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Demonstration of cytotoxic lymphocytes to virus-infected target cells in pigs inoculated with transmissible gastroenteritis virus.

Lymphocytes, cytotoxic to virus-infected target cells, were induced in pigs orally exposed to transmissible gastroenteritis virus. They were studied and experiments were carried out by using autochthonous testicle cells as target cells to avoid genetic incompatibility of effector lymphocytes and target cells. Cytotoxic lymphocytes were demonstrated in Peyer's patches, mesenteric lymph nodes, spleen, and peripheral blood on postinoculation day (PID) 7. Cytotoxic activity of lymphocytes increased thereafter and reached the maximal amount at PID 21. Lymphocyte cytotoxicity was somewhat greater in lymphocytes of peripheral blood and spleen than in those of Peyer's patches and mesenteric lymph nodes after PID 14. On the contrary, lymphocyte reactivity to the viral antigen measured by lymphocyte proliferative assay was higher in Peyer's patch and mesenteric lymph node cells than in peripheral blood and splenic cells. Lymphocyte cytotoxicity was depressed by treating effector cells with anti-porcine thymocyte serum and complement. However, lymphocyte suspensions treated with anti-porcine thymocyte serum and complement were still cytotoxic to some extent against virus-infected target cells, although T lymphocytes were completely excluded by the treatment. This suggests that cytotoxic mechanism other than the direct action of cytotoxic T lymphocytes may be involved in the cytotoxicity assay systems used in the present studies. In experiments in which allogenic cells (testicle cells of siblings) were used together with autochthonous cells as targets, lymphocyte cytotoxicity was equally expressed against both autochthonous and allogenic target cells in 2 of 3 experiments. However, lymphocyte cytotoxicity was greater against autochthonous cells than against allogenic target cells in 1 of 3 experiments.

Animals↗

Clinical evaluation of transmissible gastroenteritis virus vaccines and vaccination procedures for inducing lactogenic immunity in sows.

Two federally licensed attenuated live transmissible gastroenteritis (TGE) virus vaccines (an IM vaccine and an oral-IM vaccine) and 1 nonlicensed nonattenuated live TGE virus vaccine were evaluated and compared in sows free of TGE virus-neutralizing antibodies. Litters from the sows were challenge exposed at 3 and 5 days of age, and results were combined according to the vaccine administered to the sows. The survivability of pigs suckling sows vaccinated with the nonattenuated vaccine was significantly (P less than 0.01) greater than that of pigs suckling sows vaccinated with the IM attenuated vaccine, significantly (P less than 0.05) greater than that of pigs suckling sows vaccinated with the oral-IM attenuated vaccine, and significantly (P less than 0.05) greater than that of pigs suckling sows that had not been vaccinated. The differences, however, between survivability of litters from sows vaccinated with the IM attenuated vaccine or the oral-IM attenuated vaccine and that of litters from the sows not vaccinated were not significant (P greater than 0.10). The nonattenuated TGE vaccine, although giving a higher level of protection than the attenuated vaccine, was eventually overwhelmed. Dexamethasone did not increase the incidence of diarrhea, and levamisole did not potentiate the lactogenic immunity in sows after given their first dose of the nonattenuated vaccine. Survivability in litters suckling sows that developed diarrhea after given their first dose of the nonattenuated vaccine was not greater than that in litters suckling sows that did not develop diarrhea. The best results were obtained when 3-day-old suckling pigs were challenge exposed with virulent TGE virus.

Animals↗

Applications of peroxidase-antiperoxidase staining technique for detection of transmissible gastroenteritis virus in pigs.

The peroxidase-antiperoxidase (PAP) staining technique was used for the detection of transmissible gastroenteritis virus (TGEV) in small intestines of TGEV-infected 8-week-old pigs and in infected McClurkin pig testicle cells by means of light microscopy. The specific-positive reaction was characterized by the presence of many brown granules of various sizes in the cytoplasm of infected cells. Nonspecific granules caused by endogenous peroxidases in the cytoplasm of eosinophils stained by PAP were darker, larger, more round, and more uniform in size than were specific granules. Acetone fixation was superior to fixation with periodate-lysine-paraformaldehyde or 10% formalin. Our results indicate that the PAP staining technique is a sensitive, specific technique for detection of TGEV in the small intestines of pigs.

Animals↗

Isolation of hemagglutination-defective mutants for the analysis of the sialic acid binding activity of transmissible gastroenteritis virus.

The surface protein S of transmissible gastroenteritis virus (TGEV) has a sialic acid binding activity that enables the virus to agglutinate erythrocytes. A protocol is described that has been successfully applied to the isolation of hemgglutination-defective mutants. The potential of these mutants for the characterization of the sialic acid-binding site and the function of the binding activity is discussed.

Animals↗

Infection with porcine respiratory coronavirus does not fully protect pigs against intestinal transmissible gastroenteritis virus.

Eight nine-week-old specific-pathogen-free pigs which had been infected with the transmissible gastroenteritis virus (TGEV)-related porcine respiratory coronavirus (PRCV) and four uninfected littermates were challenged with TGEV. The previous PRCV infection failed to protect them against the enteric TGEV infection. Virus excretion in faeces was detected by an ELISA in all the pigs for three to six consecutive days after inoculation. Although little diarrhoea was observed, the infection extended through much of the small intestine of one of the previously infected pigs four days after inoculation. Challenge with TGEV caused a secondary neutralising antibody response. By using a peroxidase conjugate of a monoclonal antibody which recognises a specific antigenic site on TGEV, antibodies against TGEV could be distinguished from antibodies against PRCV in an ELISA blocking test.

Animals↗

Interferon induction in porcine leukocytes with transmissible gastroenteritis virus.

Leukocytes were harvested from the peripheral blood, mesenteric lymph node and small intestinal lamina propria from groups of three piglets before, and 1, 2 and 3 weeks after infection with virulent transmissible gastroenteritis virus (TGEV) at 2 weeks of age. The donor piglets developed clinical signs of transmissible gastroenteritis which persisted for up to 3 days, and they developed peak serum titres of TGEV-neutralizing antibodies 2 weeks post-infection. The leukocytes were cultured in the presence of pokeweed mitogen (PWM), various dilutions of purified TGEV, or control media for 3 or 5 days, and the culture supernatants were tested for antiviral activity in MDBK cells challenged with vesicular stomatitis virus. The antiviral activity was characterized as porcine interferon (IFN)-alpha or porcine IFN-tau on the basis of its stability at pH 2.0 and neutralization by anti-human IFN-alpha antibodies. Viability of the leukocytes in culture, determined by trypan blue exclusion, was highest for the peripheral blood leukocytes and lowest for the mesenteric lymph node leukocytes. There were no consistent differences in antiviral activity between cultures incubated for 3 or 5 days. Porcine IFN-alpha was found in the supernatants of the leukocyte cultures stimulated with TGEV antigen, harvested before or after infection of the donor piglets with TGEV. Porcine IFN-tau was demonstrated in the supernatants of the leukocyte cultures stimulated with PWM, more frequently when the leukocytes were harvested post-infection. This was the first demonstration of IFN induction in vitro in leukocytes from porcine gut-associated lymphoid tissue.

Animals↗

Financial impact of transmissible gastroenteritis in pigs.

The financial impact of an epizootic of transmissible gastroenteritis in pigs was evaluated in a California sow herd through estimating growth, feed, and profit functions. Two groups of pigs were studied: pigs born before and surviving the epizootic (epizootic [E] pigs), and pigs born after the epizootic (postepizootic [PE] pigs). Short-term profits were maximized at 165 days for both groups of pigs, ranging from $47.14 for female E pigs to $60.32 for male PE pigs. Accordingly, it was concluded that pigs surviving or born shortly after a transmissible gastroenteritis epizootic are profitable to raise, if raised under management conditions similar to those in the study herd.

Animal Feed↗

[Use of a microvirus-neutralizing reaction in diagnosing transmissible gastroenteritis (TGE)].

A micro virus-neutralization reaction was designed and tested to detect antibodies to the virus of transmissive gastroenteritis. Use was made of a stable cell line, SPEV, and a laboratory strain of the virus that had been adapted to it. The optimal concentration values of the cell suspension and the normal calf serum contained in it were determined. A total of 90 blood serum samples from pigs were comparatively investigated for the presence of virus-neutralising TGE antibodies, the tube test being performed trough the inoculation of the cells in suspension. On the other hand, the micro virus-neutralization test was carried out in two variants: the sera were diluted via Mikrotiter micropipettes and micropipettes of the Takachi apparatus. It was found that the micro virus-neutralization test in its two variants was not inferior in terms of sensitivity to the tube test, was more readily applicable, and was less material consuming.

Animals↗

Experience with a planned exposure program for the control of enzootic transmissible gastroenteritis in swine.

Oral inoculation of pregnant sows and gilts with a homogenate of pig intestines containing live, virulent transmissible gastroenteritis (TGE) virus was associated with significant (P < 0.01) reduction of mortality in nursery pigs in a herd affected with enzootic TGE. The mortality of weaned pigs from April through June 1981, when sows were not vaccinated or inoculated, was 9.3%. Mortality of weanling pigs from July through December 1981 was 5.7% (P < 0.01), and stayed consistently between 2.6 and 4.8% from 1982 through 1990. After beginning the exposure program, there were no clinical signs attributable to TGE in the farrowing house or nursery until 1986, when recrudescence of TGE was evident in the nursery. Although the source of the virus could not be substantiated, recrudescence suggested that TGE virus would not be eradicated as long as exposure was continued. The most plausible explanation for the decrease in the incidence of diarrhea and mortality in the nursery pigs after initiation of the planned exposure program is that a higher, longer lasting amount of lactogenic immunity was provided to suckling pigs. Presumably over time, this resulted in less TGE virus carryover into the nursery by weanling pigs, thereby reducing and confining the environmental challenge to the nursery. Strict all-in, all-out pig movement in the nursery with thorough cleaning and disinfecting presumably further reduced the environmental challenge.

Administration, Oral↗

Active and passive immune responses to transmissible gastroenteritis virus (TGEV) in swine inoculated with recombinant baculovirus-expressed TGEV spike glycoprotein vaccines.

OBJECTIVE: Baculovirus-expressed transmissible gastroenteritis virus (TGEV) spike (S) glycoprotein vaccines were inoculated parenterally in swine to determine whether such vaccines could induce serum and whey virus-neutralizing (VN) antibodies and protective lactogenic immunity for TGEV-challenge-exposed pigs. ANIMALS AND PROCEDURES: 3 recombinant baculoviruses that expressed full or partial length TGEV Miller strain S glycoproteins were inoculated SC in 17 conventionally raised 11-day-old TGEV-seronegative pigs to determine whether the recombinant S glycoproteins would elicit serum VN antibodies. Eleven TGEV-seronegative pregnant sows were inoculated SC or intramammarily with subunit vaccines (R2-2 or R3-5) or control proteins. Pigs born to 9 of the 11 sows were challenge exposed at 4 to 5 days of age with the virulent Miller strain, and passive immunity was assessed. Serum and whey antibody responses to TGEV were analyzed by VN and ELISA testing. RESULTS: Recombinant S glycoproteins (R2-2 or R3-5) containing the 4 major antigenic sites induced similar VN antibody titers to TGEV in serum and colostrum, but low (some sows) or no VN antibody titer was detected in milk. Subcutaneous inoculation of sows with R2-2 or R3-5 elicited IgG, but not IgA antibodies to TGEV in colostrum. Morbidity was 100%, and mortality ranged from 20 to 80% in TGEV challenge-exposed pigs nursing sows inoculated SC or intramammarily with TGEV S glycoprotein vaccines. CONCLUSIONS AND CLINICAL RELEVANCE: Parenterally administered TGEV S glycoprotein vaccines elicit VN antibodies to TGEV in serum and colostrum that do not fully provide active or passive immunity in swine.

Animals↗

Observations on clinical aspects of transmissible gastroenteritis of pigs in Norfolk and Suffolk, 1980-81.

Outbreaks of transmissible gastroenteritis in 56 breeding units, six maiden gilt sites and 64 fattening units were investigated during the period December 1980 to October 1981. The disease invariably occurred in classical virulent form, resulting in very high mortality in baby piglets. Reproductive sequelae were sometimes evident. Individual outbreaks continued for longer than in previous years. There was a high rate of recrudescene, particularly in large breeding units, and infection also persisted in many fattening units. The value of control measures and supportive therapy is discussed.

Abortion, Veterinary↗

Immunity to transmissible gastroenteritis virus and porcine respiratory coronavirus infections in swine.

Despite the pioneering efforts to identify correlates of passive immunity to transmissible gastroenteritis virus (TGEV), effective vaccines for the control of TGE in suckling pigs have remained elusive. The initial concept of an enteromammary immunologic axis in monogastrics originated from studies of lactogenic immunity to TGEV in swine. These studies revealed that infection of pregnant swine with virulent TGEV stimulated high titers of SIgA antibodies in milk which correlated with protection of suckling pigs against TGE; parenteral or oral inoculation with live attenuated or killed TGEV vaccines induced mainly IgG antibodies in milk which generally provided poor protection to suckling pigs. The recent appearance of PRCV infections in swine and continuing studies of TGEV infections, present a unique model for further studies of mucosal immunity. Research using these viruses has increased our understanding of the various components of the common mucosal immune system and their interactions. Although the most important consideration in designing an effective vaccine for TGEV is the stimulation of GALT through intestinal virus replication, studies addressing the contribution of BALT to immunity to TGEV and PRCV may provide insights for alternative vaccine approaches. The mechanism by which exposure to PRCV elicits a variable-degree of immunity to TGEV challenge is unknown. Virus replication in the gut or respiratory tract is a major factor affecting the magnitude of the immune response at the respective site and may be necessary for the recruitment of specific immune cells from other mucosal inductive sites, i.e., GALT to BALT and BALT to GALT migration.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Induction of the 2-5A system by interferon and transmissible gastroenteritis virus.

Interferon (IFN) treatment increased the level of 2',5'-oligoadenylate (2-5A) synthetase and inhibited replication of transmissible gastroenteritis virus (TGEV) in cultured swine testicular cells. Despite a minimal increase in TGEV titer in IFN-treated swine testicular cells, there was rapid cellular destruction. IFN-treated swine testicular cells had detectable levels of 2-5A (3.6 nM 6 h post-infection) after infection with 30 pfu TGEV/cell. Infection of neonatal pigs with a virulent strain of TGEV caused a significant increase in serum IFN and enterocyte 2-5A synthetase activity, as compared to control pigs. The level of enterocyte 2-5A synthetase in TGEV-infected pigs was increased 25-fold before viral-induced damage of the intestine consistently was present. 2-5A was not detected in enterocytes of either TGEV-infected or control pigs (less than 0.5 nM). Preparations of enterocytes contained two subpopulations of cells, one of which does not support replication of the virus. The considerable dilution of TGEV-infected cells (villous enterocytes) with uninfected cells (crypt cells) may be responsible for our inability to detect 2-5A in enterocytes from TGEV-infected pigs. These results indicate that the 2-5A system in porcine enterocytes and cultured testicular cells is modulated by TGEV infection and/or interaction with IFN.

2',5'-Oligoadenylate Synthetase↗

Site-specific alteration of transmissible gastroenteritis virus spike protein results in markedly reduced pathogenicity.

The pathogenicity of neutralization-resistant mutants of the enteric coronavirus transmissible gastroenteritis virus (TGEV) was examined in the newborn piglet. The parental virus (Purdue-115 strain), as well as several mutants selected using monoclonal antibodies (MAbs) directed to antigenic sites A and B, caused an acute enteritis with 100% mortality. By contrast, most of the site D (MAb 40.1) mutants exhibited a strongly reduced enteropathogenicity, leading to the survival of animals inoculated with up to 1000-fold the 100% lethal dose of parental virus. Such a phenotypical change was correlated with point mutations or a small deletion, all located within the S gene sequence coding for the Pro-145 to Cys-155 segment of the mature polypeptide. These observations suggest that an N-terminal subregion of the S molecule is an essential determinant for pathogenesis in TGEV infection.

Animals↗

Effect of ambient temperatures on multiplication of attenuated transmissible gastroenteritis virus in the bodies of newborn piglets.

Newborn piglets were found to be more resistant to infection with attenuated transmissible gastroenteritis virus when maintained at higher temperatures. This was attributed to a decreased rate of virus propagation and spreading in the bodies of the infected animals. The highest virus levels were detected in the tissues of piglets maintained at 8 to 12 C. In contrast, no virus was recovered from piglets maintained at 35 to 37.5 C. The virus was found only in the lymph nodes and respiratory organs in the piglets maintained at 20 to 23 C.

Animals↗

Isolation of transmissible gastroenteritis virus from pharyngeal swabs obtained from sows at slaughter.

A virologic survey was conducted to determine the frequency of transmissible gastroenteritis (TGE) virus infection in farm-raised sows. Pharyngeal swab specimens collected in an abattoir were examined for TGE virus by inoculation onto swine-testes cell culures. The virus was detected in 61 (3%) of a sample of 2,058 Iowa sows after slaughter. All TGE viral isolates, given orally to 2- or 3-day-old pigs, caused acute gastroenteritis and in some cases death. All pigs that recovered from illness had serum antibody to TGE virus.

Animals↗

Electrolyte transport in piglets infected with transmissible gastroenteritis virus. Stimulation by verapamil and clonidine.

The effects of clonidine, an alpha 2-adrenergic agonist, and verapamil, a Ca2+ channel blocker, on Na+ and Cl- absorption were studied in stripped jejunal mucosa from control and transmissible-gastroenteritis-virus-infected piglets. All infected piglets developed severe diarrhea 18-24 hours after oral inoculation. Jejunum from infected animals, as compared with control jejunum, had decreased mucosal-to-serosal, serosal-to-mucosal, and net Na+ and Cl- fluxes. Clonidine and verapamil caused a decrease in short-circuit current and stimulation of Na+ and Cl- absorption in control jejunum. In infected piglets, although the jejunum exhibited severe villus atrophy, both drugs stimulated Na+ and Cl- absorption and the magnitude of Na+ and Cl- absorption was similar in control and transmissible-gastroenteritis-infected jejunum. In contrast, D-glucose stimulated Na+ absorption, and the decrease in short-circuit current caused by verapamil and clonidine, were decreased in transmissible-gastroenteritis-infected jejunum. Such pharmacological stimulation of Na+ and Cl- absorption might be useful in the management and treatment of certain viral diarrheal diseases.

Animals↗