Mitogenic reactivity of mononuclear cells isolated from thymus, spleen and umbilical cord blood of pig foetuses

The in vitro mitogenic reactivity of mononuclear cells from the thymus, spleen and umbilical cord blood of Danish Landrace pig foetuses ranging in gestational age (GA) from 48 to 112 days was monitored by means of a microculture lymphocyte transformation test (LTT). Dose-response profiles for concanavalin A (Con A), pokeweed mitogen (PWM) and leucoagglutinin (LAG) were set up for the various age-groups and the results showed that the onset and development of mitogenic reactivity in the pig foetus is age-related. The results indicate the occurrence of mitogen responsive cells in the thymus and cord blood at 48 days GA and in the spleen at 54 days but statistically significant reactivity (p less than 0.01) for the various tissues could only be demonstrated at later stages of gestation. Thymus cells from all foetuses ranging in GA from 54 to 112 days exhibited significant reactivity to Con A, PWM and LAG. While the first detectable definite response of spleen cells was seen at 60 days GA when 50% of the foetuses exhibited significant reactivity to the 3 mitogens, spleen cells from all foetuses beyond that age responded significantly. Cord blood cells from only 50% of the foetuses of 60 and 70 days GA responded significantly to Con A and PWM but after this stage, cord blood cells from all foetuses did. The first significant response of cord blood cells to LAG was seen at 70 days GA but only in 50% of the foetuses and it was not until 100 days GA that significant reactivity to LAG was detected in all foetuses.     

Immunohistochemical studies on ontogeny of bovine lymphoid tissues.

Developing lymphoid tissues of bovine fetuses ranging from 70 to 270 days of fetal age were examined by histological and immunohistochemical procedures. In the peripheral blood, surface membrane immunoglobulin bearing cells (B-lymphocytes) and sheep red blood cell rosette forming cells (T-lymphocytes) had already appeared by 70 days of fetal age. In the lymph nodes intracytoplasmic IgM positive cells appeared at 90 days of fetal age. The cells positive for IgG appeared at 150 days of fetal age and IgA positive cells appeared at 180 days of fetal age. The spleen contained intracytoplasmic immunoglobulin positive cells at almost the same time as those in the lymph nodes. In the ileocecal region, IgM positive cells and IgG positive cells were present at 180 days of fetal age and IgA positive cells were present at 210 days of fetal age. The tonsils contained IgM positive cells and IgG positive cells at 240 days of fetal age. In the thymus, terminal deoxynucleotidyl transferase positive cells appeared at 90 days of fetal age.     

Detection of in ovo-inoculated infectious bronchitis virus by immunohistochemistry and in situ hybridization with a riboprobe in epithelial cells of the lung and cloacal bursa

In situ hybridization and immunohistochemistry were utilized to identify tissues infected in ovo with infectious bronchitis virus (IBV). Chicken embryos were inoculated in ovo (chorioallantoic sac) with the Arkansas (Ark) serotype of IBV at 18 days of age. At 24, 48, 72, and 120 hr postinfection (HPI), bursa, lung, spleen, heart, and thymus were collected, fixed in 10% neutral buffered formalin, and paraffin embedded. The digoxigenin-labeled antisense S1 riboprobe detected viral mRNA in the cytoplasm of respiratory epithelial cells in the primary bronchus at 24, 48, and 72 HPI. Viral mRNA was detected in bursa samples collected at 48 hr. Immunohistochemistry detected viral antigens in epithelial cells of the parabronchi and bursal tissues at 24 and 48 hr, respectively. No viral mRNA or antigen was detected by in situ hybridization or immunohistochemistry, respectively, in heart, thymus, or spleen at any time after inoculation. On the basis of these data, IBV apparently initially infects lung tissue, then migrates to and infects cells of the bursa. These results indicate that in situ hybridization can be useful in detection of IBV-infected chickens and in understanding the pathogenesis and virulence of IBV infection.     

Characterization and separation of bovine lymphocyte populations

Bovine lymphocyte populations were characterized by surface markers, rosette-forming ability and behaviour towards mitogens. After pre-treatment with neuraminidase 16% of the bovine blood lymphocytes and 14% of the bovine spleen cells formed spontaneous (E) rosettes with sheep erythrocytes. About 20% EAC rosette-forming cells were detected among both cell populations. Protein A receptors were detectable among 8% of the blood lymphocytes and 26% of the spleen cells. Bovine lymphocytes responded to pokeweed mitogen (PWM), phytohemagglutinin (PHA) and concanavalin A (Con A). An enrichment of bovine B and T cells was obtained by E-rosette sedimentation (81–84% B cells) and by filtration through nylon fiber columns (51–65% T cells). The T cells obtained after nylon filtration still responded to the mitogens PHA, Con A and PWM. Enriched B-cell populations responded to bacterial lipopolysaccharide (LPS). After monocyte depletion the mitogenic response of blood lymphocytes was not influenced.     

Ontogenesis of the reticulum with special reference to neuroendocrine and glial cells: a comparative analysis of the merino sheep and iberian red deer

The present study was designed to compare the differences in the ontogenesis of the reticulum in sheep (domestic ruminant) and deer (wild ruminant). A total of 50 embryos and foetuses Merino sheep and 50 Iberian deer were used, from the first pre‐natal life until birth. The appearance of the reticulum from the primitive gastric tube was earlier in the sheep (22% gestation, 33 days) than in the deer (25% gestation, 66 days). In both cases, it displayed a primitive epithelium of a stratified, cylindrical, non‐ciliary type. At around 48% gestation in the sheep (72 days) and 36% (97 days) in the deer, the reticulum was configured of four clearly differentiated layers: mucosa (with epithelial layer and lamina propria), submucosa, tunica muscularis and serosa. The stratification of the epithelial layer was accompanied by modifications in its structure with the appearance of the primitive reticular ribs. The primary ribs began to be formed first in the deer, at 117 days of pre‐natal life (40% gestation) and later in the sheep (79 days, 53% gestation). The differentiation of the corneum papillae in the primary ribs coincided with the appearance of secondary reticular ribs. These structures began to be formed first in the deer, at 142 days of pre‐natal life (51% gestation) and later in the sheep (83 days, 55% gestation). The presence of neuroendocrine cells (non‐neuronal enolase‐positive cells) in the reticular mucosa was not detected until 97 days (36% gestation) in deer and 81 days (54% gestation) in sheep. The presence of glial cells (GFAP‐positive cells) occurred at around 142 days (51% gestation) in deer and at 112 days (75% gestation) in sheep. In conclusion, the presence of neuroendocrine and glial cells was detected in deer at earlier stages than sheep.     

鸡传染性法氏囊病的病理学研究

人工接种２８日龄非免疫鸡传染性法氏囊病病毒（ＩＢＤＶ）后,对感染鸡的法氏囊、胸腺、脾、盲肠扁桃体、哈德氏腺、肝、肾进行病理组织学检查。感染后４８ｈ,法氏囊淋巴组织最早出现坏死且长久存在。其他淋巴器官的病变出现较迟,程度轻微且恢复较快。ＩＢＤＶ单抗免疫荧光检测,法氏囊及其他淋巴器官中均检测到病毒,接种后１２ｈ法氏囊中即检出病毒,持续时间也最长（攻毒后１２ｄ）,其次是盲肠扁桃体（攻毒后８ｄ）。攻毒１３ｄ以后,上述器官均未检测到病毒。法氏囊粘膜上皮的扫描电镜观察,攻毒后２ｄ,上皮细胞肿胀,微绒毛减少或消失。攻毒后３ｄ,局部上皮细胞坏死、脱落,并向整个粘膜层扩展,攻毒后１０ｄ,上皮层基本修复。     

Age Difference in Morphology and Immunohistology inthe Thymus and Spleen in Crl:CD (SD) Rats

We investigated chronological changes in immunohistochemical phenotyping in the thymus and spleen in Crl:CD rats up to the age of about one year. In the thymus, T cells increased markedly from 3 to 4 weeks of age. Proliferating cells also increased markedly at these points. B cells tended towards an increase with age. In the spleen, white pulp increased until 9 weeks of age and remained fairly stable thereafter. In the periarteriolar lymphoid sheath and marginal zone, T cells gradually increased until 9 weeks of age and became almost flat thereafter. In the lymph follicle, T cells increased with age. B cells tended towards an increase with age in all areas of the spleen. It was concluded that development of the thymus was most marked from 3 to 4 weeks of age and that both the thymus and spleen had matured by 9 weeks of age.     

Divergent selection of chickens for antibody production to sheep erythrocytes: age effect in parental lines and their crosses

Age dependency of antibody response to sheep red blood cell (SRBC) antigen was measured in lines of chickens divergently selected for this trait and in reciprocal crosses between them. At 7 days of age, there were differences among populations for frequency of responders to SRBC antigen. This qualitative pattern persisted in the quantitative context of the antibody titers of those who responded, demonstrating genetic differences in both the event and subsequent levels of antibody. Although chickens from the high line had significantly higher titers than those from the low line and cross populations, all reached serological maturity by 14 days of age. From this age, high-line chickens had higher bursa and spleen weights and lower thymus weights relative to body weight than those from the low line: relative to body weight, spleen and bursa weights increased at a faster rate through 19 and 25 days of age, respectively, and then plateaued. In contrast, there was a progressive increase in thymus weight relative to body weight through 40 days of age.     

Morphology of lacrimal gland in pig fetuses

The morphological and histological examinations of the lacrimal gland were conducted on pig fetuses coming from the 20th, 24th, 27th, 30th, 35th, 50th, 63rd, 94th and 112th day of gestation. The morphological examinations were carried out using the method of macroscopic preparation with a forehead magnifying glass and binocular (magnification 1.5-5.0x). In order to better visualize the anatomical elements, 60-80% absolute alcohol and 0.5-4% acetic acid solution were used for the examinations. On the 20th, 24th, 27th, and 30th day of gestation the whole fetuses were collected for the histological examinations. The whole eyeball with developing accessory organs was collected from the pig fetuses on the 35th day of gestation. On the 50th, 63rd, 94th and 112th day of gestation only the lacrimal gland was collected. Staining with H-E and Azan method was performed. On the 20th, 24th, 27th, 30th and 35th day of gestation ectodermal cells were not found in the collected material. On the 50th and 63rd day of gestation the connective tissue divides the gland parenchyma into indistinct lobes composed of gland cells. On the 94th day of gestation the number of lobes is substantially higher than on the 50th and 63rd day of gestation, while the number of lobules forming lobes decreases. On the 112th day of gestation each lobe is composed of 8-22 excretory ducts made up of the simple cuboid epithelium with a round nucleus arranged less or more peripherally.     

Monoclonal antibodies directed towards the two major cell populations in the bursa of Fabricius of the chicken

Two mouse monoclonal IgM antibodies, B.1 and B.2, have been produced using the mouse myeloma cell line Sp2/0-Ag 14 and spleen cells from mice immunized with chicken bursa cells. The binding of the monoclonal antibodies to cells in suspension or tissue sections was demonstrated by means of the unlabeled peroxidase-antiperoxidase method. B.1 recognizes 61% of the bursa cells, 10-14% of the cells of spleen and of the peripheral mononuclear blood leukocytes and 1% of the thymus cells. The B.1+ cells are regarded as B cells. Their location in tissue sections corresponds with the known B-dependent areas of lymphoid organs. Competitive binding and double marker experiments proved that the B.1 antigen is distinct from surface immunoglobulin (Ig). In the bursa all B.1+ cells are also Ig+, whereas in the thymus, spleen and blood only about 90% of the B.1+ cells show this conformity. B.2 mainly recognizes so called reticular epithelial and reticular cells of the bursa (36%), thymus (20%) and spleen (13%). The B.2+ cells represent the second major cell population of the bursa.     

妊娠后期母羊饲粮精料比例对羔羊生长性能、消化性能及血清抗氧化指标的影响

本试验旨在研究妊娠后期母羊饲粮精料比例对羔羊生长性能、消化性能和血清抗氧化指标的影响。试验选用66只妊娠90 d、平均体重为(44.45±2.20)kg的初产湖羊,按照体重相近原则随机分为3组,每组11个重复,每个重复2只。各组母羊妊娠期饲粮精料比例分别为50%、40%和30%,分娩后母羊饲喂相同的全混合日粮(TM R)。羔羊10日龄,每只母羊取1只羔羊断母乳,饲喂代乳粉;15日龄补饲开食料;20日龄补饲苜蓿干草,自由采食至60日龄。每10 d测定1次羔羊体重,51~60日龄进行羔羊消化代谢试验,20和60日龄采集羔羊血液测定血清抗氧化指标。结果表明:妊娠后期母羊饲粮精料比例对羔羊1、10、20、30、40、50、60日龄体重,20、60日龄体尺指标及营养物质表观消化率均无显著影响(P0.05)。随母羊饲粮精料比例降低,20日龄羔羊血清总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)活性极显著升高(P0.01),60日龄50%组显著或极显著低于其他2组(P0.05或P0.01);30%组20和60日龄羔羊血清谷胱甘肽过氧化物酶(GSH-Px)活性极显著高于其他2组(P0.01);血清中MDA的含量在20日龄时随母羊饲粮精料比例的降低而极显著降低(P0.01),60日龄时50%组显著高于30%组(P0.05)。结果提示,妊娠后期母羊饲粮的精料比例对产后早期断奶羔羊的体重、体尺、营养物质表观消化率无显著影响,但随母羊饲粮精料比例的降低,羔羊血清的抗氧化能力提高。     

Porcine interleukin 2: parameters of production and biochemical characterization

Interleukin 2 (IL2) or T cell growth factor (TCGF) has been characterized in a number of species but not in porcines. Porcine IL2 was detected in supernates (SN) of cultures of pig lymphocytes by: 1) the stimulation of the IL2-sensitive murine T cell line, CT6; 2) a costimulator assay involving porcine thymocytes; and 3) by the in vitro maintenance of antigen or mitogen-induced porcine lymphoblastoid cells. Porcine IL2 production by pig lymphocytes was induced by the mitogens Concanavalin A (Con A) Phytohemagglutiniin (PHA), and Pokeweed mitogen (PWM), but not by lipopolysaccharide (LPS). IL2 activity was demonstrated in the SN of mitogen-stimulated lymphocyte cultures as early as 24 hr after initiation of culture, reached peak levels at 48 hr, and decreased by 72 hr. Mitogens induced IL2 secretion by pig peripheral blood mononuclear cells, lymph node cells, and spleen cells, but not thymus cells. The cells responsible for IL2 production are presumptive T cells because: 1) they are nylon wool non-adherent; and 2) are non-surface-Ig bearing. In contrast, SN from cultures of surface Ig-positive cells had minimal IL2 activity. Porcine IL2 resembles rat and human IL2 in that it has an apparent molecular weight of approximately 15,000, and does not bind to DEAE-cellulose (DE-52) ion exchange columns equilibrated in 0.05 M sodium phosphate buffer (pH 7.6).     

Surface markers for characterisation of bovine blood lymphocyte populations and changes in these from birth to maturity

A sheep erythrocyte (E) rosette technique was developed for use with cattle lymphocytes. This involved the use of 17 per cent Ficoll 400 and preservative-free heparin (84 iu/ml) in the saline-erythrocyte mixture. Using this technique, 83 per cent of peripheral blood lymphocytes in cattle aged between six and 10 years were found to form E rosettes. The remaining cells (17 per cent) were B-cells, so that no cells remained unmarked. Lymphocytes from very young calves contained a population of unmarked or null cells, but these rapidly diminished as the animals matured. A peak of total lymphocytes recovered from blood, as well as E rosette-forming cells, occurred in calves aged four to six months. The non-E rosette-forming cells were mostly B-cells and it was suggested that this was associated with calf weaning. The total number of lymphocytes recovered, as well as E rosette-forming cells, gradually fell with the age of the cattle sampled. Null cells were virtually absent from the blood of cattle six years and older. Bovine T-cells could be further subdivided into Fc mu, Fc gamma and C' receptor-bearing subpopulations on the basis of overlap with R rosette-forming cells. Some further separation of these cells from B-cells was achieved using density gradient centrifugation on Percoll. Separation of E rosette-forming cells with Fc receptors from E rosette-forming cells without Fc receptors was achieved by nylon wool columns, to which the Fc receptor bearing cells were adherent. It was concluded that bovine blood lymphocytes had blood T-lymphocyte populations with markers which may correspond to the 'helper' (Fc mu ) and 'suppressor' (Fc gamma ) populations described for the human.     

Distribution patterns of mast cells in the uterus of pregnant Meishan pigs

The present study was performed to investigate the numerical distribution of mast cells (MCs) in the uteri of pregnant Meishan pigs to explore the functions of MCs in pig pregnancy. The uterine samples from pregnant (on days 15, 26 and 50 of gestation) pigs were obtained respectively and stained with toluidine blue. The results were as follows: MCs were constitutively located in the uterus of the Meishan pig, with the distribution varying with gestational stages. On days 15 and 26 of gestation, MCs were mainly distributed around the blood vessels and uterine glands within the endometrium. On day 50 of gestation, MCs were mostly distributed in the myometrium. These results indicated that uterine MCs possibly have versatile functions in pig pregnancy.     

Morphology of deep gland of the third eyelid in pig foetuses

The morphological and histological examinations of the deep gland of the third eyelid were carried out on pig foetuses coming from the 35th, 50th, 63rd, 94th and 112th day of gestation. The morphological examinations were conducted using the method of macroscopic preparation with a forehead magnifying glass and binocular (magnification 1.5-5.0x). In order to make anatomical elements more visible, 60-80% absolute alcohol and 0.5-4% acetic acid solution were used for the examinations. For the histological examinations, the whole eyeball with developing accessory organs was collected from the pig foetuses on the 35th day of gestation. On the 50th, 63rd, 94th and 112th day of gestation only the deep gland of the third eyelid was collected. Staining with haematoxylin-eosin and Azan method was performed. It was found during the examinations that the process of the formation of the deep gland of the third eyelid starts on the 35th day of gestation. On the 50th day of gestation, the gland cells are evenly distributed in the connective tissue stroma. On the 63rd day of gestation, the connective tissue divides the gland parenchyma into indistinct lobes composed of 6-15 lobules. On the 94th day of gestation, the gland lobes become visible; the efferent ducts are situated in the central part of the lobe. On the 112th day of gestation, the lobes are composed of a high number of lobules composed of two kinds of excretory ducts. The first type of the excretory ducts is lined with the simple cuboid epithelium whose nuclei are situated at the base of the cell. The other type of the excretory ducts is lined with the simple cuboid epithelium whose nuclei are round and arranged less or more peripherally.     

饲粮添加甜菜碱对巴马香猪血常规指标和器官生长的影响

本试验旨在研究饲粮添加甜菜碱对巴马香猪血常规指标及器官生长的影响。选用26头体况相近的妊娠巴马香猪,随机分为2组,其中对照组12头,饲喂基础饲粮;试验组14头,在基础饲粮中添加3.5 kg/t甜菜碱。仔猪28日龄断奶并适应7 d后,每窝选取4头接近平均体重的仔猪,随机分为2组,每组2头,对照组或试验组的每2窝中的2头仔猪合并为1栏。对照组12栏,母猪添加组7栏,均饲喂基础饲粮;母-子猪添加组7栏,在基础饲粮中添加2.5 kg/t甜菜碱。分别于仔猪95和125日龄时,每栏选择接近平均体重的巴马香猪1头,前腔静脉采血,检测血常规指标;处死后分离心脏、肝脏、脾脏、肺脏、肾脏和胃并称重并计算器官指数。结果表明,与对照组相比,母-子猪添加组95日龄时的嗜酸性粒细胞数(EOS)、体重、脾脏重和脾脏指数显著升高(P<0.05),平均血小板体积(MPV)显著降低(P<0.05);125日龄时的嗜碱性粒细胞百分比(BAS%)、体重、心脏重、肝脏重、脾脏重、肾脏重、胃重和肝脏指数显著升高(P<0.05)。与母猪添加组相比,母-子猪添加组95日龄时的体重、心脏重、肝脏重、脾脏重和肾脏重以及125日龄时的心脏重、肝脏重、肾脏重、胃重和肝脏指数显著增加(P<0.05)。由此可见,妊娠巴马香猪及其子代饲粮添加甜菜碱可影响血液中EOS、BAS%和MPV,增加体重,促进器官生长。     

酶解谷朊粉对肉仔鸡免疫功能的影响

为了探究酶解谷朊粉对肉仔鸡免疫功能的影响,本试验选用108只肉鸡,随机分为对照组、谷朊粉组和酶解组3个处理组,每组3个重复。对照组饲喂全价基础日粮,谷朊粉组饲喂添有1.2%谷朊粉的日粮,酶解组饲喂添有1.2%谷朊粉酶解液的日粮。结果表明,日粮中添加谷朊粉酶解物后,在21日龄时,酶解组肉仔鸡的胸腺、法氏囊指数较对照组均有显著提高（P〈0.05）,42日龄时,各处理组肉仔鸡的胸腺、法氏囊指数差异均不显著（P〉0.05）,而在脾脏指数方面,无论是21日龄还是42日龄,添加酶解谷朊粉组脾脏指数均呈升高趋势（P〉0.05）。对于整个试验期,胸腺、法氏囊的发育有不同程度的促进作用,其中对早期免疫器官的发育具有显著的改善作用。在肉仔鸡整个生长阶段,日粮中添加谷氨酰胺可提高新城疫HI滴度,还可促进肉鸡外周血T淋巴细胞的分裂增殖,其中以42日龄时酶解组提高最为显著。     

Immunoglobulins in porcine umbilical cord blood and maternal placenta.

Studies were made of the immunoglobulin (Ig) in serums from umbilical cord of newborn pigs and maternal placenta. The neutralization test for porcine parvovirus and Japanese encephalitis virus was carried out with the serum of the sow and that of the umbilical cord of the newborn pig. Comparative studies of the serums from the dam and the umbilical cord were also done with gel filtration. Of 20 umbilical cord serum samples, IgG was seen in 5 samples (25%), IgA in 1 sample (5%), and IgM in 9 samples (45%). The amount of any 1 of the 3 classes of Ig in the serums was between 13.5 and 28.0 mg/dl. Among the samples examined, 1 had both IgG and IgA and 1 had IgG and IgM, but none had both IgA and IgM and none had 3 classes of immunoglobulins (i.e., IgG, IgA, and IgM). Only 7 samples (35%) did not have any class of Ig. The IgG disappeared from the blood of hysterectomy-produced colostrum-deprived pigs at 3 days of age, and IgM disappeared when pigs were 7 days of age. Neutralization antibodies of porcine parvovirus and Japanese encephalitis virus in maternal serum were not transferred to the fetus through the placenta. Results of immunohistologic surveys indicated that the sow's Ig were not transferred to the fetus through the placenta. Therefore, it is believed that the Ig in the porcine fetus might be synthesized in certain cells in the placental tissue, and the degree of production of the Ig in the placental tissues may differ in each case. The component, which seems to be Ig, was observed as the obscure band of the beta- to gamma-globulin area in serum of the umbilical cord. Comparison was made, with gel filtration, of maternal serum and serum from the umbilical cord of the newborn pig originating from the same sow. Seemingly, the IgG in the umbilical cord serum is mainly in the lower molecular weight fraction, whereas IgG in the sow's serum was distributed in the high to low molecular weight fractions.     

Morphology of the third eyelid and superficial gland of the third eyelid on pig fetuses

The morphological and histological examinations of the third eyelid and superficial gland of the third eyelid were conducted in pig fetuses coming from the 20th, 24th, 27th, 30th, 35th, 50th, 63rd, 94th and 112th day of gestation. The morphological examinations were carried out by applying the method of macroscopic preparation with a forehead magnifying glass and binocular (magnification 1.5-5.0x). In order to make anatomical elements more visible, 60-80% absolute alcohol and 0.5-4% acetic acid solution were used for the examinations. On the 20th, 24th, 27th and 30th day of gestation, the whole fetuses were collected for the histological examinations. The whole eyeball with developing accessory organs was collected from the pig fetuses on the 35th day of gestation. On the 50th, 63rd, 94th and 112th day of gestation, only the superficial gland with the third eyelid was collected. Staining with haematoxylin-eosin (H-E) and Azan method was performed. On the 20th, 24th, 27th and 30th day of gestation, the primordia of the glandular epithelium were not found in the examined material. The process of the third eyelid and superficial gland formation starts on the 35th day of gestation. On the 50th and 63rd day of gestation, the gland surrounding the cartilage of the third eyelid is composed of the high amount of loose connective tissue and gland cells which give rise to excretory segments. On the 94th day of gestation, the gland lobes become visible, the efferent ducts form. On the 112th day, the cartilage of the third eyelid assumes the appearance of the mature hyaline cartilage. The excretory segments are composed of simple cuboid epithelium with a large, round nucleus arranged less or more peripherally. Their number increases 2- or even 3-fold at the end of gestation.     

Analysis of a sheep anti-pig T lymphoblast serum with specificity for E rosette-forming lymphocytes

Sheep antibodies against a pig E-rosette-forming lymphoblastic T lymphoma raised by two intravenous injections of 10(10) cells showed little lymphocytotoxic activity which could be absorbed with red cells, alveolar macrophages or kidney or liver cell homogenates. Bone marrow absorption yielded subpopulation specific antibody which binds to E rosette-forming cells (E.RFC) using either complement-mediated cytotoxicity or indirect antiglobulin rosette formation. In 30 blood lymphocyte preparations from 20 pigs with a range of approximately 20-85% E rosettes the mean E% 43.5 +/- 2.7 agreed with the % antigen+ cells by cytotoxicity mean = 42.6 +/- 2.7 and in each individual sample these figures also agreed closely. In samples of blood lymphocytes enriched and depleted for E rosettes, results of %E+ also agreed closely with % antigen+ cells. This relationship also held for thymocytes and the specific antibodies could be completely absorbed with thymocytes. These data show that the antibody identified peripheral and thymic E.RFC. Bound to lymphocytes the antibody inhibited E rosette-formation with sheep red blood cells (SRBC) in saline (S) and dextran (DS) and with pig RBC in dextran and in Ficoll, but did not affect B cells shown by immunofluorescence, direct antiglobulin rosette formation or Fc rosette-formation, either in saline or dextran, (which include T gamma cells). E rosette inhibition was dependent on antibody concentration, showing single and double sigmoid curves for S and DS rosettes respectively, consistent with differing ease of inhibition of the strong and weak rosette formation. The same spectrum of inhibition of rosette formation by antibody binding followed subsequent incubation with C'6-deficient rabbit serum, but with C'-sufficient serum resulted in loss of cells which require dextran for Fc rosette-formation (T gamma). Thus the serum reveals E rosette-forming T cells and their subpopulations, perhaps by binding to the SRBC receptor.