良型遗传学方法和蛋白质组学方法可供这些比较分析方法所利用。本综述将论述用于探测这些模型的基因组学方法中的某些方法,并将重点介绍黑色素瘤和抗传染性疾病的基因组学研究情况,论述在设计此类研究时应予以考虑的问题。本文最后将对利用基因组学方法开发控制具有最高经济价值的猪病—猪繁殖与呼吸综合征病毒(Porcine Reproductive and Respiratory Syndrome,PRRSV)—新型替代方法的可行性、和将从PRRSV研究中获得的知识应用到人传染性疾病研究中的可行性作简短讨论。     

Prevalence of megabacteria in budgerigar colonies

Objective To measure the prevalence of megabacteria in budgerigar-breeding colonies and to evaluate possible methods to reduce the prevalence.
Design A monitoring study over several years.
Sample population Two budgerigar ( Melopsittacus undulatus ) colonies with over 300 birds each.
Procedure The prevalence of megabacteria in the faeces in two budgerigar breeding colonies, colony 1 and 2, was determined by faecal examination of each bird. Following an initial survey (1990), most of the birds that were scored 2+ or more were culled and a management practice was implemented to discriminate against positive birds. Consecutive yearly surveys (1991, 1992) were conducted on the young birds bred in these colonies. The prevalence of megabacteria in colony 2 was also evaluated in 1994 and 1996 after all the birds were treated with amphotericin B administered in drinking water.
Results The prevalence of megabacteria in the two colonies was significantly (P < 0.001) different. Overall the prevalence of megabacteria adjusted for colony differences was significantly higher (P < 0.025) in males compared to females. Age was not an influencing factor. After the initial survey, the prevalence in the offspring did not significantly (P > 0.05) decrease in the following two annual breeding seasons but by inference it did significantly decrease after amphotericin B treatment.
Conclusion The practice of culling most birds with more megabacteria in faeces and discriminating against positive birds when selecting birds for breeding or culling birds on show quality does not decrease megabacteria prevalence in the offspring. However, a reduction in prevalence does occur with administration of amphotericin B. Birds may have amphotericin B-resistant organisms and these birds need to be identified and culled.     

Analyses of monoclonal antibodies reacting with porcine wCD6: Results from the Second International Swine CD workshop

Among the 57 monoclonal antibodies analyzed within the T-cell group of the Second International Swine CD Workshop, one mAb fell within cluster T14a that included the CD6 standard a38b2 (No. 175). The new mAb MIL8 (No. 082) and a38b2 both precipitated from activated T-cells a 150 kDa monomeric protein. Staining patterns on the various cell types were similar. There was no inhibition of binding of either mAb to peripheral blood T-cells with the opposite mAb. The new mAb, MIL8, reacts with a separate epitope on porcine wCD6.     

Report on the analyses of mAb reactive with porcine CD8 for the second international swine CD workshop

Based on an analysis of their reactivity with porcine peripheral blood lymphocytes (PBL), only three of the 57 mAbs assigned to the T cell/activation marker group were grouped into cluster T9 along with the two wCD8 workshop standard mAbs 76-2-11 (CD8a) and 11/295/33 (CD8b). Their placement was verified through the use of two-color cytofluorometry which established that all three mAbs (STH101, #090; UCP1H12-2, #139; and PG164A, #051) bind exclusively to CD8⁺ cells. Moreover, like the CD8 standard mAbs, these three mAbs reacted with two proteins with a MW of 33 and 35 kDa from lymphocyte lysates and were, thus, given the wCD8 designation. Because the mAb STH101 inhibited the binding of mAb 76-2-11 but not of 11/295/33, it was given the wCD8a designation. The reactivity of the other two new mAbs in the T9 cluster with the various subsets of CD8⁺ lymphocytes were distinct from that of the other members in this cluster including the standards. Although the characteristic porcine CD8 staining pattern consisting of CD8^low and CD8^high cells was obtained with the mAb UCP1H12-2, a wider gap between the fluorescence intensity of the CD8^low and CD8^high lymphocytes was observed. In contrast, the mAb PG164A, not only exclusively reacted with CD4⁻/CD8^high lymphocytes, but it also failed to recognize CD4/CD8 double positive lymphocytes. It was concluded that this mAb is specific for a previously unrecognized CD8 epitope, and was, thus, given the wCD8c designation. A very similar reactivity pattern to that of PG164A was observed for two other mAbs (STH106, #094; and SwNL554.1, #009). Although these two mAbs were not originally positioned in the T cell subgroup because of their reactivity and their ability to inhibit the binding of PG164A, they were given the wCD8c designation. Overall, five new wCD8 mAbs were identified. Although the molecular basis for the differences in PBL recognition by these mAbs is not yet understood, they will be important in defining the role of CD8⁺ lymphocyte subsets in health and disease.     

Analyses of monoclonal antibodies reacting with porcine CD3: results from the Second International Swine CD Workshop

Among the 57 monoclonal antibodies (mAb) analyzed within the T-cell group from the Second Swine CD Workshop, six mAb fell within clusters T10 and T11 (No. 088, STH164; No. 148, FY1A3; No. 149, FY2C1; No. 150, FY1H2; No. 151, FY2A11; No. 169, BB23-8E6). The mAb within these two groups gave a similar appearance on flow cytometry and stained all peripheral blood T-cells as defined by CD4 and wCD8 staining. All six mAb precipitated a 24 kDa protein. On the basis of inhibition analyses performed as part of the workshop and from published data, the mAb define at least three epitopes. There is only minimal stimulation of resting peripheral lymphocytes, but four of the mAb produce strong stimulation in the presence of PMA. With the exception of STH164, all have been shown to react with CD3-transfected COS cells. The new mAb, therefore, react with three epitopes on porcine CD3 designated CD3a (BB23-8E6, FY2A11), CD3b (FY1A3, FY2C1), and CD3c (FY1H2). mAb STH164 appears to be reactive with another epitope, however, since its reactivity with CD3 has not been confirmed it is designated as wCD3.