CD40-CD40L-mediated help from CD4 T cells is essential to induce primary CD8 T cell responses specific to the non-inflammatory cell-based antigen H60. In this study, using H60 as a model antigen, we generated recombinant vaccinia viruses (rVVs) expressing the H60 CD8 epitope and investigated whether CD4 help was required to activate the CD8 T cell response specific to the virally expressed H60. The immune response after infection with rVVs expressing H60 was similar to that after immunization with H60 congenic splenocytes, with a peak frequency of H60-specific CD8 T cells detected in the blood on day 10 post-infection. A CD8 T cell response specific for virally derived H60 was not induced in CD4-depleted mice, but was in CD40-deficient mice. These results provide insights into the characterization of the CD8 T cell response specifically for antigens originating from cellular sources compared to viral sources.
Animals ; Immunization ; Mice ; T-Lymphocytes ; Vaccinia virus

BACKGROUND: We compared vaccinia virus shedding from the vaccine inoculation site (vaccination lesion) and two sites of a dressing covering the vaccination site; the outer surface of the semipermeable dressing (outer surface) and the inner surface of the semipermeable dressing, that is, the surface of a folded gauze under the semipermeable membrane (gauze surface) MATERIAL AND METHODS: Subjects were recruited from the volunteers who participated in a clinical trial of the efficacy of a 1:10 dilution of Lancy-Vaxina? (Berna Biotech, Switzerland), and were seen every 2-3 days (days 6, 8, 10, 13, and 15 after smallpox vaccination) for scheduled dressing changes. Swab specimens were obtained from the vaccination lesion, the outer surface, and the gauze surface. Quantitative viral culture assays for these specimens were done. RESULTS: Vaccinia virus was recovered from 126 (81%) of the 156 vaccination lesion samples collected from the 40 participants. A high virus titer was recovered from the vaccination lesion (geometric mean titer (log10)=3.91 on day 8). Of the 39 swab samples obtained from the gauze surface of the gauze, 16 (41%) were positive for virus. An intermediate titer was recovered from the gauze surface (geometric mean titer (log10)=0.91 on day 8). Of the 133 swab samples obtained from the outer surface, only one (0.8%) was positive for vaccinia. No virus was recovered from the outer surface on day 8. CONCLUSION: Our findings suggest that the addition of a semipermeable dressing to the folded gauze further reduces viral shedding and therefore increases protection.
Bandages* ; Membranes ; Smallpox* ; Vaccination* ; Vaccinia virus* ; Vaccinia* ; Viral Load ; Virus Shedding ; Volunteers

BACKGROUND: We compared vaccinia virus shedding from the vaccine inoculation site (vaccination lesion) and two sites of a dressing covering the vaccination site; the outer surface of the semipermeable dressing (outer surface) and the inner surface of the semipermeable dressing, that is, the surface of a folded gauze under the semipermeable membrane (gauze surface) MATERIAL AND METHODS: Subjects were recruited from the volunteers who participated in a clinical trial of the efficacy of a 1:10 dilution of Lancy-Vaxina? (Berna Biotech, Switzerland), and were seen every 2-3 days (days 6, 8, 10, 13, and 15 after smallpox vaccination) for scheduled dressing changes. Swab specimens were obtained from the vaccination lesion, the outer surface, and the gauze surface. Quantitative viral culture assays for these specimens were done. RESULTS: Vaccinia virus was recovered from 126 (81%) of the 156 vaccination lesion samples collected from the 40 participants. A high virus titer was recovered from the vaccination lesion (geometric mean titer (log10)=3.91 on day 8). Of the 39 swab samples obtained from the gauze surface of the gauze, 16 (41%) were positive for virus. An intermediate titer was recovered from the gauze surface (geometric mean titer (log10)=0.91 on day 8). Of the 133 swab samples obtained from the outer surface, only one (0.8%) was positive for vaccinia. No virus was recovered from the outer surface on day 8. CONCLUSION: Our findings suggest that the addition of a semipermeable dressing to the folded gauze further reduces viral shedding and therefore increases protection.
Bandages* ; Membranes ; Smallpox* ; Vaccination* ; Vaccinia virus* ; Vaccinia* ; Viral Load ; Virus Shedding ; Volunteers

PURPOSE: The purpose of this study was to evaluate the anti-tumoral effect of recombinant vaccinia virus (rVV) (Thymidine kinase (-)/GM-CSF (+)) that was administered as a US guided intratumoral injection in a rabbit model of hepatic VX2 carcinoma. MATERIALS AND METHODS: VX2 carcinoma was implanted in the livers of 12 rabbits. US was performed at every week interval to detect hepatic mass after the implantation of VX2 carcinoma. The accurate tumor size and volume was evaluated with CT when the tumor was detected on US. US guided injection of rVV (109 pfu/ml) was preformed in three rabbits, intravenous injection of the same dose of rVV was done in two rabbits and another seven rabbits that were without any treatment were selected as a control group. We evaluated the change of the hepatic tumor size and extrahepatic metastasis on serial CT. Tumor specimens were harvested from rabbits that were killed at 8 weeks after VX2 implantation. These tissues were histoimmuopathologically compared to each other (the virus injection group and the control group). The differences between these groups were statistically assessed with student t-tests. RESULTS: Tumor growth was significantly suppressed in the US guided injection group compared with the intravenous injection group or the control group (p< 0.01). The intravenous injection group showed statistically significant tumor suppression compared to the control group (p< 0.01) until 2 weeks after virus injection. Quantification of the pulmonary metastatic nodules was performed in view of both the number and volume. The average number or volume of the pulmonary metastatic nodules in the US injection group was much smaller than these in the control group. Histopathologically, the tumors of the US guided injection group showed less extensive necrosis than those of the control group. Immunohistochemically, the tumor of the US guided injection group showed more prominent infiltration of CD4 (+) and CD8 (+) lymphocytes than did the tumors of the other group. CONCLUSION: rVV was markedly effective in suppressing hepatic tumor growth and extrahepatic metastasis in a rabbit model of hepatic VX2 carcinoma. US guided intra-tumoral injection was more effective than systemic intravenous injection.
Humans ; Injections, Intravenous ; Liver ; Lymphocytes ; Necrosis ; Neoplasm Metastasis ; Phosphotransferases ; Rabbits ; Vaccinia virus* ; Vaccinia*

While presenting biological characteristics of vaccinia virus and laboratory-acquired infections during related research processes, this paper focuses on benefits and risks of vaccinia virus immunization in relation to laboratory-acquired infections, describes characteristics and the adaptation of vaccinia virus vaccine, analyses the role vaccinia virus immunization plays in the prevention and control of laboratory-acquired infections, and finally proposes solutions and countermeasures to further promote and implement immune control strategies. The problem related to immune strategy and laboratory- acquired infections which is being raised, analyzed and explored plays an active and instructive role in vaccinia virus related researches and laboratory- acquired infections, and also helps to recommend and develop relevant immune strategy for future vaccine control of such infections.
Contraindications ; Humans ; Smallpox Vaccine ; adverse effects ; Vaccination ; standards ; Vaccinia ; immunology ; prevention & control ; Vaccinia virus ; immunology

DNA vaccine approaches have been applied to generate the protective immunity against various pathogens. However, the strength of immune responses induced by DNA vaccine is weak compared with conventional vaccines. The primeboost vaccination using DNA vaccine and other viral vector has been suggested as one way to circumvent this limitation. In the present study, we used in vivo CTL activity assay to determine CD8+ T cell-mediated immunity induced by prime-boost vaccination with a DNA vaccine (gB498-505 DNA) and recombinant vaccinia virus (VVgB498-505) expressing gB498-505 epitope peptide (SSIEFARL) of herpes simplex virus type 1 (HSV-1) glycoprotein B (gB). The most potent in vivo CTL activity was induced in mice received VVgB498-505 when both gB498-505 and VVgB498-505 were used at priming step and boosted with the alternative vaccine vector expressing whole antigen protein (gBw). Priming with vaccine vector expressing gBw followed by the use of VVgB498-505 at boosting step also induced strong in vivo CTL activity. We also examined in vivo CTL activity after immunization of mice with epitope-expressing vaccine vector at both priming and boosting step. Curiously, in vivo CTL activity mediated by CD8+ T cells was strongly elicited at memory stage when animals were primed with VVgB498-505 and subsequently boosted with gB498-505 DNA. Because the use of VVgB498-505 at priming followed by boosting with gB498-505 DNA induced most optimal immunity, these results suggest that the order of vaccine type should be carefully considered when used vaccine type expressing only epitope for prime-boost vaccination.
Animals ; DNA* ; Glycoproteins ; Herpesvirus 1, Human ; Immunity, Cellular ; Immunization ; Memory ; Mice ; Simplexvirus ; T-Lymphocytes* ; Vaccination* ; Vaccines ; Vaccinia virus* ; Vaccinia*

BACKGROUND: DNA vaccination represents an anticipated approach for the control of numerous infectious diseases. Used alone, however, DNA vaccine is weak immunogen inferior to viral vectors. In recent, heterologous prime-boost vaccination leads DNA vaccines to practical reality. METHODS: We assessed prime-boost immunization strategies with a DNA vaccine (minigene, gB498-505 DNA) and recombinant vaccinia virus (vvgB498- 505) expressing epitope gB498-505 (SSIEFARL) of CD8+ T cells specific for glycoprotein B (gB) of herpes simplex virus (HSV). Animals were immunized primarily with gB498-505 epitope-expressing DNA vaccine/recombinant vaccinia virus and boosted with alternative vaccine type expressing entire Ag. RESULTS: In prime-boost protocols using vvgBw (recombinant vaccinia virus expressing entire Ag) and vvgB498-505, CD8+ T cell-mediated immunity was induced maximally at both acute and memory stages if primed with vvgBw and boosted with vvgB498-505 as evaluated by CTL activity, intracellular IFN-staining, and MHC class I tetramer staining. Similarly gB498-505 DNA prime-gBw DNA (DNA vaccine expressing entire Ag) boost immunization elicited the strongest CD8+ T cell responses in protocols based on DNA vaccine. However, the level of CD8+ T cell-mediated immunity induced with prime-boost vaccination using DNA vaccine expressing epitope or entire Ag was inferior to those based on vvgBw and vvgB498-505. Of particular interest CD8+ T cell-mediated immunity was optimally induced when vvgB498-505 was used to prime and gB DNA was used as alternative boost. Especially CD8+ T cell responses induced by such protocol was longer lasted than other protocols. CONCLUSION: These facts direct to search for the effective strategy to induce optimal CD8+ T cell-mediated immunity against cancer and viral infection.
Animals ; Communicable Diseases ; DNA* ; Glycoproteins ; Immunity, Cellular* ; Immunization ; Memory ; Simplexvirus ; T-Lymphocytes ; Vaccination* ; Vaccines, DNA ; Vaccinia virus* ; Vaccinia*

OBJECTIVETo construct the virus-like parcel expressing hepatitis B virus (HBV) C gene and identify its immunogenicity.

METHODSHBV C gene was cloned into the shuttle vector pSC11, and the resulted plasmid pSC11-C was transfected into modified vaccinia virus Ankara (MVA).

RESULTSpSC11-C was correctly constructed as verified by sequence analysis and PCR, and the recombinant virus-like parcel possessed good immunogenicity.

CONCLUSIONThe MVA-C expressing HBV C gene has been successfully constructed to provide important basis for gene therapy research of chronic HBV infection.

Genes, Viral ; Genetic Vectors ; Hepatitis B Core Antigens ; genetics ; Recombination, Genetic ; Vaccinia virus ; genetics

OBJECTIVEBy analyzing the status and characteristics of vaccinia virus laboratory-acquired infections in the bibliographical information, this paper provides relevant recommendations and measures for prevention and control of vaccinia virus laboratory-acquired infections in China.

METHODSChoosing PubMed, Embase, Biosis and SCIE, SSCI, CPCI-S as well as CPCI-SSH covered by Web of Science as the data source, indexing the bibliography of vaccinia virus laboratory-acquired infections, this paper analyzes the information on whether to vaccinate, the occurrence time of symptoms, diseasedparts, symptom characteristics and the disease-causing reasons.

RESULTSThe outcome shows that 52. 9% of the cases never get vaccinated, 82.4% engaged in vaccinia virus related researches never get vaccinated in 10 years, 52. 9% get infected by the accidental needlestick in hands during the process of handling animal experiments, 70. 6% of infections occur in the hands and having symptoms after being exposed with an average of 5. 1 days.

CONCLUSIONAlthough it is still controversial that whether or not to be vaccinated before carrying out vaccinia virus related works, it should be important aspects of prevention and control of vaccinia virus laboratory-acquired infections with the strict compliance with the operating requirements of the biosafety, by strengthening personal protection and timely taking emergency measures when unforeseen circumstances occur, as well as providing the research background information to doctors.

China ; Humans ; Laboratory Infection ; prevention & control ; transmission ; virology ; Needlestick Injuries ; virology ; Occupational Exposure ; adverse effects ; Vaccinia ; etiology ; prevention & control ; transmission ; virology ; Vaccinia virus

For rapid and accurate screening of recombinant modified vaccinia virus Ankara (rMVA) that satisfied the quality standards of clinical trials, a novel shuttle vector that can delete the marker gene automatically during virus propagation was construted: pZL-EGFP. To construct the pZL-EGFP, the original shuttle vector pSC11 was modified by replacing the LacZ marker gene with enhanced green fluorescent protein (EGFP) and then inserting homologous sequences of TKL into the flank regions of EGFP. Baby hamster kidney (BHK)-21 cells were cotransfected with pZL-EGFP and MVA, and underwent ten passages and one plaque screening to obtain the EGFP-free rMVA carrying the exogenous gene. Resulting rMVA was tested by polymerase chain reaction and western blotting to verify pZL-EGFP function. A novel shuttle vector pZL-EGFP containing an EGFP marker gene which could be deleted automatically was constructed. This gene deletion had no effect on the activities of rMVA, and the exogenous gene could be expressed stably. These results suggest that rMVA can be packaged efficiently by homologous recombination between pZL-EGFP and MVA in BHK-21 cells, and that the carried EGFP gene can be removed automatically by intramolecular homologous recombination during virus passage. Meanwhile, the gene deletion had no influence on the activities of rMVA and the expression of exogenous target gene. This study lays a solid foundation for the future research.
Animals ; Biomarkers ; Cricetinae ; Epithelial Cells ; virology ; Gene Deletion ; Genetic Engineering ; methods ; Genetic Vectors ; genetics ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Recombination, Genetic ; Vaccinia ; virology ; Vaccinia virus ; genetics ; physiology ; Virus Replication