Results: 5

1.
Figure 1

Figure 1. Magnitude of positive IFN-γ ELISPOT responses to individual HSV-2 peptide pools in HSV-2+, HSV-1+/2− and HSVneg subjects. From: Development of an Interferon-Gamma ELISPOT Assay to Detect Human T Cell Responses to HSV-2.

The magnitudes of all positive IFN-γELISPOT responses to any of the 34 HSV-2 peptide pools are displayed in each group of subjects (n=2 for HSVneg, n=8 for HSV-1+/2− and n=22 for HSV-2+ subjects) separated by HSV serostatus. The median magnitude of responses to HSV-2 peptide pools in each group is shown by the bar.

Christine M. Posavad, et al. Vaccine. ;29(40):7058-7066.
2.
Figure 3

Figure 3. Response rates to individual HSV-2 peptide pools in HSV-2+, HSV-1+/2− and HSVneg subjects. From: Development of an Interferon-Gamma ELISPOT Assay to Detect Human T Cell Responses to HSV-2.

PBMC from HSV-2+ (n=26), HSV-1+/2− (n=20), and HSVneg subjects (n=35) were screened using 34 pools of overlapping peptides representing 16 HSV-2 ORFs. The percentage of HSV-2+ (A), HSV-1+/2− (B) and HSVneg subjects (C) who possessed positive responses to any HSV-2 peptide pool (Any pool) or to individual HSV-2 peptide pools is displayed.

Christine M. Posavad, et al. Vaccine. ;29(40):7058-7066.
3.
Figure 4

Figure 4. Breadth and magnitude of HSV-2-specific responses in HSV-2+ subjects. From: Development of an Interferon-Gamma ELISPOT Assay to Detect Human T Cell Responses to HSV-2.

(A–B) Breadth of the T cell response to HSV-2. The percentage of HSV-2+ subjects who possessed 1 to 8 positive IFNγ ELISPOT responses to the 16 HSV-2 proteins (A) or 1 to 10 positive IFN-γ ELISPOT responses to the 34 HSV-2 peptide pools (B) was determined from all HSV-2+ subjects who possessed HSV-2 peptide-specific responses (n=22). (C) Magnitude of the cumulative T cell response to HSV-2. Shown are cumulative quantitative T cell responses for all subjects (n=2 for HSVneg, n=8 for HSV-1+/2− and n=22 for HSV-2+ subjects) with positive IFN-γ ELISPOT responses to the 34 HSV-2 peptide pools covering 16 HSV-2 proteins.

Christine M. Posavad, et al. Vaccine. ;29(40):7058-7066.
4.
Figure 2

Figure 2. IFN-γ ELISPOT responses in HSV-2+, HSV-1+/2− and HSVneg subjects. From: Development of an Interferon-Gamma ELISPOT Assay to Detect Human T Cell Responses to HSV-2.

PBMC were screened by IFN-γ ELISPOT using 34 pools of overlapping peptides representing 16 HSV-2 ORFs. Results are from 3 HSV-2 seropositive subjects (A–C), an HSV-1+/2− subject (D) and an HSVneg subject (E). The dashed line represents the median SFC/106 PBMC from the DMSO control wells; the solid line represents the threshold for positivity (4 times the mean of the DMSO control wells and ≥55 SFC/106 PBMC). IFN-γ ELISPOT responses to PHA control wells were 1,258 SFC/106 PBMC (HSV-2+ Subject in A), 2,553 SFC/106 PBMC (HSV-2+ Subject in B), 2,329 SFC/106 PBMC (HSV-2+ Subject in C), 1,145 SFC/106 PBMC (HSV-1+/2−Subject in D) and 2,811 SFC/106 PBMC (HSVneg Subject in E).

Christine M. Posavad, et al. Vaccine. ;29(40):7058-7066.
5.
Figure 5

Figure 5. Phenotype of HSV-2 peptide-specific T cells by ICS and flow cytometry. From: Development of an Interferon-Gamma ELISPOT Assay to Detect Human T Cell Responses to HSV-2.

To confirm the identity of the HSV-2 peptides and to determine the phenotype of the responding T cells, PBMC from all subjects with positive IFN-γ ELISPOT responses were incubated with the deduced 15-mer and the CD4 or CD8 phenotype of the T cells was determined by ICS and flow cytometry. Representative plots for a CD8 (top row of graphs) and a CD4 (bottom row of graphs) epitope are displayed from 2 HSV-2+ subjects. PBMC from HSV-2+ Subject #23 were incubated with the peptide UL25405–419 and PBMC from HSV-2+ Subject #15 were incubated with the peptide gB-2136–152 (bottom row). The gating hierarchy moves from left to right starting with lymphocytes [side scatter (SSC) vs forward scatter (FSC)], live cells (side scatter vs violet viability), CD3 T cells (CD3 vs CD8), CD4 and CD8 T cells. The gates used to identify IFN-γ producing CD4 or CD8 T cells are depicted by the arrows.

Christine M. Posavad, et al. Vaccine. ;29(40):7058-7066.

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