[Clinical analysis of 12 cases of ovarian yolk sac tumor].

Objective: To investigate the diagnosis, treatment and prognosis of ovarian yolk sac tumor (OYST). Methods: The clinicopathological data and follow-up data of 12 patients with OYST admitted to the Affiliated Hospital of Qingdao University from January 2013 to December 2020 were retrospectively analyzed, and the diagnosis, treatment and prognosis of OYST patients were summarized. Results: (1) The age of 12 patients with OYST ranged from 11 to 37 years, with a median age of 20 years. At the first visit, all 12 patients had pelvic masses. Reasons for seeing a doctor: 6 cases of abdominal distension and abdominal pain, 4 cases of mass in the lower abdomen, 1 case of vaginal bleeding, and 1 case of appendicitis. International Federation of Obstetrics and Gynecology (FIGO) 2014 staging: 4 cases in stage Ⅰa, 2 cases in stage Ⅰc, 1 case in stage Ⅱc, 4 cases in stage Ⅲc, and 1 case in stage Ⅳb. (2) All 12 patients were examined by color Doppler ultrasound before operation, among which 10 cases showed unilateral adnexal masses and 2 cases bilateral adnexal masses. The median maximum diameter of tumor was 16.5 cm (range: 6.0-28.0 cm). The preoperative levels of alpha fetoprotein (AFP) in 12 patients (all >1 210 µg/L) were significantly higher than normal (<25 µg/L). Among the 11 patients with cancer antigen 125 (CA125) detection results, 9 patients showed elevated serum CA125 levels. (3) Among the 12 patients, 8 young infertile patients who needed to preserve their reproductive function underwent appendectomy, 3 infertile patients underwent staged surgery for ovarian malignant germ cell tumor, and only one bilateral lesion and infertile patient underwent unsatisfactory staged surgery for ovarian malignant germ cell tumor. Of the 12 patients, 11 patients were given combined chemotherapy regimen of bleomycin, cisplatin, and etoposide (BEP) after operation. One patient without chemotherapy developed metastasis 3 months after operation, and was given BEP chemotherapy, and her condition was controlled. (4) The deadline for follow-up was December 31st, 2022, and the median follow-up time was 60 months (range: 25-115 months). All the 12 patients survived without tumor during the follow-up period, and the median disease-free survival time was 84.5 months (range: 25-115 months). Conclusions: OYST mostly occurs in children and young women. Color Doppler ultrasound examination and serum AFP and CA125 detection have diagnostic value for OYST. Surgical treatment after diagnosis of OYST includes surgery to preserve reproductive function and timely and standardized chemotherapy after operation. The prognosis of patients is good regardless of stage.

Remote ischemic postconditioning alleviates myocardial ischemia/reperfusion injury by up-regulating ALDH2.

OBJECTIVE: The myocardial ischemia/reperfusion (I/R) injury is a significant challenge, and the clinical significance of remote ischemic postconditioning (RIPostC) in cardioprotection has been confirmed. However, the molecular mechanism remains unclear. We aimed to explore the regulatory mechanism of RIPostC in myocardial I/R. MATERIALS AND METHODS: A mouse model of myocardial I/R injury and cell model of oxygen-glucose deprivation (OGD)/re-oxygenation (OGD/R) injury were constructed. Infarct size was measured by Evans blue dye staining and TTC staining. mRNA and protein expression levels of aldehyde dehydrogenase 2 (ALDH2) were determined by RT-qPCR and Western blot analysis, respectively. Cell viability, p53 expression, apoptotic cells, expression of proteins related to apoptosis, and reactive oxygen species (ROS) generation were evaluated by CCK-8 assay, Western blot analysis, flow cytometry assay, Western blot analysis, and DCFH-DA staining, respectively. ALDH2 in H9c2 cells was knocked down, and its effects on cells treated with OGD/R and RIPostC were tested. How RIPostC affected ALDH2 expression was finally studied. RESULTS: RIPostC reduced infarct size in mice and attenuated OGD/R-induced H9c2 cell injury. Myocardial I/R-induced down-regulation of ALDH2 was abrogated by RIPostC. Moreover, the effects of RIPostC on OGD/R-treated H9c2 cells were significantly reversed by ALDH2 silence. Finally, we found RIPostC-induced up-regulation of ALDH2 in OGD/R-treated cells could be bated by activation of PI3K and/or mTOR. CONCLUSIONS: RIPostC exerted cardioprotective role against myocardial I/R both in vivo and in vitro. Up-regulation of ALDH2 might be a reason for the cardioprotection, and RIPostC might regulate ALDH2 expression via the PI3K/mTOR pathway.

Inhibition effects of acridone on the growth of breast cancer cells in vivo.

OBJECTIVE: To investigate the anti-tumor effect of acridone against breast cancer in vivo and provide a therapeutic agent for treatment of breast cancer. MATERIALS AND METHODS: The nude mice xenografted tumor model was established by MCF-7 cells. The mice were randomly divided into four groups. The mice in each group (n=6) were intraperitoneally injected with 0.1 mg/kg saline (low-dose), 0.5 mg/kg (middle-dose) and 1.0 mg/kg (high-dose) of acridone for 21 days, respectively. At the end of the animal experiment, the weight of tumors was recorded to calculate the tumor inhibition rate. The serum hormone levels in peripheral blood were determined using ELISA. Hematoxylin and eosin (HE) staining was used to analyze the histopathological changes. The expression of ABCG2 protein and mRNA were determined by Western blot and RT-PCR, respectively. RESULTS: The inhibition rates of tumor growth in the high-dose, middle-dose, and low-dose groups were 29.18%, 17.21%, and 4.27%, respectively. Compared with control and low-dose group, the tumors growth rate in high-dose and middle-dose groups were decreased significantly. Histologically, the tumors were inhibited in the growth rate, the tissue structure was broken. Estrogen in all groups with acridone treatment decreased, the progesterone in high-dose and middle-dose groups increased remarkably. The expression of ABCG2 protein and ABCG2 mRNA decreased after treatment with acridone. CONCLUSIONS: We showed that acridone could induce cell apoptosis, inhibited ABCG2 (ATP-binding cassette sub-family G member 2) protein and adjusted hormone level. The results suggested that acridone could serve as a chemotherapeutic agent for treatment of breast cancer in vivo.

Texture evolution and mechanical behaviour of irradiated face-centred cubic metals.

A physically based theoretical model is proposed to investigate the mechanical behaviour and crystallographic texture evolution of irradiated face-centred cubic metals. This model is capable of capturing the main features of irradiated polycrystalline materials including irradiation hardening, post-yield softening and plasticity localization. Numerical results show a good agreement with experimental data for both unirradiated and irradiated stress-strain relationships. The study of crystallographic texture reveals that the initial randomly distributed texture of unirradiated metals under tensile loading can evolve into a mixture of [111] and [100] textures. Regarding the irradiated case, crystallographic texture develops in a different way, and an extra part of [110] texture evolves into [100] and [111] textures. Thus, [100] and [111] textures become dominant more quickly compared with those of the unirradiated case for the reason that [100] and [111]-oriented crystals have higher strength, and their plastic deformation behaviours are more active than other oriented crystals. It can be concluded that irradiation-induced defects can affect both the mechanical behaviour and texture evolution of metals, both of which are closely related to irradiation hardening.

Ultrastructural investigation of antennae in three cutaneous myiasis flies: Melophagus ovinus, Hippobosca equina, and Hippobosca longipennis (Diptera: Hippoboscidae).

Melophagus ovinus (Linnaeus 1758), Hippobosca equina Linnaeus, 1758, and Hippobosca longipennis Fabricius, 1805 (Diptera: Hippoboscidae) are economically and medically important ectoparasites that can act as mechanic vectors of pathogens and cause myiasis in both human and domestic animals. As essential olfactory organs, antennae of these adult hippoboscids were examined using stereoscopic and scanning electron microscopes. General morphology of the antenna is provided in detail, combined with distribution, types, size, and ultrastructures of antennal sensilla. On the antennal funiculus, two types of sensilla are observed, including basiconic sensilla and coeloconic sensilla. Four common characters are shared among the three species: (1) the scape is either obsolete or fused with the fronto-clypeus; (2) branched antennal structures (branched pedicellar microtrichiae and branched arista with only one segment) are detected; (3) the enlarged antennal pedicel completely envelops the antennal funiculus; and (4) less types of sensilla on funiculus. Disparity and diversity of the antennal and sensory structures are analyzed from the phylogenetic and functional perspective. We suggest that hippoboscids are potential model for the study of the function of coeloconic sensilla in Calyptratae.

Mechanical properties for irradiated face-centred cubic nanocrystalline metals.

In this paper, a self-consistent plasticity theory is proposed to model the mechanical behaviours of irradiated face-centred cubic nanocrystalline metals. At the grain level, a tensorial crystal model with both irradiation and grain size effects is applied for the grain interior (GI), whereas both grain boundary (GB) sliding with irradiation effect and GB diffusion are considered in modelling the behaviours of GBs. The elastic-viscoplastic self-consistent method with considering grain size distribution is developed to transit the microscopic behaviour of individual grains to the macroscopic properties of nanocrystals (NCs). The proposed theory is applied to model the mechanical properties of irradiated NC copper, and the feasibility and efficiency have been validated by comparing with experimental data. Numerical results show that: (i) irradiation-induced defects can lead to irradiation hardening in the GIs, but the hardening effect decreases with the grain size due to the increasing absorption of defects by GBs. Meanwhile, the absorbed defects would make the GBs softer than the unirradiated case. (ii) There exists a critical grain size for irradiated NC metals, which separates the grain size into the irradiation hardening dominant region (above the critical size) and irradiation softening dominant region (below the critical size). (iii) The distribution of grain size has a significant influence on the mechanical behaviours of both irradiated and unirradiated NCs. The proposed model can offer a valid theoretical foundation to study the irradiation effect on NC materials.

Cross-species amplification of 92 microsatellites of Medicago truncatula.

Medicago species are important genetic sources for forage crops and nitrogen sources for various ecosystems. The ongoing genome sequencing of the model legume, Medicago truncatula, provides a wealth of genetic markers potentially useful for characterizing the population genetic structure and evolutionary history, and the potential of the wild Medicago species. Here we tested the PCR amplification of 92 microsatellites developed from M. truncatula in six other Medicago species, and found that the cross-species transferability, ranging from 53.26% to 61.96%, is comparable with those reported in other angiosperm genera. This article thus reports a number of microsatellites that are potentially useful for large-scale ecological and evolutionary genetic studies of wild Medicago species.

A genetically engineered chimeric vaccine against porcine circovirus type 2 (PCV2) improves clinical, pathological and virological outcomes in postweaning multisystemic wasting syndrome affected farms.

The present study describes the effects of a commercially available genetically engineered chimeric vaccine against porcine circovirus type 2 (PCV2) on clinical, pathological and virological features in three multi-site farms suffering from postweaning multisystemic wasting syndrome (PMWS). The vaccine product was able to reduce clinical signs, PCV2 viral load in lymphoid organs and/or sera, and overall mortality in nurseries and fattening units. This is the first time in which is shown that a PCV2 vaccine is able to decrease specifically PMWS-associated mortality. Another novelty of this study is the assessment of PMWS-like histological lesions in a large number of vaccinated and non-vaccinated pigs under field conditions.

Photosynthetic and biochemical activities in flag leaves of a newly developed superhigh-yield hybrid rice (Oryza sativa) and its parents during the reproductive stage.

Responses of net photosynthetic rates to intercellular CO(2) concentration (P (n)/C (i) curves) and photochemical characteristics were investigated in flag leaves of newly developed superhigh-yield hybrid rice (Oryza sativa L.) LiangYouPeiJiu (LYPJ) and its maternal PeiAi64S (PA64S) and paternal WuMang9311 (WM9311) lines grown in the field during the reproductive stage. The results showed that photosynthetic functions, such as the electron transport activities of photosystems and photophosphorylation, assessed in vivo from P (n)/C (i) curves under field conditions declined more or earlier than those obtained in vitro. The degradation of polypeptides of thylakoid membranes was slower than those for P (Ca=360) (light-saturated net photosynthetic rate measured at 360 mumol mol(-1)) and CE (carboxylation efficiency, obtained from the initial slope of the P (n)/C (i) curve). The initial inhibition of the PSII electron transport and oxygen-evolving activity induced by senescence occurred before the degradation of the oxygen-evolving complex. In comparison, LYPJ had intermediate photosynthetic functions in the early stage of leaf development, but greater photochemical activities in the mid and late stages. WM9311 showed a similar pattern of changes but lower values, and PA64S had higher values in the early stage but showed a faster rate of senescence than LYPJ. These findings implied that the hybrid LYPJ demonstrated intermediate photosynthetic activities between its parents in the early stage of leaf development, whereas it had higher photosynthetic activities than its parents in the mid and late stages, which may be responsible for its high yield.

Leishmune vaccine blocks the transmission of canine visceral leishmaniasis: absence of Leishmania parasites in blood, skin and lymph nodes of vaccinated exposed dogs.

Leishmune vaccine is the first licensed vaccine against canine visceral leishmaniasis. It contains the Fucose-Mannose-ligand (FML) antigen of Leishmania donovani. The potential Leishmune vaccine effect on the interruption of the transmission of the disease, was assayed by monitoring, in untreated (n=40) and vaccinated dogs (n=32) of a Brazilian epidemic area: the kala-azar clinical signs, the FML-seropositivity and the Leishmania parasite evidence by immunohistochemistry of skin and PCR for Leishmanial DNA of lymph node and blood samples. On month 11 after vaccination, untreated controls showed: 25% of symptomatic cases, 50% of FML-seropositivity, 56.7% of lymph node PCR, 15.7% of blood PCR and 25% of immunohistochemical positive reactions. The Leishmune-vaccinated dogs showed 100% of seropositivity to FML and a complete absence of clinical signs and of parasites (0%) in skin, lymph node and blood PCR samples (p<0.01). The positivity in FML-ELISA in untreated dogs significantly correlates with the PCR in lymph node samples (p<0.001) and with the increase in number of symptoms (p=0.006) being strong markers of infectiousness. The absence of symptoms and of evidence of Leishmania DNA and parasites in Leishmune-vaccinated animals indicates the non-infectious condition of the Leishmune-vaccinated dogs.

An equine protozoal myeloencephalitis challenge model testing a second transport after inoculation with Sarcocystis neurona sporocysts.

Previous challenge studies performed at Ohio State University involved a transport-stress model where the study animals were dosed with Sarcocystis neurona sporocysts on the day of arrival. This study was to test a second transportation of horses after oral inoculation with S. neurona sporocysts. Horses were assigned randomly to groups: group 1, transported 4 days after inoculation (DAI); group 2, at 11 DAI; group 3, at 18 DAI; and group 4, horses were not transported a second time (controls). An overall neurologic score was determined on the basis of a standard numbering system used by veterinarians. All scores are out of 5, which is the most severely affected animal. The mean score for the group 1 horses was 2.42; group 2 horses was 2.5; group 3 horses was 2.75; and group 4 horses was 3.25. Because the group 4 horses did not have a second transport, they were compared with all other groups. Statistically different scores were present between group 4 and groups 1 and 2. There was no difference in the time of seroconversion between groups. There was a difference between the time of onset of first clinical signs between groups 1 and 4. This difference was likely because of the different examination days. Differences in housing and handling were likely the reason for the differences in severity of clinical signs. This model results in consistent, significant clinical signs in all horses at approximately the same time period after inoculation but was most severe in horses that did not experience a second transport.

Equine vaccine for West Nile virus.

To meet the urgent need of controlling West Nile virus (WNV) infection in the equine population, we have developed a killed WNV vaccine. A dose titration study in horses was first conducted to evaluate serum neutralization antibody responses against WNV in these animals. Horses were vaccinated intramuscularly twice with the test vaccine at low, medium and high dose, three weeks apart. Serum samples were collected periodically and were measured for serum neutralizing antibody using a plaque reduction neutralization test. Significant increases in serum neutralizing antibody were detected in all three dosage groups 14 days post the second vaccination. Twelve months after the second vaccination, horses vaccinated with the medium dose of WNV vaccine and non-vaccinated control horses were experimentally challenged with WNV. Nine out of 11 (81.8%) controls developed viraemia after challenge while only one out of 19 (5.3%) vaccinates had transient viraemia, representing a 94% preventable fraction. In a separate study, the safety of the killed WNV vaccine was demonstrated under field conditions. A total of 648 horses, including 32 pregnant mares, were enrolled in the study. During the two weeks post vaccination period, no local or systemic adverse reactions were observed following 96% of the vaccinations administered while mild, transient injection site reactions were noted in a small number of horses. These results indicate that the killed WNV vaccine developed by Fort Dodge Animal Health is safe and efficacious.

Experimental induction of chronic borreliosis in adult dogs exposed to Borrelia burgdorferi-infected ticks and treated with dexamethasone.

OBJECTIVE: To develop a method to experimentally induce Borrelia burgdorferi infection in young adult dogs. ANIMALS: 22 healthy Beagles. PROCEDURE: All dogs were verified to be free of borreliosis. Twenty 6-month-old dogs were exposed to Borrelia burgdorferi-infected adult ticks and treated with dexamethasone for 5 consecutive days. Two dogs not exposed to ticks were treated with dexamethasone and served as negative-control dogs. Clinical signs, results of microbial culture and polymerase chain reaction (PCR) testing, immunologic responses, and gross and histologic lesions were evaluated 9 months after tick exposure. RESULTS: Predominant clinical signs were episodic pyrexia and lameness in 12 of 20 dogs. Infection with B burgdorferi was detected in microbial cultures of skin biopsy specimens and various tissues obtained during necropsy in 19 of 20 dogs and in all 20 dogs by use of a PCR assay. All 20 exposed dogs seroconverted and developed chronic nonsuppurative arthritis. Three dogs also developed mild focal meningitis, 1 dog developed mild focal encephalitis, and 18 dogs developed perineuritis or rare neuritis. Control dogs were seronegative, had negative results for microbial culture and PCR testing, and did not develop lesions. CONCLUSIONS AND CLINICAL RELEVANCE: Use of this technique successfully induced borreliosis in young dogs. Dogs with experimentally induced borreliosis may be useful in evaluating vaccines, chemotherapeutic agents, and the pathogenesis of borreliosis-induced arthritis.

Construction and characterization of a glycoprotein E gene-deleted bovine herpesvirus type 1 recombinant.

OBJECTIVE: To construct and characterize a recombinant glycoprotein (g)E gene-deleted bovine herpesvirus (BHV) type 1 (BHV-1). PROCEDURE: The BHV-1 gEgene-coding region and the flanking upstream and downstream sequences were cloned. The aforementioned cloned DNA was digested with suitable enzymes to release the amino terminal two thirds of that region, and was ligated to the beta-galactosidase (beta-gal) gene. The resulting plasmid DNA was cotransfected with DNA from full-length, wild-type (WT), BHV-1 Cooper strain of the virus. Recombinant viruses expressing beta-gal (blue plaques) were plaque purified and assayed further by blot hybridization for genetic characterization and by immunoblotting for reactivity against BHV-1 gE peptide-specific rabbit polyclonal antibody. One recombinant virus, gEdelta3.1IBR, was characterized in vitro and in vivo. The ability of the recombinant virus to induce BHV-1 neutralizing antibodies in infected calves was investigated by plaque-reduction tests. RESULTS AND CONCLUSIONS: The gEdelta3.1IBR virus contained a deletion in the viral gE gene-coding sequences where a stable chimeric reporter (beta-gal) gene was inserted. One-step growth kinetics and virus yield of the recombinant and parent viruses were similar, but early after infection, the recombinant virus yield was comparatively less. After intranasal inoculation, the recombinant gEdelta3.1IBR virus replicated in the upper respiratory tract of calves, but the amount of progeny viruses produced was hundredsfold reduced, and duration of virus shedding was shorter. Results of in vivo calf experiments and serum neutralization tests indicated that deleting the gE gene has little effect on inducing neutralizing antibodies against BHV-1, but is sufficient to reduce BHV-1 virulence in calves.

Field study of the safety, immunogenicity, and efficacy of an inactivated equine rotavirus vaccine.

OBJECTIVE: To determine safety, immunogenicity, and efficacy of an inactivated equine rotavirus vaccine. DESIGN: Prospective randomized controlled trial. ANIMALS: 316 pregnant Thoroughbred mares during the first year of the study and 311 during the second year. PROCEDURE: During the first year, mares received 3 doses of vaccine or placebo, IM, at 8, 9, and 10 months of gestation. Serum neutralizing antibody titers were measured before vaccination and 1 and 35 days after foaling. Antibody titers were measured in foals 1, 7, 35, 60, 90, and 120 days after birth. During the second year, mares that had been vaccinated the previous year received a single booster dose of vaccine approximately 1 month prior to parturition. Mares that had received the placebo the previous year and mares new to the study received 3 doses of vaccine or placebo. Serum neutralizing antibody titers were measured in samples taken from mares approximately 1 day after foaling and from foals approximately 1 and 60 days after birth. RESULTS: Adverse reactions were not observed. Antibody titers were significantly increased at the time of foaling and 35 days after foaling in vaccinated, compared with control, mares and for 90 days after birth in foals born to vaccinated, compared with foals born to control, mares. Incidence of rotaviral diarrhea was lower in foals born to vaccinated, compared with foals born to control, mares, but the difference was not significant. CLINICAL IMPLICATIONS: Results suggest that the equine rotavirus vaccine is safe and immunogenic and that reasonable efficacy under field conditions can be expected.

Efficacy of an inactivated feline leukemia virus vaccine.

An inactivated whole-virus FeLV vaccine, developed from a molecularly cloned FeLV isolate (FeLV-61E-A), was assessed for its ability to protect cats against homologous and heterologous virulent viral challenge. The fractions of cats that resisted the induction of persistent viremia after FeLV challenge were as follows: FeLV-61E-A vaccine, 95%; adjuvant controls, 26%; and established commercial control FeLV vaccine, 35%. The prechallenge mean neutralizing antibody titers for each group were as follows: FeLV-61E-A vaccine, 1:43; adjuvant controls, < 1:8; and commercial control FeLV vaccine, 1:12. The prototype FeLV-61E-A vaccine was developed commercially for immunization of pet cats by substitution of a proprietary adjuvant and development of stable, high antigen production cell lines. This vaccine (Fel-O-Vax) has been studied extensively, alone and in multivalent combination with other feline virus vaccines, in seven efficacy trials involving a total of 150 immunized cats. These studies yielded an FeLV-resistant fraction of 87% in vaccinated cats as compared with 8% in adjuvant controls. The duration of immunity induced by an FeLV-61E-A commercial vaccine (Fel-O-Vax-LvK IV) was also assessed. One year after vaccination, 100% of challenged vaccinated cats and none of challenged controls resisted induction of persistent viremia. The results of these studies demonstrate that an inactivated FeLV vaccine prepared from a molecularly cloned subgroup A FeLV produces a high level of protective immunity against heterologous and homologous FeLV infection. This vaccine-induced immunity is durable for at least 1 year without intervening booster immunization or exposure to virus.

Development and testing of an inactivated feline leukemia virus vaccine.

We assessed an inactivated whole virus feline leukemia virus (FeLV) vaccine developed from a molecularly cloned feline leukemia virus isolate (FeLV-61E-A) for its ability to protect cats against homologous and heterologous virulent virus challenge. The fractions of cats that resisted the induction of persistent viremia after FeLV challenge were the following: (1) FeLV-61E-A vaccine, 95%; (2) adjuvant controls, 26%; and (3) established commercial control FeLV vaccine, 35%. The pre-challenge mean neutralizing antibody titers for each group were (1) FeLV-61E-A vaccine, 1:43; (2) adjuvant controls, <1:8; and (3) established commercial control FeLV vaccine: 1:12. The commercial version of the prototype FeLV-61E-A vaccine (Fel-O-Vax, Fort Dodge Laboratories, Fort Dodge, IA) was developed through use of a proprietary adjuvant and a stable high antigen production cell lines. The efficacy and duration of immunity produced by Fel-O-Vax was studied alone and in multivalent combination with other feline virus vaccines in seven subsequent efficacy trials conducted in over 150 immunized cats. The overall FeLV-resistant fraction in these trials was 87% in vaccinated cats versus 8% in adjuvant controls. The duration of protective immunity induced by the multivalent Fel-O-Vax-LvK IV at 1 year postvaccination was 100% in challenged vaccinees versus 0% in challenged controls. The results of these studies show that an inactivated FeLV vaccine prepared from a molecularly cloned subgroup A FeLV can produce high level protective immunity against FeLV infection. This immunity is durable for at least 1 year without intervening booster immunization or virus exposure.

Protection of cats from infectious peritonitis by vaccination with a recombinant raccoon poxvirus expressing the nucleocapsid gene of feline infectious peritonitis virus.

Feline Infectious Peritonitis Virus (FIPV) is a coronavirus that induces an often fatal, systemic infection in cats. Various vaccines designed to prevent FIPV infection have been shown to exacerbate the disease, probably due to immune enhancement mediated by virus-specific immunoglobulins against the outer envelope (S) protein. An effective vaccine would be one that induces cell-mediated immunity without disease enhancing antibodies. In this report, we describe the use of a recombinant raccoon poxvirus that expresses the gene encoding the nucleocapsid protein of FIPV (rRCNV-FIPV N) as an effective vaccine against FIPV-induced disease. Cats were parenterally or orally vaccinated twice, three weeks apart. Cats were then orally challenged with Feline Enteric Coronavirus (FECV), which induces a subclinical infection that can cause enhancement of subsequent FIPV infection. Three weeks later, cats were orally challenged with FIPV. The FIPV challenge induced a fatal infection in 4/5 (80%) of the controls. On the other hand, all five cats vaccinated subcutaneously with rRCNV-FIPV N showed no signs of disease after challenge with FIPV. Four of the five subcutaneous vaccinates survived an additional FIPV challenge. Vaccination with rRCNV-FIPV N induced serum IgG antibody responses to FIPV nucleocapsid protein, but few, if any, FIPV neutralizing antibodies. In contrast to the controls, protected vaccinates maintained low FIPV serum neutralizing antibody titers after FIPV challenge. This suggests that the protective immune response involves a mechanism other than humoral immunity consisting of FIPV neutralizing antibodies.

Efficacy of an inactivated vaccine against clinical disease caused by canine coronavirus.

Canine Coronavirus (CCV) is a causative agent of diarrhea in dogs. The reproduction of severe clinical disease with experimental CCV infection has been difficult. We have recently developed a CCV challenge model which reproduced clinical signs of disease in susceptible dogs. The following study was designed to determine whether immunization with an inactivated CCV vaccine would protect dogs from clinical disease induced using this model. Dogs (n = 13) were vaccinated with an inactivated CCV vaccine. Vaccinates and controls (n = 5) were orally inoculated with virulent CCV virus and treated with dexamethasone on days 0, 2, 4, and 6 after virus challenge. Control dogs developed clinical signs including diarrhea, dehydration, anorexia, depression, and nasal and ocular discharge. Diarrhea was noted in 80% of the controls and 60% progressed to a severe watery or bloody diarrhea that persisted for multiple days. Conversely, only 2/13 (15%) vaccinates developed mild diarrhea and none developed bloody diarrhea. The control dogs averaged 10.8 days of diarrhea compared to 1.4 days for vaccinates over the 21 day observation period. In addition to reduced clinical signs, the number of days of virus shedding and the level of CCV in feces was different for controls (100% shed virus) and vaccinates (38% shed virus). This study demonstrates that vaccination with an inactivated CCV vaccine can significantly reduce not only viral replication, but the occurrence of clinical disease following a virulent CCV infection.