In vitro modulating effects of porcine immunoglobulin G on mitogens-induced lymphocyte response in precolostral, suckling and weaned piglets

The influence of allogeneic IgG on in vitro reactivity of peripheral blood lymphocytes (PBL) of neonatal colostrum-deprived piglets as well as of suckling and weaned piglets was studied. PBL were preincubated with purified allogeneic IgG for 24 h before their ability to respond to PHA, Con A or PWM was tested. PBL of precolostral piglets pretreated with allogeneic IgG exhibited higher response to PHA (P less than 0.01) than untreated control cells. An increased response of PBL treated with IgG was also observed in suckling piglets as compared to their respective control cells (P less than 0.01). Responsiveness of PBL treated with IgG to PWM was suppressed. No differences in response to Con A regardless of the sources of lymphocytes was observed as compared to IgG untreated controls. The results suggest that pretreatment of lymphocytes of piglets with allogeneic IgG modulates their reactivity to mitogens, suppressing the response to PWM and stimulating the response to PHA, respectively.     

Transmissible gastroenteritis virus antibody production in vitro by porcine peripheral blood leukocytes.

The purpose of this study was to demonstrate the in vitro production of transmissible gastroenteritis virus (TGEV)-specific antibodies by peripheral blood leukocytes (PBL) harvested from piglets infected with TGEV. Piglets were infected with the virulent Purdue strain of TGEV and at intervals postinfection their PBL were cultivated in the presence of TGEV antigen, control antigen or pokeweed mitogen (PWM). The culture supernatants were tested for TGEV antibodies by a fixed cell enzyme immunoassay. Antibodies were never found in the supernatants of unprimed PBL cultures from control piglets, nor in cultures stimulated with control antigen, and antibodies were produced more frequently in response to stimulation of primed PBL with viral antigen than with PWM. In PBL cultures stimulated with viral antigen, TGEV antibodies of the IgG class were produced more frequently than IgA class antibodies. Optimal antibody responses were produced by PBL harvested two weeks after infection and cultivated at a concentration of 10(7) cells/mL for five days.     

The role of interferon in spontaneous cell-mediated cytotoxicity in pigs

Specific release of 51Cr and the production of interferon (IFN) increased in parallel in a spontaneous cell-mediated cytotoxicity (SCMC) assay in which uninfected PK-15 cells or PK-15 cells persistently infected with transmissible gastroenteritis virus (PK-15-TGE cells) were used as targets, and peripheral blood lymphocytes (PBL) from a young adult pig were used as effector cells. Higher levels of both specific 51Cr release and IFN were obtained in the assays containing PK-15-TGE cells. Co-cultivation of PBL from newborn piglets with PK-15-TGE cells yielded similar levels of IFN to those produced by co-cultivation of adult PBL and PK-15-TGE cells, but lower levels of IFN were produced by co-cultivation with uninfected PK-15 cells. Pretreatment of adult PBL with IFN augmented their SCMC effector activity for both PK-15 and PK-15-TGE cells in the 51Cr release assay. Pretreatment of the PK-15-TGE target cells with IFN did not affect their release of 51Cr in the SCMC reaction, while IFN pretreatment of PK-15 targets protected them against SCMC. In a single cell cytotoxicity assay the effects of IFN pretreatment on the effector adult PBL and on the PK-15 and PK-15-TGE target cells were confirmed, and SCMC incompetent PBL from neonatal piglets were rendered cytotoxic by pretreatment with IFN. PBL from newborn piglets bound to either target cell with the same frequency as PBL from SCMC competent adult pigs, and IFN pretreatment of either effector or target cells had no effect on target-binding frequency.     

Long term substitution and specific immune responses after transfer of bovine peripheral blood lymphocytes into severe combined immunodeficient mice.

The long term immune responsiveness of bovine peripheral blood lymphocytes engrafted into severe combined immunodeficient mice (bovine PBL SCID mice) was analyzed. After intraperitoneal transfer (i.p.) of 2x10(7) bovine PBL into SCID mice, FACS analysis revealed successful engraftment of bovine CD4 and CD8+ T cells in the peritoneal cavity, the peripheral blood, spleen, lymph nodes, bone marrow, and thymus of reconstituted mice for up to 13 weeks. As shown by immunocytochemistry in sections of spleens from SCID mice 16 weeks after substitution, bovine T and B cells were localized perivasculary forming pseudofollicular structures. Nevertheless, histopathology of spleen and liver from bovine PBL SCID mice revealed pathological alterations indicating rejection of xenogenic cells or graft versus host disease (GVHD). On the functional level, i.p. transfer of bovine PBL into SCID mice induced increasing levels of bovine IgM and IgG in the sera of recipients. Bovine Ig could be detected up to 20 weeks. Immunization of SCID mice reconstituted with PBL of normal donors with dinitrophenol (DNP)-edestin induced a weak specific bovine antibody response in recipient mice. In contrast, a secondary specific bovine IgG response was observed after antigen restimulation of SCID mice reconstituted with PBL from calves preimmunized either with DNP-edestin or keyhole limpet hemocyanin (KLH) showing functional T cell-independent and -dependent antibody responses of bovine PBL SCID mice. Our data demonstrate that transfer of bovine PBL into SCID mice leads to a long term engraftment of bovine cells in lymphatic and non-lymphatic organs inducing a functional substitution of T and B cell immune response of SCID mice. Therefore, bovine PBL SCID chimera can serve as a small animal model for the analysis of bovine lymphopoiesis and infectious diseases of cattle.     

Responses of enriched populations of feline T and B lymphocytes to mitogen stimulation.

Lymphocytes from healthy adult cats were separated into T and B cell-enriched subfractions by centrifuging rosetted cells on sodium metrizoate/Ficoll gradients. The responsiveness of unseparated lymphocytes (T + B), and T and B cell-enriched subfractions to stimulation with mitogens (phytohemagglutinin-P (PHA-P), concanavalin A (ConA), and pokeweed mitogen (PWM) was tested. Cultures of unseparated lymphocytes and those enriched in T cells showed similar responsiveness of PHA, ConA, and PWM stimulation; however, only a weak response to ConA and PWM was observed in B cell-enriched cultures. The mitogenic effects of PHA-P, ConA, and PWM on feline lymphocytes appeared to be due primarily to T-cell activation.     

Phenotypic comparison of ileal intraepithelial lymphocyte populations of suckling and weaned calves

Ileal intraepithelial lymphocyte (IEL) suspensions from suckling calves (1-3 weeks old) and weaned calves (3-6 months old) were phenotyped to determine whether there were differences in the lymphocyte populations consistent with postnatal maturation of the mucosal immune system. Flow cytometric comparisons of IEL from the two age groups revealed the presence of significantly larger proportions of CD4+ T lymphocytes and CD8+ T cells in the weaned animals. In contrast, there was a significantly larger proportion of B-B2+ IEL in the suckling calves. Freshly isolated IEL from both groups of calves expressed mRNA for TNF-alpha and IFN-gamma, but not IL-4 or IL-10. The B-B2+ IEL population was more closely examined by flow cytometry. These cells co-expressed IgM and CD21. However, they did not express IgA, IgG1, nor any of several additional leukocyte differentiation molecules. Immunohistochemical data confirmed the presence of IgM+ lymphocytes, and the paucity of IgA+ and IgG1+ lymphocytes in suckling calf ileum. However, substantial numbers of IgA+ and IgG1+ cells were observed in weaned calf ileum. Together, the data are consistent with ongoing postnatal maturation of the gut mucosal immune system.     

Activation of natural killer cells in newborn piglets by interferon induction

Natural killer (NK) cell activity in the peripheral blood lymphocytes (PBL) of newborn piglets, normally negligible, was stimulated by in vitro treatment with porcine type I interferon (IFN), and the NK activity of PBL from weaned piglets was augmented by the same treatment. Binding of the PBL to the PK-15 targets used in the single cell cytotoxicity assay for NK activity was not affected by age or by IFN treatment. When newborn piglets were treated with a single intravenous dose at 2 days of age of 0.5 mg/kg of polyinosinic:polycytidylic acid complexed with poly-L-lysine and carboxymethylcellulose (poly ICLC), a synthetic IFN inducer, their IFN levels peaked at 6 h post-induction, and NK activity in their PBL peaked at 24 h post-induction at the level normally found in weaned piglets. The NK activity then declined until 7 days post-induction, when it increased again in a similar manner to that in untreated control piglets. Target-binding of the PBL was not affected by poly ICLC treatment of the piglets. Newborn piglets treated with poly ICLC and subsequently exposed to infection with transmissible gastroenteritis (TGE) virus showed a delay in onset of clinical signs of TGE compared with untreated control piglets. It was concluded that NK cells in newborn piglets can be activated by treatment of the piglets with poly ICLC, and that the presence of active NK cells is associated with some increase in resistance to challenge with TGE virus.     

Effect of dietary supplementation of oregano essential oils to sows on colostrum and milk composition, growth pattern and immune status of suckling pigs

This study evaluated the effects of supplementing sow diets with oregano essential oils (OEO) during gestation and lactation on sow colostrum and milk composition and on the growth pattern and immune status of suckling pigs. A total of 70 second-parity sows were randomly assigned to 1 of 2 gestation dietary treatments within 24 h after service: control (CON) or CON + 250 mg/kg of OEO (OREG). In lactation, sows were again assigned to either the CON or OREG dietary treatment. Thus, the lactation treatments were CON-CON, CON-OREG, OREG-CON, and OREG-OREG. Colostrum and blood samples were collected from 6 sows per lactation dietary treatment. Thymus lymphocyte (T lymphocyte) subpopulations (γδ, cluster of differentiation 8, and 32 cluster of differentiation 4) were enumerated in blood and mammary secretions along with IGF-1, IgG, and IgA concentrations. Piglet growth rate were determined from 18, 17, 17, and 18 litters from the CON-CON, CON-OREG, OREG-CON, and OREG-OREG lactation dietary treatments, respectively. Growth rates were determined in 630 piglets, and piglets were individually identified and weighed on 1, 5, 9, 12, 16, and 19 d of age. Oregano essential oil supplementation during gestation or lactation had no effect (P > 0.05) on GE, CP, GE:CP, GE:fat, and IGF-1 in sow milk. Reductions of the fat percentage in milk on d 7 (P < 0.05) and d 14 (P = 0.07) were found in sows supplemented with OEO during lactation compared with those in the CON treatment. Milk from sows supplemented with OEO during lactation had the greatest number of T lymphocytes compared with those in the lactation CON treatment on d 14 of lactation (P < 0.01). The number of T lymphocytes in milk was greater for sows in the CON-OREG treatment compared with those other treatments on d 14 of lactation (P < 0.05). Energy intake was greater on d 1 to 5 in piglets from sows fed OEO during gestation than those from sows in the CON treatment (P < 0.05). A trend (P = 0.10) for greater milk intake was observed in piglets from sows supplemented with OEO during gestation compared with those from sows in the CON treatment. Similarly, a tendency for an increase in ADG on d 1 to 5 was found in piglets from sows supplemented with OEO during gestation compared with those from sows in the CON treatment (P = 0.10). Insulin-like growth factor-1 at birth and on d 7 and 14 of lactation did not differ among piglets from sows assigned to the different dietary treatments. Oregano essential oil supplementation of sow diets did not affect (P > 0.05) immunoglobulin concentrations in piglets after suckling. Supplementing sow diets with OEO during gestation or lactation did not affect (P > 0.05) the T lymphocytes, percentage of T-lymphocyte subpopulations, and natural killer cell activity of piglets during lactation. Supplementing sow diets with 250 mg/kg of OEO during gestation and lactation did not affect the growth potential of and immune responses in suckling piglets.     

The comparative profile of lymphoid cells and the T and B cell spectratype of germ-free piglets infected with viruses SIV,PRRSV or PCV2

Lymphocyte subsets isolated from germ-free piglets experimentally infected with swine influenza virus (SIV), porcine reproductive and respiratory syndrome virus (PRRSV) or porcine circovirus type 2 (PCV2) were studied and the profile of these subsets among these three infections was monitored. Germ-free piglets were used since their response could be directly correlated to the viral infection. Because SIV infections are resolved even by colostrum-deprived neonates whereas PRRSV and PCV2 infections are not, SIV was used as a benchmark for an effectively resolved viral infection. PRRSV caused a large increase in the proportion of lymphocytes at the site of infection and rapid differentiation of B cells leading to a high level of Ig-producing cells but a severe reduction in CD2^—CD21⁺ primed B cells. Unlike SIV and PCV2, PRRSV also caused an increase in terminally differentiated subset of CD2⁺CD8α⁺ γδ cells and polyclonal expansion of major Vβ families suggesting that non-specific helper T cells drive swift B cell activation. Distinct from infections with SIV and PRRSV, PCV2 infection led to the: (a) prevalence of MHC-II⁺ T cytotoxic cells, (b) restriction of the T helper compartment in the respiratory tract, (c) generation of a high proportion of FoxP3⁺ T cells in the blood and (d) selective expansion of IgA and IgE suggesting this virus elicits a mucosal immune response. Our findings suggest that PRRSV and PCV2 may negatively modulate the host immune system by different mechanisms which may explain their persistence.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0091-x) contains supplementary material, which is available to authorized users.     

Distribution of T and B lymphocytes in mammary dry secretions, colostrum and blood of adult dairy cattle

The distribution of lymphocyte subpopulations from dry secretions, colostrum and blood from 10 healthy adult Hostein-Fresian cows was studied using the TH21A and B26A mouse monoclonal antibodies (MAb) to adult bovine B and T lymphocytes, respectively. The mammary gland lymphocytes (MGL) were isolated from composite sample of all four quarters by density centrifugation over discontinuous gradient of ficoll-diatrizoate. The peripheral blood lymphocytes (PBL) were purified using the ficoll-thrombin method. Isolated PBL and MGL were analyzed using the two fluorochromes method (TFM) and laser flow cytometry (LFC). The mean viability of isolated PBL and MGL from dry secretions and colostrum after the TFM and LFC were 92.4% +/- 3.2%, 91.4% +/- 6.0% and 87.1% +/- 6.1%, respectively. There was a good correlation between the two MAbs and the percentage of surface immunoglobulin (SIg) positive cells in the peripheral blood using the TFM. The PBL yielded a mean percentage of 21.2% B cells, 66.4% T cells and 9.4% "Null cells" (TH21A+; SIg-). The TFM on MGL from dry secretions and colostrum indicated two distinct patterns (group I and II) of SIg and reactivity to MAb markers (p less than 0.001). The MGL data included in group I and group II were gathered from both colostral and dry secretions. In comparison to the distribution of lymphocyte subsets within peripheral blood the mean percentages of B cells, T cells and "Null cells" in the mammary gland were respectively, 2.8%, 88.1% and 5.4% for group I and 3.5%, 89.0% and 15.1% for group II. In the mammary secretions, the use of SIg alone was not considered to be a good marker for B cells; in four animals a mean percentage of 15.6% (13.9/89.0 X 100) of the mammary gland T lymphocytes were also SIg+. Of the TH21A+ MGL, only 18.8% were SIg+ in group II compared with 34.1% for MGL from group I and 69.3% for the PBL. Marked differences in cell size distribution and cell surface antigen density were found when PBL and MGL from dry secretions were compared by LFC using the B26A MAb. The results of this study demonstrate a difference in the percentages of peripheral blood and mammary gland B and T lymphocytes and confirm previous findings in which the T lymphocytes were found to represent the major subpopulation of lymphocytes in bovine mammary secretions. This may represent an essential event in the adoptive transfer of cellular immunity through the colostrum in cattle.     

B and T lymphocytes in the bovine mammary gland: Rosette formation and mitogen response

Characterization of lymphocyte subpopulations in the bovine mammary gland was accomplished using cells obtained from dry secretions. Correlation of cell surface properties with functional capacity was attempted by assaying the ability to form erythrocyte-antibody (EA) rosettes, erythrocyte-antibody-complement (EAC) rosettes, and sheep erythrocyte (E) rosettes and the ability to respond to phytohemagglutinin (PHA), Concanavalin A (Con A), and pokeweed mitogen (PWM) in the lymphocyte stimulation test. Results were compared with those obtained for peripheral blood lymphocytes (PBL) from the same animals. Mammary gland lymphocytes (MGL) formed significantly fewer (p < .01) EA and EAC rosettes, but significantly greater (p < .01) E rosettes compared to PBL. MGL were significantly less responsive (p < .05) to mitogens than were PBL. MGL contained a large proportion of T lymphocytes, which do not respond to T lymphocyte mitogens in culture.     

Effects of prenatal stress on cellular and humoral immune responses in neonatal pigs

In the present study, we investigated the effects of a daily 5 min restraint stress of pregnant sows in the last five gestational weeks on the development and reactivity of the immune system of the offspring. Maternal stress resulted in significant decreased serum immunoglobulin G (IgG) concentrations in suckling piglets at 1 and 3 days of age. Furthermore, the stress treatment of the sows had an immunosuppressive effect on lymphocyte proliferation in response to the T-cell mitogen concanavalin A (ConA) at postnatal days 1 and 7. A suppressive effect was also found in response to the B-cell mitogens lipopolysaccharid (LPS) at days 1 and 35 and pokeweed mitogen (PWM) at day 1 of life, whereas natural killer (NK) cell cytotoxicity was not altered by prenatal stress. The relative thymus weights were significantly reduced in prenatally stressed piglets on the first and 35th day of life and the morbidity and mortality during the suckling period were significantly increased in prenatally stressed litters, as shown by a higher frequency of diseased and died piglets per litter. In addition, the ConA-, LPS- and PWM-stimulated lymphocyte proliferation at the age of 7, 21 and 35 days, and the NK cell cytotoxicity at the age of 21 and 35 days decreased in prenatally stressed and in control piglets 1h after a corticotropin (ACTH) injection. However, the cellular immunity was always higher in the control piglets which might be a result of the weaker stress hormone reactivity in prenatally stressed animals. In conclusion, the results provide first experimental evidence that prenatal maternal stress during late gestation is able to impair both humoral and cellular immune function in suckling piglets. The data also suggest that gestational stress in pigs may affect the ontogeny of the foetal immune system with consequences on the susceptibility to diseases and immune responsiveness to stressful stimuli of the offspring.     

妊娠后期母猪和仔猪补饲外源精胺对初生和28日龄仔猪肠道形态结构及二糖酶活性的影响

本文通过研究妊娠后期母猪和仔猪补饲外源精胺对初生和28日龄仔猪肠道形态结构和二糖酶活性的影响,初步探讨干预“仔猪早期断奶综合征”的技术措施.试验第1阶段,选择6头体重和膘情相近、胎次为3、已怀孕91 d的健康“长×大”母猪,随机分为Ⅰ、Ⅱ和Ⅲ3个组,每个组设2个重复,每个重复1头母猪.Ⅰ、Ⅱ和Ⅲ组妊娠母猪饲粮中外源精胺的添加量为0、1.5和3.0 mg/kg,饲喂至分娩结束;第2阶段,分娩后母猪采食同一种不含外源精胺的饲粮,Ⅰ、Ⅱ和Ⅲ组哺乳仔猪于7日龄起相应补饲外源精胺添加量为0、3.0和6.0 mg/kg的哺乳仔猪饲粮至28日龄.在仔猪初生和28日龄时,分别从每窝仔猪中选1头接近平均体重的健康仔猪进行屠宰,用于胃肠发育指标的测定.结果表明:与未添加外源精胺相比,妊娠母猪饲粮添加3.0 mg/kg外源精胺显著提高了初生仔猪十二指肠和回肠的柱状细胞数量、十二指肠和空肠的杯状细胞数量(P＜0.05),显著增加了空肠隐窝深度(P＜0.05),显著提高了麦芽糖酶和蔗糖酶比活力(P＜0.05);哺乳仔猪饲粮添加6.0 mg/kg外源精胺显著增加了28日龄仔猪空肠黏膜重(P＜0.05),显著提高了十二指肠和空肠的麦芽糖酶和蔗糖酶比活力(P＜0.05),极显著增加了空肠柱状细胞和杯状细胞数量(P＜0.01),极显著增加了十二指肠和空肠隐窝深度(P＜0.01).因此,在妊娠后期母猪和哺乳仔猪饲粮中添加外源精胺有利于初生和28日龄仔猪肠道形态结构的改善和二糖酶活性的提高.     

Immunologic Profiles of Peripheral Blood Leukocytes and Serum Immunoglobulin G Concentrations in Perinatal Mares and Neonatal Foals (Heavy Draft Horse)

The purpose of this study was to examine sequential changes in the immunologic parameters of perinatal mares and neonatal foals of the heavy draft horse. Blood samples were collected from clinically healthy pregnant mares and their newborn foals every week from 1 month before the expected foaling date, and 1 hour, 1 day (24-48 hours), and 1, 2, 3, and 4 weeks after foaling. Peripheral blood samples were used to examine total leukocyte counts (n = 20), differential leukocyte counts (n = 20), lymphocyte subpopulations (n = 13), lymphocyte responses to mitogens (n = 10), neutrophil phagocytic function (n = 12), and serum immunoglobulin G (IgG) concentrations (n = 10). In perinatal mares, remarkable changes observed included increased neutrophils, decreased lymphocytes, decreased CD4⁺ T lymphocytes, and decreased lymphocyte responses to mitogens at delivery. These changes were speculated to be the result of physical stress associated with delivery. In neonatal foals, increase in the phagocytic function of neutrophils, and increase in serum IgG concentration after suckling colostrum and increase of lymphocytes accompanied by physiologic growth were observed. Compared to dams, foals showed lower phagocytic function of neutrophils before suckling and fewer lymphocytes and lower lymphocyte responses to mitogens within 1 day after birth. This study revealed immunologic dynamics in perinatal mares and neonatal foals. Immunologic functions are suppressed in foaling mares and are immature in neonatal foals, especially before colostral intake. We expect these data will be useful for further studies in the field of clinical immunology, and preventive medicine.     

Cellular interactions regulating the in vitro response of bovine lymphocytes to ovalbumin.

We examined the contribution of MHC class II-restricted T cells (CD4+), MHC class I-restricted T cells (CD8+), gamma/delta T cell receptor (TCR)+ T cells, B cells and macrophages to the development and control of in vitro proliferative responses of bovine lymphocytes to ovalbumin (OA). Cell populations for in vitro assay were obtained from peripheral blood (peripheral blood leukocytes, PBL) of OA-primed cattle. Specific cell populations were depleted or purified from PBL by staining with monoclonal antibodies (MAbs) against the appropriate differentiation antigens and sorting on a Fluorescence Activated Cell Sorter (FACS). OA-specific in vitro responses of in vivo primed PBL were dependent on the presence of CD4+ T cells. Their presence could not be replaced by the inclusion of T cell growth factor (TCGF) in the culture system, indicating that CD4+ T cells probably actively proliferate in response to antigenic stimulation. Bovine CD8+ T cells and gamma/delta TCR+ T cells appeared to exert a suppressive effect on proliferative responses. No proliferation was observed in PBL after the depletion of MHC class II+ cells. In this case, the response could be restored by the addition of macrophages or LPS-activated B cells to the MHC class II- population.     

Effects of Staphylococcus aureus mastitis on bovine mammary gland plasma cell populations and immunoglobin concentrations in milk

Bovine mammary tissue and milk samples were examined to determine effects of chronic Staphylococcus aureus mastitis on the humoral immune response. Parenchymal and teat end tissues from lactating bovine mammary glands were stained immunohistochemically to determine distribution of immunoglobulin (Ig) G1-, IgG2-, IgA-, and IgM-producing plasma cells. Numbers of all Ig-producing plasma cells tended to be higher in tissues from S. aureus infected quarters compared with controls, but most differences were not statistically different. Numbers of IgG1-producing plasma cells at the Furstenberg's rosette area of infected quarters were significantly (P less than 0.05) higher than uninfected quarters. There were no significant differences in concentrations of Ig isotypes in milk from S. aureus infected and uninfected quarters. Data suggest that the antigenic effect of chronic S. aureus infection on the humoral immune response of the bovine mammary gland is minimal. Persistency of S. aureus infection may result, in part, from suboptimal stimulation or immunosuppression of the mammary immune system.     

Immunosuppression of in vivo and in vitro lymphocyte responses in swine induced by Trichinella spiralis or excretory-secretory antigens of the parasite.

The in vivo and in vitro effects of Trichinella spiralis excretory-secretory (ES) antigens on porcine peripheral blood lymphocyte (PBL) responses induced with mitogens (phytohemagglutinin, PHA; concanavalin A, Con A; pokeweed mitogen, PWM) or unrelated antigen (Protein A) were studied to determine whether ES antigens depress lymphocyte responses in experimental swine trichinosis, and/or if this response was manifested after lymphocytes from infected pigs had been pretreated with ES antigens. Additionally, the range of inhibition of lymphocyte responses was tested in parasite-free pigs using different doses of ES antigens and compared with the responsiveness of control cultures from the same animals. The responses of lymphocytes from pigs inoculated with 4 x 10(3) muscle larvae (ML) were strongly depressed (P < 0.05) at post-inoculation days (PID) 7 (after stimulation with PHA), 14, 35 (Con A or PWM), and 49 (PWM). At PID 56 and 63 the lymphocytes from T. spiralis-infected pigs responded better (P < 0.05) to all three mitogens than those from non-infected controls. After 7 weeks post-inoculation, PBL which were pretreated with 10 or 250 micrograms ml-1 of ES antigens showed significantly weaker (P < 0.05, P < 0.001) responses to PWM or PHA, respectively, than those from non-infected animals. The responsiveness of lymphocytes from both groups of pigs to Protein A was not affected by the pretreatment with ES antigens in vitro. The responses of lymphocytes from the parasite-free pigs induced by PHA, PWM or Protein A were strongly depressed (P < 0.01) after in vitro pretreatment regardless of the dose of ES antigens (5, 10, 15, or 20 micrograms ml-1) applied.     

Use of alpha-naphthyl acetate esterase staining to identify T lymphocytes in cattle

Peripheral blood lymphocytes (PBL), lymph nodes, and/or spleens from clinically normal cattle were examined for cytochemical staining of alpha-naphthyl acetate esterase (ANAE). Two types of positive-staining patterns in ANAE staining resulted. By a combination of ANAE staining and latex-ingesting test, diffuse ANAE-positive cells were considered as mononuclear phagocytic cells. Using erythrocyte rosettes, erythrocyte antibody complement rosettes, nylon-wool column technique, surface immunoglobulin (SIg) staining, and the ANAE staining technique, granular ANAE-positive lymphocytes were shown to be T lymphocytes. The frequency of T and B lymphocytes in PBL, spleens, and lymph nodes of clinically normal cattle was measured, using ANAE staining and SIg staining. In PBL, 47.7% were ANAE-positive and 26.9% were SIg-positive; in spleens, 22.4% were ANAE-positive and 53.7% were SIg-positive; and in lymph nodes, 38.5% were ANAE-positive and 28.3% were SIg-positive. The frequencies of T and B lymphocytes in PBL, spleens, and/or lymph nodes from cattle with enzootic bovine leukosis (EBL) and cattle with persistent lymphocytosis and in tumor cells from cattle with EBL were measured. When compared with those of clinically normal cattle, PBL, spleens, and lymph nodes of cattle with EBL and the PBL of cattle with persistent lymphocytosis contained numerous SIg-positive cells and few ANAE-positive cells. Tumor cells from cattle with EBL contained 7.3% ANAE-positive and 78.0% SIg-positive cells.     

Effect of corticosteroid on porcine leukocytes: age-related effects of corticosteroid inhibition on porcine lymphocyte responses to mitogens

The effects of prednisolone sodium succinate on the responses of porcine lymphocytes to phytohemagglutinin (PHA) and pokeweed mitogen (PWM) were investigated. Sensitivity of peripheral blood lymphocytes (PBL) to the synthetic glucocorticoid, prednisolone, was related to age of the lymphocyte donor. The greatest sensitivity was found in PBL from animals less than one week old; PBL from animals between 2 to 4 months retained some glucocorticoid sensitive cells; whereas, PBL from animals older than 6 months were exceptionally resistant to steroid. Similar age-associated sensitivities were found for lymphocytes from lymph node, spleen and thymus. Significant differential sensitivities among the various lymphoid organs were found with the thymic lymphocyte possessing the greatest sensitivity to steroid and the PBL lymph node and splenic lymphocytes possessing the highest resistance to the suppressive effects of steroid. The age related differences in sensitivity to steroid did not appear to be caused by differences in the number of steroid receptors because lymphocytes from susceptible and resistant animals had similar numbers of receptors. The results suggest that the age related sensitivity may be associated with a higher percentage of sensitive thymic-derived lymphocyte in the PBL, lymph node and spleen of the younger animals. Results of this study also suggest that the adult pig (6 months) should be classified as a steroid resistant species.     

Studies on immunoglobulins and trypsin inhibitor in colostrum and milk from sows and in serum of their piglets.

The concentrations of IgG, IgM, IgA and the specific sow colostrum trypsin inhibitor (SCTI) were measured by radial immunodiffusion in colostrum and milk samples from sows and in serum samples from their offspring during the suckling period. A clear time dependence was found for all the measured variates in both whey and serum. Statistically significant positive correlations were found between, on the one hand, concentrations of IgG and IgA, but not IgM, in sera from 39 suckling piglets 1 and 3 days old, and, on the other hand, concentrations of the same immunoglobulins and of the trypsin inhibitor in maternal colostrum (n = 7). Multiple regression analyses showed that at day 1 and day 3 the levels of both IgG and IgA in serum samples from the suckling piglets were positively influenced by both the SCTI and the IgG or IgA contents in maternal colostrum.