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PubMed1985

Immunologic response of sheep to inactivated and virulent bluetongue virus.

Humoral and cellular immune responses of sheep to inactivated and virulent bluetongue virus (BTV) were studied. All sheep inoculated with inactivated BTV developed BTV group-specific nonneutralizing antibodies, as determined by agar-gel immunodiffusion. The development of group-specific, nonneutralizing, complement-fixing antibodies was variable and appeared to be dependent on immunizing BTV serotype, sheep breed, and individual variation. Virus-neutralizing antibodies were never detected after inoculation with the inactivated BTV. In vitro lymphocyte stimulation to BTV soluble antigen was observed with cells from all inoculated Warhill sheep and with cells from 1 of 3 inoculated Suffolk cross sheep. Complement-fixation titers did not appear to correlate with the degree of protection observed, ie, duration of postchallenge-exposure viremia. The development of postchallenge-exposure neutralizing antibody titer was inversely correlated to protective immunity. The development of a response to BTV antigen in the lymphocyte-stimulation test associated most closely with protection. Warhill sheep were afforded better protection, by inoculation with inactivated BTV, to live virus challenge exposure than were the Suffolk cross sheep. Approximately 30% of the inoculated Suffolk cross sheep responded to challenge exposure with intensified clinical signs of blue-tongue, compared with the challenge-exposed control sheep of the same breed.

Animals↗
PubMed1965

Preliminary bluetongue Transmission with the sheep ked Melophagus ovinus (L.).

Five experiments indicated that the sheep ked MELOPHAGUS OVINUS (L.), can transmit bluetongue virus (BTV) in sheep. It was not determined whether these were mechanical or biological transmissions, although the results suggested mechanical transmission. Sheep keds were manually transferred from a BTV-host sheep to 18 susceptible test sheep. Of these, 10 were positive (5 with mild reactions), 6 questionable, and 2 negative for BTV. Three of the mildly reacting sheep and 3 of the questionable sheep had highly intensified reactions on challenge inoculation. Five of the positive sheep were immune on challenge inoculation. Blood from 2 positive reactors was subpassaged into susceptible sheep, which reacted with typical disease signs.

Animals↗
PubMed1981

Anaplasma ovis in Idaho sheep.

Blood samples from 16 anaplasmosis complement-fixation (CF) positive sheep from a flock near Cambridge, Idaho, were inoculated into 3 intact sheep and 3 splenectomized calves. The 3 sheep became CF positive, and 2 developed parasitemia. None of the 3 calves had signs of anaplasmosis, but all became ill when they were subsequently challenge exposed with blood from known carriers of Anaplasma marginale. After a 75-day observation period, blood from the recipient intact sheep was subinoculated into 3 splenectomized sheep. These splenectomized sheep developed clinical anaplasmosis and became CF positive. In only 1 sheep was there a positive rapid card agglutination test. Therefore, the rapid card agglutination test was not considered to be diagnostic for anaplasmosis carrier status in sheep. Electron microscopic studies were performed on infected sheep erythrocytes to determine the morphologic structure of the organism involved. The causative organism was determined to be Anaplasma ovis.

Anaplasmosis↗
PubMed1994

An assessment of the toxicity of parenteral treatment with copper EDTA and copper heptonate in sheep.

The toxicity of 2 parenteral copper (Cu) supplements was investigated. Di-sodium copper ethylene diamino tetra acetate (Cu EDTA) and Cu heptonate were administered to sheep (n = 9) by a single subcutaneous injection at a concentration of 0,2, 1 and 2 mg Cu/kg each (Trial 1.) Nine sheep were untreated and served as controls. The same treatments were applied to 2 sheep each (Trial 2) with the addition of 3 mg Cu/kg live body mass as Cu heptonate, and Cu heptonate administered intravenously at rates of 0,2, 0,4 and 0,6 mg Cu/kg live body mass. In Trial 1, 67% of the sheep treated with Cu EDTA at 2 mg Cu/kg live body mass died within 3 to 17 d after treatment, while no mortalities occurred in sheep where Cu heptonate was administered at the same dosage rate and even at 3 mg Cu/kg live body mass (P < or = 0,01). Post-mortem examination suggested acute Cu toxicity in all cases. Liver Cu concentrations were markedly increased (P < or = 0,05) by both supplements in groups of 3 treated sheep slaughtered over a 3-month period compared to control animals. The liver Cu concentrations of sheep that succumbed to Cu toxicity were within the normal range of 100 to 450 mg/kg DM. Results from Trial 2 suggested that the 2 sheep treated with 2 mg Cu/kg live body mass as Cu EDTA, experienced a haemolytic crisis between 5 and 11 d after treatment, resulting in the death of one of these sheep. The haemolytic crisis was characterised by a severe decrease in haemoglobin concentration and haematocrit.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Sheep as maintenance host for Leptospira interrogans serovar hardjo subtype hardjobovis.

Transmission of Leptospira interrogans serovar hardjo subtype hardjobovis from naturally infected sheep to uninfected sheep and calves was studied. A microscopic agglutination test and ELISA were used to determine specific antibody responses in serum. Polymerase chain reaction was used to detect bacterial shedding in urine. Six sheep were derived from a dairy farm where cows were infected with L hardjobovis. Three of these sheep were seropositive for L hardjobovis, and 1 also shed leptospires in the urine. The other 2 sheep shed leptospires in the urine 7 days after the first observation date. The 6 sheep were placed on an isolated pasture together with a second group of 6 noninfected sheep. During the observation period of 140 days, 1 sheep of the second group became infected with L hardjobovis. On 5 consecutive days, a urine mixture from the 4 infected sheep was sprayed on the heads of 4 noninfected calves. Within 56 days, all calves that had been sprayed with urine shed L hardjobovis in the urine and became seropositive for L hardjobovis.

Animals↗
PubMed1996

Immunomodulative effects of bovine immunodeficiency-like virus (BIV)-infection and mixed infection of BIV and bovine leukemia virus on sheep.

Experimental bovine immunodeficiency-like virus (BIV)-infection and mixed infection of BIV and bovine leukemia virus (BLV) were performed on sheep. BIV proviral DNA and anti-BIV antibodies were persistently detected in all BIV-inoculated sheep. A slight increase in lymphocyte counts was observed in BIV-infected sheep, but the percentages of CD4+ and CD8+ cells in sheep peripheral blood mononuclear cells (PBMCs) were not significantly changed. A transient decrease in lymphocyte blastogenic response to concanavalin. A was observed in two of three BIV-infected sheep at 3-6 months after inoculation. From 6 months after BLV-inoculation to sheep which were previously infected with BIV, the numbers of lymphocytes expressing a tumor-associated antigen (TAA) of bovine leukosis were increased compared to those of a sheep inoculated with BLV alone. The BLV titers in PBMCs and the antibody titers against BLV from sheep infected with both BIV and BLV were higher than those of a sheep inoculated with BLV alone.

Adjuvants, Immunologic↗
PubMed1990

Molecular cloning of sheep T cell receptor gamma and delta chain constant regions: unusual primary structure of gamma chain hinge segments.

The primary structure of sheep T cell receptor (TcR) gamma and delta chain constant (C) regions has been determined by cDNA cloning. A comparison of the nucleotide and deduced amino acid sequences of the sheep chains with known human and mouse sequences shows that the primary structure of the immunoglobulin, transmembrane and cytoplasmic C gamma domains and all of the C delta region has been substantially conserved. However, the hinge or connector region of sheep gamma chains differs significantly from all known TcR chains. Clones representing two different sheep C gamma genes were isolated and both contain additional sequence in this region, making them the longest TcR chains so far identified. The hinge region of both sheep C gamma sequences contains two additional cysteine residues and a motif of five amino acids (TTESP or TTEPP) which has been triplicated in one of the clones. Other repetitive segments of 13-17 amino acids could also be identified suggesting that, as in the human C gamma 2 gene, this region of the sheep genes could have arisen from an exon duplication or triplication event. Southern blot analysis of sheep DNA confirmed the presence of one C delta gene and at least two C gamma genes. A restriction fragment length polymorphism that is probably associated with allelic sequence variation in the sheep C delta gene was detected in DNA from different animals. Although the essential structure of the gamma/delta TcR appears well conserved through evolution, the marked heterogeneity evident in the hinge region of gamma chains both within and between species, and particularly the presence of additional cysteine residues in the sheep sequences, may be of structural and functional importance.

Amino Acid Sequence↗
PubMed1998

The roles of CYP3A and CYP2B isoforms in hepatic bioactivation and detoxification of the pyrrolizidine alkaloid senecionine in sheep and hamsters.

The roles of cytochrome CYP3A and CYP2B isozymes in the bioactivation and detoxification of the pyrrolizidine alkaloid (PA) senecionine (SN) have been investigated in vitro with sheep and hamster hepatic microsomes. Our results show that the rate of SN activation measured by (+/-)-6, 7-dihydro-7-hydroxy-1-hydroxymethyl-5H-pyrrolizine (DHP) formation greatly exceeded the rate of SN N-oxide formation (detoxification) in hamsters. In contrast, SN N-oxide, a detoxification product, was the major metabolite in sheep with much lower DHP production. Immunoinhibition studies with anti-sheep CYP3A and CYP2B antibodies show that members of CYP3A subfamily play the major role in the conversion of PA to pyrrolic metabolites in both species (over 90% in sheep; 68% in hamster). These enzymes also contribute 38.8 and 41. 3% of SN N-oxidation in sheep and hamsters, respectively. In contrast, CYP2B isoforms have a limited capacity toward DHP formation in both species (47% in sheep; 32% in hamster), while these enzymes catalyzed only 24.6 and 35.4% SN N-oxidation in sheep and hamster, respectively. Using triacetyloleandomycin (TAO) and gestodene, two highly selective chemical inhibitors of CYP3A isoforms, our data show that 90% of DHP formation was inhibited by either inhibitor in sheep. Gestodene appeared to be more efficient than TAO in the inhibition of DHP production in hamsters. Testosterone 6beta-hydroxylase activity, a functional marker of CYP3A, was significantly inhibited by TAO and gestodene in sheep liver microsomes and by gestodene (100 microM) in hamster liver microsomes. These results suggest that CYP3A isozymes have important roles in bioactivation and detoxification of PA in both species, whereas CYP2B subfamily members are less efficient in biotransformation of PA.

Animals↗
PubMed2000

Arsenic metabolism in seaweed-eating sheep from Northern Scotland.

Cation exchange and anion exchange liquid chromatography were coupled to an ICP-MS and optimised for the separation of 13 different arsenic species in body fluids (arsenite, arsenate, dimethylarsinic acid (DMAA), monomethylarsonic acid (MMAA), trimethylarsine oxide (TMAO), tetramethylarsonium ion (TMA), arsenobetaine (AsB), arsenocholine (AsC), dimethylarsinoyl ethanol (DMAE) and four common dimethylarsinoylribosides (arsenosugars). The arsenic species were determined in seaweed extracts and in the urine and blood serum of seaweed-eating sheep from Northern Scotland. The sheep eat 2-4 kg of seaweed daily which is washed ashore on the most northern Island of Orkney. The urine, blood and wool of 20 North Ronaldsay sheep and kidney, liver and muscle from 11 sheep were sampled and analysed for their arsenic species. In addition five Dorset Finn sheep, which lived entirely on grass, were used as a control group. The sheep have a body burden of approximately 45-90 mg arsenic daily. Since the metabolism of arsenic species varies with the arsenite and arsenate being the most toxic, and organoarsenic compounds such as arsenobetaine the least toxic compounds, the determination of the arsenic species in the diet and their body fluids are important. The major arsenic species in their diet are arsenoribosides. The major metabolite excreted into urine and blood is DMAA (95 +/- 4.1%) with minor amounts of MMAA, riboside X, TMA and an unidentified species. The occurrence of MMAA is assumed to be a precursor of the exposure to inorganic arsenic, since demethylation of dimethylated or trimethylated organoarsenic compounds is not known (max. MMAA concentration 259 microg/L). The concentrations in the urine (3179 +/- 2667 microg/L) and blood (44 +/- 19 microg/kg) are at least two orders of magnitude higher than the level of arsenic in the urine of the control sheep or literature levels of blood for the unexposed sheep. The tissue samples (liver: 292 +/- 99 microg/kg, kidney: 565 +/- 193 microg/kg, muscle: 680 +/- 224 microg/kg) and wool samples (10470 +/- 5690 microg/kg) show elevated levels which are also 100 times higher than the levels for the unexposed sheep.

Animals↗
PubMed2006

PrP(Sc) deposition in nervous tissues without lymphoid tissue involvement is frequently found in ARQ/ARQ Sarda breed sheep preclinically affected with natural scrapie.

The pathogenesis of natural scrapie in Sarda breed sheep was investigated in 1050 asymptomatic and 49 sick sheep from scrapie-affected flocks. Central and peripheral nervous system, along with lymphoreticular system (LRS) tissues, were subjected to immunohistochemistry (IHC) and Western-blotting (WB) for detection of pathological isoform of the prion protein (PrP(Sc)). A total of 69 of the 1050 clinically healthy sheep were found to be infected with scrapie, with PrP(Sc) being detected in both the central nervous system (CNS) and enteric nervous system (ENS) plexuses of 60 of the sheep, while IHC and WB yielded evidence of (PrP(Sc)) deposition only in lymphoid tissues of the remaining 9 clinically healthy sheep. PrP(Sc) was also detected in the CNS, as well as in ENS plexuses from all of the 49 clinically affected sheep. Nevertheless, 18 of the 69 clinically healthy animals (26%, 17 ARQ/ARQ and 1 ARQ/AHQ sheep), along with 3 ARQ/ARQ sheep (6%) of the clinically affected group, showed no IHC or WB evidence of PrP(Sc) in lymphoid tissues, but PrP(Sc) could be still detected in their CNS and ENS plexuses. The study demonstrates dual CNS and ENS PrP(Sc) deposition in Sarda sheep with scrapie, in spite of an apparent lack of lymphoid tissue involvement in a number of cases.

Animals↗
PubMed2002

Search for healthy carriers of scrapie: an assessment of subclinical infection of sheep in an Icelandic scrapie flock by three diagnostic methods and correlation with PrP genotypes.

Subclinical infection in scrapie of sheep, characterized by a long incubation period, may be of importance for the spread of the disease. We screened brain samples from all 65 sheep in a scrapie-affected flock for subclinical infection and correlated with results of PrP genotyping, which is of relevance for the epidemiology and the question, whether by breeding for resistant genotypes one would be breeding for healthy carriers. The sensitivity of three methods was compared, i.e. histopathological examination for vacuoles (HP), immunohistochemical staining (IHC) and Western blotting (WB) for PrP(Sc). Five sheep showed definite clinical signs and histological scrapie lesions, and signs of infection were detected in 25 of 60 asymptomatic sheep, by HP and/or IHC and WB. The IHC was slightly more sensitive than HP and WB. Sheep with subclinical infection were, with one exception, either homo- or heterozygotes for 136-V, as were four of the five sheep with clinical scrapie. The incidence of the VRQ allelic variant in the flock was unusually high compared to the Icelandic sheep population probably contributing to the high prevalence of both clinical and subclinical infection in the flock. Neither sheep with definite scrapie nor detectable subclinical infection, were of the resistant AHQ genotype, indicating that Icelandic AHQ sheep are not healthy carriers of scrapie infection.

Age Factors↗
PubMed1983

The production of developmental defects of enamel in the incisor teeth of penned sheep resulting from induced parasitism.

Thirty-four sheep were infected with nematode parasites (Trichostrongylus spp. and Ostertagia circumcincta) during central incisor formation. The tooth enamel was examined for defects of mineralization at eruption and compared with that of 14 control sheep. The type and number of parasites given, the duration of the infection and the age of the sheep were varied. Sheep receiving high doses of parasites e.g. 200,000 T. vitrinus + 20,000 O. circumcincta showed severe systemic effects (debilitation, scouring and weight loss) which were of short duration (7-10 days) as the infections were terminated by anthelmintic treatment. Hypoplastic enamel (a break in continuity) was induced in all 5 sheep infected at 8 1/4-9 months of age and demarcated opacities (an alteration in the translucency) in 3 out of 4 sheep infected at 10 months of age. Sheep given 150,000 or 40,000 T. colubriformis showed less obvious systemic effects although the weight loss continued in some sheep for up to 54 days. Although opacities of the enamel in the form of parallel lines, sometimes with localized increase in width or with a diffuse opacity superimposed, were seen by microscopic examination (X 10) of teeth in this group, many of the teeth from the control sheep showed similar lesions. Only the hypoplastic lesions can be attributed with confidence to the experimental manipulations.

Animals↗
PubMed1984

Histological, macroscopic and microhardness observations of fluoride-induced changes in the enamel organ and enamel of sheep incisor teeth.

When aged 8.5 months, 10 sheep born in the same week were given 4 mg fluoride (F)/kg body weight orally for 26 days. Three sheep received no F. Sheep were killed at the end of the treatment period and later at selected stages of tooth development. The macroscopic changes in the enamel of one incisor were related to the cellular changes in the enamel organ of the contralateral tooth. A break in enamel continuity, hypoplasia, was seen on the labial enamel of 9 of the 10 F-treated sheep. Pitting of the enamel was associated with shortening of some ameloblasts and aggregations of cysts affecting cells late in their secretory phase in the first-killed sheep. In sheep killed later, these changes were associated with cells which had progressed into their maturation phase. A more extensive absence of enamel with ledge formation cervically, seen in one sheep, was associated with displacement or death of almost all the cells in their secretory phase during F treatment and consequent retention of the organic matrix. The hypoplastic lesions resulted from secretory-cell reaction during the period of F dosing. Diffuse patchy opacities, characterized by an irregular hypomineralized surface zone, were only apparent in the enamel of the later-killed sheep and were associated in one sheep with abnormal ameloblast regression in the contralateral tooth. These defects possibly resulted from the long-continued release of F stored in the bones during the period of F dosing.

Animals↗
PubMed1985

Selective exposure and analysis of the sheep tracheal lobe as a model for toxicological studies of respirable particles.

A conscious sheep model, recently developed to study sequentially the bronchoalveolar milieu, was further refined to use in the rapid in vivo assessment of the biological effects of respirable particles. In this model, the anatomically isolated tracheal lobe was selectively exposed to either 100 ml phosphate buffered saline (PBS) (control group of 12 sheep), 100 mg of 0.1-micron latex beads in 100 ml PBS (latex group of 12 sheep), or 100 mg of UICC Canadian chrysotile fibers in 100 ml PBS (asbestos group of 12 sheep). Bronchoalveolar lavages (BAL) of the tracheal lobe were obtained prior to exposure and at Days 1, 8, 15, 21, 29, 45, and 60 after exposure. Whole-lung detailed pulmonary function tests (PFT) were performed at the same times and the histopathology of the lobe was examined in six sheep in each group at Days 29 and 60. In the control sheep, there were no significant changes over time in any of the measures, and lung morphology remained normal. In the latex group, there was no significant change in PFT, the BAL analyses documented early transient increase in cellularity (macrophages and neutrophils at Day 1) and only macrophages after; lung histology documented an early macrophagic alveolitis which decreased to less than 10% of the initial inflammatory reaction at Day 60, without other distortion of the lung and airway architecture. In the asbestos sheep, the only change in whole-lung PFT was a 10-torr fall in arterial O2 pressure. BAL analyses documented persistent increases in macrophages, neutrophils, and lactate dehydrogenase as well as increasing gamma-globulins. Lung histology revealed a macrophagic and neutrophilic peribronchiolar alveolitis at Day 30, which regressed substantially by Day 60, but persistent peribronchiolar alveolitis, early fibrosis, and severe distortion of the small airways, lesions comparable to those of early asbestosis in sheep or humans. Thus selective exposure and sequential analyses of the sheep tracheal lobe in terms of BAL histomorphology should be valuable for rapid in vivo assessment of toxicity of respirable particles.

Air Pollutants↗
PubMed1989

Generation of a sheep x mouse heterohybridoma cell line (1C6.3a6T.1D7) and evaluation of its use in the production of ovine monoclonal antibodies.

A stable aminopterin-sensitive sheep X mouse heterohybridoma cell line (1C6.3a6T.1D7) for use in the generation of sheep monoclonal antibodies is described. The line was first constructed by fusing the mouse myeloma line, NSO, to normal sheep lymphocytes obtained from the efferent lymphatic vessel of a cannulated popliteal lymph node. The line was rendered sensitive to aminopterin through a combination of irradiation and treatment with the anti-metabolite drug 6-thioguanine. Characterisation of the cloned cell line showed that it did not secrete sheep immunoglobulin (Ig) molecules, express Ig on the surface membrane, or express normal sheep B or T cell surface markers. The 1C6.3a6T.1D7 line has remained stable in tissue culture for over 2 years, showing no signs of reversion to aminopterin resistance. The 1C6.3a6T.1D7 cells have been used as fusion partners with lymphocytes from antigen primed sheep to generate sheep monoclonal antibodies to human chorionic gonadotropin (hCG) or a synthetic peptide analogue of the VP1 capsid protein of foot and mouth disease virus (FMDV). To optimise the efficiency of heterohybridoma generation, comparisons were made of peripheral blood, efferent lymph or excised lymph nodes as sources of antigen-stimulated lymphocytes for fusion. The results showed that lymphocytes prepared from either efferent lymph or lymph node on the fourth day following antigenic stimulation gave similar high fusion efficiencies, and both were vastly superior to peripheral blood lymphocytes. Results were also obtained which showed that the blast cells present in lymphoid tissues due to antigenic stimulation were the major cell types involved in the generation of viable antibody-secreting sheep X sheep X mouse heterohybridomas.

Aminopterin↗
PubMed1977

Mapping of mitochondrial DNA of individual sheep and goats: rapid evolution in the D loop region.

Mitochondrial DNA (mtDNA) from sheep and goat was compared by restriction endonuclease analysis and heteroduplex mapping in the electron microscope. The fragment patterns produced by endonuclease Hae III from three individual sheep and two goat mtDNAs all differed from each other. The three sheep mtDNAs had identical Eco RI and Hind III fragments, but the two goat mtDNA patterns differed from each other as well as from sheep mtDNA. We estimate that each sheep mtDNA differs from each other by 0.5-1% of its nucleotide sequences, the two goat mtDNAs by 1-2%, and there is a 6-11% sequence difference between sheep and goat mtDNAs. We have mapped the Eco RI and Hind III sites of goat and sheep mtDNA and determined the positions of the D loop, which marks the replication origin, relative to the restriction map. The D loops are at homologous positions on the mtDNAs from both species, but the goat D loop is only 75% as long as the sheep D loop. Regions with a high degree of sequence divergence occur at both ends of the D loop. We suggest that a duplication of about 150 base pairs has occurred in the region where the sheep and goat D loops differ in length. We discuss mtDNA evolution in terms of divergence of isolated "mitochondrial DNA clones."

Animals↗
PubMed1992

Topical anesthetic-induced methemoglobinemia in sheep: a comparison of benzocaine and lidocaine.

Benzocaine induces methemoglobin (MHb) in several species, whereas lidocaine may increase MHb in cats and human. Elevated MHb (greater than 20%) in sheep after benzocaine exposure was recently recognized. MHb decreases blood oxygen-carrying capacity which can complicate interpretation of experimental data. Sheep are used in research which requires tracheal intubation and blood gas analysis. Since benzocaine and lidocaine are used to provide local anesthesia prior to intubation, we compared MHb production by sheep after exposure to these drugs. A dose-response relationship between benzocaine and MHb was investigated. Eight crossbred Dorset ewes were dosed intranasally with benzocaine for 2 sec or with 40 mg of lidocaine in a randomized crossover design. Sheep with elevated MHb after the 2-sec benzocaine dose were later dosed with benzocaine intranasally for 10 sec. MHb levels were measured periodically on a CO-Oximeter. A quantitative MHb response to an indirect MHb former, p-aminopropiophenone (PAPP), by each sheep was determined 15 min after PAPP (0.6 mg/kg iv). MHb levels remained at baseline (1-2%) after lidocaine exposure in all sheep, as well as in four sheep (nonresponders) after the 2-sec benzocaine dose. Four sheep (responders) demonstrated 16.5-26.4% MHb after the 2-sec benzocaine dose. The responders formed 38.2-50.5% MHb after the 10-sec benzocaine dose. All responders developed high MHb after PAPP, while nonresponders developed slightly elevated MHb after PAPP. An N-hydroxy metabolite of benzocaine is the likely active MHb-forming substance. Benzocaine should be replaced by lidocaine when local anesthesia of the nasal or oropharyngeal region in sheep is required.

Administration, Topical↗
PubMed1996

Immunochemical and sub-structural characterization of sheep lung cytochrome P450LgM2.

Sheep lung cytochrome P450LgM2 belonging to gene subfamily 2B, was obtained in highly purified form and antibodies against sheep lung cytochrome P450LgM2 were produced in rabbits by using the previously developed methods in our laboratory. Immunological and enzymatic studies showed that antibodies against lung cytochrome P450LgM2 inhibited benzphetamine N-demethylation, ethylmorphine N-demethylation and aniline 4-hydroxylation reactions in sheep lung microsomes about 99, 80 and 62%, respectively. Benzphetamine N-demethylation reaction in sheep lung microsomes was only catalyzed by cytochrome P450LgM2 isozyme while the other isozymes of P450 as well as P450LgM2 are also involved in the metabolism of ethylmorphine and aniline. Similar to lung microsomes, benzphetamine N-demethylase activity of the reconstituted systems containing purified sheep lung cytochrome P450LgM2 or phenobarbital (PB)-treated rabbit liver cytochrome P450LM2(2B4) was also inhibited by P450LgM2 antibodies about 95 and 82%, respectively. A 50% inhibitory effect of sheep P450LgM2 antibodies was also observed in ethylmorphine N-demethylase activity of the reconstituted system containing purified sheep lung cytochrome P450LgM2. SDS-PAGE peptide maps obtained following the partial proteolysis of purified sheep lung cytochrome P450LgM2 and PB-rabbit liver P450LM2(2B4) isozymes, using chymotrypsin and papain, were similar in general. However they showed some differences both qualitatively and quantitatively, suggesting that some differences exist among the amino acid sequences of sheep lung cytochrome P450LgM2 and rabbit liver cytochrome P4502B4.

Animals↗