ABSTRACT
Canine parvovirus (CPV2) is one of the most virulent virus causing acute hemorrhagic enteritis and myocarditis in dogs. Infection mainly caused by the ingestion of virus through the mucosal route. Therefore, induction of mucosal immunity is essential in prevention of Canine Parvovirus (CPV2) infection. For safe and effective delivery of viral antigens to the mucosal immune system, a novel surface antigen display system for lactic acid bacteria using the poly-gamma-glutamic acid synthetase A protein (pgsA) of Bacillus subtilis as an anchoring matrix was applied in order to display CPV2 antigen on the surface of the recombinant L. casei. Recombinant fusion proteins comprised of pgsA and the capsid protein (VP2-S1) showed stable expression in Lactobacillus casei. Surface localization of the fusion protein was verified by cellular fractionation analyses. Oral and nasal inoculations of recombinant L. casei into mice resulted in high levels of serum immunoglobulin G (IgG) and mucosal IgA, as demonstrated by ELISA using recombinant VP2-S1 proteins. Mice receiving intranasal immunization mounted higher antibody response than those receiving oral immunization. These results indicate that mucosal immunization with recombinant L. casei expressing CPV2 VP2-S1 protein on its surface provides an effective means for elicitation of strong antibody responses against CPV 2 VP2-S1.
Subject(s)
Animals , Dogs , Mice , Antibody Formation , Antigens, Surface , Antigens, Viral , Bacillus subtilis , Bacteria , Capsid Proteins , Capsid , Eating , Enteritis , Enzyme-Linked Immunosorbent Assay , Immune System , Immunity, Mucosal , Immunization , Immunoglobulin A , Immunoglobulin G , Lactic Acid , Lacticaseibacillus casei , Lactobacillus , Ligases , Myocarditis , Parvovirus, Canine , Proteins , Recombinant Fusion Proteins , VirusesABSTRACT
Calcium (Ca) is an essential element to maintain body homeostasis. However, many factors disturb calcium absorption. Aspartic acid chelated calcium (AAC) was synthesized by new methods using calcium carbonate and aspartic acid. This study was carried out to investigate the bioavailability of AAC in Ca-deficient rats. The experimental groups were as follows: NC; normal diet control group, CD-C; untreated control group of Ca-deficient (CD) rats, CD-CaCO3; CaCO3 treated group of CD rats, CD-AAC; AAC treated group of CD rats, and CD-SWC; and seaweed-derived Ca treated group of CD rats. The Ca content of various types of Ca was held constant at 32 mg/day, and the four CD groups were fed for 7 days after randomized grouping. Ca content in serum, urine, and feces within feeding periods were analyzed to confirm Ca absorption. Serum Ca content was significantly higher in the CD-AAC (11.24 mg/dL) and CD-SWC (10.12 mg/dL) groups than that in the CD-C (8.6 mg/dL) group 2 hours following the first administration. The Ca content in feces was significantly lower in the CD-AAC (35.4 mg/3 days) and CD-SWC (71.1 mg/3 day) groups than that in the CD-CaCO3 (98.7 mg/3 days) group (p > 0.05). AAC had a 2.3-fold higher absorption rate of Ca than that of SWC. No differences in fibula length were observed in the NC and CD groups. The fibula weights of the CD-AAC (0.33 g) and CD-SWC (0.33 g) groups increased compared to those in the CD-C (0.27 g) group; however, no significant difference was observed between the CD groups. We conclude that bioavailability of AAC is higher than that of seaweed-derived Ca or inorganic Ca. Thus, these findings suggest the AAC has potential as a functional food material related to Ca metabolism.
Subject(s)
Animals , Rats , Absorption , Adsorption , Aspartic Acid , Biological Availability , Calcium , Calcium Carbonate , Diet , Feces , Fibula , Functional Food , Homeostasis , Weights and MeasuresABSTRACT
The ORF5 gene encodes a major envelope glycoprotein (GP5), which is one of the three major proteins of porcine reproductive and respiratory syndrome virus (PRRSV). The GP5 protein has been known to be a 24.5-26kDa N-glycosylated envelope protein. The GP5 is involved in inducing neutralizing antibodies. For this reason, the GP5 is primary candidate for the PRRSV subunit vaccine. To produce the native form of GP5 in mammalian cells, we have cloned the ORF5 gene from PRRSV CNV-1 into the Semliki Forest virus (SFV)-based expression vector, resulting in recombinant pSFV-ORF5. By the infection with recombinant pSFV-ORF5 to BHK-21 cells, the GP5 expression was confirmed by immunocytochemistry and immunoblotting assay. The recombinant virus particle harboring ORF5 gene was infectious to BHK-21 and MARC-145. The RNA synthesis and expression of GP5 in the infected cell was also confirmed by RT-PCR.
Subject(s)
Animals , Base Sequence , DNA Primers , Genes, Viral , Plasmids/genetics , Porcine respiratory and reproductive syndrome virus/genetics , Restriction Mapping , Reverse Transcriptase Polymerase Chain Reaction , Semliki forest virus/genetics , Swine , Viral Envelope Proteins/genetics , Viral Proteins/genetics , Virology/methodsABSTRACT
Angiogenesis is recognized as a critical factor in the growth of tumor cells and plays a key role in the tumor metastasis. Recent studies for antiangiogenic substances are getting popular. The angiostatin, one of the antiangiogenic substances, leads to the increased apoptosis of the tumor cells by inhibiting the neovascularization of the tumor. The angiostatin was identified as the internal fragments of the plasminogen which has no antiangiogenic activity. By hydrolysis of the plasminogen, the angiostatin can be produced. In this study, we constructed the SFV-derived DNA vector by employing the cytomegalovirus immediate early enhancer/ promoter (CMV). This vector makes it possible to transfect the cells with DNA without the in vitro transcription process. The C-myc epitope and polyhistidine residue sequences were placed in downstream of the angiostatin gene to make it eligible to detect the expressed protein. The murine Ig kappa-chain V-J2-C signal sequence was placed in upstream to secrete the expressed protein from the cells. We confirmed the expression of angiostatin in the BHK-21 cells using DNA-based SFV replicon.
Subject(s)
Animals , Cricetinae , Angiostatins/analysis , Base Sequence , Cell Line , DNA Primers , Gene Expression Regulation, Viral , Immunohistochemistry , Kidney , Plasmids , Replicon/genetics , Semliki forest virus/genetics , TransfectionABSTRACT
During the period from January to December of 2001, a total of 3,391 swine sera were submitted to our laboratory from 256 farms for the diagnosis of porcine reproductive and respiratory syndrome (PRRS). The antibody to porcine reproductive and respiratory syndrome virus (PRRSV) was tested by the indirect immunofluorescent antibody (IFA) test. Of the 256 farms tested, 230 farms (89.8%) were positive for the PRRSV antibody. The overall seroprevalence of the PRRSV antibody was 52.1% (1765/3391). Most of the pigs seemed to be infected with PRRSV at around 50 to 60 days old. The seroprevalence of the antibody became higher with age, and peaked at around 100 days old. More than one-third of the adult pigs, including boars, gilts, and sows, was positive for the PRRSV antibody. The infection of PRRSV was chronic and confined to growers and/or finishers in most farms. However, the antibody was detected in all production phases at some farms.
Subject(s)
Animals , Female , Male , Age Factors , Antibodies, Viral/blood , Fluorescent Antibody Technique, Indirect/veterinary , Korea/epidemiology , Porcine Reproductive and Respiratory Syndrome/diagnosis , Porcine respiratory and reproductive syndrome virus/immunology , Seroepidemiologic Studies , Sex Factors , SwineABSTRACT
In Sprague Dawley (SD) rats, antibodies against strains of Orinentia tsutsugamushi, Kato, Karp and Gilliam, were produced in order to investigate their longevity and cross-reactivities to their corresponding homologous and heterologous antigens. By immunofluorescence assay (IFA) of IgG and IgM, it was shown that the immunity to the homologous strains persisted at a higher level (longevity of at least 34 weeks with higher IFA titers). On the other hand, the immunity to the heterologous strains persisted at a lower level (longevity of 10 to 34 weeks with lower IFA titers). Since infection with one strain of O. tsutsugamushi does not preclude reinfection with other strains, understanding of the antigenic diversity of O. tsutsugamushi and duration of the immunity to both homologous and heterologous strain is very important in diagnosis of scrub typhus.
Subject(s)
Animals , Rats , Antibodies , Antigenic Variation , Antigens, Heterophile , Diagnosis , Fluorescent Antibody Technique , Hand , Immunoglobulin G , Immunoglobulin M , Longevity , Orientia tsutsugamushi , Rats, Sprague-Dawley , Scrub TyphusABSTRACT
We evaluated Immunochromatographic assay kit to screen HBsAg in human serum. When the reference HBsAg was applyed to ICA, HA and EIA kits, the limit of detection for HBsAg were found out to be 4, 2 and 0.25 ng/ml respectively. But ICA kit required 5 minutes to read the result whereas HA and EIA kit more than one hour. The sensitivity was 97% (29 of 30 samples) and the specificity 100% (45 samples) compared with conventional EIA. The ICA kit needs no instrument or machine to perform the test contrary to the conventional methods. Therefore, this rapid and sensitive ICA kit can be used for HBsAg-screening, especially in the emergency room and in the scene of the accident.