Research ArticleInfectious Disease

Maternal immunization confers protection against neonatal herpes simplex mortality and behavioral morbidity

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Science Translational Medicine  10 Apr 2019:
Vol. 11, Issue 487, eaau6039
DOI: 10.1126/scitranslmed.aau6039
  • Fig. 1 Human maternal HSV-specific antibody is transferred to offspring and is neutralizing.

    Human maternal and paired cord and neonate sera were probed for HSV-1– and HSV-2(gG2)–specific antibodies via multiplex. Antibodies found in (A) maternal-cord pairs and (B) maternal-neonate pairs are plotted by glycoprotein binding (maximum binding, 1; minimum binding, 0). Regression lines for each glycoprotein are shown in corresponding colors. The dotted line of identity refers to a 1:1 transfer ratio between maternal and cord or neonate samples. Above the dotted line represents a higher corresponding cord or neonatal value, whereas below represents a higher corresponding maternal value. gD’ refers to the ectodomain of HSV-1 gD. (C) Cord samples were binned into tertiles based on mean antibody binding in (A); high responder (above second tertile) and low responder (below first tertile) groups were assayed for neutralizing antibodies against HSV-1 via a serum neutralization assay. Serum from HSV-1 seronegative adults was used as a biological negative control. Statistical significance was determined using one-way analysis of variance (ANOVA) with Dunnett’s multiple comparisons. In (A) and (B), regression lines were compared to the line of identity. In (C), bars represent the mean. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

  • Fig. 2 Antibodies in sera and TGs from latently infected mice neutralize HSV-1.

    Antibody responses to HSV-1 and control antigens were defined via multiplexed antibody assay for matched (A) sera and (B) TG extract samples. Plaque reduction neutralization assays of (C) sera and (D) diluted TG extracts from latently infected B6 (red), naïve B6 (blue), and latently infected muMT (black) mice. SN50 (dotted line) refers to the serum dilution that neutralized 50% of the virus. Statistical significance for (A) and (B) was determined using an unpaired t test between sera/extracts from latently infected animals and those of naïve animals for each antigen. Statistical significance for (D) was determined by one-way ANOVA with Tukey’s multiple comparisons. MFI, median fluorescence intensity. Error bars indicate SD (A and B). **P < 0.01, ***P < 0.001, and ****P < 0.0001. Serum and TG extracts were pooled from 10 to 18 mice in each group, and data are representative of two independent experiments.

  • Fig. 3 Maternal passive antibody treatment protects against neonatal herpes.

    Pooled sera or purified IgG from latently infected (immune) or naïve mice was administered to muMT dams before parturition, and pups were infected intranasally with 105 PFU (plaque-forming units) (A), 104 PFU (B and C), or 103 PFU (D) of HSV-1. Viral titers were determined in pups 3 days after infection. Data are shown as viral titers in individual pups from dams treated with immune sera/IgG (red) or naïve sera/IgG (blue). Statistical significance was determined by multiple t tests (A and C) and log-rank test (B and D). **P < 0.01, ***P < 0.001, and ****P < 0.0001. Data are representative of at least three independent experiments.

  • Fig. 4 Immunization with HSV-2 dl5-29 allows maternal transfer of HSV-specific antibodies.

    (A) B6 females were vaccinated with HSV-2 dl5-29 virus or mock lysate and boosted and bred 21 days later. i.m., intramuscularly. (B) Sera from immunized dams and (C) their naïve offspring were assessed for HSV-1 specificity. gD’ refers to the ectodomain of HSV-1 gD. (D) Immunofluorescence of TG tissue from naïve neonates of mock-immunized dams (left) and that of dl5-29–immunized dams (right), stained with anti-mouse IgG (green) and 4′,6-diamidino-2-phenylindole (DAPI) (blue). Scale bar, 500 μm. Statistical significance in (C) was determined by unpaired t test for each antigen. Data are representative of two to three independent experiments *P < 0.05, ***P < 0.001, and **** P <0.0001.

  • Fig. 5 Maternal immunization with dl5-29 protects against neonatal herpes.

    B6 females were immunized and boosted as shown in Fig. 4A. Neonates P1-2 from dams immunized with dl5-29 (red) or from mock-immunized dams (blue) were challenged with HSV. (A) Viral titers in perfused organs from neonates infected with 104 PFU of HSV-1. Survival of neonates challenged with 103 PFU of HSV-1 (B) or HSV-2 (C). (D) Offspring from mock- or dl5-29–immunized dams were removed immediately at birth from their birth dam, fostered by equivalent or reciprocal (mock- or dl5-29–immunized) dams, and challenged P2 with 103 PFU of HSV-1 to assess milk (green) and placental (purple) protective contributions. Statistical significance was determined by multiple t test (A) or log-rank test (B to D). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. Data are representative of at least two independent experiments.

  • Fig. 6 Neonatal herpes causes anxiogenic behavior, which can be prevented by maternal immunization.

    Mice challenged with 100 PFU of HSV-1 as neonates (P1-2) were analyzed in the OFT 5 weeks after infection. (A) Open field behavior in offspring of latently infected and mock-infected dams is shown by movement tracking (top) and heat map (bottom). Location preferences (B) can be quantified as time spent in the inner region (within red lines) and outer periphery or (C) as thigmotaxis, a ratio of time spent in the outer perimeter over total time. Similarly, offspring of dl5-29– and mock-immunized dams (D) were assessed for location preference (E) and thigmotaxis (F). As a control, thigmotaxis of age-matched, untreated, naïve mice was quantified (G). Data are represented as individual animals, and statistical significance was determined by unpaired t test. Error bars represent SD. ***P < 0.001 and ****P < 0.0001. Data are representative of two to three independent experiments.

Supplementary Materials

  • www.sciencetranslationalmedicine.org/cgi/content/full/11/487/eaau6039/DC1

    Materials and Methods

    Fig. S1. Maternal antibody is not present in 18-month-old children.

    Fig. S2. Measurement of IgG staining in TG tissues from Fig. 4D.

    Fig. S3. Long-term (≥245 days) protection by maternal immunization with dl5-29.

    Fig. S4. Neonatal mice do not show signs of infection with a low HSV-1 dose.

    Fig. S5. Total ambulation of HSV-challenged and naïve mice in the OFT.

    Data file S1. Primary data.

    References (8589)

  • The PDF file includes:

    • Materials and Methods
    • Fig. S1. Maternal antibody is not present in 18-month-old children.
    • Fig. S2. Measurement of IgG staining in TG tissues from Fig. 4D.
    • Fig. S3. Long-term (≥245 days) protection by maternal immunization with dl5-29.
    • Fig. S4. Neonatal mice do not show signs of infection with a low HSV-1 dose.
    • Fig. S5. Total ambulation of HSV-challenged and naïve mice in the OFT.
    • References (8589)

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    Other Supplementary Material for this manuscript includes the following:

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