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Pilot Study on the Use of DNA Priming Immunization to Enhance Y. pestis LcrV-Specific B Cell Responses Elicited by a Recombinant LcrV Protein Vaccine.

Li W, Wang S, Lu S - Vaccines (Basel) (2013)

Bottom Line: However, there is limited information on the mechanism of this effect.Previously, we demonstrated that a DNA vaccine expressing LcrV antigen can protect mice from lethal mucosal challenge.The finding that DNA immunization can enhance antigen-specific B cell responses is highly significant and will help guide similar studies in other model antigen systems.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, University of Massachusetts Medical School, Worcester, MA 01605, USA. wei.li@umassmed.edu.

ABSTRACT
Recent studies indicate that DNA immunization is powerful in eliciting antigen-specific antibody responses in both animal and human studies. However, there is limited information on the mechanism of this effect. In particular, it is not known whether DNA immunization can also enhance the development of antigen-specific B cell development. In this report, a pilot study was conducted using plague LcrV immunogen as a model system to determine whether DNA immunization is able to enhance LcrV-specific B cell development in mice. Plague is an acute and often fatal infectious disease caused by Yersinia pestis (Y. pestis). Humoral immune responses provide critical protective immunity against plague. Previously, we demonstrated that a DNA vaccine expressing LcrV antigen can protect mice from lethal mucosal challenge. In the current study, we further evaluated whether the use of a DNA priming immunization is able to enhance the immunogenicity of a recombinant LcrV protein vaccine, and in particular, the development of LcrV-specific B cells. Our data indicate that DNA immunization was able to elicit high-level LcrV antibody responses when used alone or as part of a prime-boost immunization approach. Most significantly, DNA immunization was also able to increase the levels of LcrV-specific B cell development. The finding that DNA immunization can enhance antigen-specific B cell responses is highly significant and will help guide similar studies in other model antigen systems.

No MeSH data available.


Related in: MedlinePlus

lcrV-DNA and V-protein immunization groups and vaccine components. Each mouse received 2 immunizations: prime at Week 0 and boost at Week 4 using designated codon optimized V DNA vaccine (V-DNA), V protein (V-Prot), or empty DNA vaccine vector (Vector) as indicated.
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vaccines-02-00036-f001: lcrV-DNA and V-protein immunization groups and vaccine components. Each mouse received 2 immunizations: prime at Week 0 and boost at Week 4 using designated codon optimized V DNA vaccine (V-DNA), V protein (V-Prot), or empty DNA vaccine vector (Vector) as indicated.

Mentions: Female BALB/c mice of 6–8 weeks old were purchased from Taconic Farms (Germantown, NY, USA) and housed in the animal facility managed by the Department of Animal Medicine at the University of Massachusetts Medical School (UMMS) in accordance with IACUC approved protocol. Mice (5/group) received two immunizations at Weeks 0 and 4 with designated vaccination regimens listed in Figure 1. Each mouse received codon optimized lcrV DNA vaccine (V-opt) (X2), V protein alone (X2), V-protein formulated with Incomplete Freund Adjuvant (IFA) (X2), V-opt DNA prime followed by V protein/IFA boost, or DNA vector alone immunization as the negative control. DNA immunizations were conducted via gene gun using a Helios gene gun (Bio-Rad). V.opt or the pSW3891 vector plasmid was coated onto 1.0-micron gold beads at 2 μg DNA/mg gold. Each shot delivered 1 μg of DNA and a total of six non-overlapping shots were delivered to shaved abdominal skin at each immunization after animals were anesthetized. Protein immunizations were done by intramuscular (i.m.) injection at the quadriceps, one injection site at one leg each with a dose of 1 μg/site (X2 sites). Sera were collected prior to and at two weeks after each immunization and at additional time points as indicated in Figure 1. At Week 16, animals were euthanized and splenocytes and bone marrow cells were isolated for B cell assays.


Pilot Study on the Use of DNA Priming Immunization to Enhance Y. pestis LcrV-Specific B Cell Responses Elicited by a Recombinant LcrV Protein Vaccine.

Li W, Wang S, Lu S - Vaccines (Basel) (2013)

lcrV-DNA and V-protein immunization groups and vaccine components. Each mouse received 2 immunizations: prime at Week 0 and boost at Week 4 using designated codon optimized V DNA vaccine (V-DNA), V protein (V-Prot), or empty DNA vaccine vector (Vector) as indicated.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494201&req=5

vaccines-02-00036-f001: lcrV-DNA and V-protein immunization groups and vaccine components. Each mouse received 2 immunizations: prime at Week 0 and boost at Week 4 using designated codon optimized V DNA vaccine (V-DNA), V protein (V-Prot), or empty DNA vaccine vector (Vector) as indicated.
Mentions: Female BALB/c mice of 6–8 weeks old were purchased from Taconic Farms (Germantown, NY, USA) and housed in the animal facility managed by the Department of Animal Medicine at the University of Massachusetts Medical School (UMMS) in accordance with IACUC approved protocol. Mice (5/group) received two immunizations at Weeks 0 and 4 with designated vaccination regimens listed in Figure 1. Each mouse received codon optimized lcrV DNA vaccine (V-opt) (X2), V protein alone (X2), V-protein formulated with Incomplete Freund Adjuvant (IFA) (X2), V-opt DNA prime followed by V protein/IFA boost, or DNA vector alone immunization as the negative control. DNA immunizations were conducted via gene gun using a Helios gene gun (Bio-Rad). V.opt or the pSW3891 vector plasmid was coated onto 1.0-micron gold beads at 2 μg DNA/mg gold. Each shot delivered 1 μg of DNA and a total of six non-overlapping shots were delivered to shaved abdominal skin at each immunization after animals were anesthetized. Protein immunizations were done by intramuscular (i.m.) injection at the quadriceps, one injection site at one leg each with a dose of 1 μg/site (X2 sites). Sera were collected prior to and at two weeks after each immunization and at additional time points as indicated in Figure 1. At Week 16, animals were euthanized and splenocytes and bone marrow cells were isolated for B cell assays.

Bottom Line: However, there is limited information on the mechanism of this effect.Previously, we demonstrated that a DNA vaccine expressing LcrV antigen can protect mice from lethal mucosal challenge.The finding that DNA immunization can enhance antigen-specific B cell responses is highly significant and will help guide similar studies in other model antigen systems.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, University of Massachusetts Medical School, Worcester, MA 01605, USA. wei.li@umassmed.edu.

ABSTRACT
Recent studies indicate that DNA immunization is powerful in eliciting antigen-specific antibody responses in both animal and human studies. However, there is limited information on the mechanism of this effect. In particular, it is not known whether DNA immunization can also enhance the development of antigen-specific B cell development. In this report, a pilot study was conducted using plague LcrV immunogen as a model system to determine whether DNA immunization is able to enhance LcrV-specific B cell development in mice. Plague is an acute and often fatal infectious disease caused by Yersinia pestis (Y. pestis). Humoral immune responses provide critical protective immunity against plague. Previously, we demonstrated that a DNA vaccine expressing LcrV antigen can protect mice from lethal mucosal challenge. In the current study, we further evaluated whether the use of a DNA priming immunization is able to enhance the immunogenicity of a recombinant LcrV protein vaccine, and in particular, the development of LcrV-specific B cells. Our data indicate that DNA immunization was able to elicit high-level LcrV antibody responses when used alone or as part of a prime-boost immunization approach. Most significantly, DNA immunization was also able to increase the levels of LcrV-specific B cell development. The finding that DNA immunization can enhance antigen-specific B cell responses is highly significant and will help guide similar studies in other model antigen systems.

No MeSH data available.


Related in: MedlinePlus