Bovine antibody dependent cell-mediated cytotoxicity (ADCC) effector cells

ADCC effector cells from bovine blood were separated by centrifugation, adherence and rosetting techniques. Each enriched cell population, peripheral blood mononuclear cells (PBM), null lymphocytes, monocytes and neutrophils, was then examined for its capacity to mediate ADCC. Utilizing heterologous sensitizing antisera it was found that monocytes had approximately twice the ADCC activity of null lymphocytes and that neutrophils had essentially no activity. However, when homologous sensitizing antisera were used it was found that neutrophils possessed the greatest activity followed by monocytes and null cells. Results confirm the existence of an ADCC active null lymphocyte in the bovine.     

Inflammatory cytokines and antigen presenting cell activation

Immune responses are stimulated in response to threats against health. In animals, defense against infectious agents, particularly rapidly growing viruses and bacteria, requires an immediate response to limit growth and dissemination, and then stimulation of a more prolonged, specific immunity to prevent re-infection. The process by which animals meet the dual needs of an immediate response to danger and initiation of long-term protection is substantially influenced by inflammatory cytokines produced primarily by macrophages and professional antigen presenting cells (APCs). Inflammatory cytokines mobilize the immune system in response to danger and increase the efficiency of an immune response as effectors of APC function. Here we review the evidence for the involvement of inflammatory cytokines in immune induction and as mediators of APC activity, with a particular emphasis on swine and on the induction of immunity at mucosal surfaces. The vast majority of infections occur at mucosal surfaces of the enteric, respiratory and reproductive tracts, and induction of protective immunity at these sites is particularly challenging. Induction of immunity at mucosal surfaces of the small intestine is greatly facilitated by the oral adjuvant, cholera toxin (CT). CT potentiates inflammatory cytokine and costimulatory molecule expression in macrophages, and stimulates humoral and cell-mediated immune responses both locally and systemically. These observations are consistent with the hypothesis that activation of APCs is a key step in the induction of antigen-specific immunity, and that inflammatory cytokine expression is a hallmark of activated APC function. The efficacy of vaccine adjuvants, particularly in the context of mucosal immunity, may be determined by their ability to induce a controlled inflammatory response in gut-associated lymphoid tissue, characterized by the expression of various costimulatory molecules and inflammatory cytokines. Thus, elucidation of the patterns of inflammatory cytokine expression and features of APC activation will help to facilitate the rational development of more efficacious vaccines.     

Detection of Infectious Bovine Rhinotracheitis (IBR) by Immunofluorescence

Infectious bovine rhinotracheitis can be readily and accurately diagnosed by direct fluorescent antibody techniques on infected bovine embryo kidney monolayers. Non-specific fluorescence was encountered in direct observation of bovine nasal secretions and cotyledon impressions.     

Bovine respiratory syncytial virus (BRSV): a review

Bovine respiratory syncytial virus (BRSV) infection is the major cause of respiratory disease in calves during the first year of life. The study of the virus has been difficult because of its lability and very poor growth in cell culture. However, during the last decade, the introduction of new immunological and biotechnological techniques has facilitated a more extensive study of BRSV as illustrated by the increasing number of papers published. Despite this growing focus, many aspects of the pathogenesis, epidemiology, immunology etc. remain obscure. The course and outcome of the infection is very complex and unpredictable which makes the diagnosis and subsequent therapy very difficult. BRSV is closely related to human respiratory syncytial virus (HRSV) which is an important cause of respiratory disease in young children. In contrast to BRSV, the recent knowledge of HRSV is regularly extensively reviewed in several books and journals. The present paper contains an updated review on BRSV covering most aspects of the structure, molecular biology, pathogenesis, pathology, clinical features, epidemiology, diagnosis and immunology based on approximately 140 references from international research journals.     

Influence of different tumour types on natural cytotoxicity (NK cell activity) and mitogen-induced lymphocyte proliferation in isolated blood lymphocytes from 110 dogs with tumours

The cell-mediated immune response of blood lymphocytes from 110 untreated dogs with different tumours was evaluated. The influence of different tumour types on the cellular immune system was examined by assessing the percentage of isolated large granular lymphocytes (LGL), in vitro natural cytotoxicity and mitogen-induced lymphocyte proliferation. Although the overall natural cytotoxicity of dogs with different tumours was decreased, the overall difference from control values was not statistically significant. However, mitogen-induced lymphocyte proliferation was significantly depressed in dogs with tumours in comparison with the controls. Dogs with mammary carcinomas showed significantly lower natural cytotoxicity than controls and dogs with myeloid neoplasms showed significantly lower mitogen-induced lymphocyte proliferation. Abnormalities exist not only in natural cytotoxicity but also in mitogen-induced lymphocyte proliferation. For the dog, this is the first study to assess the influence of different tumours using a combined evaluation of natural cytotoxicity and mitogen-induced lymphocyte proliferation in such a large number of animals.     

Bovine interleukin 2 (IL-2) production and activity on bovine and murine cell lines

Long-term growth of T cell cultures requires addition of Interleukin 2 (IL-2). In order to maintain bovine cultures, optimal conditions for bovine IL-2 production were defined using peripheral blood mononuclear cells (PBM). Irradiation and preculture enhanced IL-2 production possibly by reducing suppressor activity. IL-2 activity was also detected in Bovine Herpesvirus Type 1-stimulated cultures. Unlike mitogen-stimulated cultures, a wide variation in IL-2 activity was seen between supernatants produced by virus-stimulated cells from different animals indicating the clonal nature of antigen specific cells from individuals. Bovine IL-2-dependent cells used to quantitate IL-2 activity were characterized as: PNA, esterase negative, H4+ (anti Ia-like), B29+ (anti-pan T cell), and C5- (anti-monocyte). The observations that bovine IL-2 can maintain activated murine cells, CTLL-20 and HT-2, could lead to the replacement of rat IL-2 with bovine IL-2 in long-term murine cultures. Conditions described here result in large volumes of active medium.     

Bovine herpesvirus type 4: a special herpesvirus (review article)

This paper summarizes the history of and information on bovine herpesvirus type 4 (BoHV-4) from the first isolation to the most recent results. For almost twenty years BoHV-4 has been considered a typical herpes 'orphan' virus, which infects several species but causes no illness. The latest experiments revealed the close relationship of this virus with the immune system and other tissues. The virus was even considered as a possible candidate for a vector vaccine. BoHV-4 as a strange herpesvirus has several features which are not characteristic of other herpesviruses, such as several latency sites, persistence in serum, dividing cells necessary for virus replication, and the wide host range. In addition to describing the main features of the virion, replication, clinical signs, nomenclature problems, this review intends to concentrate on the new and strange results coming out from several laboratories worldwide. It is also suggested that because the virus combines several properties of various herpesvirus subfamilies and because of its close relationship with the immune system, it may deserve further attention as a representative of a potentially new genus within the Gammaherpesvirinae subfamily.     

In vitro Fertilization of Bovine Oocytes Exposed to Bovine Herpesvirus 1 (BHV-1)

Contents: Cumulus-oocyte complexes were matured for 24 hours in the presence or absence of bovine herpes virus-1 (BHV-1, 10⁶ TCID₅₀/ml), fertilized in vitro and then co-cultured on oviductal cells to the blastocyst stage. The percentages of oocytes which cleaved were 48% (n = 905) for oocytes matured in the presence of BHV-1, and 51% (n = 1004) for the control oocytes. Comparing BHV-1-treated to control, the percentages of cleaved oocytes which developed to the blastocyst stage were 29% and 31%, respectively (p > 0.05). Embryos produced from the experimental group of oocytes tested positive for BHV-1.     

Bovine Viral Diarrhoea Virus is Internalized by Clathrin‐dependent Receptor‐mediated Endocytosis

Bovine viral diarrhoea virus (BVDV) is a pestivirus within the family Flaviviridae. In contrast to the members of the genus flavivirus, nothing is known about the viral entry route for pestiviruses. In this study, the process of BVDV infection following attachment to the cell surface was examined. BVDV clearly co‐localizes with clathrin, with early endosome antigen‐1 (EEA‐1), an early endosome marker, and also with lysosomal‐associated membrane protein‐2 (LAMP‐2), a lysosomal marker. BVDV internalization is inhibited by compounds that block clathrin‐ but not caveolae‐dependent endocytosis. These findings demonstrate that BVDV enters the cells via the clathrin‐coated pit pathway.     

抗体形成细胞介导的溶血空斑和溶血试验

研究了用绵羊红细胞免疫大鼠,其脾脏中抗体形成细胞中,免疫后第４天平均有８２３个抗体形成细胞。免疫后第３天细胞介导的溶血平均为２２．９９％,免疫后第４天为９２．４５％。讨论了溶血空斑法的条件并与溶血法作了比较。     

利用性别决定基因(SRY)进行牛早期胚胎性别鉴定的研究

利用牛性别决定基因(SRY)和酪蛋白αs1基因(CSN1S1)分别设计雄性特异性引物和内标引物,两对引物均各设计一对嵌套式引物进行多重PCR和多重巢式PCR,用于牛早期胚胎性别鉴定。SRY基因和CSN1S1基因外、内引物扩增片断长度分别为357bp、232bp、528bp和367bp。实验结果表明,这两个引物组合建立的多重PCR扩增体系均有较高的扩增稳定性和准确率,但在常规PCR体系中,它们的灵敏度均不高(约50pg,相当于16个细胞的DNA量),这在一定程度上限制了其在胚胎性别鉴定中的应用。而由二者建立的巢式PCR灵敏度可达到5pg,故可以很好地满足胚胎性别鉴定的需要。     

Bovine babesiosis (Babesia bovis infection) in Zambia

Summary

Two cases of Babesia bovis, a parasite associated with the tick Boophilus microplus, are reported for the first time from the central part of Zambia. It is concluded that infected B. microplus ticks are occasionally introduced into central Zambia by tick‐infested cattle from the north‐eastern part of the country where B. bovis is endemic. The spread of B. microplus in Southern Africa in a westward direction is discussed and related to the epidemiology of bovine babesiosis in Zambia.     

Restraint effects on stress-related hormones and blood natural killer cell cytotoxicity in pigs with a mutated ryanodine receptor

A mutation in the ryanodine receptor gene (RYR1) of the calcium release channel is responsible for increased stress susceptibility in pigs. In the present study, the relation of a mutation in RYR1 with the neuroendocrine (stress-related hormone) response and the immune defense represented by natural killer cell cytotoxicity (NKCC) during a 4-h restraint and recovery phase in 60 male pigs was investigated. Blood samples were collected from pigs previously divided into RYR1 genotypes (nn, Nn, NN), based on PCR amplification and restriction analyses. The blood samples collected during the restraint and recovery phases of the experiment were used to determine NKCC (⁵¹Cr-release assay), large granular lymphocyte number (hematologic method), and plasma concentrations of prolactin (PRL), GH, ACTH, and cortisol (COR) (by specific RIA). The greatest degree of NKCC response (P < 0.05) to restraint stress relative to controls was observed for the stress-susceptible homozygote group (nn). Measures of stress-related hormones were positively correlated with NKCC during the entire experimental period (P < 0.001 for all investigated hormones) in the nn group. Immunostimulatory effects in the early (0–60 min) phase of restraint were associated with increased hormone responses, especially PRL and GH. In the late (180–240 min) phase of stress and the recovery phase (480 min), a decrease in immune response was accompanied by an elevated COR response in all RYR1 genotypes. Moreover, divergent responses of both PRL (greatest in nn, P < 0.001) and GH (greatest in NN, P < 0.001) to the 4-h restraint were observed. Our results suggest that stress-susceptible RYR1-mutated homozygotes develop a greater level of immune defense, including cytotoxic activity of NK cells, and accompanied by more pronounced stress-induced changes in neuroendocrine response than stress-resistant heterozygous (Nn) and homozygous (NN) pigs.     

shRNA介导chTERT基因沉默抑制MDCC-MSB1细胞增殖

几乎所有类型的人类肿瘤都可以检测到端粒酶的活性,通过抑制端粒酶逆转录酶基因(TERT)表达降低端粒酶活性可能是进行抗肿瘤治疗的一个新靶点。本试验通过构建靶向端粒酶TERT基因的shRNA(Sh1,Sh2和Sh3)表达质粒载体转染MDCC-MSB1细胞抑制chTERT基因表达,从而降低端粒酶活性及细胞增殖。Real-time PCR结果显示,Sh1和Sh3在质粒载体转染细胞后48,72,96h显著抑制chTERT基因表达(P<0.05),且呈持续效果;TRAP法端粒酶活性分析显示,Sh1和Sh3在质粒转染细胞72h显著降低端粒酶活性;流式细胞分析显示,Sh3在转染后72h显著降低了S期细胞的比例(P<0.01),G2/M期细胞比例显著上升(P<0.01),G0/G1期细胞比例没有明显变化(P>0.05),抑制了MDCC-MSB1细胞的增殖。结果提示,shRNA通过抑制TERT基因表达可有效降低端粒酶活性,阻滞肿瘤细胞的增殖,为抗癌症治疗提供了新的备选方案及具有参考价值的基础科学数据。     

Bovine anaplasmosis transmission studies conducted under controlled natural exposure in a Dermacentor andersoni = (venustus) indigenous area of eastern Oregon

In Anaplasma marginale transmission studies conducted on the high semi-arid range of eastern Oregon during the 1974 and 1975 vector season, A marginale-susceptible calves (principals) were maintained on 2 raised tick-proof platforms. Anaplasmosis-susceptible control calves of approximately the same age and latent-infected cows grazed the area surrounding the platforms. One latent-infected steer spent the entire 1975 vector season on a platform with the principals. The 28 principals did not develop anaplasmosis, whereas 15 of 30 (50%) controls became infected. The disease was not transmitted from the latent-infected cattle to the principals exposed only to flying hematophagous insects, whereas 50% of the controls exposed to the Rocky Mountain wood tick Dermacentor andersoni) = (venustus) developed the disease. Dermacentor andersoni appears to be the principal vector on this range.