作者：
Dong, Wang Nai;Jun, Li Jing;Bing, Wang Ai;Zhe, Zhu;Yan, Chang;...
期刊：
POLISH JOURNAL OF VETERINARY SCIENCES,2016年19(3):471-476 ISSN：1505-1773
通讯作者：
Bang, Deng Zhi
作者机构：
[Bing, Wang Ai; Dong, Wang Nai; Yu, Mao; Feng, Wang Zan; Jun, Li Jing; Zhe, Zhu; Yang, Zan; Yan, Chang; Bang, Deng Zhi] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.
通讯机构：
[Bang, Deng Zhi] H;Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.
关键词：
Life Sciences;Zoology;Life Sciences, other;Medicine;Veterinary Medicine
摘要：
In order to investigate whether PCV2b can be transmitted across the placenta in sows thereby causing vertical infection of fetuses, six sows in 75 day of pregnancy were either intramuscularly inoculated with a PCV2b culture supernatant (n = 4) or mock infected with cell culture supernatant (n = 2). At parturition, 3 newborn piglets from each litter were randomly selected and euthanatized prior to suckling. Samples of the mesenteric lymph nodes, spleens, kidneys, hearts and lungs were collected for PCR, histopathological and immunohistochemical assays. The results showed that the newborn piglets from PCV2b-infected sows had histopathological lesions in the tested tissues. Moreover, PCV2b antigen and DNA were detected as well. These findings therefore suggested that porcine circovirus type 2b can be transmitted across the placenta of sows, thereby leading to PCV2b vertical infection of the fetuses.
通讯机构：
[Xue, Liqun] H;Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.
关键词：
DBY;antibody;spermatozoa;gender selection
摘要：
DEAD (Asp-Glu-Ala-Asp) box polypeptide 3, Y-linked (DBY or Ddx3y) is a candidate gene for male-specific antigen. The DBY gene detected in capacitated mouse sperm codes putative ATP-dependent RNA helicase. The objective was to produce anti-predicted DBY multi-epitope fusion protein antibody, which could be used to determine male specificity of DBY. Epitope prediction is to aid the design of molecules that can mimic structure and function of a genuine epitope, is a useful tool in protein molecule design. This study predicted the DBY epitopes, prepared rabbit poloclonal antibody against DBY multi-epitope fusion protein, then investigated its immunoreactivity. The fusion protein used as the antigen consisted of three regions of DBY with greatest divergence from other family members, cloned together in-frame (with a His tag to facilitate purification). The resulting antibody recognized both the DBY1-2-3 fusion protein and an endogenous DBY protein of the same size. Furthermore, DBY protein was present (Western blot) in testis, male mouse splenocytes and brain, whereas a weaker band was present in the female brain and splenocytes, and finally, ovary produced only a barely visible protein band. Optical density of DBY protein was higher for males versus corresponding tissues from females. Finally, positive signals of DBY1-2-3 antibody were present on only similar to 60% of mature murine sperm (based on immunofluorescent staining and flow cytometry), in accordance with the expected proportion of Y-bearing sperm. We hypothesized that our antibodies recognized a specific epitope present in subpopulations of mouse sperm. Therefore, we concluded that anti-DBY1-2-3 antibody could be an alternative way of producing antibodies to DBY protein. Furthermore, this novel DBY antibody against a multi-epitope artificial antigen has potential for both investigating male-specific binding of DBY and as a new method of sex selection.
作者机构：
[Naidong Wang; Yang Zhan; Aibing Wang; Zhe Zhu; Zhanfeng Wang] College of Veterinary Medicine, Hunan Agricultural University, Changsha, China;[Zhibang Deng; Yi Yang] College of Veterinary Medicine, Hunan Agricultural University, Changsha, China zbangd@hunau.edu.cn yiyang@hunau.edu.cn
通讯机构：
[Deng, Zhibang] C;College of Veterinary Medicine, Hunan Agricultural University, Changsha, China yiyang@hunau.edu.cn
摘要：
High-titer serologically detected male (SDM) antibody fragments are essential for specific binding to the SDM antigen and promoting its application. The A8 clone previously obtained from an original phage antibody library was further affinity-matured by light- and high-chain shuffling respectively, to generate the end product B9 clone. The binding capacity of B9 phage Fabs to male splenocytes doubled the value of its parental A8 clone (determined using ELISA). Based on immunofluorescent staining, B9-Fabs mainly bound to the surface antigen of male splenocytes and recognized testicular cells. The resulting B9-Fabs detected a single protein (approximately 40 kDa determined using Western blot analysis of male splenocytes and testis); its high SDM antigen binding ability might have been because of mutation sites and varied lengths of the amino acid sequences in the complementarity determining regions-3 of the kappa and Fd chains. The new recombinant clones of Fab that were phage-enhanced using chain shuffling were candidate molecules for investigating molecular mechanisms of SDM antigens specific binding and applications. (C) 2013 Elsevier Inc. All rights reserved.
作者：
Deng Zhibang*;Gong Qianlong;Wang Naidong;Yuan Anwen;Yu Xing Long
期刊：
TURKISH JOURNAL OF VETERINARY & ANIMAL SCIENCES,2013年37(4):475-478 ISSN：1300-0128
通讯作者：
Deng Zhibang
作者机构：
[Gong Qianlong; Yu Xing Long; Deng Zhibang; Wang Naidong; Yuan Anwen] Hunan Agr Univ, Coll Vet Med, Changsha, Hunan, Peoples R China.
通讯机构：
[Deng Zhibang] H;Hunan Agr Univ, Coll Vet Med, Changsha, Hunan, Peoples R China.
关键词：
PCV2b DNA-positive semen;artificial insemination;PCV2b transmission to fetuses;reproductive failure;Kunming mice
摘要：
In order to investigate whether porcine circovirus 2b (PCV2b) can be replicated in the spermatids of mice and whether semen from PCV2b-infected male mice can induce reproductive failure in female mice, 10 male Kunming mice were intraperitoneally inoculated with PCV2b, and 10 female Kunming mice were artificial inseminated with semen from the PCV2b-inoculated male mice. Semen from inoculated male mice and newborn mice or dead fetuses from inseminated female mice were tested for PCV2b by polymerase chain reaction, transmission electron microscopy, and immunofluorescent histochemistry. The results showed that PCV2b was capable of being replicated in spermatids in Kunming mice and could be transmitted from semen to fetuses by artificial insemination, causing reproductive failure in pregnant Kunming mice and PCV2b vertical infection to the newborn mice.
作者机构：
[Gong, Qian Long; Xue, Li Qun; Wang, Nai Dong; Yu, Xing Long; Deng, Zhi Bang; Luo, Wei; Yuan, An Wen] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.
通讯机构：
[Xue, Li Qun] H;Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.
关键词：
Porcine circovirus;PCV2b transmission;contact and vertical infection;Kunming mice
摘要：
To investigate porcine circovirus type 2b (PCV2b) transmission by contact and vertical infection in Kunming mice (an outbred mouse stock deriving from Swiss albino mice with a high ratio of gene heterozygosis), four mice in cage 6 were inoculated with PCV2b and 25 mice without any treatment were placed into cages 1 to 5 (five mice in each cage). Seven days after being infected, the PCV2-binoculated mice were co-mingled with non-inoculated mice from cages 1 to 5 successively at 7, 14, 21, 28 and 35 days post infection (dpi), respectively, for 3 days. In addition, eleven pregnant mice were injected with PCV2b. Samples were collected from non-inoculated mice and three newborn mice from each litter for PCV2b detection by polymerase chain reaction (PCR) and immunohistochemistry (IHC). The PCR results showed that PCV2b transmission rate among mice in cages 1, 2, 3, 4 and 5 was 0/5, 2/5, 5/5, 5/5 and 1/5, respectively. PCV2b antigen signals generally appeared in most organs of the non-inoculated mice in which viruses were detected by PCR. PCV2b DNA was also detected in newborn mice of PCV2b-infected litters, and viral antigen signals were observed in their organs as well. PCV2b was transmitted in Kunming mice by contact, and it also caused vertical infection through the placenta.
摘要：
雄性特异性组织相容性抗原（male specific minor histompatibility antigens,H-Y抗原）是由Y染色体上的基因编码,在雄性动物细胞中普遍表达（包括胚胎和滋养层细胞）。H-Y抗原不仅能引起基因型相同的雌性动物排斥雄性组织,也能导致人白细胞抗原匹配干细胞移植术后出现移植抗宿主性疾病（GVH）。细胞毒性T淋巴细胞检测到几个不同的H-Y抗原表位,这些肽是从胞内蛋白分离出来,由主要组织相容性复合分子结合呈递在细胞表面。H-Y抗原肽与人白细胞抗原Ⅰ类和Ⅱ类分子特异结合参与免疫反应,从而影响移植结果。近年来越来越多的文献报道了H-Y抗原的相关研究,作者主要综述编码H-Y抗原的相关候选基因,并对它在疾病等方面的前景作出了一些展望。
通讯机构：
[Xue, Li-Qun] H;Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.
关键词：
Mouse model;PCV-2b;Histopathological lesions;Viral distribution
摘要：
The viral distribution and lesions in Kunming mice experimentally infected with porcine circovirus type 2b (PCV-2b) were investigated. Seventy special pathogen free mice were divided into 2 groups with 35 mice in each group. The test group (TG) was infected with PCV-2b, the control group (CG) was inoculated with sterile cell cultures. Five mice in each group were sacrificed at 3, 7, 14, 21, 28, 35 and 42 dpi (day post infection), respectively. Necropsies were performed on all mice and tissues were collected for testing by histopathology, immunohistochemistry, transmission electron microscope (TEM) and polymerase chain reaction (PCR). Apoptosis and necrosis in lymphoid organs were observed in virus-infected mice, and became severe from 14 to 28 dpi. The proportion of PCV-2b antigen-positive cells was moderate in lung, heart, thymus, liver or kidney, and low in brain from TG. In spleen and cervical lymph node, the proportions of PCV-2b antigen-positive cells were low to high from 7 to 28 dpi, and moderate from 35 to 42 dpi. PCV-2b DNA was detected in all tissues examined in TG from 7 to 42 dpi. Viral inclusion bodies presented in the cytoplasm of lymphocytes, macrophages, hepatocytes, podocytes, neurocytes, spermatids and uterine epithelial cells in TG. In CG, no viruses and viral lesions were detected. PCV-2b could replicate in mice, and PCV-2b associated lesions in mice were similar to those observed in pigs. The present results indicate that it is possible to use Kunming mouse as an animal model for PMWS research.
