Antigen In Cattle Research Articles

Serological response over time to recombinant Neospora caninum antigens in cattle after a neosporosis-induced abortion.

Recombinant Neospora caninum tachyzoite antigens were evaluated in an enzyme-linked immunosorbent assay (ELISA) for recognition by serum antibodies (Ab) from Neospora-infected cattle. Serum samples were obtained every 2 to 3 weeks for 8 to 15 months from 10 cows with histories of Neospora-associated abortion. Serum samples were also obtained from offspring of these animals and from a large number of cows that had aborted a fetus, due to infection by Neospora or other organisms, at various times during gestation. All 10 cows had positive ELISA Ab titers to both recombinant N. caninum tachyzoite antigens after abortion, during subsequent gestation, and after parturition. In three cows, there was a noticeable peak in Ab titers early in gestation. Calves born to Neospora-infected cows also had positive titers of Ab to the recombinant tachyzoite antigens, and these titers remained elevated for at least 4 months after birth. A portion of the serum immunoglobulin in calves may have been derived from colostrum of infected cows. A calf born from a seronegative mother had a positive ELISA titer only after being fed colostrum from a seropositive cow. However, precolostral titers in calves born from Neospora-infected cows were high at birth, suggesting that the parasite was transmitted to the fetus via the placenta and induced a humoral immune response therein. The recombinant tachyzoite antigens were also useful for corroborating clinical diagnoses of Neospora-induced abortion. A significant difference (P < 0.05) between anti-recombinant antigen Ab titers in cows that aborted due to Neospora and those in cows that aborted from other causes was found.

The sensitizing effect of a Brucella abortus antigen in cattle after repeated intradermal inoculations

An antigen prepared from a mucoid strain of B. abortus was repeatedly injected intradermally into cattle to determine whether it sensitizes cattle so that they test false positive with the skin delayed-type hypersensitivity (SDTH) test. Cattle ( n = 14) that were experimentally inoculated with microorganisms that serologically cross-react with B. abortus, and uninfected cattle ( n = 15) were tested with the antigen 2 to 5 times over a period of 16 to 17 weeks. An increase in skinfold thickness of ≥ 2.0 mm on the injection site was considered a reaction elicited by the antigen. The sera from the cattle were tested with the serum agglutination test, complement fixation test, and an enzyme-linked immunosorbent assay for antibodies against B. abortus. Results showed that none of the animals had an increase in skinfold thickness of ≥ 2.0 mm on the injection site of the antigen, which is considered a positive reaction. However, cattle experimentally inoculated with microorganisms other than B. abortus tested seropositive for brucellosis after one SDHT test only. We conclude that the B. abortus antigen inoculated intradermally does not sensitize cattle after repeated inoculations, and therefore does not interfere with the subsequent use of the SDTH test in the diagnosis of brucellosis.

6.20 A new non-lineage specific antigen with an Mr of 115 kDa and 39 kDa present on bovine leukocytes identified by monoclonal antibodies within BoWC10

Five mAbs out of a total of 189 submitted to the Second Workshop were shown to be similar when statistical analyses of flow cytometry data were performed. For the duration of the workshop the five mAbs were assigned to temporary cluster TC35. The mAbs precipitated two polypeptides by SDS-PAGE under reducing conditions. One had an M r of 115 kDa, the other an M r of 39 kDa. The antigen recognized was present on thymocytes, a subpopulation of CD2 + T cells and a major subpopulation of B cells. The antigen is upregulated after long-term activation with concanavalin A. It is proposed that the mAbs in TC35 should be regarded as a single cluster identifying a novel leukocyte differentiation antigen in cattle. These mAbs were accepted as a new workshop cluster BoWC10.

Bovine babesionsis: Failure to induce interferon gamma production in response to Babesia bovis antigens in cattle

The immune response to Babesia bovis infection or vaccination ws evaluated by measuring antibody and interferon gamma (IFN-γ) production to protective recombinant and crude native B. bovis antigens. Cells from vaccinated or infected cattle failed to produce detectable IFN-γ when stimulated with B. bovis antigens in vitro. In contrast, antibody was induced by protective recombinant B. bovis antigens. These findings are consistent with the argument that immunity to B. bovis infection is correlated most strongly with humoral rather than cell-mediated immune responses.

Development and immunophenotyping of the pharyngeal tonsil (adenoid) in cattle

Immunoperoxidase staining and electron and light microscopy were used to characterize the development of the pharyngeal tonsil in 98 cattle aged between 30 days of gestation and 12 years. The rugae of the pharyngeal tonsil were poorly formed before 95 days of gestation. Microvillous (M) cells associated with intra-epithelial leucocytes (lympho-epithelium) were scattered among ciliated and goblet cells covering most of the surface in post-natal animals. Intra-epithelial leucocytes were rare in fetuses, but ciliated and M cells could be distinguished. Leucocytes of the lamina propria started to accumulate at approximately 120 days of gestation. A loose accumulation of mononuclear cells progressed into a B-cell rich upper and T-cell rich lower layer, with typical lymphoid tissue organization in post-natal animals and lymphoid involution in aged cattle. Primary lymphoid follicles formed at 5 months of gestation, but germinal centres did not form until 2 to 4 weeks after birth. Except for null cells, the relative number of cells staining for each leucocyte phenotype or MHC class II antigen increased with age, especially during the neonatal period. The early development, strategic location and specialized structure of the pharyngeal tonsil suggest an important role in modulating inhaled antigens in cattle. Fetal and neonatal calves had minimal lymphoid tissue priming, as indicated by lack of secondary follicles, low MHC class II expression and few intra-epithelial leucocytes. The phenotypic differences may be relevant to the increased susceptibility of calves to infectious diseases shortly after birth.

Serological reactivity to Mycobacterium bovis protein antigens in cattle

The serological response to 12 purified Mycobacterium bovis antigens were examined in an ELISA assay. These antigens included the majority of M. bovis protein antigens described to date and in most cases they were very similar to the M. tuberculosis antigens of the same molecular mass. The purified antigens were tested against sera from M. bovis infected cattle, M. bovis culture-negative cattle from infected herds and animals infected with related microorganisms, mainly other mycobacterial species. All the antigens gave strong reactions with at least some sera from the M. bovis infected group and showed cross-reactivity with some of the sera from the other two groups. The antigen with the highest specificity reacted strongly with only 60% of the M. bovis infected sera. Antigens that reacted with most or all of the M. bovis infected sera also gave the highest cross-reactivity with sera from the other two groups. These results indicate that a serological test based on any one or a combination of these antigens, without removal of the cross-reacting epitopes, would be unsatisfactory.

Cell-mediated immune responses to Babesia bovis merozoite antigens in cattle following infection with tick-derived or cultured parasites

Peripheral blood mononuclear cells from cattle experimentally infected with Babesia bovis were examined for parasite-specific cell-mediated immune responses. Unfractionated merozoites and soluble and membrane fractions derived from merozoites were all antigenic for immune cattle, although the membrane fraction was the most stimulatory. Cattle responded to different antigenic fractions in a differential manner, and only that animal immunized with autologous cultured parasites responded to parasitized erythrocyte culture supernatants. Plastic-adherent cells (presumably monocytes/macrophages) were required for a proliferative response to babesial antigens but not to the T-cell mitogen concanavalin A, suggesting that babesial proteins are not simply mitogenic for T cells. Lymphocyte responses directed against a different hemoparasite from Mexico, Babesia bigemina, indicate that this parasite shares cross-reactive T-cell epitopes with B. bovis. These studies define a system whereby T lymphocytes from babesia-immune cattle can be used in proliferation assays to identify babesial merozoite antigens which are immunogenic for T cells. Because identification of helper T-cell epitopes is important for the design of a babesial subunit vaccine which will evoke anamnestic responses, the studies described here provide a basis for such experiments.

Polymorphism of the CD4 and CD5 differentiation antigens in cattle

Monoclonal antibodies (mAbs) specific for bovine CD4 and CD5 antigens have been found to identify polymorphic determinants on these molecules. In the case of CD5, mAb IL-A67 recognises one allotypic form of the antigen while four other CD5-specific mAbs in the workshop (CC17, CC29, BLT-1 and 8C11) recognise a second allotype. The CD4-specific mAbs submitted to the workshop reacted with the cells of all animals tested. However, a further two mAbs (CC26 and IL-A18) specific for CD4 were found to react with cells only from about 85% of animals tested. Sequential immuno-precipitation experiments together with family studies showed that the allotypes of CD4 and CD5 are both inherited in a simple Mendelian manner and are co-dominantly expressed. One of the CD5 allotypes was not detected in Bos taurus animals while the gene frequency of the second allotype was only about 10% in the B. indicus animals tested. The gene frequency of the CD4 allotype detected by CC26 and IL-A18 was similar in the two sub-species.

Characterisation of lymphocyte populations in African buffalo ( Syncerus caffer) and waterbuck ( Kobus defassa) with workshop monoclonal antibodies

In order to measure different lymphocyte populations in buffalo ( Syncerus caffer) and waterbuck ( Kobus defassa), we analysed the monoclonal antibodies from the 1st International Workshop on Leukocyte Antigens in Cattle, Sheep and Goats for suitable cross-reactive reagents. Peripheral blood mononuclear cells from three buffalo and three waterbuck were tested with the whole panel of monoclonal antibodies (mAbs) together with some additional antibodies against MHC and Ig. In some clusters almost all antibodies cross-reacted (CD2, CD8), in others almost none cross-reacted (CD4, CD5) and in cluster CD6, mAbs only reacted with buffalo but not waterbuck. Double staining experiments were performed on buffalo PBM with the cross-reacting antibodies, to confirm that they detected similar cell populations as in bovine PBM. This was shown with reagents against CD2, CD4, CD6, CD8, CD11, WC1, WC3 and Ig. The molecular weights of the buffalo antigens correlated well with those of the homologous cattle antigens. In the CD5 cluster, only one mAb reacted with the two wild species, and defined an unusual CD2 + CD5 − cell population in buffalo. Also mAbs cross-reacting with buffalo MHC class II detected unusual expression on resting T cells. From the results presented, it is clear that the workshop panel contains mAbs against the most important T and B cell antigens of buffalo and probably waterbuck, which will allow us to compare functional lymphocyte populations in cattle and wild ruminants.

Summary of workshop findings for leukocyte antigens of cattle

Summary of workshop findings for leukocyte antigens of cattle

Additional information on the linear order of the genes encoding the red blood group C‐system antigens in cattle

A linear model for the genes controlling the system C blood group factors was presented by Bouw et al. (1974) and confirmed by Guerin et al. (1981). The relative order of the R and W genes was not established by either of these studies. A recently observed recombination in the C system is reported here, providing evidence for the positioning of the R and W genes.

Characterization of a bovine thymic differentiation antigen analogous to CD1 in the human.

Three monoclonal antibodies (MoAb), TH97A, CC13, and CC14, define a thymic differentiation antigen in cattle. The antigen is expressed on 50-60% of bovine thymocytes, located mainly in the cortical areas, but is not expressed on peripheral blood mononuclear cells (PBMC). In cryostat sections of lymph node, the antibodies react with large dendritic-like cells in the paracortical regions. They also react with a proportion of the large 'frilly' cells in afferent lymph and with dendritic-like cells in the dermis. The antibodies apparently do not react with cells in the epidermis. Biochemical analysis of the antigen recognized by MoAb TH97A reveals two bands of 44 kDa and 12 kDa under reducing conditions. These polypeptides are distinct from bovine class I major histocompatibility complex molecules reactive with the MoAb w6/32. The tissue distribution of positive cells together with results of biochemical analyses indicate that the antigen recognized by these MoAb is the bovine analogue of the human CD1.

Synteny mapping of the genes for 21 steroid hydroxylase, alpha A crystallin, and class I bovine leukocyte antigen in cattle.

Synteny mapping of the genes for bovine alpha A crystallin (CRYA-1), steroid 21 hydroxylase (21OH), and a class I major histocompatibility locus bovine leukocyte antigen (BoLA) was accomplished by Southern blot analysis of DNA from bovine-hamster hybrid somatic cells. Comparison of the distribution patterns of CRYA-1, 21OH, and BoLA sequences among the hybrid cells revealed that the genes are located on two different chromosomes in the bovine genome. CRYA-1 mapped with superoxide dismutase (SOD1 in syntenic group U10, while 21OH and BoLA were concordant with glyoxylase (GLO1) in syntenic group U21. Therefore, the syntenic array of CRYA-1, 21OH, and BoLA in cattle is different from that observed in rodents, where the genes are closely linked to each other and to GLO1, and consistent with reported linkages in man.

The development and analysis of species specific and cross reactive monoclonal antibodies to leukocyte differentiation antigens and antigens of the major histocompatibility complex for use in the study of the immune system in cattle and other species

We examined the potential of developing a set of species specific and cross reactive monoclonal antibodies (MoAbs) for use in the study of the phylogenetic and functional relation of class I and Class II antigens of the major histocompatibility complex (MHC) and leukocyte differentiation antigens in cattle and other species. Comparing immunization strategies demonstrated the number of hybrids producing cross reactive antibodies can be increased by hyperimmunization of mice with lymphoid cells from multiple species. Comparing various methods of assay (antibody-complement mediated cytotoxicity [CT], enzyme linked immunosorbent assay [ELISA] and flow microfluorimetry [FMF]), revealed FMF is the most useful technique for the primary assay of hybridomas producing MoAbs of potential interest. By using dual parameter and dual fluorescence analysis, we could determine whether a given MoAb reacted with mononuclear cells (lymphocytes and monocytes) and/or granulocytes, and also whether any two MoAbs of different isotype and specificity recognized antigens present on identical or separate populations of leukocytes. Comparing the patterns of MoAb reactivity with leukocytes obtained from cows, goats, sheep, pigs, horses and humans, as well as comparing the patterns of reactivity with a panel of lymphoid cell lines derived from cattle (with enzootic bovine leukemia) and humans (with various forms of leukemia), revealed sets of MoAbs reactive with unique antigenic determinants present on BoLA class I (15 MoAbs) and class II (9 MoAbs) antigens, and also MoAbs reactive with determinants present on leukocyte differentiation antigens (36 MoAbs). Dual fluorescence analysis demonstrated the antigens detected by some MoAbs are predominantly expressed on one lineage of leukocytes while others are expressed on two or more lineages of leukocytes. Dual and single fluorescence analysis also demonstrated the PNA receptor(s) is: 1) expressed on T cells, granulocytes and class II antigen monocytes and 2) absent or expressed in low amount on sIgM + B cells and a newly defined Non T/Non B population of cells. The strategies described for identifying and analyzing the specificity of MoAbs demonstrate the feasibility of developing a set of cross reactive MoAbs for identifying homologous molecules in multiple species and delineating their functional and phylogenetic relation.

An Optimized Enzyme-Linked Immunosorbent Assay for Quantitative Diagnosis of Bovine Fascioliasis

An optimized immunoassay for detection of antibody to Fasciola hepatica antigen in cattle was developed through the adaptation of a kinetics-dependent, enzyme-linked immunosorbent assay (k-ELISA) to a microplate format. Enhanced sensitivity and a strict quantitative nature were achieved with the utilization of enzyme kinetics. With this k-ELISA, significant (P less than 0.01) elevations in anti-F. hepatica antibody could be detected as early as 2 wk post-infection in experimentally infected calves. Furthermore, fluke-burden related differences in anti-F. hepatica antibody levels between 3 different levels of fluke infection were evident.

Development of serum antibody activity as determined by enzyme-linked immunosorbent assay to Psoroptes ovis (Acarina: Psoroptidae) antigens in cattle infested with P. ovis

Calves infested with Psoroptes ovis (Hering, 1838) for the first time (naive) or previously infested calves were examined for serum antibody activity to P. ovis (obtained from rabbits) antigens by enzyme-linked immunosorbent assay (ELISA) to determine the temporal appearance of specific serum antibody activity. The development of the serum antibody activity to P. ovis antigens was then correlated with the development of lesions (% scab) and with changes in the number of P. ovis. The serum antibody activity [ELISA OD 414 value (EODV)>0.290] to P. ovis antigens in naive calves, in most cases, is first detected at the same time or slightly after the detection of mites and mite-caused lesions; and the development of specific serum antibody activity paralleled the increase in the P. ovis population and the percentage scab until 7 weeks post-infestation (PI). In previously infested calves, if serum antibody activity to P. ovis antigens was not already present from the previous infestation, specific serum antibody activity was detected at the same time and developed in a similar manner as in the naive calves. The serum antibody activity to P. ovis antigens could be detected after calves were relieved of their P. ovis burdens by pesticide treatment or after the P. ovis population began to decline when the calves were allowed to groom themselves. Serum antibody activity to P. ovis antigens was not detected in any of the control calves during the test period.

Immunochemical characterization of major histocompatibility antigens in cattle.

Immunochemical characterization of major histocompatibility antigens in cattle.

Diagnosis of clinical leukosis in cattle and its correlation with bovine leukemia virus antigens using the agar gel immunodiffusion (AGID) test

During an 18-month period a total of 317 bovine sera were tested for antibody to bovine leukemia virus (BLV) using the agar gel immunodiffusion (AGID) test. Of these field submissions, 130 (41%) were positive for antibody to BLV and 34 (26%) had a clinical diagnosis of bovine leukosis. A second (AGID) precipitin line was found in tests with sera from five of these cattle. Four cows had grossly enlarged palpable lymph nodes or subcutaneous masses. The two antigens identified in the sera of the 5 cattle were the glycoprotein gp envelope antigen, typically found in cattle exposed to BLV, and the p24 antigen which is part of the virus core. We found that in these 5 cattle with antibodies to both antigens the concentration of antibody to the gp antigen was greater than that of the p24 antigen. The presence of antibody to the p24 viral antigen in cattle with clinical disease and its implication in the confirmation of a diagnosis of bovine leukosis is discussed.

Occurrence of Antibodies to Group Specific Chlamydia Antigen in Cattle in Various Areas of Finland

The occurrence of group specific complement fixing antibodies was studied in 361 cattle sera from 36 herds in 6 areas in Finland. Sixty-two (17.2%) were positive. The antibody frequency increased significantly from the south to the north and the frequency was significantly higher in forest than in field pastures. The reasons for the differences are discussed. The tick, Ixodes ricinus, perhaps has no significant epidemiological role in chlamydial epidemiology in Finland.

Immune response to virus-infection-associated (VIA) antigen in cattle repeatedly vaccinated with foot-and-mouth disease virus inactivated by formalin or acetylethyleneimine

The results of experiments to investigate antibody to 'virus infection associated' (VIA) antigen in cattle repeatedly vaccinated with formalin- or acetylethyleneimine- (AEI) inactivated foot-and-mouth disease (FMD) vaccines under laboratory conditions are reported. Results are also presented from some vaccinated animals subsequently exposed to FMD infection. Antibody against VIA was not detected before and after the first vaccination with formalin or AEI-inactivated vaccine but did develop in all animals after the second formalin vaccination and persisted throughout the experiment. After the second AEI vaccination, 4 of 12 animals developed antibody which persisted for at least 37 days. This transient response in some cattle was repeated after successive vaccinations but, in general, more animals responded as the number of vaccinations increased. After exposure to infection a transient VIA antibody response was occasionally observed in immune AEI-vaccinated animals. Some immune repeatedly AEI-vaccinated cattle did not develop detectable VIA antibody after challenge despite the persistence of virus in oesophageal-pharyngeal (O/P) fluid. The presence of antibody to VIA antigen is not conclusive proof that vaccinated animals have been exposed to infection and field data must be interpreted with caution.