Changes in the generation cycle of duodenal crypt cells in chickens infected with Eimeria acervulina

Changes in the duration of the progenitor cycle and its four phases were determined for duodenal crypt cells in chickens infected with Eimeria acervulina. Metaphase curves were constructed using percent labelled metaphase nuclei in duodenal crypt cells at short intervals after the injection of [3H]thymidine. The duration of the progenitor cycle and its four phases were calculated using the synthetic index and data obtained from the metaphase curves. The cycle time was reduced from 14 h in control birds to 10.2 h at 2 days and 10.6 h at 4 days postinfection. The change was attributable entirely to a reduction in G1 or the presynthetic phase. In addition, the population of dividing cells within each duodenal crypt was almost doubled in infected birds. These increases in cell production precedes all the histological changes observed earlier in the intestines of E. acervulina infected chickens. At least in this instance, changes in crypt morphology seems, therefore, to result from an induced change in the functional activity of the crypt.     

In situ characterization of leucocyte subpopulations after infection with Eimeria tenella in chickens

We characterized the leucocyte subpopulations after infection with Eimeria tenella in both naive and immune chickens. Immunocytochemical staining was used to characterize the cells in situ, so that the interaction between host and parasite could be studied. More leucocytes were detected in the lamina propria of immune chickens, and leucocytes infiltrated the ceca more rapidly than in naive chickens, but the infiltration was less pronounced than in naive chickens. In naive chickens, most infiltrated leucocytes were macrophages and T cells. Two days after inoculation the number of CD4+ cells had increased greatly. In immune chickens, mainly T cells (CD4+ and CD8+) infiltrated the lamina propria, and in contrast to naive chickens, the number of CD8+ cells exceeded the number of CD4+ cells. Furthermore, we characterized which cells contained a parasite and which cells were detected next to the parasites, because these cells are probably involved in the arrested development of the parasites. In naive chickens, sporozoites were significantly more often located within or next to macrophages than in immune chickens. In immune chickens, sporozoites were significantly more often located within or next to CD3+, CD8+, and TCR2+ cells. In conclusion, the marked increase of CD4+ cells after primary infection suggests that these cells are involved in the induction of the immune response, whereas the increase of CD8+ cells after challenge infection suggests that these cells act as effector cells.     

A preliminary study of the nature of infection and immunity in chickens given an attenuated line of Eimeria acervulina

The Houghton (H) strain of Eimeria acervulina was attenuated by serial passage through chickens of the first oocysts produced during infection. This selection pressure resulted in a reduction in the pre-patent period of the parasite, shown to be due to the selection of a line predominantly with only 3 instead of 4 generations of schizonts. The precocious line had a reproductive potential much lower than that of the parent strain and it was significantly less pathogenic. Chickens given oocysts of the precocious line were almost completely immune to challenge with the Houghton strain.     

Kinetics study and characterisation of target excreted/secreted antigens of immunoglobulin G, M, A and E antibodies from mice infected with different strains of Toxoplasma gondii

A kinetics study and characterisation of target excreted/secreted antigens of immunoglobulin (Ig) G, M, A and E antibodies were realised by Western blotting with immune sera of mice inoculated with three strains of Toxoplasma gondii: RH, C56 and S3. IgG antibodies of the immune sera recognised the major proteins of the three excreted/secreted antigen preparations with molecular masses of 30, 45, 63 and 77 kDa. IgM antibodies recognised proteins revealed by IgG antibodies but with variable intensities; some proteins were revealed during a short period. IgA antibodies did not recognise the 35-kDa antigen or the antigens inferior to 28 kDa. The RH excreted/secreted antigens were revealed with the highest intensity. The IgE antibodies were briefly detected in trace amounts during period from the 20th to the 35th day. The RH strain with its excreted/secreted antigens had the best antigenicity and is a good model for immunoprotection studies.     

Effect of betaine on the growth performance of chicks inoculated with mixed cultures of avian Eimeria species and on invasion and development of Eimeria tenella and Eimeria acervulina in vitro and in vivo

At 7 d postinoculation (DPI) with a mixed culture of avian Eimeria species, 21-d-old chicks maintained in batteries and floor pens on a diet containing 0.15% (3 lb/ton) betaine plus 66 ppm (60 g/ton) salinomycin were significantly heavier and had significantly lower feed conversion ratios and mortality than chicks fed diets containing 0.15% betaine or 66 ppm salinomycin alone, or the control diet. At 31 DPI, when the chicks were 45 d old, the differences between the diet groups were not as great as at 7 DPI. In vitro, except at high concentrations, betaine was nontoxic to sporozoites of Eimeria tenella or Eimeria acervulina and had little effect on their invasion and development in cultured cells. In vivo, invasion by E. tenella and E. acervulina sporozoites was significantly reduced in all chicks fed diets containing betaine or salinomycin compared with that in control chicks. There was a significant interaction between betaine and salinomycin that impacted on invasion by both species. Overall development of E. tenella did not appear to be adversely affected by addition of betaine to diets containing salinomycin. Conversely, development of E. acervulina was reduced in chicks fed diets containing 0.075% (1.5 lb/ton) betaine plus 66 ppm salinomycin as compared with that in chicks fed salinomycin alone.     

Comparison between effects of standard feed and whole wheat supplemented diet on experimental Eimeria tenella and Eimeria maxima infections in broiler chickens

Fas is a surface receptor that can transmit signals for apoptosis. Using retroviral cDNA library-based functional cloning we identified a gene, toso, that blocks Fas-mediated apoptosis. Toso expression was confined to lymphoid cells and was enhanced after cell-specific activation processes in T cells. Toso appeared limited to inhibition of apoptosis mediated by members of the TNF receptor family and was capable of inhibiting T cell self-killing induced by TCR activation processes that up-regulate Fas ligand. We mapped the effect of Toso to inhibition of caspase-8 processing, the most upstream caspase activity in Fas-mediated signaling, potentially through activation of cFLIP. Toso therefore serves as a novel regulator of Fas-mediated apoptosis and may act as a regulator of cell fate in T cells and other hematopoietic lineages.     

Immunocytochemical colocalization of specific immunoglobulin A with sendai virus protein in infected polarized epithelium

Thrombocytopenia is often the dose-limiting toxicity for radionuclide therapy. Prediction of platelet counts after therapy is important for treatment planning. Simple prediction methods based on linear correlation between radiation dose and blood count nadir have been insufficient because they have not considered time, because of the complicated hierarchical structure of the hematopoietic system in which platelets are not directly injured by low dose rate radiation and because of changing radiation dose rates to marrow with time. This study addresses these problems using a cell kinetics model. METHODS: The model consists of compartments for progenitor cells, megakaryocytes, platelets and stromal cells. A linear quadratic formula was used for progenitor cell survival. Stromal cells were described by a model based on a maximum likelihood estimate for cellular damage, repair and proliferation. Reported values for murine cellular turnover rates and radiosensitivity of progenitor cells were used in the model calculations. Experimental mice received 4 Gy of external beam radiation for tumor implantation and 12.4-23.3 MBq 67Cu-2-iminothiolane-BAT-Lym-1 (BAT = 6-[p-(bromoacetamido) benzyl]-1,4,8,11-tetra-azacyclotetradecane-N,N',N',N'-tetraacet ic acid) 19-30 days later. Blood counts were measured three times each week. RESULTS: The model predicted the severity of thrombocytopenia, and the time of the nadir corresponded to measured values in mice. For a dose of 14.2 MBq 67Cu-2-iminothiolane-BAT-Lym-1 that induced a platelet nadir of 20% of baseline (Grade II), the model predicted that at least 20 days were needed before a second 14.2-MBq injection if a subsequent nadir of <10% of baseline (Grade IV) was to be avoided. CONCLUSION: The nadir and duration of thrombocytopenia predicted by the model were similar to those observed in the mice. Predicted information could be useful for planning the dose and timing of fractionated radionuclide therapy. This model provides a stepping stone for future development of a predictive model for patients.     

Alkaline phosphatase activities in intestinal mucosa from calves infected with Cooperia punctata and Eimeria bovis

Thirty calves 12 weeks of age raised under essentially parasite-free conditions were used to determine the effects of Cooperia punctata and Eimeria bovis (in single and combination infections) on mucosal alkaline phosphatase activities at 8 locations in the small intestine. In experiment 1, 5 calves infected with C punctata 3 weeks previously had highly significant (P less than 0.01) reductions in mucosal alkaline phosphatase activities compared with those in 5 noninfected control calves. These reductions were greatest in the 2 locations of the intestine closest to the pylorus. Infected calves had a mean of 44,356 C punctata adults present. In experiment 2, 5 calves infected with E bovis 2 weeks previously had significant (P less than 0.05) reductions in mucosal alkaline phosphatase activities compared with those in 5 noninfected control calves. These reductions were present in the caudal half of the intestine. Numerous 1st-generation E bovis schizonts were present in the caudal third of the intestine. In experiment 3, 5 calves infected with C punctata and E bovis of the same durations as in experiments 1 and 2 had highly significant (P less than 0.01) reductions in mucosal alkaline phosphatase activities in the cranial half of the intestine compared with those in the controls. These reductions were much larger than in either of the monospecific infections (experiments 1 and 2). A mean of 31,968 adult C punctata were recovered from the infected calves, and numerous E bovis schizonts were observed in the caudal third of the intestine.     

Eimeria acervulina: influence of corticosterone-induced immunosuppression on oocyst shedding and production characteristics in broilers, and correlation with a computer simulation model

An experiment was conducted to investigate the effects of immune responsiveness on excretion of oocysts after E. acervulina infection and subsequent effects on production characteristics of broilers (Gallus domesticus). These effects were determined in broilers repeatedly infected with 2.85 x 10(3) oocysts of E. acervulina and treated with various dosages of corticosterone in the diet (0, 10, 20 and 30 p.p.m.). Corticosterone treatment did not have an effect on the peak oocyst excretion, although it was administered from 4 days before initial infection. The number of oocysts excreted shortly after the peak and the length of the excretion period were increased in corticosterone-treated groups. The absence of a difference in peak oocyst excretion was ascribed to the existence of a time-lag between first contact with the parasite and rate of development of protective immunity. In a recently developed computer simulation model this period was assumed to be 5 days. Assuming that immunosuppression, through corticosterone, is only effective when protective immunity is in operation, the results indicate a time-lag of at least a few days, which supports the inclusion of such a time-lag in the computer simulation model. General immunosuppressive effects of the corticosterone treatment, monitored by antibodies and mitogen-induced lymphocyte stimulation confirmed that immunosuppression occurred shortly after medication started. Infection did not have a significant influence on production characteristics in animals without dietary corticosterone. However, with increasing corticosterone levels the negative effects of infection on production also increased.     

Kinetic analysis of T cells and antibody production in chickens infected with Marek's disease virus

In chickens inoculated with a Marek's disease (MD) vaccine and subsequently with virulent MD virus (MDV), CD4+ T cell population was drastically decreased following a transient increase at 21 days after hatching (16 days after MDV infection). To elucidate the immune response after the decrease of CD4+ T cell population, the antibody production against sheep red blood cells (SRBC) was examined in these chickens. Chickens challenged with a virulent MDV after MD vaccination produced lower titers, of anti-SRBC antibody than untreated control chickens. Antibody production against SRBC was also lowered in vaccinated chickens or chickens challenged with a virulent MDV.     

Prediction of myocardial infarction in dyslipidemic men by elevated levels of immunoglobulin classes A, E, and G, but not M

BACKGROUND: Immune mechanisms have been suggested to play an important role in the development of coronary atherosclerosis and its thrombotic complications. We evaluated the predictive value of the levels of various serum immunoglobulin classes in middle-aged men at increased risk of myocardial infarction. METHODS: Using nested case-control design and logistic regression analysis, we estimated the association between serum immunoglobulins and the risk of coronary end points (nonfatal or fatal myocardial infarction or sudden cardiac death) in dyslipidemic men (levels of non-high-density lipoprotein cholesterol >5.2 mmol/L [>201 mg/dL]) participating in the Helsinki Heart Study. The cases consisted of 135 subjects in whom a coronary end point occurred during the 5-year observation period of the study, and the controls were 135 subjects who did not suffer coronary end points during this period. Levels of IgA, IgE, IgG, and IgM were determined in serum samples collected at study entry. RESULTS: Levels of IgA, IgE, and IgG, but not IgM, were significantly higher in cases than in controls. After adjustment for other risk factors, such as age, smoking, and blood pressure, the risk of coronary disease showed a significant relation to the levels of IgA, IgE, and IgG. The risk in the highest quartile of each distribution as compared with the lowest quartile was 2.2-fold for IgA (95% confidence interval, 1.0-4.5), 2.8-fold for IgE (1.3-5.9), and 2.8-fold for IgG (1.3-5.9). Hypertriglyceridemia and a low level of high-density lipoprotein cholesterol were associated with increased risk of a coronary end point only if the levels of IgA, IgE, or IgG were also elevated. CONCLUSION: Elevated levels of IgA, IgE, and IgG are associated with myocardial infarction and cardiac death in men with dyslipidemia. The present data suggest that, for dyslipidemia to cause coronary atherothrombosis, an immune response reflected by elevated levels of these immunoglobulin classes is an important determinant.     

Significance of concentrations and ratio of immunoglobulin G and M in the evaluation of anti-Toxoplasma IGM

The Authors have tested the significance of the correlation of the mean total IgM concentration, the IgG: IgM ratio and the presence of specific anti-Toxoplasma gondii IgM, determined with different serological reactions. The piece of research has been carried out on 72 samples of subjects with active toxoplasmic infections.     

Evaluation of the efficacy of human antimeningococcal immunoglobulin G in infant rats experimentally infected with Neisseria meningitidis group B

Infants rats, a well known model for the experimental reproduction of bacterial meningitis, were used by us to test the protective potential of antibodies developed in humans who had been vaccinated with the Cuban antimeningitis vaccine (VA-MENGOCBC). Newborn rats were inoculated by the intraperitoneal and intranasal routes with suspensions of Neisseria meningitidis group B bacteria. Bacteremia kinetics were evaluated from blood and brain-spinal fluid cultures. Samples of the central nervous system were taken and smears of backbone fluids prepared for histopathologic evaluations. Characterization of bacteremia evolution, as well as the mean lethal dose of germs and histopathologic features, were determined. After standardization of the model, therapeutic schemes were applied using passive immunization pre- and post-infection with N. meningitidis. A significant level of protection was obtained in relation to control animals that received the same challenge doses.     

Development and comparison of assays for measuring immunospecific antibody response in serum of chickens and rabbits immunized with human immunoglobulin G

The antibody response development during polyclonal antibody production is a relevant parameter to monitor during the immunization period to be able to optimize the immunization protocol and to determine the optimal antibody harvest time. Although rabbits and other mammals are most often used for polyclonal antibody production, the chicken is a relevant alternative. There are both scientific reasons, economic reasons, and animal welfare reasons to consider when choosing the chicken instead of a mammal for this purpose, because antibodies in generous quantities can be harvested from the egg yolk. This study compared different assays for measuring antibody response in rabbit and chicken serum. An inhibition liquid phase absorption assay (ILPAA), a rocket immunoelectrophoresis (RIE) assay, and a line immunoelectrophoresis (LIE) assay were compared to ELISAs. The ELISA proved to be the most useful assay for routine use, as it was less time-consuming and because the assay could easily be adapted to both serum antibody types. However, electrophoretic assays were the most useful as combined analytical and quantitative tools and must be considered essential when analyzing specificities of polyclonal antibody preparations.     

Persistent elevation of immunoglobulin G titer against the C region of recombinant group A streptococcal M protein in patients with rheumatic fever

To analyze the immune response to the C region of group A streptococcal M protein in patients with rheumatic fever (RF), we cloned the structural gene for the C region of type 12 M protein and produced recombinant C region of M protein. IgG titers against the C region of M protein were measured by ELISA from sera of patients with RF (n = 10), uncomplicated streptococcal pharyngitis (n = 26), and age-matched controls (n = 49). IgG titers against the C region were significantly higher in patients with RF than in controls or patients with uncomplicated streptococcal pharyngitis (43 versus 1.5 or 1.8 micrograms/mL, p < 0.01). Studies using overlapping synthetic peptides of the C region demonstrated that increased IgG reactivity was observed against the amino-terminal halves of the C repeat blocks (C1, C2) in RF, indicating that these domains are the main immunodominant epitopes in the C region.     

Host protective antibodies and serum immunoglobulin isotypes in mice chronically infected or repeatedly immunized with the nematode parasite Nematospiroides dubius

The nematode parasite Nematospiroides dubius survives to give a chronic primary infection in mice. However, mice subjected to weekly infections of 125 larvae, interspersed by treatment with an anthelmintic to prevent the accumulation of lethal numbers of adult worms in the intestine, develop host-protective antibodies in their serum. The protective effect of these antibodies was demonstrated by passive transfer to naive recipients or to mice already adoptively immunized with immune mesenteric lymph node cells (IMLNC). Sera were first shown to exhibit protective activity during the third and fourth weeks of the multiple immunizing infection, reaching a peak level by week six beyond which there was no further increase in protective activity. This increase was correlated with a ten-fold, concurrent rise in serum IgG1 levels. None of the other immunoglobulin isotypes underwent comparable changes in concentration nor could they be correlated with the pattern of appearance of host-protective antibodies in the sera of donor mice. This suggested that host protective antibodies were of the IgG1 class. CFLP and C57BL10 mice (the latter is a weak responder strain) both had high levels of host-protective antibodies in their serum. However when the sera from NIH mice (a strong responder strain) were compared, they exhibited far less protective activity on passive transfer to recipient mice, although when given together with IMLNC, serum from multiply-immunized NIH mice enhanced the protective effect of IMLNC synergistically. When primary infection serum was assayed in this passive/adoptive transfer model, no host-protective antibodies could be demonstrated, even with pools of primary infection serum taken 10 and 17 weeks after infection. These results are discussed with respect to the possible mechanisms by which N. dubius evades the host immune system to give rise to long-lasting primary infections in mice.