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Eight murine monoclonal antibodies (mAb) were used to identify the equine CD8 alpha or CD8 beta chains and to define the expression of these chains on lymphocytes from various lymphoid tissues. CD8 alpha was a 39 kDa protein and CD8 beta was a 32 kDa protein. Both chains were expressed on most of the CD8+ T lymphocytes in the peripheral blood, spleen, thymus, mesenteric lymph nodes and ileal intraepithelial lymphocytes (IEL), however, in each lymphoid compartment a percentage of lymphocytes expressed only the CD8 alpha chain. The largest percentage of CD8 alpha alpha expressing T lymphocytes was 37.7% of the IELs. Purified T lymphocytes from the ileum expressing CD8 alpha beta co-expressed the alpha beta T cell receptor (TCR). In contrast, purified CD8+ T lymphocytes from the PBMC co-expressed either the alpha beta or gamma delta TCR by RT-PCR. Use of pooled anti-CD8 alpha mAb of the murine IgG2a isotype and rabbit complement resulted in lysis of the entire CD8 expressing population in peripheral blood mononuclear cells (PBMC). These results indicated that CD8 dimer usage by equine T lymphocytes is similar to other species and that the mAb described can be further used to separate equine CD8+ T lymphocyte subsets from the lymphoid tissues to define their function in protection against viral and other infections.

A 14- to 26-kilodalton fraction of Staphylococcus aureus exoproteins isolated by molecular sieve chromatography and electroelution from polyacrylamide gels was shown to specifically react with antibodies in milk of naturally infected dairy cows. Silver staining of the antigen preparation electrophoresed in polyacrylamide gels showed the strongest reactivity in the 24- to 26-kilodalton region with lesser staining at lower apparent molecular sizes. An enzyme-linked immunosorbent assay was developed to differentiate infected from uninfected cows for diagnostic purposes. Samples from S. aureus-infected cows reacted in the assay, and samples from uninfected cows did not. There was no correlation between numbers of somatic cells in the samples and reactivity to the antigens. Samples from cows infected with coagulase-negative staphylococci did not react with the antigens. It was found, however, that some samples from uninfected cows that were recently postpartum or producing low amounts of milk contained antibodies which bound the antigens. This was believed to be due to transport from blood to the mammary gland of antibodies which were generated by previous intramammary infections or infections at other sites.

Foals with combined immunodeficiency had normal levels of purine salvage pathway enzymes, including adenosine deaminase, nucleoside phosphorylase, and xanthine oxidase.

Concentrations of immunoglobulin (Ig) G1 were measured in 590 sera from 24- to 36-hour-old lambs, and failure in passive transfer (FPT), less than 6 mg/ml, occurred in 20 lambs. Of the 20 FPT lambs, 45% died before 3 weeks of age, whereas only 5% of the 570 lambs with adequate passive transfer died before 3 weeks of age. The low percentage of FPT was attributed to management practices ensuring suckling by the lambs and possibly to influences from several years of selecting ewes on the basis of weaned lamb production. The correlation between the concentration of IgG1 in 257 postpartum, presuckle ewe colostrum samples and the IgG1 concentrations in 362 lamb sera from those ewes was low (r = 0.32). However, the mean serum IgG1 concentration in 20 lambs from ewes with the lowest postpartum, presuckle colostrum IgG1 concentrations (less than 30 mg/ml) was significantly lower (P less than 0.001) than mean serum IgG1 concentration in 24 lambs from ewes with the highest postpartum, presuckle colostrum IgG1 concentrations (greater than 110 mg/ml). Postpartum, presuckle colostrum IgG1 was measured in 7 ewes whose lambs had FPT, and the IgG1 values varied throughout the colostrum IgG1 range. Colostrum IgG1 concentrations could not be used to explain FPT or to identify ewes likely to have lambs with FPT.

Variable surface antigens of cloned populations of African trypanosomes were studied using 2-dimensional gel electrophoresis and monoclonal antibodies. Two-dimensional gel maps showed that the only major differences in protein profiles of the non-nuclear materials from different clones of solubilized trypanosomes were attributable to the variable surface antigens and that purification doesn't alter these molecules, at least with respect to charge and apparent m.w. Purified variable surface antigens were used as immunogens and a number of monoclonal antibodies were derived using cell-fusion technology. In radioimmunoassay, each of the monoclonal reagents was shown to be specific for the immunizing antigen, and in immunofluorescence tests using acetone-fixed trypanosomes, each bound specifically to the clone from which the antigens were purified. Only 20% of the monoclonal reagents bound to living trypanosomes, however, providing evidence for both exposed and nonexposed antigenic sites on the variable surface antigens. Those reagents derived to a single antigen bound to different nonoverlapping antigenic sites, which were on the protein portion of the molecules and not the carbohydrate moieties. The data show that monoclonal antibodies are ideal probes for studying localization of antigenic sites on the antigen molecules and for studying antigen synthesis, glycosylation, and architecture in relation to the cell surface. In addition, the finding of exposed and nonexposed antigenic sites on trypanosome variable antigens allows a possible explanation for the role of the antigens in pathogenesis of African trypanosomiasis.

Serum IgG1 concentrations of calves less than 3 weeks old and dying from infectious disease were significantly lower (P less than 0.01) than those of clinically normal calves. Fifty percent of the dead calves had serum IgG1 concentrations that were more than 2 standard deviations below the normal mean, and an additional 35% had IgG1 concentrations that were more than 1 standard deviation below the normal mean. Low IgG1 concentrations were attributed to failures in passive transfer of colostral immunoglobulin. The few calves dying of noninfectious causes generally had normal serum immunoglobulin concentrations. The results of this study emphasize the importance of adequate colostral intake and absorption to the neonatal calf. In view of the large numbers of calves that die from neonatal infection each year, it may be assumed that failure in passive transfer, as reflected by low serum immunoglobulin concentrations, is one of the most important factors influencing neonatal calf mortality.

Presuckle colostral samples and lamb serum samples taken 36 h postpartum were assayed for immunoglobulin G1 (IgG1) concentration (mg/ml) using single radial immunodiffusion. Breeds sampled included Polypay (P), Rambouillet (R), Targhee (T), Columbia (C), Finnish Landrace (F) and Finn crosses (Fx). Sources of variation examined in IgG1 concentration in colostrum (dam trait) included dam's sire breed, dam's sire, age of ewe and number of lambs born. All sources of variation were statistically significant. Least-squares means of IgG1 levels for sire breed were 80, 64, 67, 64, 72 and 69 mg/ml for P, R, T, C, F and Fx breed groups, respectively. A fetal stimulus may exist to increase the mass of IgG1 in colostrum available for multiple births (61, 69 and 77 mg/ml for single, twin and triplet, respectively). Ewe age was a significant source of variation because of a high mean concentration of IgG1 in the yearling's colostrum (100 mg/ml), whereas only slight differences occurred between the other age groups (65 to 67 mg/ml), except for the 7-yr older group (53 mg/ml). Sources of variation examined in IgG1 concentration of lamb serum at 36 h postpartum (lamb trait) included lamb's sire breed, lamb's sire, age of dam, birth type and sex, with dam's colostral IgG1 concentration and day born as covariates. Sire within breed, birth type and the two covariates were significant. Least squares means for sire breed were 36, 32, 33, 32, 31 and 32 mg/ml of serum for P, R, T, C, F and Fx groups, respectively. Lamb serum IgG1 decreased as birth type increased. The heritability of IgG1, estimated by paternal half-sib analyses, was .19 +/- .12 for colostrum and .18 +/- .06 for lamb serum.

Combined immunodeficiency (CID), a defect in both B- and T-lymphocytes, was found to occur in 2.3% of 257 foals of Arabian breeding. All affected foals died by 5 months of age. The belief that CID is transmitted as an autosomal recessive genetic defect was supported by results from matings of dams and sires that had previously produced affected foals. Based on a prevalence of 2.3%, the proportion of carriers of the CID trait among the adult population surveyed was estimated to be 25.7%. Recent descriptions of other immunologic defects in foals emphasized the need for careful differential diagnosis. Disorders that could be confused with CID include failure in passive transfer of immunoglobulins from dam to foal, via colostrum; transient hypogammaglobulinemia; agammaglobulinemia (associated with B-lymphocyte deficiency and normal T-lymphocytes), and selective IgM deficiency.

The capacity of cells from thymus, liver, spleen, mesenteric lymph nodes, peripheral blood, and bone marrow to respond to in vitro phytolectin and allogeneic lymphocyte-stimulation was determined in 16 pony fetuses 61 to 200 days old (gestational age). Phytolectin-responsive cells were detected in the thymus at the 80th gestational day, peripheral blood at 120 days, lymph node at 160 days, and spleen at 200 days. Mixed lymphocyte culture-responsive cells were detected in thymus at 100 days and in the spleen at 200 days (gestational age). Immunoglobulins (Ig) M and IgG were quantitated by radioimmunoassay. They were detected in fetuses prior to 200 days of age. All of 50 normal newborn foals had detectable quantities of IgM (165 +/- 56 micrograms/ml of serum). Quantities of IgG in normal newborn foal serum were lower and more variable. The minimal-maximal concentrations of IgG were 2 to 170 micrograms/ml of serum with a mean and SD of 51 +/- 49 micrograms/ml. The results indicate that (1) functional T lymphocytes are present in the fetus by 100 days (gestational age), (2) functional B lymphocytes are present by 200 days, and (3) that foals are immunocompetent before birth.

Newborn Saanen goats were orally infected with two distinct biologically cloned isolates of the caprine arthritis-encephalitis lentivirus (CAEV), designated CAEV(63) (Crawford TB, Adams DS, Cheevers WP, Cork LC: Science 207:997, 1980) and CAEV (Co) (Cork LC, Hadlow WJ, Crawford TB, Gorham JR, Piper RC: J Infect Dis 129:134, 1974; Narayan O, Clements JE, Strandberg JD, Cork LC, Griffin DE: J Gen Virol 50:69, 1980). All infected goats seroconverted for antibodies to CAEV, and there were no serologic reversions. Histologic lesions consisted of chronic inflammation of variable incidence and severity in joints and mammary gland. Clinical arthritis, estimated by enlargement of carpi, correlated with severe inflammation of radiocarpal synovium. Statistical evaluations of clinical and histologic parameters indicated that chronic joint disease was significantly more frequent and more severe in the group of goats infected with CAEV(63) than in the group infected with CAEV (Co). Thus, the severity of chronic arthritis is determined in part by the relative pathogenicity of individual CAEV isolates. Analysis of the temporal development of carpal arthritis during the first 3 years of infection indicated that (a) the extent of eventual chronic disease in individual joints was apparent relatively early after infection, (b) joints with severe chronic lesions were clinically enlarged at irregular intervals and (c) the pattern of recurrent arthritis was unique for each joint. Thus, chronic arthritis develops with a progressive course which may be promoted by recrudescent inflammation. Infectious virus was recovered 3 years after infection from selected tissues of 12 of 17 CAEV(63)-infected goats and 11 of 18 CAEV(Co)-infected goats. Virus recovery was independent of the virus isolate used for experimental infection and did not correlate with the severity of lesions.

The results of a study on the induction of IFN-alpha, IFN-beta, and IFN-gamma in normal and SCID foals showed a deficiency of IFN-gamma but not IFN-beta in SCID foals. The ability of SCID mononuclear cells to produce IFN-alpha in response to poly I:C but not to NDV may indicate a partial deficiency of IFN-alpha in SCID foals. The deficiency of IFN-gamma and presence of IFN-beta in SCID foals supports the classification of IFN-gamma and IFN-beta as immune and nonimmune interferons, respectively. Furthermore, the deficiency of IFN-gamma in SCID foals may in part explain the high susceptibility to infections observed in SCID foals.

Persistence of virus and a marked hypergammaglobulinemia are consistent features observed in mink with Aleutian disease virus infection. These events may result from altered lymphocyte function in infected mink. To test this possibility we determined the ability of lymphocytes from infected mink to respond to phytolectins (PHA and Con A), and a specific antigen (KLH) in a lymphocyte transformation test and quantitated the number of lymphocytes bearing surface immunoglobulin. Infected mink had a significantly lower response than normal mink to PHA and Con A (P less than 0.025), and the difference in the responses increased progressively with time post-infection. Infected mink immunized with KLH also had a decreased ability to respond to this antigen when compared to normal immunized mink (P less than 0.001). The number of lymphocytes bearing surface immunoglobulin was significantly elevated in three of 10 mink infected for 10 weeks, and in all of seven mink infected 32 weeks (P less than 0.001).

Caprine arthritis-encephalitis is a retrovirus-induced disease resulting in lymphoproliferative lesions of the CNS and joints. Peripheral blood leukocytes of chronically infected goats were analyzed for the types of cells present and for their reactivity to viral antigen and polyclonal stimulants. Two of 9 infected goats had abnormal numbers of B lymphocytes--one elevated and the other deficient. Lymphocyte reactivity to viral antigens was transiently detectable by a lymphoblastogenic assay in 5 of the 9 goats. The reactive cells were peanut agglutinin-negative T lymphocytes. Concanavalin A induced more division in T lymphocytes of infected goats than in lymphocytes of noninfected goats, whereas the reactions to phytohemagglutinin, pokeweed mitogen, and bacterial lipopolysaccharide were no different in the 2 goat groups. It is concluded that goats infected by the caprine arthritis-encephalitis virus have antigen-reactive T lymphocytes and that infection promotes the response to a nonspecific T-cell stimulant.

Selective immunoglobulin M deficiency was diagnosed in 5 foals, on the basis of reduced serum immunoglobulin M concentrations (more than 2 standard deviations below the normal mean). All 5 foals had clinical signs or lesions involving the respiratory tract. Lkebsiella sp was isolated from joint fluids, bronchial lymph nodes, or tracheal washings from the 3 foals in which such samples were available for microbiologic evaluation.

Between January 1973 and September 1979, 2,092 horses and ponies were evaluated for immunologic disorders. A total of 418 abnormalities were detected in 416 (20%) of the animals tested. Disorders encountered were failure or partial failure of colostral immunoglobulin transfer from mare to foal (228 cases), combined immunodeficiency (159 cases), selective immunoglobulin M deficiency (19 cases), agammaglobulinemia (3 cases), transient hypogammaglobulinemia (2 cases), and lymphosarcoma (7 cases). Four conclusions were drawn from the study. (1) Immunologic abnormalities occur commonly in horses and ponies. (2) Failure and partial failure of passive transfer were the most common disorders, involving 19.7% of surveyed foals at risk. (3) Combined immunodeficiency remains a disease limited to Arabian horses. (4) Considering the high frequency of immunologic disorders in horses and the availability of diagnostic tests for the disorders, older animals with recurrent infections as well as all newborn foals should be evaluated for immune disorders.

Forty-eight-hour serum concentrations of immunoglobulin G1 (IgG1) were determined for 123 female calves born on 1 farm during a 12-month period and fed 2.8 L of colostrum by esophageal feeder within 2 hours of birth. Mean monthly serum IgG1 concentrations were lowest in the winter, and increased during the spring and early summer to reach their peak in September, after which they decreased. Eight of 64 calves (12.5%) born from June through October had failure of passive transer of IgG1 (serum IgG1 less than 8 mg/ml), whereas 27 of 58 (46.5%) born from November through May had failure of passive transfer.

Measurement of serum immunoglobulins in 46 foals less than 2 weeks old revealed 9 foals with hypogammaglobulinemia. The hypogammaglobulinemia was attributed to failure in transfer of immunoglobulins from dam to foal via colostrum. Three of the affected foals did not nurse at all, or only slightly, and 2 of these died of infections within a few days after birth, whereas the 3rd foal did not grow as well as normal foals. Six of the affected foals nursed in an apparently normal manner, and 5 of these had nonfatal respiratory infections between 2 and 5 weeks of age. Analysis of serum samples from surviving foals demonstrated that immunoglobulins were eventually produced. One other foal examined had hypogammaglobulinemia at 57 days of age, an age when the foal should have produced large amounts of immunoglobulin independent of passive transfer. This foal had simultaneous infections and hypogammaglobulinemia, but eventually produced normal amounts of immunoglobulin. Cellmediated immunity was normal at 3 months of age. This condition was designated transient hypogammaglobulinemia and was thought to be due to a temporary inability to make immunoglobulins.

The transfer of immunoglobulin G (IgG) and immunoglobulin M (IgM) from ewe to lamb was quantitated to determine the occurrence of failure in passive transfer. Concentrations of IgG and IgM in ewe serum did not correlate with those in the colostrum. Colostrum from all ewes contained abundant amounts of immunoglobulins when compared with serum values, with IgG being selectively concentrated over IgM. Absorption through the intestinal tract of the lamb appeared to be a nonselective process, lacking predilection for IgG and IgM. All lambs tested 24 hours after birth absorbed colostral immunoglobulins to some extent; however, 13 (14%) of 91 clinically normal lambs demonstrated some failure of passive transfer. In contrast, failure of passive transfer was found in 27 (46%) of 59 lambs dying of natural causes between 24 hours and 5 weeks of age. Evidence presented emphasizes the importance of absorption of adequate amounts of immunoglobulins to enable the newborn lamb to survive the first few weeks of life.