Complement activation and fibrinolysis during infection with Theileria parva (East Coast fever) in cattle

The role played by complement and fibrinolysis in the pathogenesis of pulmonary oedema and the disseminated, subendothelial, petechial haemorrhages in cattle infected with $\text{Theileria parva}$ was investigated. There was a marked reduction in total haemolytic complement in animals that died of this disease whereas the survivors showed only transient changes. The reduction in C3 levels in animals with severe disease, however, was equivalent to that in the survivors. Neither circulating soluble immune complexes nor free antibody against macroschizonts were detected in sera from cattle with severe infection. In addition to the complement changes, high levels of fibrinogen degradation products (FDPs) were detected in sera from the infected animals. The rate of production of FDPs and reduction in total haemolytic complement paralleled the clinical course of the disease. It is, therefore, suggested that the pulmonary oedema and disseminated petechial haemorrhages seen in East Coast fever may be the direct result of the combined effects of complement activation and consumption of fibrinogen and fibrin.     

Development of immune responsiveness to Ascarissuum antigens in pigs vaccinated with ultraviolet-attenuated eggs

Pigs fed $\text{Ascaris}$$\text{suum}$ eggs attenuated by short wave ultraviolet-radiation developed resistance to challenge infection. Per os inoculation of pigs on three successive weeks with 10,000 eggs irradiated to total exposures of 150,100 and 75 μW-min/cm², respectively, resulted in an 88% reduction in the number of larvae recovered from the lungs 7 days after challenge with 10,000 infective eggs. Peripheral blood lymphocytes (PBL) from vaccinated pigs were specifically stimulated in vitro to incorporate tritiated thymidine by egg hatching fluid (Ea) and by excretory-secretory products obtained from cultures in defined-media of second-stage larvae (L2) developing to third-stage (L2–3) and from cultures of third-stage larvae developing to fourth-stage (L3–4). PBL were also specifically stimulated by living L2. Ea and L2 stimulated pig PBL significantly at 7 days after the first inoculation; responses to L2–3 and L3–4 developed 7 days after a second inoculation. The antigen-responsive cells in the PBL population were non-immunoglobulin-bearing lymphocytes. Antibodies to Ea and L2–3 were not detected in the serum of vaccinated pigs, and only 3 of 7 pigs had low concentrations of serum antibodies to L3–4.     

Brucella abortus antigen-induced lymphocyte reactivity in guinea pigs

A whole blood lymphyocyte transformation (WBLT) assay was used to detect anti-brucella lymphocyte reactivity in guinea pigs. Brucella antigens stimulated an antigen-specific lymphoproliferative response in WBLT assays from $\text{Brucella abortus}$ infected guinea pigs. The response was best detected from 6 to 16 weeks after challenge inoculation with viable $\text{B. abortus}$ 2308. Lymphocytes were not stimulated by unrelated bacterial antigens and control animals did not respond to the Brucella antigens. The responding cell population was characterized as mostly T lymphocytes. The WBLT assay was found to be specific for the detection of anti-brucella lymphocyte reactivity. However, a negative response was not definitive, which indicated a need for repeated testing to establish that a guinea pig did not have anti-brucella lymphocyte sensitization.     

Presence of antigen sensitized leukocytes in carp (Cyprinus carpio L) following bath immunization against Flexibacter columnaris

Bath immunization of carp (Cyprinus carpio L) resulted in protection of fish at natural challenge. Stimulation of leukocytes derived from thymus, spleen, anterior kidney and mid-kidney of fish immunized with Flexibacter columnaris bacterin revealed the presence of antigen sensitized cells in all lymphoid tissues except the anterior kidney. After 28 days a response was obtained in thymus and spleen leukocyte cultures.     

Bovine babesiosis: The immunization of cattle with fractions of erythrocytes infected with Babesia bovis (syn B. argentina)

Soluble antigen which protected susceptible cattle against challenge with $\text{Babesia bovis}$ was extracted from $\text{B. bovis}$-infected erythrocytes by sonic disintegration and separation of the soluble from the insoluble matter by ultracentrifugation. The material was then fractionated by the precipitation of fibrinogen-like proteins. The precipitate contained the babesial antigens that were located on the stroma of the infected erythrocytes. Antigen originally located on the parasite remained in solution. Both fractions conferred protection on splenectomized calves against challenge with $\text{B. bovis}$. However, the fraction containing the parasite antigens appeared to have more potential for development as a killed vaccine because it was not heavily contaminated with antigenic material from bovine erythrocytes.     

An ELISA test to detect antibody to Bordetella bronchiseptica

An enzyme-linked immunosorbant assay (ELISA) was developed to measure antibody to $\text{Bordetella bronchiseptica}$ in dogs. The ELISA test was more rapid and sensitive and required 50 to 150 times less antigen than the amount of antigen required for the conventional tube agglutination test. A survey of 50 canine serum samples using ELISA suggested that 8% of all sera had titers greater than 1:64, 56% had titers of 1:8 to 1:64, and 36% had titers of less than 1:8. The mean titer of survey sera was 1:46 and the median titer was 1:16. Serum antibody responses in dogs inoculated with a commercially available bacterin were compared with responses in dogs inoculated with experimental endotoxin depleted bacterin.     

Application of indomethacin as a potentiator of lymphocyte blastogenesis in Brucella abortus exposed cattle

Lymphocytes from $\text{Brucella abortus}$ field strain infected, strain 19 vaccinated, non-exposed and field strain infected, but immunologically unresponsive cattle were incubated with $\text{B. abortus}$ antigen and indomethacin. There were significant increases (P < 0.005) in the blastogenic responses, as measured by [³H] thymidine uptake, in cultures with indomethacin as compared to cultures without indomethacin. Lymphocyte blastogenic responses to $\text{B. abortus}$ antigen were potentiated by indomethacin in both $\text{B. abortus}$ exposed and non-exposed cultures. However, potentiation of sensitized lymphocyte blastogenic responses by indomethacin was significantly greater (P < 0.005) than that in non-exposed lymphocytes. Additionally, indomethacin significantly potentiated Brucella-induced lymphocyte blastogenic responses in lymphocytes from anergic cattle.     

Some immunopathological aspects of aged Praomys (Mastomys) natalensis

$\text{Praomys (Mastomys) natalensis}$ develops a wide variety of autoantibody specificities with age; of these only those to thyroid colloid were associated with a histopathological lesion e.g. thyroiditis. An inverse relationship was found between the mean number of autoantibodies and the occurrence of lymphoreticular tumors. It appears that $\text{Mastomys}$ is a better model for studies on the general mechanisms of autoimmunity than for investigating specific autoimmune diseases per se.     

Intestinal mast cell response in thymectomised and normal mice infected with Trichinella spiralis.

In NIH mice, expulsion of $\text{Trichinella spiralis}$ from the small intestine and increase in intestinal mast cells were each dependent on the presence of T-lymphocytes. Both changes were deficient in thymectomised mice but could be largely restored by reconstitution of thymectomised mice with syngeneic mesenteric lymph node cells. In both NIH and BALB/C mice the majority of the increased number of mast cells occurred within the intestinal epithelium. In NIH mice increase in the number of intestinal mast cells coincided roughly with expulsion of the parasites.In BALB/C mice increase in the numbers of intestinal mast cells did not appear to be connected with the location or expulsion of parasites and it is concluded that mast cell proliferation, accumulation and discharge $\text{per se}$ do not result in or from worm expulsion.     

Coccidiosis: T-lymphocyte-dependent effects of infection with Eimeria nieschulzi in rats

The intestinal pathology caused by infection with $\text{Eimeria nieschulzi}$ was investigated and comparisons were made between the effects in athymic nude (rnu/rnu) rats and their heterozygous (rnu/+) litter-mates. Most of the changes noted, i.e. increase in gut weight, partial villous atrophy and increased numbers of mast, goblet and pyroninophilic cells were shown to be largely or wholely thymus dependent. The numbers of intraepithelial lymphocytes were decreased in both groups during the period of study. The peripheral blood leucocyte response was similar in both groups of rats during a primary infection but differed after a challenge inoculum, indicating that the secondary type of response which occurred in the rnu/+ rats was thymus dependent, as is resistance to reinfection.     

Solid-phase enzyme immunoassay of bovine antibody responses following immunization against and natural infection with Pasteurella haemolytica

A solid-phase enzyme immunoassay (EIA) was developed in order to monitor bovine antibody responses following immunization against and natural infection with $\text{Pasteurella haemolytica}$ serotype A:1. Non-ionic surfactants, used in many antibody EIAs to reduce non-specific immunoglobulin binding, had to be avoided because they inhibited specific binding of bovine antibodies to $\text{P. haemolytica}$ antigens. Calves were immunized with a KSCN extract of $\text{P. haemolytica}$. Subcutaneously immunized animals developed a significantly higher humoral antibody response than did intranasally vaccinated animals. Intranasally immunized calves developed a slightly, but not significantly higher nasal antibody response than did calves vaccinated subcutaneously. Field study results based on bacterial isolation and EIA detection of antibodies to $\text{P. haemolytica}$ indicate that cattle can generate carrier states where bacteria are present in the upper respiratory tract, yet no humoral antibody response is induced. The converse was also found where cattle were free from $\text{P. haemolytica}$ in the upper respiratory tract, yet possessed a good humoral antibody response to $\text{P. haemolytica}$.     

Effects of ionizing radiation on in vitro proliferative responses of porcine peripheral blood lymphocytes

The radiosensitivity of $\text{in vitro}$ proliferative responses of porcine peripheral blood lymphocytes (PBL) was assessed. PBL were stimulated by Con-A, PHA, culture supernates from mitogen-stimulated porcine lymphocytes, or in the case of antigen-primed swine, specific antigens. The resulting levels of proliferation were assessed by a determination of the level of incorporation of tritiated thymidine $\text{in vitro}$, and in some cases by the presence of blast cells in the cultures. Porcine PBL were found to be more radioresistant than either mouse PBL or mouse spleen cells. Irradiation levels of greater than 3000 rads were necessary to arrest Con-A or PHA-induced proliferative responses. Proliferation induced by lymphokines in the form of supernates from mitogen-stimulated lymphocyte cultures was arrested in PBL that had received 3000 rads prior to culture. Antigen-induced proliferative responses in primed porcine PBL populations were the most radiosensitive, in that a previous irradiation with 500 rads was sufficient to completely abolish a secondary $\text{in vitro}$ proliferative response.     

The immune response of goats vaccinated with low and high doses of Brucella melitensis Rev 1

The serological response, lymphocyte reactivity, and dermal hypersensitivity reactions of goats vaccinated with high and low doses of $\text{Brucella melitensis}$ Rev 1 organisms to Brucella antigens were studied. Antibodies to soluble antigen A2 were detectable by immunoelectrophoresis (IE), appeared later than agglutinating antibodies but disappeared faster in most vaccinated animals. Anti-A2 antibodies appeared earlier in goats which received the high dose. Antibodies to polysaccharide B antigen were not detected. Lymphocytes reactive to Brucella antigens in lymphocyte transformation assays appeared at variable times after vaccination. In contrast to the humoral response to antigen A2, the appearance of circulating, reactive lymphocytes was not dependent on vaccine doses. All vaccinated animals demonstrated dermal hypersensitivity reactions to one of the antigenic extracts. Skin reactions peaked at 24 hrs post inoculation. The reactions were elicited when all but one goat had undetectable anti-A2 antibodies and no circulating, reactive lymphocytes as measured in whole blood lymphocyte transformation assay.     

Antibodies to Brucella abortus in sera from strain 19 vaccinated and non-vaccinated cows as determined by enzyme linked immunosorbent assay and conventional serologic methods

The sensitivity of an enzyme linked immunosorbent assay was compared with 4 other serologic methods for detecting antibodies in $\text{B. abortus}$ vaccinated and non-vaccinated cows. Antibodies were detected by ELISA in sera from more than 93% of the culture positive cows, however, less than 55% of the infected animals were identified by any other serologic method. Similarly, antibodies were detected in 82% of the culture negative cows by the ELISA method.     

The use of a cathodic antigen in the immunoelectrophoretic serodiagnosis of Echinococcus granulosus in sheep

An antigen was prepared from purified sheep hydatid-cyst fluid by gelfiltration on Sephadex G-200 followed by chromatography on DEAE-cellulose. In each case the first-peak material was used. This antigen, which migrated cathodically, was concentrated and used in immunoelectrophoretic analyses of 4X concentrated sera from sheep experimentally and naturally infected with $\text{Echinococcus granulosus}$ and from uninfected sheep. Of 34 sheep with $\text{E. granulosus}$ infection 31 were positive with the cathodic antigen while of 85 sheep without $\text{E. granulosus}$ 8 were (falsely) positive. Many false positives appeared to be associated with heavy infections of $\text{Taenia ovis}$ or $\text{T. hydatigena}$ larvae. The “arc 5” immunoelectrophoresis test, which is the most specific immuno-diagnostic test for echinococcus infection in humans, was not able to specifically identify $\text{E. granulosus}$ infections in sheep.     

The irradiation of Babesia bovis. II. The immunogenicity of irradiated blood parasites for intact cattle and splenectomised calves

Intraerythrocytic forms of $\text{B. bovis}$ were exposed to 350 Grays (Gy) γ irradiation and were then injected intravenously into intact two and three year old Hereford steers. One of 15 steers died on initial infection and subsequently six steers were given a virulent heterologous challenge three weeks after recovery; all six animals were highly immune. The remaining eight animals were kept under quarantine conditions for 10 months and were then challenged with a different virulent heterologous strain of $\text{B. bovis}$. Seven of eight were highly immune, but one animal died. Subsequently a further 12 steers were injected intravenously with 1 × 10⁸ irradiated organisms. All showed only mild transient clinical signs. After 12 months quarantine in a tick-free area these animals were then challenged with a virulent heterologous strain and all 12 were shown to be highly immune. Irradiation reduced the infective dose from 1 × 10⁸ to 2.5 × 10³ parasites. These parasites multiplied at the same rate, and achieved the same maximum parasitaemia as the parent non-irradiated strain, but the disease produced by them was not severe. A dose of 2.5 × 10³ non-irradiated paasites was lethal to all of the four animals which received it. It was concluded that irradiation had produced a predominantly avirulent parasite population.     

mRNA from porcine colostral and milk cells coding for specific antibodies in response to orally administered Escherichia coli strains

The effect of feeding live $\text{Escherichia coli}$ bacteria to sows on the lymphocyte populations in colostrum and milk was studied by $\text{in ovo}$ translation of the mRNA of these cells (Kortbeek-Jacobs and van der Donk, 1978). Four $\text{E. coli}$ strains were tested: one wild type K88 producing strain, two conjugated K12 strains producing K88 antigen and one wild type K12 strain. Only after feeding the wild type K88 bacteria, colostral lymphocytes contained mRNA coding for anti-K88 antibodies, predominantly of the IgA and IgM isotypes. Milk lymphocytes of the sows receiving one of the other strains were capable of producing more anti-K88 antibodies than the sows that were fed the wild type K88 strain. Antibodies were predominantly of the IgM class. The reactivity of the milk lymphocytes is ascribed to the strain with which the piglets were challenged. The results support the phenomenon of sensitized gut lymphocytes that home to the mammary gland.     

The role of serum and biliary antibodies and cell-mediated immunity in the clearance of S. typhimurium from chickens

The development of three parameters of immunity in response to a non-lethal infection of $\text{Salmonella typhimurium}$ in adult chickens has been examined. Intravenous inoculation of 1 × 10⁶ organisms established infection in the liver, spleen and intestinal tract of all birds; the organism persisted in these sites until day 9 of the infection, after which it was cleared rapidly from all sites. High levels of agglutinating and haemagglutinating antibodies were found in serum and bile 5 days after infection; they peaked at days 7 to 10, and detectable antibody was still present in both fluids 6 weeks after infection. The presence of this antibody did not appear to cause a significant reduction in organism numbers in any of the sites examined. Cell mediated immunity was detected at day 14. It is suggested that cell mediated immunity is responsible for clearance of the organisms from the tissues.     

Nippostrongylus brasiliensis infection in the rat: 1. Production of bile lgA and serum lgG antibodies by adult rats

Bile lgA and serum lgG antibody responses were assayed by a micro ELISA technique. Antibodies of both classes were produced by 14 days after infection against whole worm antigens and worm metabolic product antigens, by adult rats infected once with $\text{Nippostrongylus brasiliensis}$. Rats were reinfected on day 28 and titres of both classes of antibody against both antigens were greater 7 and 14 days after reinfection than after the initial infection. Bile lgA antibodies were produced against phosphorylcholine after a single infection with the parasite but no increased response occurred after reinfection. The possible significance of lgA and lgG antibody responses in relation to immunity to intestinal nematode infections is discussed.     

The early phase of the immune response to live and killed Staphylococcus aureus vaccines in sheep — Cellular and immunoglobulin parameters

Various cellular and protein parameters in efferent lymph were monitored during early stages of immune responses in the draining popliteal lymph nodes of sheep following injection with either live or killed $\text{Staphylococcus aureus}$ vaccines. Significantly higher outputs of leucocytes and lymphoblasts were measured in lymph from animals injected with the live vaccine (Group 1) compared with animals given the killed vaccine (Group 2) and non-immunised controls (Group 3). At 48 h post-injection the mean output of leucocytes in lymph for Group 1 was 9.5 × 10⁷ cells/h, and for Groups 2 and 3 comparable mean outputs were 5.2 × 10⁷ and 2.2 × 10⁷ respectively.There was a marked increase in IgM-containing cells in Group 1 animals, significant differences between Group 1 and 2 occurring 96 h post-injection. A difference was observed between the two immunised groups in the kinetics of output of cells containing antibody to a staphylococcal antigen extract. There was a slower increase in antibody-containing cells for Group 1 than for Group 2 animals. The mean proportion of antibody-containing cells reached 0.09% (of total leucocytes) at 72 h post-injection for Group 2 and 0.10% at 120 h post-injection for Group 1.Rapid increases in flow rate of lymph were recorded in animals in Group 1. These results were in contrast to lower corresponding values observed in sheep in Groups 2 and 3. There was a twofold increase in the lymph: serum (L:S) concentration ratio for IgG2 in animals in Group 1 and the evidence suggested that this was due to local synthesis of IgG2 by lymphoid cells in the abscess at the site of injection and/or in the draining popliteal lymph node. This change in L:S ratio for IgG2 was not recorded for sheep in Groups 2 and 3.