scholarly journals Strong humoral response elicited by a DNA vaccine targeting gastrin-releasing peptide with optimized adjuvants inhibits murine prostate carcinoma growth in vivo

2009 ◽  
Vol 16 (4) ◽  
pp. 1171-1184 ◽  
Author(s):  
Yong Lu ◽  
Didier J L Mekoo ◽  
Kedong Ouyang ◽  
Xiangbing Hu ◽  
Yanhua Liu ◽  
...  
Keyword(s):  
Dna Vaccine ◽  
Cell Epitope ◽  
Humoral Response ◽  
Plasmid Vector ◽  
Antitumor Effects ◽  
Gastrin Releasing Peptide ◽  
Humoral Immune ◽  
Mycobacterial Hsp70 ◽  
Linear Alignment

Previous studies demonstrated that the elevated expression and receptor binding of gastrin-releasing peptide (GRP) in various types of cancer suggest that GRP might be a putative target for immunotherapy in neoplastic diseases. DNA vaccine for hormone/growth factor immune deprivation represents a feasible and attractive approach for cancer treatment; nevertheless, there is still a need to increase the potency of the DNA vaccine. Here, based on six copies of the B cell epitope GRP18–27 in a linear alignment as an immunogen, we designed several anti-GRP DNA vaccines containing different combinations of immunoadjuvants, such as HSP65, tetanus toxoid830–844 (T), pan HLA-DR-binding epitope (PADRE) (P), and mycobacterial HSP70407–426 (M), on a backbone of pCR3.1 plasmid vector with eight 5′-GACGTT-3′ CpG motifs and the VEGF183 signal peptide (VS). The effects of these immunoadjuvants in enhancing GRP-specific humoral immune response were then evaluated by comparing the respective immunogenicity and antitumor effects. Immunization of mice with pCR3.1-VS-HSP65-TP-GRP6-M2 elicited much higher levels of specific anti-GRP antibodies and more effectively inhibited the growth of a GRP-dependent tumor RM-1 in vivo. Interestingly, plasmids encoding for 2HSP70407–426, but not the one with 1 or 3HSP70407–426 showed stronger immune stimulatory potential as well as impressive antitumor activity, suggesting that 2HSP70407–426 is an efficient molecular adjuvant for developing self-epitope vaccines. The highly immunogenic, potent anti-tumorigenic and antiangiogenesis activities of the anti-GRP DNA vaccine offered a novel immunotherapeutic approach in the treatment of GRP-dependent tumors and their complications.

scholarly journals Specific Antibodies Elicited by a Novel DNA Vaccine Targeting Gastrin-Releasing Peptide Inhibit Murine Melanoma Growth In Vivo

2009 ◽  
Vol 16 (7) ◽  
pp. 1033-1039 ◽  
Author(s):  
Jing Fang ◽  
Yong Lu ◽  
Kedong Ouyang ◽  
Guojun Wu ◽  
Huiyong Zhang ◽  
...  
Keyword(s):  
Amino Acids ◽  
Dna Vaccine ◽  
Tandem Repeats ◽  
Plasmid Vector ◽  
T Cell Epitopes ◽  
Specific Antibodies ◽  
Gastrin Releasing Peptide ◽  
Mycobacterial Hsp70

ABSTRACT The elevated expression and receptor binding of gastrin-releasing peptide (GRP) in various types of cancer, especially in malignant melanoma of the skin, suggest that GRP might be a putative target for immunotherapy in neoplastic diseases. We have therefore constructed a novel DNA vaccine coding for six tandem repeats of a fragment of GRP from amino acids 18 to 27 (GRP6) flanked by helper T-cell epitopes for increased immunogenicity, including HSP65, a tetanus toxoid fragment from amino acids 830 to 844 (T), pan-HLA-DR-binding epitope (PADRE) (P), and two repeats of a mycobacterial HSP70 fragment from amino acids 407 to 426 (M). The anti-GRP DNA vaccine (pCR3.1-VS-HSP65-TP-GRP6-M2) was constructed on a backbone of a pCR3.1 plasmid vector with eight 5′-GACGTT-3′ CpG motifs and the VEGF183 signal peptide (VS). Intramuscular (IM) injections of anti-GRP vaccine in mice stimulated the production of high titers of specific antibodies against GRP and suppressed the growth of subcutaneous tumors of B16-F10 melanoma cells. Parallel results were obtained in vitro, showing inhibition of B16-F10 cell proliferation by GRP antisera. IM injections of the DNA vaccine also significantly attenuated tumor-induced angiogenesis associated with intradermal tumors of B16-F10 cells. In addition, lung invasion of intravenously injected cells was highly diminished, suggesting potent antimetastatic activity of the DNA vaccine. These findings support the highly immunogenic and potent antitumorigenic activity of specific anti-GRP antibodies elicited by the anti-GRP DNA vaccine.

scholarly journals Intradermal DNA Electroporation Induces Cellular and Humoral Immune Response and Confers Protection against HER2/neu Tumor

2015 ◽  
Vol 2015 ◽  
pp. 1-10 ◽  
Author(s):  
Alessia Lamolinara ◽  
Lorenzo Stramucci ◽  
Albana Hysi ◽  
Manuela Iezzi ◽  
Cristina Marchini ◽  
...  
Keyword(s):  
Dna Vaccine ◽  
Mononuclear Cells ◽  
Humoral Response ◽  
Inflammatory Cells ◽  
Cell Types ◽  
Control Group ◽  
Intradermal Vaccination ◽  
Humoral Immune ◽  
Tumor Onset

Skin represents an attractive target for DNA vaccine delivery because of its natural richness in APCs, whose targeting may potentiate the effect of vaccination. Nevertheless, intramuscular electroporation is the most common delivery method for ECTM vaccination. In this study we assessed whether intradermal administration could deliver the vaccine into different cell types and we analyzed the evolution of tissue infiltrate elicited by the vaccination protocol. Intradermal electroporation (EP) vaccination resulted in transfection of different skin layers, as well as mononuclear cells. Additionally, we observed a marked recruitment of reactive infiltrates mainly 6–24 hours after treatment and inflammatory cells included CD11c+. Moreover, we tested the efficacy of intradermal vaccination against Her2/neu antigen in cellular and humoral response induction and consequent protection from a Her2/neu tumor challenge in Her2/neu nontolerant and tolerant mice. A significant delay in transplantable tumor onset was observed in both BALB/c (p≤0,0003) and BALB-neuT mice (p=0,003). Moreover, BALB-neuT mice displayed slow tumor growth as compared to control group (p<0,0016). In addition, while in vivo cytotoxic response was observed only in BALB/c mice, a significant antibody response was achieved in both mouse models. Our results identify intradermal EP vaccination as a promising method for delivering Her2/neu DNA vaccine.

scholarly journals Conjugation of lymphoma idiotype to CD40 antibody enhances lymphoma vaccine immunogenicity and antitumor effects in mice

Blood ◽  
2012 ◽  
Vol 119 (9) ◽  
pp. 2056-2065 ◽  
Author(s):  
Jennifer Carlring ◽  
Marika J. Szabo ◽  
Robert Dickinson ◽  
Evy De Leenheer ◽  
Andrew W. Heath
Keyword(s):  
T Cells ◽  
Immune Responses ◽  
Lipid A ◽  
Keyhole Limpet Hemocyanin ◽  
Potential Candidate ◽  
Antitumor Effects ◽  
Gm Csf ◽  
Humoral Immune ◽  
Monophosphoryl Lipid A

AbstractPersonalized immunotherapy of lymphoma based on tumor idiotype (Id) has shown anti-idiotype humoral immune responses in 40%-50% and cellular immune responses in 50%-75% of follicular lymphoma patients, indicating that this therapy can be clinically successful. We have developed a novel vaccine against lymphoma consisting of an anti-CD40 Ab (ADX40) chemically conjugated to the tumor idiotype A20 and tested it in a murine lymphoma model. BALB/c mice were immunized with 2 doses of immunogen alone or in conjunction with additional adjuvants before tumor challenge. ADX40-Id vaccination resulted in significantly retarded tumor growth and reduced mouse morbidity. Moreover, similar mouse survival was obtained with 2 injections of ADX40-Id as with 8 injections using the standard therapy of keyhole limpet hemocyanin Id + GM-CSF. Co-administration of ADX40-Id with 3-O-deacyl-4′-monophosphoryl lipid A further significantly enhanced vaccine efficacy, resulting in an increased overall survival. Anti-Id–specific Abs were detected at elevated levels after ADX40-Id immunization; however, in vivo depletion of CD4 and/or CD8 T cells before challenge showed that CD8 effector T cells were the major mediators of tumor protection. The results of the present study show that the ADX40-Id conjugate vaccine is a potential candidate as a stand-alone vaccine or in combination with currently licensed adjuvants for lymphoma immunotherapy.

scholarly journals Induction of specific immune response and suppression of fertility by B-cell-epitope-based mimovirus vaccine

Reproduction ◽  
2011 ◽  
Vol 142 (5) ◽  
pp. 659-666 ◽  
Author(s):  
Zi-gang Shen ◽  
Wei He ◽  
Ji Zhang ◽  
Hai-yang He ◽  
Xia Yang ◽  
...  
Keyword(s):  
B Cell ◽  
Cell Epitope ◽  
Reproductive Function ◽  
Serum Testosterone ◽  
Antigen Delivery ◽  
Dna Encoding ◽  
Igg Antibody ◽  
Humoral Immune ◽  
Contraceptive Vaccines

SPINLW1 (previously known as eppin (epididymal protease inhibitor)) is a target under intense scrutiny in the study of male contraceptive vaccines. B-cell-dominant epitopes are now recognized as key parts of the induction of humoral immune responses against target antigens. The generation of robust humoral responses in vivo has become a crucial problem in the development of modern vaccines. In this study, we developed a completely novel B-cell-dominant-epitope-based mimovirus vaccine, which is a kind of virus-size particulate antigen delivery system. The mimovirus successfully self-assembled from a cationic peptide containing a cell-penetrating peptide of TAT49–57 and a plasmid DNA encoding both three SPINLW1 (103–115) copies and adjuvant C3d3. The male mice were immunized with the epitope-based mimovirus vaccine, which resulted in a gradual elevation of specific serum IgG antibody levels. These reached a peak at week 4. Mating for the fertility assay showed that the mimovirus vaccine had accomplished a moderate fertility inhibition effect and investigation into the mechanism of action showed that it did so by interfering with the reproductive function of the sperm but that it did not damage the structures of the testes or cause serum testosterone to decline. Our results suggest an ideal protocol for suppressing fertility in mice by an engineered mimovirus vaccine.

scholarly journals A DNA Vaccine Coding for the Brucella Outer Membrane Protein 31 Confers Protection against B. melitensis and B. ovis Infection by Eliciting a Specific Cytotoxic Response

2005 ◽  
Vol 73 (10) ◽  
pp. 6537-6546 ◽  
Author(s):  
Juliana Cassataro ◽  
Carlos A. Velikovsky ◽  
Silvia de la Barrera ◽  
Silvia M. Estein ◽  
Laura Bruno ◽  
...  
Keyword(s):  
T Cells ◽  
Outer Membrane ◽  
Dna Vaccine ◽  
Cytotoxic T Lymphocyte ◽  
Protective Efficacy ◽  
Outer Membrane Proteins ◽  
Humoral Response ◽  
T Cell Subsets

ABSTRACT The development of an effective subunit vaccine against brucellosis is a research area of intense interest. The outer membrane proteins (Omps) of Brucella spp. have been extensively characterized as potential immunogenic and protective antigens. This study was conducted to evaluate the immunogenicity and protective efficacy of the B. melitensis Omp31 gene cloned in the pCI plasmid (pCIOmp31). Immunization of BALB/c mice with pCIOmp31 conferred protection against B. ovis and B. melitensis infection. Mice vaccinated with pCIOmp31 developed a very weak humoral response, and in vitro stimulation of their splenocytes with recombinant Omp31 did not induced the secretion of gamma interferon. Splenocytes from Omp31-vaccinated animals induced a specific cytotoxic-T-lymphocyte activity, which leads to the in vitro lysis of Brucella-infected macrophages. pCIOmp31 immunization elicited mainly CD8+ T cells, which mediate cytotoxicity via perforins, but also CD4+ T cells, which mediate lysis via the Fas-FasL pathway. In vivo depletion of T-cell subsets showed that the pCIOmp31-induced protection against Brucella infection is mediated predominantly by CD8+ T cells, although CD4+T cells also contribute. Our results demonstrate that the Omp31 DNA vaccine induces cytotoxic responses that have the potential to contribute to protection against Brucella infection. The protective response could be related to the induction of CD8+ T cells that eliminate Brucella-infected cells via the perforin pathway.

scholarly journals Design, Construction and Immunogenicity Assessment of pEGFP-N1-KMP11-GP96 (Fusion) as a DNA Vaccine Candidate against Leishmania major Infection in BALB/c Mice

2020 ◽  
Author(s):  
Abdolhossein DALIMI ◽  
Vahid NASIRI
Keyword(s):  
Dna Vaccine ◽  
Leishmania Major ◽  
Protective Efficacy ◽  
Cutaneous Lesion ◽  
Plasmid Vector ◽  
Humoral Immune ◽  
Pcr Products ◽  
Immunogenicity Assessment ◽  
Cellular Immune ◽  
Design Construction

Background: KMP-11 (Kinetoplastid membrane protein-Π) exists in all species of kinetoplastid family. It is fully conserved and the protein produced by this gene can induce a very high cellular immune response. We aimed to design a suitable construction for a Leishmania major DNA vaccine and evaluate the protective efficacy of it as a candidate for DNA vaccine against cutaneous leishmaniasis in BALB/c mice. Methods: This experimental study was conducted in Tehran City, Iran, between April 20, 2015 and May 30, 2016. KMP-11 gene of L. major (MRHO/IR/75/ER, Iranian strain) and NT-GP96 of Xenopus GP96 DNA from a pBluescript-GP96 plasmid were amplified by PCR and the purified PCR products were cloned into the pJET1.2/blunt plasmid vector, then, subcloned into pEGFP-N1 plasmid as an expression vector. Finally, the KMP-11 gene was fused with GP96 and afterward the combination cloned in pEGFP-N1. All the cloned genes confirmed by enzyme digestions. Then, four groups of mice were immunized with PBS, pEGFP-N1, pEGFP-N1-KMP, and pEGFP-N1-fusion. Four weeks after immunization, all animals were challenged with L. major virulent promastigotes. Results: The constructed fusion potentially showed an ability to elicit Th1 responses that led to cutaneous lesion healing. Interestingly, the group received KMP11-GP96 –GFP showed the highest ratio of IFN- γ /IL-4 and IgG2a/IgG1 compare to other groups. No side effect was observed after using the fusion in the mice. Conclusion: The constructed fusion could well stimulate both the cellular and humoral immune systems that led to cutaneous lesion healing in mice.

scholarly journals Enhanced magnitude and breadth of neutralizing humoral response to a DNA vaccine targeting the DHBV envelope protein delivered by in vivo electroporation

Virology ◽  
2012 ◽  
Vol 425 (1) ◽  
pp. 61-69 ◽  
Author(s):  
Ghada Khawaja ◽  
Thierry Buronfosse ◽  
Catherine Jamard ◽  
Sylviane Guerret ◽  
Fabien Zoulim ◽  
...  
Keyword(s):  
Dna Vaccine ◽  
Envelope Protein ◽  
Humoral Response ◽  
In Vivo Electroporation

scholarly journals Influence of introduction of interleukin-2 and interleukin-12 into experimental marker DNA-vaccine

2019 ◽  
Vol 25 ◽  
pp. 160-165
Author(s):  
Ia. O. Pokholenko ◽  
T. P. Gulko ◽  
V. A. Kordium
Keyword(s):  
Immune Response ◽  
Dna Vaccine ◽  
Humoral Immune Response ◽  
Interleukin 2 ◽  
Classical Swine Fever ◽  
Interleukin 12 ◽  
Chimeric Proteins ◽  
Humoral Immune

Aim. To determine the effects of the combined administration of recombinant expression vectors containing genes of murine interleukin-2 and interleukin-12 on humoral immune response, elicited by the experimental marker DNA-vaccine against classical swine fever. Methods. Expression of chimeric proteins in vitro and in vivo was determined by western-blot analysis. The antibodies specific to target antigens in blood serum have been detected by ELISA. Results. A series of recombinant plasmid vectors containing the genes of murine interleukin-2 and chimeric murine interleukin-12 have been developed. It has been shown that target chimeric proteins were expressed from the developed vectors both in vitro in HEK293 and in vivo in murine muscle tissue. The use of combined administration of murine interleukin-2 and interleukin-12 genes resulted in significant enhancement of titer of the anti-E2 and anti-β-galactosidase IgG, induced by vaccination with experimental marker DNA-vaccine against CSF, and model DNA-vaccine respectively. Conclusions. The data obtained show that the introduction of recombinant expressing vectors containing genes of murine interleukin-2 and interleukin-12 into vaccine preparations enhances humoral immune response elicited by the experimental marker DNA-vaccine against CSF and modelDNA-vaccine. Keywords: DNA-vaccine, humoral immune response, interleukin-2, interleukin-12, classical swine fever.

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