PubMed HealthSearch

Biomedical subjects

P G Reddy

Publications and source records attributed to P G Reddy.

At least 19 recordsLinked to original sources

Interleukin 2 treatment of Staphylococcus aureus mastitis.

A study was conducted in dairy cows to evaluate the efficacy of recombinant bovine interleukin 2 (rBoIL-2) as an adjunct to antibiotic therapy in Staphylococcus aureus mastitis. In normal, non-mastitic cows, intramammary infusion of rBoIL-2 caused a tenfold increase in somatic cell counts (SCC) in milk. Co-administration of 2 mg of rBoIL-2 and sodium cephapirin in cows with established S. aureus mastitis decreased SCC and shedding of S. aureus compared with values from cows that were given only sodium cephapirin or 10 mg rBoIL-2 with sodium cephapirin. Cows in the 2 mg rBoIL-2 group cleared the infection earlier and at 2 weeks after treatment had not relapsed with staphylococcal mastitis. These data suggest that rBoIL-2 may be useful as an immunotherapeutic agent in controlling mastitis.

Animals

Repeated restraint and isolation stress alters adrenal and lymphocyte functions and some leukocyte differentiation antigens in lambs.

Lambs were used to evaluate the effect of repeated restraint and isolation stress (RIS) on secretion of cortisol, lymphocyte proliferative responses to mitogens, production of interleukin-2, and expression of leukocyte differentiation antigens. Differentiation antigens evaluated included cluster of differentiation antigens 2, 4, and 8 (CD2, CD4, and CD8, respectively); B cells; and major histocompatibility complex (MHC) class II, DQ, and DR. Lambs were assigned to either control (CON; n = 12) or to RIS treatment (n = 12) then were stanchioned in environmentally controlled rooms at 18 degrees C and constant light for 11 d before jugular vein catheters were inserted on d 0 of the experiment. On d 12, 13, and 14, lambs in the RIS treatment were removed to another location, restrained, and isolated from visual and tactile contact with other lambs for 6 h on each day. Following the 6-h stress treatment, lambs were returned to their home stanchions. The CON lambs remained in their stanchions. Samples of serum were obtained from all lambs, beginning before RIS (0 h) and at .5-h intervals until the completion of stress (6 h) on d 12 and 14. In addition, samples of whole blood were obtained at 0 and 6 h on d 12, 13, and 14 for evaluation of immune function characteristics. Fitted polynomial curves describing the cortisol response in RIS and CON lambs differed (P less than .005) on both d 12 and 14, reflecting the unmistakable increase in cortisol in response to the stressor.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Response of young dairy calves with lasalocid delivery varied in feed sources.

Growth, age at weaning, fecal scores, and blood metabolites of young dairy calves were measured to determine the most effective method of lasalocid administration. Forty Holstein bull calves were blocked by date of birth and assigned randomly to one of four treatment groups: no lasalocid; lasalocid in starter; lasalocid in prestarter and starter; or lasalocid in milk, prestarter, and starter for a 12-wk period. Calves were fed milk twice daily until they consumed 227 g/d of prestarter, at which time the p.m. milk feeding was discontinued, and starter was offered for ad libitum intake as a mixture with the 227 g/d of prestarter. When total dry feed consumption reached 1.3% of birth weight, the calf was weaned. When the calf was 5 wk of age, the prestarter was discontinued. Daily gain tended to be greatest during the first 6 wk for the calves receiving lasalocid in milk, prestarter, and starter. These calves also were weaned with less variation in days to weaning. By wk 8 through 12, there were no differences in gain among the treatment groups.

Animal Feed

Neutrophil and lymphocyte response to supplementation with vitamins C and E in young calves.

Calves, beginning at 3 d of age, were bottle-fed milk replacer unsupplemented, or supplemented with 10 g of vitamin C, or supplemented with 10 g of vitamin C plus 57 IU of vitamin E/kg of dry milk replacer (10 calves per treatment). Neutrophils from wk 2 and 4 blood samples were assayed for Staphylococcus aureus phagocytosis and neutrophil-mediated, antibody-dependent cellular cytotoxicity, and lymphocytes from those samples were assayed for concanavalin A-induced, phytohemagglutinin-induced, or pokeweed mitogen-induced proliferation. Ocular and nasal discharges of calves supplemented with vitamins C or C and E were less than those of controls for wk 1 to 8. Calves receiving supplementation with vitamin C and E tended to have higher mitogen-induced lymphocyte proliferative responses at wk 2. Neutrophil-mediated phagocytosis and antibody-dependent cellular cytotoxicity were lower at wk 2 and 4 for calves supplemented with vitamin C than for controls. Neutrophil function of calves supplemented with vitamins C and E together was near, or slightly higher than, that of unsupplemented calves for wk 2 and 4, suggesting that the addition of vitamin E negated the adverse effects that vitamin C alone had on neutrophil functions.

Animals

Effect of recombinant fibroblast interferon and recombinant immune interferon on growth and the antigenic phenotype of multidrug-resistant human glioblastoma multiforme cells.

To study the effect of drug resistance on the response of stage IV astrocytomas to interferon, a human glioblastoma multiforme cell line, GBM-18, was transfected with an expression-vector plasmid containing a human multidrug resistance (MDR) gene (pHaMDR1/A), and clones surviving in colchicine were isolated. GBM-18 multidrug-resistant subclones displayed cross-resistance to other chemotherapeutic agents, including vincristine, doxorubicin, and dactinomycin. The multidrug-resistant phenotype was reversible when GBM-18 multidrug-resistant cells were cultured in colchicine and the calcium-channel blocker verapamil. The level of the MDR1 gene (also known as PGY1) message was increased in GBM-18 multidrug-resistant cells selected for increased resistance to colchicine, and this effect was not correlated with an amplification of the MDR1 gene. In both parental GBM-18 and GBM-18 multidrug-resistant cells, growth was suppressed to a greater degree when cultures were treated with the combination of fibroblast interferon (IFN-beta) and immune interferon (IFN-gamma). Parental cells and multidrug-resistant subclones varied in their de novo and/or interferon-modulated expression of HLA class I and class II antigens, a high-molecular-weight melanoma-associated antigen, and intercellular adhesion molecule 1 (ICAM-1). Of the antigens tested, ICAM-1 and HLA class I antigens were the most sensitive to enhanced expression induced by IFN-beta and IFN-gamma when used alone or in combination. The results of the present study indicate that multidrug-resistant human glioblastoma multiforme cells retain their increased sensitivity to the antiproliferative activity of the combination of IFN-beta plus IFN-gamma, and differences in antigenic phenotype are apparent in independent multidrug-resistant glioblastoma multiforme clones.

Antibodies, Monoclonal

Rat urate oxidase: cloning and structural analysis of the gene and 5'-flanking region.

The structural gene (UOX) encoding rat urate oxidase (UOX) spans at least 23 kb and is composed of eight exons and seven introns. All of the exon-intron splice junction sequences conformed to the GT/AG consensus established for eukaryotic genes. The transcription start point (tsp) was determined using S1-type nuclease protection riboprobe, and assigned to an adenine 54 nucleotides (nt) upstream of the ATG start codon. A 456-bp 5'-terminal fragment, starting at the ATG codon, carries a putative TATA (ATAAAA) sequence at -32, and two putative 'CAAT box' sequences at -62 and -71 bp upstream from the tsp. No sequence resembling 'GC' box hexanucleotides (GGGCGG or CCGCCC) was found. The structural features of the 5'-flanking region of the UOX gene are distinct from the 5'-flanking sequences of peroxisomal beta-oxidation system genes which contain one or more 'GC' box elements but lack TATA- and CAAT-like features [Osumi et al., J. Biol. Chem. 262 (1987) 8138-8143; Ishii et al., J. Biol. Chem. 262 (1987) 8144-8150]. The 5'-flanking region of the UOX gene reveals a sequence, TTAGTAATT at nt -276 from the tsp, which appears to be complementary to the underlined part of the liver-specific LF-B1/HNF-1 consensus sequence, GTTAATNATTAAC (where N = A, C, T, G or no nt).

Amino Acid Sequence

Induction of immune response to bovine herpesvirus-1 with anti-idiotypic antibodies.

Previously, we prepared rabbit anti-idiotypic (anti-Id) antibodies against murine monoclonal antibodies (MAbs) specific for the major bovine herpesvirus-1 (BHV-1) envelope glycoproteins. Glycoprotein III (gIII) contains neutralization epitopes and may be the virus attachment protein. Anti-Id antibodies to a neutralizing MAb that reacts with gIII were purified by sequential immunoaffinity chromatography. Immune responses to the purified anti-Id reagent and BHV-1 were compared in mice. Both groups of mice produced BHV-1-specific neutralizing antibodies. However, lymphocyte proliferative responses and interferon and interleukin-2 production were specific for the respective immunizing antigens. These results suggest that the anti-Id reagent may bear an internal image of a B-cell-stimulating epitope of glycoprotein gIII; however, this epitope does not stimulate a virus-specific cellular immune response in mice.

Animals

Interrelationship between hypersensitivity to soybean proteins and growth performance in early-weaned pigs.

The objective of this growth trial was to determine the interrelationship between immunological criteria, gut morphology, and performance of starter pigs fed soybean proteins processed by different methods. One hundred twenty-five pigs were orally infused with 6 g/d of either dried skim milk, soybean meal (48% CP), soy protein concentrate, extruded soy protein concentrate, or experimental soy protein concentrate from 7 to 11 d of age and then fed a diet containing the same protein sources from weaning (d 21) to 35 d of age. All pigs were fed a corn-soybean meal diet containing 10% dried whey, 1.25% lysine, and 3% soybean oil for the remaining 21 d of the experiment. Xylose absorption and anti-soy immunoglobulin G (IgG) titers were measured on d 6 postweaning, and skin-fold thickness after intradermal injection of protein extracts was measured on d 7 postweaning. A total of 25 pigs (five pigs/treatment) was euthanatized on d 7 postweaning. Villus height and crypt depth from duodenum samples were measured. These measurements were obtained to elucidate a relationship between the hypersensitivity responses to soybean products and growth performance of baby pigs. Pigs fed diets containing soybean meal had a lower (P less than .05) rate of gain (d 0 to 14) and villus height, higher (P less than .01) serum anti-soy IgG titers, and increased skin-fold thickness (d 6 and 7 postweaning) after intradermal injection compared with those fed dried skim milk. Pigs fed other soy proteins also had lower ADG from d 0 to 14 postweaning; however, pigs fed moist extruded soy protein concentrate tended (P less than .09) to have higher ADG and improved feed utilization when compared with pigs fed soybean meal (d 0 to 14).(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed

Removal of nocturnal secretion of melatonin fails to reduce antibody synthesis and interleukin-2 production of lambs.

Crossbred ewe and wether lambs were used to evaluate the effects of a normal, nocturnal elevation in the concentration of melatonin in the serum on immunological functions. The nocturnal elevation in melatonin was eliminated by exposing half the lambs to constant light (LL), whereas the remainder received a 12-h light, 12-h dark cycle (LD). Immune function was challenged by treating half the lambs in LL and half of the lambs in LD with dexamethasone (DEX; .04 mg/kg); the remainder of the lambs received only a saline vehicle (SAL). The resulting treatment combinations were designated LD+SAL (n = 5), LD+DEX (n = 5), LL+SAL (n = 5), and LL+DEX (n = 5). Lambs were stanchioned individually in environmental rooms; photoperiod treatments commenced on that day (d -14). Also on d -14, lambs were given 1 mg ovalbumin/lamb in adjuvant. Lambs were given a booster injection of .5 mg ovalbumin/lamb on d 0. Treatments with DEX and SAL also began on d 0 and were repeated every 48 h through d 14. Catheters were placed in the jugular vein of all lambs on d 12; samples of plasma and serum were collected hourly from 0800 on d 14 to 0800 on d 15; plasma was assayed for adrenocorticotropic hormone (ACTH) and serum was assayed for cortisol and melatonin. In addition, samples of serum obtained at 0800 on d 15 were used to evaluate antibody titers to ovalbumin. Samples of whole blood also were obtained at 0800 on d 15, and total and differential leukocyte numbers and production of interleukin-2 (IL-2) by lymphocytes were determined.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenocorticotropic Hormone

Repeated restraint and isolation stress in lambs increases pituitary-adrenal secretions and reduces cell-mediated immunity.

Rambouillet crossbred ewe lambs were used to evaluate the effect of repeated restraint and isolation stress (RIS) on endocrinological and immunological functions. Lambs were blocked by weight and assigned to either RIS (n = 6) or to control (CON; n = 6) treatments. All lambs were tethered in environmentally controlled rooms at 22 degrees C with constant light and, at this time (d 0), were given 1 mg of ovalbumin in adjuvant. On d 12, catheters were placed nonsurgically into the jugular vein of all lambs, and they were reimmunized with .5 mg of ovalbumin in incomplete adjuvant. Each lamb in the RIS treatment group was removed from its home stanchion, isolated from visual and tactile (but not auditory and olfactory) contact with other lambs, and restrained for 6 h on d 14, 15, and 16. Lambs in CON treatment remained undisturbed in their home stanchions. Lymphocyte blastogenic function and production of interleukin-2 (IL-2) were assessed in samples of blood collected before and at the conclusion (0 and 6 h) of each of the three stress bouts. In addition, ACTH and cortisol secretion in response to RIS was evaluated in samples of plasma and serum collected at .5-h intervals on the first and last days of stress (d 14 and 16). Finally, antibody production in response to immunization against ovalbumin was assessed in samples of serum obtained 14 d after reimmunization on d 12 of the study. Polynomial curves fit to the ACTH and cortisol data differed (P less than .005) on both d 14 and 16 between RIS and CON treatments.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenocorticotropic Hormone

Measuring suitability of soybean products for early-weaned pigs with immunological criteria.

Two trials were conducted to determine the suitability of soybean products for baby pigs. Weanling pigs (n = 40 and 48 in Trials 1 and 2, respectively) were infused orally (6 g/d) with dried skim milk, soybean meal (SBM, 48% CP); soy protein concentrate, moist extruded soy protein concentrate, or soy protein isolate from d 7 to 12 of age. Pigs were then fed a diet containing the same protein source for 1 (Trial 1) or 2 (Trial 2) wk after weaning (d 21 of age). To avoid exposure of pigs to soybean proteins, the dams of pigs were fed a corn-corn gluten meal-based diet supplemented with lysine and tryptophan from d 109 of gestation. All pigs in Trial 1 were killed at 28 d of age, and samples of ileal digesta and small intestine were obtained. In Trial 2, the soy protein isolate was not included, and all pigs were fed a diet containing 4% soybean oil and 1.25% lysine for the last 3 wk of the trial. Growth performance, skin-fold thickness, after intradermal injection of extracts of the corresponding proteins, and anti-soy immunoglobulin G (IgG) titers were measured. Results indicated that pigs fed diets containing SBM had lower (P less than .05) villus height and xylose absorption but higher (P less than .05) serum anti-soy IgG titers and increased skin-fold thickness compared with the mean of pigs given milk and all other soy treatments.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed

Transient hypersensitivity to soybean meal in the early-weaned pig.

An experiment was conducted to determine whether baby pigs develop hypersensitivity to dietary soybean proteins. Thirty-two pigs were orally infused with either dried skim milk (5 g/d; control) or soybean meal (48% CP; 5 g/d) from d 7 to 14 after birth. Sows were fed a corn-corn gluten meal-based diet supplemented with lysine and tryptophan to avoid exposure of pigs to soybean proteins. Pigs were weaned at 21 d of age and fed diets containing either soybean meal or milk proteins until d 56. One half of the pigs were killed at 28 d of age and the rest at 56 d of age. Segments of small intestine were collected, and intraepithelial lymphocytes were isolated. At 28 d of age, pigs fed diets containing soybean meal had lower (P less than .05) villus height (221 vs 298 microns) and rate of gain (86 vs 204 g/d) than control pigs did. Pigs fed a diet containing soybean meal had higher (P less than .05) immunoglobulin G (IgG) titers to soybean protein than did pigs fed a milk protein-based diet. Blood and intestinal lymphocytes collected on d 28 and 56 did not exhibit any proliferative response when cultured with purified soy proteins (2.5 or 5 microns/ml). Phytohemagglutinin- and pokeweed mitogen-induced lymphocyte proliferations were higher (P less than .05) at d 56 than at d 28, but there were no differences attributable to protein source. There were no differences (P greater than .05) in skin-fold thickness measurements following intradermal injection with soy or milk proteins. Decreased villus height and increased serum IgG titers to soybean proteins coinciding with inferior performance of early weaned pigs fed diets containing soybean meal indicate that conventionally processed, commercial soybean meal may retain some antigens that can cause transient hypersensitivity in piglets.

Animals

Adjuvanticity of recombinant bovine interleukin-1 beta: influence on immunity, infection, and latency in a bovine herpesvirus-1 infection.

Recombinant bovine interleukin-1 beta (rBoIL-1 beta) was administered to calves in conjunction with a bovine herpesvirus-1 (BHV-1) vaccine. All calves were immunized against BHV-1 and three groups received rBoIL-1 beta at 33, 100, or 330 ng/kg on days 1 and 15; control animals received physiological saline. All calves were challenged with BHV-1 on day 22. Total leukocytes were increased by rBoIL-1 beta, primarily by causing neutrophilia and monocytosis; CD4/CD8 ratios tended to be increased in rBoIL-1 beta-treated animals. Serum neutralizing antibody titers and cytotoxic responses against BHV-1-infected bovine kidney fibroblasts were increased and virus excretion was decreased in rBoIL-1 beta-treated calves. On days 58 and 59, control and 100 ng/kg rBoIL-1 beta-treated calves were injected with dexamethasone (.04 mg/kg). Virus excretion was less and clinical signs of BHV-1 infection were lower in rBoIL-1 beta-treated calves after dexamethasone injection. These data suggest that rBoIL-1 beta may be an effective adjuvant to BHV-1 immunization.

Adjuvants, Immunologic

Bovine recombinant granulocyte-macrophage colony-stimulating factor enhancement of bovine neutrophil functions in vitro.

Neutrophils were purified from blood of dexamethasone-treated (0.04 mg/kg of body weight) and untreated calves. Cells were untreated (controls) or cultured in media containing 5 or 10 ng of bovine recombinant granulocyte-macrophage colony-stimulating factor (rbGM-CSF)/ml for 10 to 12 hours before being tested for various functions. Dexamethasone treatment of calves decreased luminol-dependent chemiluminescence, decreased phagocytosis of Pasteurella multocida and several Staphylococcus spp by various degrees, and decreased antibody-dependent cell-mediated cytotoxicity against bovine herpesvirus-infected cells by 26 to 32%. The percentage phagocytosis of coagulase-positive S aureus and S intermedius was higher than that of coagulase-negative S epidermidis for neutrophils from all calves. Culture of neutrophils with rbGM-CSF significantly increased (P less than 0.05) all of the aforementioned functions, compared with control neutrophils; however, rbGM-CSF-induced increases in function tended to be higher in neutrophils from dexamethasone-treated calves than in neutrophils from untreated calves.

Animals

Bovine recombinant interleukin-2 augments immunity and resistance to bovine herpesvirus infection.

The in vivo administration of bovine recombinant interleukin-2 (rIL-2) was evaluated in calves vaccinated and then challenged with bovine herpesvirus-1 (BHV-1). In Experiment 1, 24 calves were allotted to four groups: control; bovine rIL-2; BHV-1 vaccine (modified-live); and bovine rIL-2 + BHV-1 vaccine. Serum neutralizing antibody titers to BHV-1 were increased sixfold, and virus shedding was fourfold less in calves vaccinated and treated with rIL-2 (25 micrograms/kg, intramuscularly) when compared to calves that received vaccine only. Treatment with rIL-2 induced lymphokine-activated killer activity that was eliminated by pretreating effector cells with complement and a monoclonal antibody (B26A) specific for the sheep red blood cell receptor. The rIL-2 treatment in BHV-1-vaccinated calves increased the calves' ability to withstand a BHV-1 challenge. However, during treatment with rIL-2, calves developed diarrhea and mild fever that abated after IL-2 treatment was stopped. A second experiment was then conducted to determine a dose of rIL-2 that would enhance immunity to BHV-1 without causing adverse side effects. Twenty-five calves were allotted to five groups that received injections of rIL-2 at 0.0, 25.0, 2.5, 0.25, or 0.025 micrograms kg-1 day-1 for 5 days. All calves received a modified-live BHV-1 vaccine. Calves treated with 25.0 micrograms kg-1 day-1 showed similar adverse side effects as in the first experiment but all other calves were normal. Compared to control calves, those treated with 25.0, 2.5, and 0.25 micrograms kg-1 day-1 of rIL-2 had higher (P less than 0.05) serum antibody titers to BHV-1 and following challenge lower (P less than 0.05) BHV-1 titers in nasal secretions; additionally, clinical disease as evidenced by nasal and ocular discharge was less severe (P less than 0.05). In vitro cytotoxic responses against BHV-1-infected bovine kidney cells were increased (P less than 0.05) in calves treated with rIL-2 in a dose dependent manner. These data suggest that bovine rIL-2 at 2.5 to 0.25 micrograms/kg may be an effective adjuvant to immunization.

Adjuvants, Immunologic

Cloning and expression in Escherichia coli of an esterase-coding gene from the oil-degrading bacterium Acinetobacter calcoaceticus RAG-1.

A putative esterase gene (est) from Acinetobacter calcoaceticus RAG-1 has been cloned into Escherichia coli. Esterase-positive clones exhibited high levels of esterase activity even in intact cells. In addition, expression of the est gene conferred on E. coli the ability to grow on simple triglycerides such as triacetin (TAC). The original esterase-positive plasmid pRA17 carried a 2.2-kb insert from a partial MboI digest of RAG-1 DNA, which gave a single band with RAG-1 DNA following Southern hybridization. By subcloning and sequencing the est gene was found to contain a sequence of 870 bp which could be translated to yield a protein of Mr 32,700. In support of the sequencing results was the finding that when pRA17 was expressed in minicells, a unique peptide of Mr 32,500 was identified. This peptide was not found in minicells transformed with esterase-negative plasmids, such as pRA176, which contained a Tn5 insertion in the est gene. The fact that the production of active esterase depended on the orientation of the est gene within the vector suggested that transcription proceeded from the tet promoter in pBR322.

Acinetobacter

The nucleotide sequence of a full length cDNA clone encoding rat liver urate oxidase.

Recently we reported the sequence of a cDNA clone (pUOX-1), isolated from a lambda gt11 cDNA library, which encoded for rat liver urate oxidase (EC 1.7.3.3), but this clone lacked the nucleotide sequences encoding the N-terminal region for this enzyme. Using the cDNA insert from the pUOX-1 clone as a probe, we have now isolated a full length cDNA clone, pUOX-2, from a lambda gt10 library by plaque hybridization. Nucleotide sequence analysis of the pUOX-2 clone showed that it has 1379 base pairs with an open reading frame coding for 303 amino acid residues corresponding to a molecular mass of 34,931 daltons. In addition to the open reading frame the pUOX-2 contains 439 bp of 3'-untranslated and 41 bp of 5'-untranslated sequences. The consensus polyadenylation signal AATAAA precedes a stretch of poly(A)+ residues at the 3' end.

Amino Acid Sequence