This experiment was performed to determine whether cultured rabbit corneal cells were infected with HSV-1. The direct immunofluorescent method was used to detect the presence of the herpes simplex antigen in HSV-1 infected corneal epithelial cells, keratocytes, and endothelial cells in vitro with fluorescein-labeled antiserum to HSV-1. Immunofluorescent localization of the HSV-1 antigen was expressed in all three types of corneal cells.
Animals ; Antigens, Viral/*analysis ; Cells, Cultured ; Cornea/*microbiology ; Herpesviridae Infections/*microbiology ; Rabbits ; Simplexvirus/*immunology

OBJECTIVETo explore the etiology and clinical characters of hepatitis caused by non-hepatotropic virus.

METHODS68 non-hepatotrophic viral hepatitis patients with negative anti-HAV-anti-HEV were diagnosed by detecting antibodies of anti-HSV IgM, anti-EBV IgM, anti-CMV IgM, anti-CSV IgM and anti-ANA, anti-mitochondrion antibody. Their clinical symptoms and signs were compared with that of acute viral hepatitis patients at the same time.

RESULTSAmong the 68 patients, 9 were infected by HSV, 12 by EBV, 8 by CMV, 14 by CSV, and the other 13 patients and 12 patients were positive for anti-ANA and anti-mitochondrion antibody, respectively. 35 of 43 non-hepatotrophic viral hepatitis patients were infected in winter and spring season. Their clinical symptoms and signs were milder than that of acute viral hepatitis patients.

CONCLUSIONLiver damage and dysfunction may be the prominent phenomenon during HSV, EBV, CMV and CSV infection, just like that of acute viral hepatitis but with milder clinical symptom and signs.

Adult ; Antibodies, Viral ; blood ; Cytomegalovirus ; immunology ; Female ; Hepatitis, Viral, Human ; etiology ; virology ; Herpesvirus 4, Human ; immunology ; Humans ; Immunoglobulin M ; blood ; Male ; Simplexvirus ; immunology

BACKGROUND: The dramatic increase in use of the IgG test for toxoplasma, rubella, cytomegalovirus (CMV), and herpes simplex virus (HSV) [TORCH] has led to the requirement for a high-efficiency method that can be used in the clinical laboratory. This study aimed to compare the results of BGI-Array ELISA TORCH IgG (BGI-GBI, China) screening method to those of Virion/Serion TORCH IgG ELISA (Virion/Serion, Germany). METHODS: Serum specimens (n=400) submitted for routine IgG testing by Virion/Serion ELISA were also tested using the BGI-Array ELISA method. The agreements of these two kinds of method were analyzed by kappa-coefficients calculation. RESULTS: Following repeat testing, the BGI-Array ELISA TORCH IgG assays demonstrated agreements of 99.5% (398/400 specimens), 98% (392/400 specimens), 99% (396/400 specimens), and 99.5% (398/400 specimens), respectively. The BGI-Array ELISA IgG assays provided results comparable to Virion/Serion ELISA results, with kappa-coefficients showing near-perfect agreement for the HSV (kappa=0.87), rubella (kappa=0.92) and CMV (kappa=0.93) and substantial agreement for the toxoplasma (kappa=0.80) IgG assays. The use of the BGI-Array ELISA TORCH IgG assays could reduce the turnaround time (1.5 hr vs. 5 hr by Virion/Serion ELISA for 100 specimens) and were easy to use. CONCLUSIONS: BGI-Array ELISA TORCH IgG shows a good agreement with Virion/Serion ELISA methods and is suitable for clinical application.
Antibodies, Viral/blood ; Cytomegalovirus/immunology/*metabolism ; *Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin G/*analysis/blood ; Protozoan Infections/diagnosis ; Reagent Kits, Diagnostic ; Rubella virus/immunology/*metabolism ; Sensitivity and Specificity ; Simplexvirus/immunology/*metabolism ; Toxoplasma/immunology/*metabolism ; Virion/*immunology/metabolism ; Virus Diseases/diagnosis

Cholera toxin, which has been frequently used as mucosal adjuvant, leads to an irreversible activation of adenylyl cyclase, thereby accumulating cAMP in target cells. Here, it was assumed that beta2-adrenergic agonist salbutamol may have modulatory functions of immunity induced by DNA vaccine, since beta2-adrenergic agonists induce a temporary cAMP accumulation. To test this assumption, the present study evaluated the modulatory functions of salbutamol co-administered with DNA vaccine expressing gB of herpes simplex virus (HSV) via intranasal (i.n.) route. We found that the i.n. co-administration of salbutamol enhanced gB-specific IgG and IgA responses in both systemic and mucosal tissues, but optimal dosages of co-administered salbutamol were required to induce maximal immune responses. Moreover, the mucosal co-delivery of salbutamol with HSV DNA vaccine induced Th2-biased immunity against HSV antigen, as evidenced by IgG isotypes and Th1/Th2-type cytokine production. The enhanced immune responses caused by co-administration of salbutamol provided effective and rapid responses to HSV mucosal challenge, thereby conferring prolonged survival and reduced inflammation against viral infection. Therefore, these results suggest that salbutamol may be an attractive adjuvant for mucosal genetic transfer of DNA vaccine.
Adjuvants, Immunologic/*pharmacology ; Adrenergic beta-Agonists/immunology/*pharmacology ; Albuterol/immunology/*pharmacology ; Animals ; Antibodies, Viral/immunology ; Cercopithecus aethiops ; Cytokines/immunology ; Dose-Response Relationship, Drug ; Dose-Response Relationship, Immunologic ; Herpes Simplex/immunology/*prevention & control ; Herpes Simplex Virus Vaccines ; Immunity, Mucosal/*drug effects/immunology ; Immunoglobulin A/immunology ; Immunoglobulin G/immunology ; Mice ; Simplexvirus/*immunology ; Th1 Cells/immunology ; Th2 Cells/immunology ; Vaccines, DNA/*immunology/pharmacology ; Vero Cells ; Viral Envelope Proteins/immunology

Cytokines ; secretion ; Cytotoxicity, Immunologic ; Graft vs Leukemia Effect ; Humans ; Immunophenotyping ; Lymphocyte Activation ; Retroviridae ; genetics ; Simplexvirus ; enzymology ; T-Lymphocytes ; immunology ; Thymidine Kinase ; genetics ; Transfection

OBJECTIVETo study the effection of suppression murine melanoma growth by Intratumor injection of recombinant attenuated salmonella carrying heat shock protein 70 and herpes simplex virus thymidine kinase genes.

METHODSPlasmids PCMV-mtHSP70-IRES-TK were electro-transferred into salmonella typhimurium SL7207 to construct recombinant salmonella typhimurium. In vivo, Recombinant bacteria were injected into the mouse melanoma and the antitumor effection was observed. The survival period was recorded and safety analysis for this vaccine in each group.

RESULTSIn vivo, the mtHSP70/HSV-tk recombinant bacteria can suppress tumor growth significantly and extend survival. After recombinant Salmonella, 10(9) CFU/mL, was administered as an intratumoral injection, No diarrhea were observed. During therapy, body weight did not change markedly.

CONCLUSIONResults of the animal experiment suggests intratumor injection of recombinant attenuated salmonella typhimurium containing mtHSP70 and HSV-tk genes, has targeting ability against B16 tumor cell and could significantly inhibit tumor growth .

Animals ; Bacterial Proteins ; genetics ; immunology ; Cancer Vaccines ; genetics ; immunology ; pharmacology ; Genetic Therapy ; methods ; HSP70 Heat-Shock Proteins ; genetics ; immunology ; Melanoma, Experimental ; microbiology ; pathology ; therapy ; Mice ; Mice, Inbred C57BL ; Mycobacterium tuberculosis ; genetics ; Salmonella typhimurium ; genetics ; immunology ; Simplexvirus ; enzymology ; genetics ; Skin Neoplasms ; therapy ; Thymidine Kinase ; genetics ; immunology ; Vaccines, Attenuated ; genetics ; immunology ; pharmacology ; Vaccines, DNA ; genetics ; immunology ; pharmacology

OBJECTIVEThis study was designed to investigate the value of serologic examination in the diagnosis of childhood herpes simplex virus (HSV) infection.

METHODSSerum samples were collected from 2 436 outpatients and inpatients. The samples were divided into two groups. Group 1 consisted of 321 samples which were assayed for HSV-1 IgG, HSV-1 IgM, HSV-2 IgG or HSV-2 IgM antibody using herpes simplex virus antibody kits between January 2003 and November 2005. Group 2 consisted of 2115 samples which were assayed for HSV-2 IgG and IgM antibodies by TORCH testing between October 2004 and November 2005.

RESULTSIn Group 1, the total seroprevalence of HSV infection was 44.6%, with 38.9% being HSV-1 positive and 15.9% HSV-2 positive; HSV-IgM positivity was found in 41.1% and 25.5% were HSV-IgG positive; HSV-1 seroprevalence significantly increased with age (P < 0.05). In Group 2 the seroprevalence of HSV-2 infection was 1.9%; all of the samples were HSV-2 IgG positive.

CONCLUSIONSHSV serologic examination is useful in the diagnosis of HSV infection in children.

Adolescent ; Antibodies, Viral ; blood ; Child ; Child, Preschool ; Female ; Herpes Simplex ; diagnosis ; immunology ; Humans ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Infant ; Infant, Newborn ; Male ; Retrospective Studies ; Serologic Tests ; Simplexvirus ; immunology

OBJECTIVETo study the effect of lentiviral vector mediated herpes simplex virus-thymidine kinase/ganciclovir (HSV-TK/GCV) on graft- versus-host disease (GVHD) after allogeneic bone marrow transplantation (allo- BMT) in mice.

METHODSDonor splenic lymphocytes from C57BL/6 which were infected by lentiviral vectors carrying HSV-TK were transplanted into 60Co gamma ray irradiated recipient mice with donor bone marrow cells. GCV 25 mg x kg(-1) x d(-1) was administered in 3 groups on day 0, +7, +12 respectively after transplant for 7 days by intraperitoneal injection. Survival time, severity of GVHD, incidence of GVHD, T lymphocytes immune reconstruction and of allogeneic chimerism ratio were detected after allo-BMT.

RESULTSThe average survival times for GCV 0 day, +7 day and +12 day group were (30. 10 +/- 5.21) d, (36.40 +/- 5.28) d and (28.20 +/- 4.82) d respectively, being significantly longer than that in the control group [(15.10 +/- 0.43) d] (P < 0.05). The 50 d-survival rate for TK/GCV + 7 day group was 60%. While for 0 day and +12 day group was 40% and 30% respectively. The incidence of grade III approximately IV GVHD in the control group was 100%, and the dead mice in experimental groups showed pathological changes of II approximately III GVHD. Long-term alive recipient mice only developed grade I approximately II GVHD after allo-BMT. The number of CD4+ lymphocytes in experimental groups was higher than that in control group (P <0.05), but CD8+ lymphocytes was lower on day +5, +10, +15 day (P <0.05). Allogeneic chimerism rate of recipient mice on +30 d was 100%.

CONCLUSIONSHSV-TK/GCV induced by the lentiviral vectors has a definite effect in prevention of GVHD after allo-BMT. GCV administrated from 7 days post-transplantation showed the best effects.

Animals ; Bone Marrow Transplantation ; immunology ; Ganciclovir ; pharmacology ; Genetic Vectors ; Graft vs Host Disease ; prevention & control ; Lentivirus ; genetics ; Lymphocyte Transfusion ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Simplexvirus ; enzymology ; Thymidine Kinase ; genetics ; Transfection ; Transplantation, Homologous

OBJECTIVETo construct the localization system involving anti-TfR monoclonal antibody (McAb) and AFP promoters and assess its effect on human hepatoma cell lines.

METHODSThe conjugate of anti-TfR McAb and polylysine (PLL) was made by SPDP and purified by molecular screen chromatography. DNA blocking test determined that the ratio of one pEBAF/tk to six Ab-PLL was the most suitable to couple them. The pEBAF/tk recombinant plasmid bearing HSV-TK gene was coupled to Ab-PLL by noncovalent bond. The pEBAF/tk was transferred into human hepatoma cell line HepG2, SMMC7721 and pulmonary cancer cell line A549 by receptor-mediated gene delivery (Ab-PLL-DNA) and liposome procedure. The growth inhibitory rates of HepG2, SMMC7721 and A549 cells were measured by MTT assay.

RESULTSThe inhibitory rates of HepG2/tk in 100 mg/L and 1 mg/L of GCV were 60.5% and 24.3%, respectively. The inhibitory rate of GCV to SMMC7721 was 23.2% in 3 days. The pulmonary cancer cell A549, A549/tk (Ab) and A549 /tk (lipo) could not be inhibited by the addition of GCV.

CONCLUSIONThe localization system employed in this paper has high specificity, effectiveness and safety for gene therapy. It would be a promising strategy for gene therapy.

Antibodies, Monoclonal ; therapeutic use ; Carcinoma, Hepatocellular ; therapy ; Cell Line, Tumor ; Ganciclovir ; therapeutic use ; Genetic Therapy ; Humans ; Liver Neoplasms ; therapy ; Receptors, Transferrin ; immunology ; Simplexvirus ; enzymology ; Thymidine Kinase ; genetics ; alpha-Fetoproteins ; genetics