scholarly journals A Single Intramuscular Injection of Recombinant Plasmid DNA Induces Protective Immunity and Prevents Japanese Encephalitis in Mice

2000 ◽  
Vol 74 (9) ◽  
pp. 4244-4252 ◽  
Author(s):  
Gwong-Jen J. Chang ◽  
Ann R. Hunt ◽  
Brent Davis
Keyword(s):  
Dna Vaccine ◽  
Japanese Encephalitis ◽  
Plasmid Dna ◽  
Recombinant Plasmid ◽  
Culture Media ◽  
Early Gene ◽  
Effective Vaccine ◽  
Enzyme Linked Immunosorbent Assay ◽  
Old Mice

ABSTRACT Plasmid vectors containing Japanese encephalitis virus (JEV) premembrane (prM) and envelope (E) genes were constructed that expressed prM and E proteins under the control of a cytomegalovirus immediate-early gene promoter. COS-1 cells transformed with this plasmid vector (JE-4B clone) secreted JEV-specific extracellular particles (EPs) into the culture media. Groups of outbred ICR mice were given one or two doses of recombinant plasmid DNA or two doses of the commercial vaccine JEVAX. All mice that received one or two doses of DNA vaccine maintained JEV-specific antibodies 18 months after initial immunization. JEVAX induced 100% seroconversion in 3-week-old mice; however, none of the 3-day-old mice had enzyme-linked immunosorbent assay titers higher than 1:400. Female mice immunized with this DNA vaccine developed plaque reduction neutralization antibody titers of between 1:20 and 1:160 and provided 45 to 100% passive protection to their progeny following intraperitoneal challenge with 5,000 PFU of virulent JEV strain SA14. Seven-week-old adult mice that had received a single dose of JEV DNA vaccine when 3 days of age were completely protected from a 50,000-PFU JEV intraperitoneal challenge. These results demonstrate that a recombinant plasmid DNA which produced JEV EPs in vitro is an effective vaccine.

Assay of Gcdfp-15 by Elisa: An Available Method for in Vitro Studies of Functional Differentiation in Human Breast Cancer

1988 ◽  
Vol 74 (6) ◽  
pp. 669-674
Author(s):  
Françoise Revillion-Carette ◽  
Louis Hornez ◽  
Brigitte Vandewalle ◽  
Jean Lefebvre
Keyword(s):  
Breast Cancer ◽  
Human Breast Cancer ◽  
Culture Media ◽  
Functional Differentiation ◽  
In Vitro Studies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Breast ◽  
Breast Cyst Fluid ◽  
Breast Cyst

An enzyme-linked immunosorbent assay (ELISA) was applied to a light protein, isolated from human breast cyst fluid (BCF) termed « gross cystic disease fluid protein - 15 Kda » (GCDFP-15), a potential differentiation marker in in vitro human breast cancer studies. The detection limits of this procedure, performed in microtiter plates, were 0.5 to 250 ng/well corresponding to 10 ng/ml to 5 ng/ml of sample or antigen solution. Possible cross-reaction with various antigens, especially those found in culture media, were investigated. The correlation coefficient between enzymoassay and radioimmunoassay was 0.978. The results showed that quantification of GCDFP-15 by ELISA is a specific and highly sensitive method. This procedure may be of interest in in vitro studies on the functional differentiation of breast cancer cells.

Vaxfectin® enhances both antibody and in vitro T cell responses to each component of a 5-gene Plasmodium falciparum plasmid DNA vaccine mixture administered at low doses

Vaccine ◽  
2010 ◽  
Vol 28 (17) ◽  
pp. 3055-3065 ◽  
Author(s):  
Martha Sedegah ◽  
William O. Rogers ◽  
Maria Belmonte ◽  
Arnel Belmonte ◽  
Glenna Banania ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
T Cell ◽  
Dna Vaccine ◽  
Plasmid Dna ◽  
T Cell Responses ◽  
Low Doses ◽  
Cell Responses ◽  
Plasmid Dna Vaccine

Recombinant Plasmid DNA Construct Encoding Combination of vegf165 and bmp2 cDNAs Stimulates Osteogenesis and Angiogenesis In Vitro

2016 ◽  
Vol 7 (2) ◽  
pp. 288-293 ◽  
Author(s):  
M. N. Zhuravleva ◽  
M. R. Khaliullin ◽  
R. F. Masgutov ◽  
R. V. Deev ◽  
A. A. Rizvanov
Keyword(s):  
Plasmid Dna ◽  
Recombinant Plasmid

scholarly journals EFFECT OF IN VITRO CULTIVATION ON THE PATHOGENICITY OF WEST NILE VIRUS

1946 ◽  
Vol 84 (2) ◽  
pp. 181-190 ◽  
Author(s):  
Hilary Koprowski ◽  
Edwin H. Lennette
Keyword(s):  
West Nile Virus ◽  
Culture Media ◽  
Lethal Effect ◽  
In Vitro Cultivation ◽  
Suspended Cell ◽  
West Nile ◽  
Cell Culture Media ◽  
Full Capacity ◽  
Old Mice

The West Nile virus was cultivated in suspended cell culture media employing several different tissue components, and it has been observed to survive in culture for at least 32 days. Continued propagation of the virus in vitro resulted in a change in its pathogenicity. The change lay in a marked reduction or a complete loss of the ability of the virus to produce fatal infections in mice and in hamsters on peripheral inoculation, although there was no obvious simultaneous alteration in the lethal effect of the virus by the cerebral route. In mice, the extent to which invasiveness was lost depended upon the passage level of the virus and the age of the test animals. The younger (and more susceptible) the mice, the greater the number of passages which was required to diminish the virulence of the virus by peripheral routes; after 68 passages, the virus still retained its full capacity to kill 3-day-old mice, while its ability to kill 8-day-old mice was reduced and its ability to kill mice 14 or more days of age was essentially abolished. How soon the loss of pathogenicity occurs in hamsters has not been determined. Prolonged cultivation rendered the virus avirulent for hamsters by the intraperitoneal route.

scholarly journals Comparison of Three Antigenic Extracts of Eurotium amstelodami in Serological Diagnosis of Farmer's Lung Disease

2009 ◽  
Vol 17 (1) ◽  
pp. 160-167 ◽  
Author(s):  
Sandrine Roussel ◽  
Gabriel Reboux ◽  
Bénédicte Rognon ◽  
Michel Monod ◽  
Frédéric Grenouillet ◽  
...  
Keyword(s):  
Lung Disease ◽  
Culture Media ◽  
Area Under The Curve ◽  
Serological Diagnosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Characteristic Analysis ◽  
Farmer’S Lung ◽  
Farmer's Lung ◽  
Fungal Particles

ABSTRACT In France and Finland, farmer's lung disease (FLD), a hypersensitivity pneumonitis common in agricultural areas, is mainly caused by Eurotium species. The presence of antibodies in patients' serum is an important criterion for diagnosis. Our study aimed to improve the serological diagnosis of FLD by using common fungal particles that pollute the farm environment as antigens. Fungal particles of the Eurotium species were observed in handled hay. A strain of Eurotium amstelodami was grown in vitro using selected culture media; and antigen extracts from sexual (ascospores), asexual (conidia), and vegetative (hyphae) forms were made. Antigens were tested by enzyme-linked immunosorbent assay (ELISA), which was used to test for immunoglobulin G antibodies from the sera of 17 FLD patients, 40 healthy exposed farmers, and 20 nonexposed controls. The antigens were compared by receiver operating characteristic analysis, and a threshold was then established. The ascospores contained in asci enclosed within cleistothecia were present in 38% of the hay blades observed; conidial heads of aspergillus were less prevalent. The same protocol was followed to make the three antigen extracts. A comparison of the results for FLD patients and exposed controls showed the area under the curve to be 0.850 for the ascospore antigen, 0.731 for the conidia, and 0.690 for the hyphae. The cutoffs that we determined, with the standard deviation for measures being taken into account, showed 67% for sensitivity and 92% for specificity with the ascospore antigen. In conclusion, the serological diagnosis of FLD by ELISA was improved by the adjunction of ascospore antigen.

scholarly journals Prophylactic potential of cytolethal distending toxin B (CdtB) subunit of typhoid toxin against Typhoid fever

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Reena Thakur ◽  
Preeti Pathania ◽  
Navneet Kaur ◽  
Vattan Joshi ◽  
Kanthi Kiran Kondepudi ◽  
...  
Keyword(s):  
Typhoid Fever ◽  
Bacterial Load ◽  
Lethal Dose ◽  
Active Immunization ◽  
Effective Vaccine ◽  
Enzyme Linked Immunosorbent Assay ◽  
Morphological Alterations ◽  
Log Reduction ◽  
Typhoid Toxin

AbstractTyphoid fever caused by Salmonella enterica serovar Typhi (S.Typhi) continues to be a major problem, especially in developing countries. Due to the rapid emergence of multi-drug-resistant (MDR) strains, which limits the efficacy of conventional antibiotics as well as problems associated with the existing vaccines, efforts are being made to develop effective prophylactic agents. CdtB subunit of typhoid toxin was selected for assessing its vaccine potential due to its high conservation throughout the Typhi strains. In-vitro assessment of DNase activity of cloned and purified CdtB protein showed a significant decrease in the band intensity of DNA. The measure of metabolic activity and morphological alterations assessed using different cell lines in the presence of CdtB protein showed no significant signs of toxicity. These observations were further strengthened by cell cycle analysis, assessed by flow cytometry. Keeping these observations in mind, the immunoprotective potential of CdtB was assessed using S.Typhi induced mouse peritonitis model. A significant titer of IgG antibodies (>128000) against CdtB protein was recorded in the immunized mice by enzyme-linked immunosorbent assay (ELISA), which was also validated by immunoblotting. Active immunization with the protein protected 75% mice against a lethal dose of S.Typhi Ty2. The data indicated a significant (up to 5 log) reduction in the bacterial load in the spleen and liver of immunized-infected mice compared to control (unimmunized-infected) mice which might have resulted in the modulation of histoarchitecture of spleen and liver and the levels of cytokines (IL-6, TNF-α and IL-10) production; thereby indicating the effectiveness of the subunit. The observations deduced from the study give the proof of concept of immunogenic potential of protein. However, further studies involving the immunoreactivity of CdtB with the statistically significant number of sera samples obtained from the human patients would be helpful in establishing the relevance of CdtB protein in humans and for making the strategies to develop it as an effective vaccine candidate.

The Fraction of the Snake Venom, its Leishmanicidal Effect, and the Stim-ulation of an Anti-Leishmania Response in Infected Macrophages

2020 ◽  
Vol 20 ◽  
Author(s):  
Saeideh Nikpour ◽  
Fatemeh Tabatabaie ◽  
Iraj Sharifi ◽  
Mahshid Mostafavi ◽  
Razieh Tavakoli Oliaee ◽  
...  
Keyword(s):  
Snake Venom ◽  
High Performance ◽  
Inhibitory Effect ◽  
Control Measures ◽  
Effective Vaccine ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Th2 Response ◽  
Wide Range

Background and aims:: Due to the lack of an effective vaccine and complexity of the control measures against vectors and reservoir hosts, the control of leishmaniasis depends primarily on chemotherapy. This study was aimed to assess the snake venom, Naja naja oxiana fraction 11(NNOV-F11) on L. infantum and its broad mode of action. Methods:: A wide range of in vitro advanced assays including high-performance liquid chromatography (HPLC), MTT (3-[4, 5-Dimethylthiazol-2-yl]-2, 5diphenyltetrazolium bromide; Thiazolyl blue) and macrophage assays, quantitative real-time polymerase chain reaction (qPCR), flow cytometry and enzyme-linked immunosorbent assay (ELISA) on L. infantum pro-mastigote and amastigote stages were used. IC50 values of L. infantum stages, CC50 value and apoptosis were also analyzed. Results:: The NNOV-F11 demonstrated strong antileishmanial activity against L. infantum stages in a dose-dependent man-ner compared to the untreated control group. Interleukin (IL)-12, TNF-α and iNOS genes expression as the indicators of T helper(h)1 response significantly increased; in contrast, the expression level of IL-10, as the representative of Th2 response significantly decreased (p ˂ 0.001). Reactive oxygen species (ROS) detection showed statistically a significant increase (p ˂ 0.001) after treatment with different concentrations of NNOV-F11, unlike arginase (ARG) activity which displayed a signif-icant reduction (p < 0.001). Conclusion:: NNOV-F11 possesses a potent inhibitory effect on L. infantum stages with the multifunctional and broad mode of actions which promoted the immunomodulatory role, induced ROS production, stimulated apoptotic–like mechanisms and inhibited L-ARG activity which collectively led to the parasite death. Further studies are crucial to assess the effect of the fraction NNOV-F11 on animal models or clinical settings.

scholarly journals Salmonella enterica Serovar Typhimurium Infection of Dendritic Cells Leads to Functionally Increased Expression of the Macrophage-Derived Chemokine

2005 ◽  
Vol 73 (3) ◽  
pp. 1714-1722 ◽  
Author(s):  
Guo Fu ◽  
Odilia L. C. Wijburg ◽  
Paul U. Cameron ◽  
Jason D. Price ◽  
Richard A Strugnell
Keyword(s):  
Dendritic Cells ◽  
T Cell ◽  
Salmonella Enterica ◽  
Fluorescent Protein ◽  
Culture Media ◽  
Expression Patterns ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serovar Typhimurium

ABSTRACT Gene expression in murine dendritic cells (DCs) infected with green fluorescent protein-expressing Salmonella enterica serovar Typhimurium BRD509 was studied by mRNA differential display. Infected DCs were sorted from uninfected cells by flow cytometry. The mRNA expression patterns of infected and uninfected cells revealed a number of differentially expressed transcripts, which included the macrophage-derived chemokine (MDC). Up-regulation of MDC transcription in infected DCs was confirmed by Northern blotting, and the kinetics of MDC expression was examined by real-time reverse transcription-PCR, with which 31- and 150-fold increases were detected at 2 and 6 h postinfection, respectively. The increased release by DCs of MDC into culture media was detected by an enzyme-linked immunosorbent assay. The biological activity of MDC was investigated in in vitro and in vivo assays. In vitro, supernatants from S. enterica serovar Typhimurium-infected DCs were chemoattractive to T cells, and neutralization of MDC in these supernatants inhibited T-cell migration. Passive transfer of anti-MDC antibody to mice infected with BRD509 revealed that neither growth of the bacterium nor resistance of the mice to reinfection was affected and that in vivo inhibition of MDC did not affect T-cell responses, as measured by the gamma interferon ELISPOT method 3 days after challenge infection.

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