Longitudinal studies of immune function in cattle experimentally infected with bovine immunodeficiency-like virus and/or bovine leukemia virus

The effects of single or dual infection with bovine immunodeficiency-like virus (BIV) and/or, bovine leukemia virus (BLV) on bovine immune function were examined over a 4 year period. Holstein calves were infected with BIV (four calves), BLV (five calves), BIV and BLV (five calves), or sham inoculated (three calves). Lymphocyte blastogenesis to mitogens, seven tests of neutrophil function, and mononuclear cell subset analysis by flow cytometry (BoCD4, BoCD8, BoCD2, BoWC1, sIgM+, and monocytes) were performed at regular intervals to 49 months post-infection. These data were analyzed for main effects of each virus and interaction as a 2 x 2 factorial. BIV infected cattle had lower neutrophil antibody-dependent cell-mediated cytotoxicity and iodination responses during 2 of the 4 years post-infection (P < 0.05). BIV infection was not associated with any long-term significant changes in lymphocyte blastogenesis to mitogens or changes in mononuclear cell subset numbers in blood. There was a tendency for animals infected with BIV alone to have decreased lymphocyte blastogenic responses to mitogens, but this was not statistically significant. BLV infection caused an increase in total mononuclear cells with no dramatic shift in the relative proportions of the various subsets. Co-infection with BIV and BLV did not consistently cause a different response than either virus did individually. One BIV infected animal died of non-BLV lymphosarcoma 7 months after infection. All other animals had no unusual clinical signs. In summary, infection with BIV caused a significant, temporary decrease in neutrophil function with no consistent statistically significant alteration in lymphocyte blastogenesis or mononuclear cell numbers during the first 4 years after infection. BLV infection caused an increase in lymphocyte numbers, and there appeared to be no synergism between the viruses.     

Zinc oxide and amino acids as sources of dietary zinc for calves: effects on uptake and immunity

Calf starter diets were formulated to contain 60 ppm of Zn, 150 or 300 ppm of Zn in the form of Zn-Met and Zn-Lys, or 300 ppm of Zn in the form of ZnO to compare relative bioavailability and effects on immunity. Holstein heifer calves were weaned at wk 5 and fed experimental starter diets from wk 6 to 12. Feed intake, body weight, Zn concentrations in liver and serum fractions, and mineral concentrations in serum were measured to determine the effects of treatment. In addition, peripheral blood lymphocyte blastogenesis, interleukin-2 production, cytotoxic activity, and the ability of blood neutrophils to phagocytose and kill bacteria were assessed at wk 0, 2, 4, and 6 of the trial. Feed intakes and body weight gains were similar among calves. Concentrations of Zn in serum were elevated in calves fed 300 ppm of Zn as Zn-Met and Zn-Lys but not in calves fed ZnO. Concentrations of Zn in liver were significantly elevated by 300 ppm of Zn in the form of Zn-Met and Zn-Lys (360 micrograms/g) but not by the other Zn treatments or by the control (245 micrograms/g). No treatment had an effect on the concentrations of Lys and Met in serum; however, concentrations of Lys did decrease in serum as the age of the calves increased. There was no significant treatment effect on mitogen-induced lymphocyte blastogenesis, interleukin-2 production, lymphocyte cytotoxicity, or phagocytic and intracellular killing ability of blood neutrophils. These data indicated greater absorption and retention of Zn when administered in the form of Zn-Met and Zn-Lys than that when ZnO was administered to young calves. However, there was no advantage to the immune function of extra dietary Zn.     

Effect of pentachlorophenol on immune function

The organochlorine compound, pentachlorophenol, was evaluated for effects on immune system function in male Fisher 344 rats. Pentachlorophenol was prepared in an olive oil vehicle and was administered by oral gavage twice weekly for 28 days at a dose of 2.0 mg/kg per treatment. Exposure to pentachlorophenol increased body weight gains (P=0.024) during the treatment period. Liver (P=0.034) and kidney (P=0.012) body weight ratios were also increased. Pentachlorophenol exposure enhanced T-lymphocyte blastogenesis induced by concanavalin A (Con A)(P=0.0001) and phytohemagglutinin (PHA)(P=0.048) evaluated using stimulation indices. Corresponding B-lymphocyte blastogenesis induced by lipopolysaccharide/dextran (LPS/dex)(P=0.0034) was also enhanced by pentachlorophenol exposure. Pentachlorophenol suppressed the antibody response against sheep red blood cells (SRBCs) by 39% when the response was expressed per viable spleen cell (P=0.006). This suppression was not evident when the response was expressed per spleen (P=0.22), suggesting that a compensatory mechanism or extramedullary splenic hemopoiesis was occurring minimizing the overall impact on humoral immunity. The enhanced B- and T-lymphocyte blastogenesis may also reflect compensatory or hemopoietic activity. Pentachlorophenol exposure had no effect on peritoneal macrophage phagocytosis (P=0.31) or lymphocyte cell surface antigen expression. The observed alterations in lymphocyte blastogenesis and humoral immunity subsequent to pentachlorophenol exposure do not appear to be associated with phagocytosis or lymphocyte cell surface antigen expression.     

Pharmacokinetics of doxycycline after parenteral administration in the Houbara bustard (Chlamydotis undulata)

Human seminal plasma contains a protein with an estimated molecular weight of 16 kd that binds serum immunoglobulin gamma (IgG) and is named IgG binding factor (IgBF). Purified IgBF specifically suppressed pokeweed mitogen-induced lymphocyte blastogenesis, having little or no effect on lymphocyte blastogenesis stimulated with phytohemagglutinin or Concanavalin A; antibody-dependent cell-mediated cytotoxicity; natural killer cell activity; or complement-dependent cytotoxicity of antibodies against sperm. It would appear that IgBF may suppress activation of B cells in the male and female genital tract.     

Quantification of acute phase reactants after muscle biopsy

Blood concentrations of six acute phase reactants (ESR, neutrophil count, fibrinogen, haptoglobin, alpha 1-antitrypsin, and ferritin), parameters of muscle necrosis (myoglobin, CK, ALT, and AST) as well as hemopexin, iron, and TIBC were determined before and for 7 consecutive days after muscle biopsy in patients and in a control group. A muscle biopsy was chosen as a standardized surgical procedure that induces a mild transient inflammatory response. After muscle biopsy, a significant increase occurred in five (ESR, neutrophil count, fibrinogen, haptoglobin, and alpha 1-antitrypsin) of the six acute phase reactants. The concentration of serum ferritin did not show a significant change. A significant decrease was noted in the serum iron concentration and a significant increase occurred with CK and myoglobin secondary to the muscle biopsy. Thus the inflammation of a muscle biopsy produces a significant acute phase reaction.     

Morphological criteria for the therapeutic effectiveness of antirheumatic drugs

Sixteen healthy adults had serial studies of delayed-type skin test reactivity and in vitro lymphocyte blastogenesis to several antigens over a period of 7 months. In many subjects blastogenesis varied broadly from month to month without apparent cause. Responses to all antigens usually increased or decreased together on sequential testing. Blastogenesis to coccidioidin appeared to result largely from cross-reaction with histoplasmin. Humoral factors were not demonstrably responsible for these changes. Blastogenesis rose consistently and non-specifically in subjects following revaccination to vaccinia virus. These studies reflect the lymphocyte blastogenesis reaction as a dynamic equilibrium, subject to spontaneous variation, and responding non-specifically to stimuli such as vaccination. Whatever the causes for these changes, it is clear that serial determinations of blastogenesis response to various antigens do not carry the apparent consistency of the skin test response to that antigen, and single tests must be cautiously interpreted.     

A Cryptosporidium parvum oocyst low molecular mass fraction evokes a CD4+ T-cell-dependent IFN-gamma response in bovine peripheral blood mononuclear cell cultures

T-Cell antigens that induce the in-vitro interferon-gamma response during Cryptosporidium parvum infection of neonatal calves were identified. A total oocyst extract was separated into a high and a low Mr fraction by a microfiltration technique. Both the high and low Mr fractions evoked an in-vitro interferon-gamma response in naturally infected animals, although strong individual differences between the hosts were observed. Using a complement-mediated technique CD4+ T-cells or WC1+gammadelta T-cells were depleted, whereupon the remaining lymphocyte cultures were stimulated with the different antigen preparations. It was shown that the in-vitro interferon-gamma response of Cryptosporidium-infected calves is CD4+ T-cell-dependent.     

Potassium-reduced lung preservation solutions: a screening study

PROBLEM: The direct immunosuppressive and suppressive inducing capacities of supernatants from human trophoblastic choriocarcinoma cell lines (HCS) are well investigated in several former studies. The responsible factor is not yet determined. METHOD OF STUDY: We first confirmed those data and we purified a 3-5-kDa suppressor-inducer factor from HCS by using high performance liquid chromatography (HPLC) on both DEAE and gel filtration columns, followed by ultrafiltration. We then tested the activities of such isolated fractions on in vitro immune responses from human cells and in vivo by its effects in a murine local graft-versus-host (GVH) assay (popliteal lymph node assay, PLN). RESULTS: A single fraction induces both "direct suppression" in vitro as well as in vitro suppressor cell activation/development in human peripheral blood lymphocyte cultures as assessed by suppression of cells cultured in such a fraction containing culture medium of the mixed lymphocyte reaction. Furthermore the very same fraction suppresses in vivo murine allogeneic immune responses as assessed by a local GVH reaction (PLN assay). CONCLUSIONS: We have isolated a suppressive fraction, whose activities suggest that it might be of interest not only in reproductive immunology, but also in transplantation systems.     

The primary structure of rabbit serum amyloid A protein isolated from acute phase serum

Serum amyloid A (SAA) protein, a sensitive acute phase protein and the precursor of protein AA in secondary amyloid, was purified from pooled acute phase rabbit serum using two different methods: isolation of protein SAA directly by octyl-Sepharose chromatography of total serum, and dissociation and isolation of apoSAA from acute phase high density lipoprotein (HDL). The protein SAA fraction obtained was further purified using gel filtration and ion exchange chromatography. Rabbit protein SAA has 104 amino acid residues, like human SAA, and has a partially blocked N terminus. The highly conserved region from position 33 to position 63 found in SAA from all species studied was confirmed also in rabbit SAA. No microheterogeneities were observed. The amino acid sequence showed extensive N-terminal homology with the rabbit amyloid A protein, except for the microheterogeneity in position 12 in protein AA. It also showed identical amino acid sequence with that deduced from the rabbit cDNA clone pSAA 55. Complete homologies were found with clone SAA 2, except for positions 22 and 78, clone SA8-1, except for positions 22 and 79 and clone SA7-3, except for position 22. This pSAA 55/SA7-3/SA8-1/SAA2-like protein was the only SAA isotype found both in total serum and in the HDL fraction. Isotypes corresponding to other SAA-like genes could not be found in this pool of acute phase rabbit sera.     

Helicobacter pylori: antibodies in sera of pigs and calves

64 sera of pigs and 22 sera of calves were investigated in a Helicobacter pylori-ELISA. The test system was validated with positive and negative reference sera. In contrast to other investigations it was tried to exclude false positive results by absorbing the sera with Wolinella succinogenes, Campylobacter jejuni subsp.jejuni, E. coli and Proteus mirabilis. This proved to be necessary, as the extinctions of some sera were reduced considerably. Using this technique sera of 5 pigs and 6 calves could be identified, which fulfilled the following criteria: 1. The extinctions were statistically significant above the mean value of the corresponding group after absorption with Campylobacter jejuni subsp.jejuni. 2. After absorbing the sera of pigs with 3 and the sera of calves with 4 bacteria the extinctions were above 50% of that of the human positive serum. 3. The extinctions of these sera were-like the positive reference sera-nearly not influenced by the absorption. These results indicate the presence of serum antibodies in pigs and calves, which react with epitops from Helicobacter pylori.     

Function and expression of a human idiotypic network in Schistosomiasis japonica

The cross-reactive idiotype (Hu-SJ-CRIM) is defined by polyclonal human anti-idiotypic antibodies derived from chronically S. japonicum infected patients. The present study shows that serum levels of Hu-SJ-CRIM expressed by antibodies to S. japonicum soluble egg antigen (SEA) are associated with acute infection and hepatosplenic disease. Xenogeneic anti-idiotypic antisera (anti-Hu-SJ-CRIM) suppressed human lymphocyte blastogenesis to SEA in vitro by 47-82% (P < 0.05). These anti-idiotypic antibodies also suppressed in vitro granuloma formation induced by SEA coated heads in a dose dependent manner. This immunosuppression was antigen specific in that mitogen (PHA) or non-related antigen (PPD) induced blastogenic responses were not suppressed. Surprisingly, anti-idiotypic antibodies (anti-SJ-CRIM), which describe the mouse correlate CRIM were not suppressive in the human blastogenesis or in vitro granuloma formation assays. These data indicate a dichotomy in the function and specificity of the idiotype/anti-idiotype human and murine immune networks in S. japonicum infection. Thus, only the patient derived molecules and serology form the basis for an immunoregulatory network in Schistosomiasis japonica.     

Collecting duct carcinomas of the kidney: a comparative loss of heterozygosity study with clear cell renal cell carcinoma

Lyophilizing was compared to freezing as a method of colostrum storage. Eight lots of colostrum from the first milking were divided into two equal parts; one was frozen, and the other was lyophilized. Twenty-two newborn calves were divided into two groups and fed either 2 L of frozen and thawed colostrum or 2 L of reconstituted lyophilized colostrum. The calves were bled at 12, 18, 24, and 72 h after feeding, and levels of the immunoglobulins IgG1, IgG2, IgM, and IgA were determined with a radial immunodiffusion assay, in colostrum and sera. The mean concentration of individual immunoglobulin isotypes in the sera of calves fed either frozen or lyophilized colostrum did not differ significantly. Calves fed from the same lots of colostrum had similar immunoglobulin concentrations in their sera, irrespective of the method of storage. All immunoglobulin isotypes were absorbed with equal efficiency from frozen and lyophilized colostrum as determined by calculation of the absorption coefficient.     

Acute paresis of extraocular muscles associated with IgG anti-GQ1b antibody

There have been several case reports of acute ophthalmoparesis without ataxia subsequent to infection or immunization. The nosological position and therapy for acute ophthalmoparesis have yet to be established. Sera from patients are reported to have IgG anti-GQ1b antibody, which is frequently found in sera from patients with Fisher's syndrome. To establish the etiology of acute ophthalmoparesis, I tested sera from 8 patients with acute ophthalmoparesis for anti-GQ1b antibody. High IgG anti-GQ1b antibody titers were present in sera from patients in the acute phase of the illness. I describe the successful treatment with plasmapheresis and intravenous immunoglobulin. Some cases of acute ophthalmoparesis following infection or immunization may be categorized as an auto-immune disease related to Fisher's syndrome.     

Regions of the haptoglobin 5' flanking gene sequence show different binding affinities to nuclear matrix proteins during the acute phase response

The effect of the acute phase response on the affinity of binding between nuclear matrix proteins and the rat haptoglobin (Hp) gene region was examined. Nuclear matrices isolated from acute phase livers were enriched with the 5' Hp gene flanking region (-705/+159), but not with the spliced, protein-coding sequence. Reassociation experiments with isolated nuclear protein matrix spheres and end-labeled fragments I (-146/+156), II (-146/-541), and III (-541/-705) revealed that the matrix proteins displayed an increased binding potential during the acute phase response for all of the examined regions, this being most pronounced for fragment II. BAL 31 digestion of fragment II showed that the sequence element that was responsible for the increased association with nuclear matrix proteins during the acute phase response was a tract of 38 adenine bases. The DNA region established stable associations with nuclear lamin B (67 kDa, pI 5.7) in the controls, and with lamins A (69 kDa, pI 7.0), B, isoforms of lamin C (62 kDa, pI 6.55-6.95), and a 55-kDa (pI 5.9) polypeptide during the acute phase response. Sequence ABC (-165/-56), which overlaps fragments I and II and represents the Hp cis-acting element, did not bind to the non-histone nuclear matrix proteins.     

PHA response and methylazoxymethanol acetate

Several reports have indicated the mutagenicity of the cycasin aglycone, methyl azoxymethanol (MAM). Van Den Berg and Ball (1972) have demonstrated the inhibitory effects of MAM acetate (MAMac), a more stable form of the aglycone, on DNA synthesis and cell proliferation in HeLa cells. The purpose of this study was to observe the effects of MAMac on blastogenesis in the human short-term leucocyte culture system. A depression in blastogenesis by MAMac was observed cytologically derived from two individuals. The same effect was observed utilizing [3H] thymidine as an indicator of blastogenesis in a series of cultures from 11 male individuals exposed to varying doses of MAMac, ranging from 5 through 800 mug/ml.     

Relationship of tuberculosis course in adolescents with acute phase blood proteins and genetic types of haptoglobin

The phenotype of haptoglobin (Hp) was determined and Hp and alpha 1-antitrypsin (alpha 1-AT) were many times measured in the sera of 131 adolescents with first identified tuberculosis. The phenotype of Hp was found to produce no effects on tuberculosis morbidity among adolescents, but Hp2-2 carriers fall ill more frequently if they have a baseline gastrointestinal abnormality. In tuberculosis patients with Hp2-2, resolution of infiltrative changes in the lung tissue occurs less rapidly than in those with other Hp variants. Great increases in the serum levels of alpha 1-AT, which reflects the severity of tissue damages, suggest a less favourable prognosis. By the end of sixth-month continuous combined drug therapy, an inflammatory reaction diminishes and the synthesis of acute phase reactants (APR) becomes normal in patients without baseline lung tissue decay. In the presence of the latter, the intensity of inflammation considerably decreases in the first 3 months of combined drug therapy, then becomes steady-state at a definite level, which is potentiated by continuous invasion of proteolytic enzymes. APR monitoring more accurately assesses the time course of therapy-induced changes in the process.     

Immunologic monitoring in lung allograft recipients

To identify patients with increased risk of chronic lung allograft rejection, we assessed the utility of an in vitro biopsy-derived lymphocyte growth assay and serum anti-HLA antibody screening as a complement to currently available methods of monitoring lung allograft recipients. Lymphocyte growth assay was performed on bronchoscopic fragments of tissue cultured in medium with rIL-2. Seventy-nine biopsies from 31 lung transplant recipients were tested by lymphocyte growth assay, and results were correlated with histopathology findings. Positive lymphocyte growth was found in 12/26 (46%) episodes of acute rejection, 5/44 biopsies without rejection (11%), and 0/9 episodes of bronchitis. Positive lymphocyte growth was seen in 7/16 (44%) grade A1 rejections and in 5/10 (50%) grade A2 rejections, as opposed to only 5/44 (11%) grade A0 (no rejection) biopsies (P < 0.01 for both A1 and A2 with respect to A0). Actuarial probability of remaining free from obliterative bronchiolitis (OB)* tended to be higher in patients who did not exhibit lymphocyte growth in biopsies. Sequential samples of sera obtained at the time of the biopsy were screened for lymphocytotoxic anti-HLA antibodies. Twenty-two of 44 recipients (50%) developed anti-HLA antibodies during the first postoperative year, exhibiting greater than 10% reactivity to an HLA reference panel of lymphocytes in four or more consecutive serum samples. Actuarial survival of lung allograft recipients with anti-HLA antibodies (n = 22) was lower than in those without anti-HLA antibodies (n = 22; P = 0.03). Of the 22 antibody producers, 7/12 died as a consequence of OB. Of the 22 non-antibody-producers, 1/2 deaths occurred as a consequence of OB. Anti-HLA antibodies were present in 9/11 instances of OB (82% sensitivity) and in 13/33 patients without OB (61% specificity; P = 0.03). These data indicate that lung transplant recipients with positive lymphocyte growth and anti-HLA antibodies are at an increased risk of chronic allograft rejection.     

Treatment of carbon monoxide poisoning with hyperbaric oxygen

This report describes differences in humoral immune response of acute and chronic phases of human Chagas disease. The reactivities of IgG, IgM, and IgA anti-Trypanosoma cruzi antibodies in serum samples from both groups of patients were compared by enzyme-linked immunosorbent assay (ELISA) employing either one of four antigenic fractions: mouse laminin (LAM), which reacts through Gal alpha 1-3Gal epitopes expressed on trypomastigote surface: whole intact trypomastigotes (TCT); trypomastigotes excreted/secreted antigens (TESA); and epimastigote alkaline extract (EAE). The selection of T. cruzi antigen preparations was based on their relative content of surface and internal antigens found in trypomastigote forms. The proportion of IgG reactive to carbohydrate epitopes was assessed through the decay of IgG reactivity from acute and chronic sera after m-periodate oxidation of solid-phase bound antigens. Trypomastigote and TESA antigens recognized by IgG from acute and chronic sera were also compared by immunoblotting. ELISA and immunoblotting data showed that: (1) the proportion of IgG directed to trypomastigote surface antigens was higher in acute than in chronic sera, whereas the opposite was found for internal antigens, (2) acute sera contained a higher percentage of IgG reactive to trypomastigote carbohydrate epitopes than chronic sera, and (3) anti-T. cruzi IgA was found exclusively in acute sera and led to 100% positivity when LAM, TCT, and TESA were employed as antigens. IgA ELISA with these antigens and IgG immunoblotting pattern with TESA could be useful as serological markers for the acute phase of human Chagas disease.     

The effect of vitamin E on cortisol and lactate levels and on the acid-base equilibrium in calves exposed to transportation stress

The objective of this study was to examine the effect of vitamin E administration on plasma levels of cortisol and lactate, and on acid base balance in transported calves. In the study, eight calves, aged approx. ten days, divided into two groups, were used. 20 mg of tocopherol-acetate per kg body weight were administered orally to each of the four experimental calves 24 hours before loading. The calves were transported by road for 3 hours. Blood samples collected before and after the transportation were examined for acid base balance, lactate, and plasma vitamin E and cortisol levels. The administration of vitamin E led to a decrease of cortisol levels in 24 hours (from 7.6 +/- 9.5 to 4.2 +/- 0.2 nmol/l) as well as to a significant increase (p < 0.05) of plasma vitamin E levels 26 h after administration (from 2.52 +/- 1.36 to 12.28 +/- 6.14 mumol/l). There was no difference between the groups in cortisol response due to transportation stress (Tab. III). The transportation caused typical stress changes in lactate levels and acid base balance (lactacidaemia and the tendency to acidosis, Tab. III, IV). There was approx. threefold increase in plasma lactate concentrations due to transportation (from 2.49 +/- 0.69 to 6.35 +/- 3.75 mmol/l). The results of the present study demonstrated metabolic changes which has been reported to be typical of mild physiological stress reaction. In the present study, vitamin E had no significant effect on plasma levels of cortisol, and lactate, and acid-base balance.     

Changes in serum levels of sialoglycoproteins in mice exposed to UV-B radiation

In the acute phase reaction, hepatocytes in the liver are activated and increase the plasma levels of acute phase reactants. Our previous study has shown that plasma sialic acid, an acute phase reactant, was increased following exposure of mice to UV-B radiation. Plasma sialic acid is derived from many plasma components. To clarify the type of plasma sialic acid that is increased by exposure to UV-B radiation, we performed two-dimensional electrophoresis and staining for sialic acid. Consequently, the increases in haptoglobin and hemopexin were marked and 90% or more of the increased sialic acid was derived from these two glycoproteins after exposure to UV radiation. The increase in alpha1-acid glycoprotein levels was slight and did not contribute to the total increase in plasma sialic acid after exposure to UV radiation. Plasma levels of several proteins including antichymotrypsin (ACT), were reduced following exposure to UV radiation. The discrepancy between our results and published ones regarding ACT levels are discussed in terms of the type of cytokine.