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Herpes simplex type 2 virus deleted in glycoprotein D protects against vaginal, skin and neural disease.

Petro C, González PA, Cheshenko N, Jandl T, Khajoueinejad N, Bénard A, Sengupta M, Herold BC, Jacobs WR - Elife (2015)

Bottom Line: Subunit vaccines comprised of glycoprotein D (gD-2) failed to prevent HSV-2 highlighting need for novel strategies.The antibodies elicited cell-mediated cytotoxicity, but little neutralizing activity.These studies demonstrate that non-neutralizing Fc-mediated humoral responses confer protection and support advancement of this attenuated vaccine.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Albert Einstein College of Medicine, New York, United States.

ABSTRACT
Subunit vaccines comprised of glycoprotein D (gD-2) failed to prevent HSV-2 highlighting need for novel strategies. To test the hypothesis that deletion of gD-2 unmasks protective antigens, we evaluated the efficacy and safety of an HSV-2 virus deleted in gD-2 and complemented allowing a single round of replication on cells expressing HSV-1 gD (ΔgD(-/+gD-1)). Subcutaneous immunization of C57BL/6 or BALB/c mice with ΔgD(-/+gD1) provided 100% protection against lethal intravaginal or skin challenges and prevented latency. ΔgD(-/+gD1) elicited no disease in SCID mice, whereas 1000-fold lower doses of wild-type virus were lethal. HSV-specific antibodies were detected in serum (titer 1:800,000) following immunization and in vaginal washes after intravaginal challenge. The antibodies elicited cell-mediated cytotoxicity, but little neutralizing activity. Passive transfer of immune serum completely protected wild-type, but not Fcγ-receptor or neonatal Fc-receptor knock-out mice. These studies demonstrate that non-neutralizing Fc-mediated humoral responses confer protection and support advancement of this attenuated vaccine.

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Characterization of vaginal wash and serum antibodies.(A) Isotypes of anti-HSV-2 antibodies in vaginal washes obtained at day 4 and day 8 post intravaginal challenge in mice that were immunized with HSV-2 ΔgD−/+gD−1 (ΔgD) or VD60 cell lysate (Control). Antibody responses are shown as means ± SD (n = 5 per group) and were compared by two-way ANOVA; *p < 0.05, **p < 0.01, ***p < 0.001. (B) Western blots with sucrose gradient-purified ΔgD−/+gD−1 and ΔgD−/− viruses (equivalent particle numbers based on a Western blot for VP5) probed with an anti-gD mAb (mAb (+)), sera from HSV-2 ΔgD−/+gD−1-vaccinated mice day 7 post-boost (ΔgD) or sera from VD60 lysate-vaccinated mice day 7 post-intravaginal challenge with HSV-2(4674) (Ctrl + Challenge). (C) Western blots with purified recombinant glycoprotein B-1 (2 μg per lane) probed with an anti-gB mAb (mAb (+)), sera from VD60 lysate -vaccinated mice (Ctrl), HSV-2 ΔgD−/+gD−1-vaccinated mice day 7 post boost (ΔgD) or VD60 lysate-vaccinated mice day 7 post-intravaginal challenge with HSV-2(4674) (Ctrl + Challenge).DOI:http://dx.doi.org/10.7554/eLife.06054.012
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fig6s1: Characterization of vaginal wash and serum antibodies.(A) Isotypes of anti-HSV-2 antibodies in vaginal washes obtained at day 4 and day 8 post intravaginal challenge in mice that were immunized with HSV-2 ΔgD−/+gD−1 (ΔgD) or VD60 cell lysate (Control). Antibody responses are shown as means ± SD (n = 5 per group) and were compared by two-way ANOVA; *p < 0.05, **p < 0.01, ***p < 0.001. (B) Western blots with sucrose gradient-purified ΔgD−/+gD−1 and ΔgD−/− viruses (equivalent particle numbers based on a Western blot for VP5) probed with an anti-gD mAb (mAb (+)), sera from HSV-2 ΔgD−/+gD−1-vaccinated mice day 7 post-boost (ΔgD) or sera from VD60 lysate-vaccinated mice day 7 post-intravaginal challenge with HSV-2(4674) (Ctrl + Challenge). (C) Western blots with purified recombinant glycoprotein B-1 (2 μg per lane) probed with an anti-gB mAb (mAb (+)), sera from VD60 lysate -vaccinated mice (Ctrl), HSV-2 ΔgD−/+gD−1-vaccinated mice day 7 post boost (ΔgD) or VD60 lysate-vaccinated mice day 7 post-intravaginal challenge with HSV-2(4674) (Ctrl + Challenge).DOI:http://dx.doi.org/10.7554/eLife.06054.012

Mentions: HSV-specific Ab responses were consistently observed with serum dilutions as high as 1:800,000 7 days post-boost (Figure 6A) and were detected in vaginal washes within 4 days of challenge, indicating transport of HSV Abs into the vaginal lumen (Figure 6B). The serum HSV-specific Abs exhibited low levels of neutralizing activity (1:5 dilution) (Figure 6C), but did display ADCC activity, which was reduced in the presence of anti-FcγR Ab (Figure 6D). The HSV-specific Abs in serum and vaginal washes were comprised predominantly of IgG2a and IgG2b (Figure 6E and Figure 6—figure supplement 1A) and recognized multiple viral proteins on Western blots with HSV-2-infected cell lysates as the immunogen (Figure 6F). Serum from control-immunized and subsequently HSV-2 infected mice (obtained at day 8 post–challenge) recognized a more restricted set of proteins. Western blots with purified HSV-2 ΔgD−/+gD−1 or HSV-2 ΔgD−/− indicate that the predominant antigen recognized by control-immunized, HSV-2-challenged, but not ΔgD−/+gD−1-vaccinated mice, is gD (Figure 6—figure supplement 1B). Blots with recombinant glycoprotein B-1 identify the ∼100 kDa band recognized by the ΔgD−/+gD−1-vaccinated immune serum as gB (Figure 6—figure supplement 1C).10.7554/eLife.06054.011Figure 6.Vaccination with HSV-2 ΔgD−/+gD−1 induces protective mucosal antibodies targeting multiple HSV proteins with ADCC activity.


Herpes simplex type 2 virus deleted in glycoprotein D protects against vaginal, skin and neural disease.

Petro C, González PA, Cheshenko N, Jandl T, Khajoueinejad N, Bénard A, Sengupta M, Herold BC, Jacobs WR - Elife (2015)

Characterization of vaginal wash and serum antibodies.(A) Isotypes of anti-HSV-2 antibodies in vaginal washes obtained at day 4 and day 8 post intravaginal challenge in mice that were immunized with HSV-2 ΔgD−/+gD−1 (ΔgD) or VD60 cell lysate (Control). Antibody responses are shown as means ± SD (n = 5 per group) and were compared by two-way ANOVA; *p < 0.05, **p < 0.01, ***p < 0.001. (B) Western blots with sucrose gradient-purified ΔgD−/+gD−1 and ΔgD−/− viruses (equivalent particle numbers based on a Western blot for VP5) probed with an anti-gD mAb (mAb (+)), sera from HSV-2 ΔgD−/+gD−1-vaccinated mice day 7 post-boost (ΔgD) or sera from VD60 lysate-vaccinated mice day 7 post-intravaginal challenge with HSV-2(4674) (Ctrl + Challenge). (C) Western blots with purified recombinant glycoprotein B-1 (2 μg per lane) probed with an anti-gB mAb (mAb (+)), sera from VD60 lysate -vaccinated mice (Ctrl), HSV-2 ΔgD−/+gD−1-vaccinated mice day 7 post boost (ΔgD) or VD60 lysate-vaccinated mice day 7 post-intravaginal challenge with HSV-2(4674) (Ctrl + Challenge).DOI:http://dx.doi.org/10.7554/eLife.06054.012
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fig6s1: Characterization of vaginal wash and serum antibodies.(A) Isotypes of anti-HSV-2 antibodies in vaginal washes obtained at day 4 and day 8 post intravaginal challenge in mice that were immunized with HSV-2 ΔgD−/+gD−1 (ΔgD) or VD60 cell lysate (Control). Antibody responses are shown as means ± SD (n = 5 per group) and were compared by two-way ANOVA; *p < 0.05, **p < 0.01, ***p < 0.001. (B) Western blots with sucrose gradient-purified ΔgD−/+gD−1 and ΔgD−/− viruses (equivalent particle numbers based on a Western blot for VP5) probed with an anti-gD mAb (mAb (+)), sera from HSV-2 ΔgD−/+gD−1-vaccinated mice day 7 post-boost (ΔgD) or sera from VD60 lysate-vaccinated mice day 7 post-intravaginal challenge with HSV-2(4674) (Ctrl + Challenge). (C) Western blots with purified recombinant glycoprotein B-1 (2 μg per lane) probed with an anti-gB mAb (mAb (+)), sera from VD60 lysate -vaccinated mice (Ctrl), HSV-2 ΔgD−/+gD−1-vaccinated mice day 7 post boost (ΔgD) or VD60 lysate-vaccinated mice day 7 post-intravaginal challenge with HSV-2(4674) (Ctrl + Challenge).DOI:http://dx.doi.org/10.7554/eLife.06054.012
Mentions: HSV-specific Ab responses were consistently observed with serum dilutions as high as 1:800,000 7 days post-boost (Figure 6A) and were detected in vaginal washes within 4 days of challenge, indicating transport of HSV Abs into the vaginal lumen (Figure 6B). The serum HSV-specific Abs exhibited low levels of neutralizing activity (1:5 dilution) (Figure 6C), but did display ADCC activity, which was reduced in the presence of anti-FcγR Ab (Figure 6D). The HSV-specific Abs in serum and vaginal washes were comprised predominantly of IgG2a and IgG2b (Figure 6E and Figure 6—figure supplement 1A) and recognized multiple viral proteins on Western blots with HSV-2-infected cell lysates as the immunogen (Figure 6F). Serum from control-immunized and subsequently HSV-2 infected mice (obtained at day 8 post–challenge) recognized a more restricted set of proteins. Western blots with purified HSV-2 ΔgD−/+gD−1 or HSV-2 ΔgD−/− indicate that the predominant antigen recognized by control-immunized, HSV-2-challenged, but not ΔgD−/+gD−1-vaccinated mice, is gD (Figure 6—figure supplement 1B). Blots with recombinant glycoprotein B-1 identify the ∼100 kDa band recognized by the ΔgD−/+gD−1-vaccinated immune serum as gB (Figure 6—figure supplement 1C).10.7554/eLife.06054.011Figure 6.Vaccination with HSV-2 ΔgD−/+gD−1 induces protective mucosal antibodies targeting multiple HSV proteins with ADCC activity.

Bottom Line: Subunit vaccines comprised of glycoprotein D (gD-2) failed to prevent HSV-2 highlighting need for novel strategies.The antibodies elicited cell-mediated cytotoxicity, but little neutralizing activity.These studies demonstrate that non-neutralizing Fc-mediated humoral responses confer protection and support advancement of this attenuated vaccine.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Albert Einstein College of Medicine, New York, United States.

ABSTRACT
Subunit vaccines comprised of glycoprotein D (gD-2) failed to prevent HSV-2 highlighting need for novel strategies. To test the hypothesis that deletion of gD-2 unmasks protective antigens, we evaluated the efficacy and safety of an HSV-2 virus deleted in gD-2 and complemented allowing a single round of replication on cells expressing HSV-1 gD (ΔgD(-/+gD-1)). Subcutaneous immunization of C57BL/6 or BALB/c mice with ΔgD(-/+gD1) provided 100% protection against lethal intravaginal or skin challenges and prevented latency. ΔgD(-/+gD1) elicited no disease in SCID mice, whereas 1000-fold lower doses of wild-type virus were lethal. HSV-specific antibodies were detected in serum (titer 1:800,000) following immunization and in vaginal washes after intravaginal challenge. The antibodies elicited cell-mediated cytotoxicity, but little neutralizing activity. Passive transfer of immune serum completely protected wild-type, but not Fcγ-receptor or neonatal Fc-receptor knock-out mice. These studies demonstrate that non-neutralizing Fc-mediated humoral responses confer protection and support advancement of this attenuated vaccine.

Show MeSH
Related in: MedlinePlus