Heritable characters of the red blood cells of sheep

Extract

From the start I want to make it clear that the research work described here is the work of one of us (J.V.E.), and the reason why I am presenting his work is that Evans comes from New Zealand and presented a paper on this subject to the annual meeting of the British Association for the Advancement of Science last September in Sheffield, so that it seemed appropriate that his work should also be presented to this Society.     

Genetic architecture of antibody responses of chickens to sheep red blood cells

SUMMARY: Two lines of White Leghorn chickens selected divergently for high (HH) or low (LL) antibody response 5 days after an injection with 0.1 ml of 0.25% suspension of sheep red blood cell (SRBC) antigen were used to produce parental, reciprocal F(1) , F(2) and backcross progeny. At 36 days of age males and females of the various progeny types were injected with SRBC suspension and antibody titres measured at 5 and 12 days later. Progeny of the high antibody line had higher titres at both 5 and 12 days after inoculation with SRBC than those of the low line. Reciprocal effects for SRBC titres were important only for female progeny suggesting sex-linked effects of the Z chromosome. Titres for F(1) progeny were intermediate and different from the parental lines at both 5 and 12 days after inoculation. Antibody titres 5 days after inoculation exhibited heterosis which emanated from the homogametic sex. Although maternal effects generally had no influence on antibody titres, maternal heterosis in the selected trait was due to sex-linkage. Recombination effects were negligible for both traits. ZUSAMMENFASSUNG: Genetische Architekture der Antik?rperreaktion von Hühnern auf Schaferythrozyten Zwei Wei?e Leghorn-Linien, gegens?tzlich selektiert auf starke (HH) oder niedrige (LL) Antik?rperreaktion 5 Tage nach Injektion von 0,1 ml einer 0,25% Schaferythrozytensuspension (SRBC), wurden zur Erzeugung von parentalen, reziproken F(1) , F(2) und Rückkreuzungsnachkommen herangezogen. In Alter von 36 Tagen wurde den Tieren eine SRBC Suspension injiziert und Antik?rpertiter 5 und 12 Tage sp?ter bestimmt. Nachkommen der HH Linie hatten an beiden Tagen h?here Titer als LL Nachkommen. Reziproke Effekte für SRBC Titer waren nur bei weiblichen Nachkommen wichtig, bedingt wohl durch geschlechtsgekoppelte Z-Chromosom Einflüsse. Die Titer der F(1) Nachkommen waren zwischen denen der Elternlinien. Antik?rpertiter 5 Tage nach Injektion zeigten Heterosis, die im homogametischen Geschlecht auftrat. Obwohl im allgemeinen maternale Wirkungen keinen Einflu? zeigten, war maternale Heterosis beim selektierten Merkmal auf Geschlechtskopplung zurückzuführen. Rekombinations-wirkungen waren vernachl?ssigbar.     

Cryopreservation of sheep red blood cells. 3. Complement titrations with frozen sensitized cells

The effect of different suspending and washing procedures for recovery of sensitized sheep erythrocytes (EA) after freezing at −196°C was investigated. Best results were obtained using gelatin-veronal-buffered saline-sucrose containing 0.15 mM-Ca and 1 mM-Mg (GVBSM⁺⁺-sucrose) as the suspending and first washing buffer. The cryoprotective agents tested were polyvinylpyrrolidone (PVP), neutralized PVP, purified PVP and a gum product, Avelex 1030. All PVP preparations tested gave good results as cryoprotectants in terms of cell recovery after thawing whereas Avelex 1030 was less satisfactory. The EA cells frozen in the presence of untreated PVP showed, however, increased susceptibility to the hemolytic action of complement, whereas cells frozen with purified or neutralized PVP gave titers similar to that obtained with fresh cells. Good results were also obtained with Avelex 1030. Complement titrations with frozen EA cells were more reproducible than titrations with fresh cells.     

The immune response of the chick following viral vaccinations and immunization with sheep red blood cells

The effects of viral vaccinations and immunization with sheep red blood cells (SRBC) on the humoral response of pullets were investigated. Pullets were vaccinated with Marek's disease virus, Newcastle disease virus (NDV), infectious bronchitis virus (IBV), and infectious bursal disease virus at appropriate ages used in commercial practice. At seven weeks, the pullets were intramuscularly immunized with SRBC. NDV and IBV antibodies were detected by hemagglutination-inhibition tests. Hemagglutination (HA) titers were established against SRBC. IBV antibody titers were not affected by vaccination or by immunization with SRBC. NDV antibody titers were significantly increased by vaccination and by immunization with SRBC. The SRBC agglutinin response was also positively affected by vaccination. The HA titer increase consisted of a rise in 2-mercaptoethanol (2-ME)-sensitive antibodies and a fall in 2-ME-resistant antibodies.     

Suppression of immune response to sheep red blood cells in rats experimentally infected with Trypanosoma brucei gambiense

A direct haemagglutination assay for antibodies to sheep red blood cells (SRBC) was used to assess the response of rats infected with Trypanosoma brucei gambiense. Whereas uninfected rats showed an efficient primary and secondary immune response to SRBC, trypanosome-infected rats displayed depressed antibody response starting about six days after infection. Infected rats failed to respond to a challenge dose of SRBC given 14 days after infection while uninfected control animals responded with an increased level of antibody production. These observations showed that T. b. gambiense infection inhibited both primary and secondary immune response to SRBC in rats. The result of this experiment is very important with regard to serological methods used to detect increasing levels of antibody production for diagnosis of diseases caused by bacterial and viral pathogens. In a concurrent trypanosome infection such increasing antibody levels would not be observed, leading to inaccurate diagnosis. Thus trypanosomiasis infection should be excluded under field conditions before the value of a serological diagnosis can be fully utilized.     

Cryopreservation of sheep red blood cells. 1. Experiments with polyvinylpyrrolidone and other protective agents

The protection of sheep erythrocytes at freezing temperatures was investigated using glycerol, dimethylsulfoxide (DMSO), glucose and four different types of polyvinylpyrrolidone (PVP) as cryoprotective agents. Depending on type (molecular weight) and concentration good protection was obtained with PVP, whereas glycerol, DMSO and glucose were unsatisfactory. Recovery of cells after thawing was most successful when the cells had been frozen at a concentration of 1–2 × 10⁹ cells/ml. No cells tolerated freezing at −20 °G. Best results were obtained when the cells were frozen directly in liquid nitrogen (−196°G).     

Effect of malignant catarrhal fever virus infection on the immune response of rabbits to sheep red blood cells

Rabbits infected with African Malignant catarrhal fever virus mounted a depressed antibody response to sheep red blood cells compared to the antibody response of uninfected rabbits. Immunodepression was observed in rabbits immunised 0, 3, 5, 7 and 10 days after infection. Antibody response was not depressed in the rabbits immunised 1 day after infection. Despite the depressed antibody response to SRBC, the rabbits died with rising virus neutralising antibody to the virus. The possible causes of the immunodepression are discussed.     

Effect of infections with swine fever virus on immune functions. III. Antibody response to lipopolysaccharide and sheep red blood cells

In order to determine the effect of infections with low-virulent swine fever virus (SFV) on antibody responses, pigs were administered lipopolysaccharide (LPS) or sheep red blood cells (SRBC), 2 days after infection. Infected pigs showed an enhanced primary response to LPS late during infection. The secondary response to LPS seemed to be unaffected. Both the primary and secondary antibody response to SRBC appeared to be enhanced rather than depressed in infected pigs. These in vivo findings suggest that pigs infected with low-virulent SFV do not develop a depression of B lymphocyte function.     

Selection for antibody response against sheep red blood cells and layer age affect egg quality

1. After 22 generations of divergent selection for antibody response against sheep red blood cells (SRBC), hatchability differed between the selected lines. Whether there is a relationship between hatchability and egg traits in these lines is not clear. 2. The aim of the present study was to investigate whether eggs of selected lines differed in shell and albumen characteristics. 3. Fresh eggs were collected at layer ages of 25, 29, 33, 37, 41, 45, 49, 55 and 59 weeks. In total 776 eggs from three lines (high antibody response (H), control (C) and low antibody response (L) against SRBC) were examined. 4. Albumen height decreased with layer age, but this decrease differed between lines. 5. Eggs of the C line were heaviest, followed by the L line and finally the H line (59.44 vs 55.50 vs 54.15 g, respectively). Eggshell thickness, eggshell percentage, albumen height and albumen pH were lowest in the L line, and highest in the H line, whereas the C line was intermediate. 6. From this study, we concluded that selection on antibody response to SRBC affected egg characteristics (external and internal). This may have consequences for hatchability and furthermore for chick quality. When chick quality partly underlies the immune status of an animal, differences in immune responses among selected lines may be due to differences in egg characteristics. This implies inadvertent selection for chick health at an early stage of life. Whether this is the case needs to be investigated.     

策勒黑羊、和田羊和卡拉库尔羊血液蛋白多态性研究

血液蛋白多态性可以作为一个遗传标记因子来反映动物的遗传变异情况。不同品种间的不同位点血液蛋白具有一定差异,所以由多种血液蛋白多态性所标记的多基因型,可用来标记品种特征（张细权,1997）。本研究利用血液蛋白多态性,对塔里木盆地周边的策勒黑羊、和田羊、卡拉库尔羊3个绵羊品种进行遗传多样性分析,对种质资源保护和科学开发利用以及品种的起源、进化研究提供科学依据。     

Cryopreservation of sheep red blood cells. 2. Purified polyvinylpyrrolidone and hydrolyzed starch as protective agents

The protection of sheep erythrocytes against damage at low temperature (−196°C) was investigated using purified polyvinylpyrrolidone (PVP) and hydrolyzed starch as cryoprotective agents. Identical results were obtained with untreated PVP, neutralized PVP and PVP purified by chromatography. With hydrolyzed starch the cryoprotection was dependent on the type and concentration of the starch used and on the extent of hydrolysis of the starch prior to use. Cell recovery with some starch types was the same (80–85 %) as that obtained with PVP.     

Factors affecting the capacity of Theileria annulata sporozoites to invade bovine peripheral blood lymphocytes

The number of T. annulata sporozoites invading bovine peripheral blood lymphocytes (PBL) under different conditions (in vitro) was determined. Heat-inactivation of T. annulata sporozoites for 45 min, in a thermostatically controlled, shaking water bath preset and stabilised at 60 degrees C resulted in an almost total lack of invasion of fresh, normal PBL by the sporozoites, indicating that the interiorization process is parasite-effected. The mean number of T. annulata sporozoites interiorization (per 1000 lymphocytes) in cultures set up using sporozoites and PBL, mixed and incubated at 0 degrees C for 1 h in melting ice, was highly significantly reduced (P less than 0.01), indicating the invasion of bovine lymphocytes by T. annulata sporozoites is an active process dependent on active metabolism which is markedly affected by temperature. Pre-treatment of PBL with trypsin significantly reduced the number of invading sporozoites thus incriminating proteins or glycoproteins as constituents of receptors involved in sporozoite-lymphocyte recognition.     

Rosetting of bovine T-lymphocytes with autologous and allogeneic red blood cells

Certain bovine peripheral blood lymphocytes (PBL) and foetal thymocytes were shown to bind autologous and allogeneic red blood cells (RBC). When autologous RBC were treated with dextran, approximately 10% of peripheral blood lymphocytes and about 30% of thymocytes were found to form rosettes. Cells forming autologous rosettes appear to be a population of T-lymphocytes because (1) more rosette formation occurred with thymocytes than with PBL, (2) autologous rosette formation was increased in PBL cultures enriched in T cells and was decreased in cultures depleted of T cells, (3) very few rosette forming cells had surface immunoglobulin and (4) peripheral blood mononuclear cell cultures depleted of monocytes did not show a decreased autologous rosette formation. It appears that the cells forming rosettes with autologous and allogeneic RBC belong to the same sub-population of T-cells.