anticd8 pe cy7 Search Results


cd8-pe-cy7  (Becton Dickinson)
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cd8  (Abcam)
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Abcam cd8
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phycoerythrin–cyanine (pe–cy)7-conjugated anti-cd8  (Becton Dickinson)
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fluorescein isothiocyanate (fitc)–conjugated anti-cd14  (Becton Dickinson)
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cd8-pe-cy7 (557746)  (Becton Dickinson)
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Cd8 Pe Cy7 (557746), supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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pe-cy7-conjugated anti-cd8 (sk1, mouse igg1)  (Becton Dickinson)
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cd8 53-6.7) pe-cy7  (Becton Dickinson)
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Becton Dickinson cd8 53-6.7) pe-cy7
Schematic representation showing the relative location within HSV-1 VP11/12 of the potential <t> CD8 </t> + T cell epitopes studied.
Cd8 53 6.7) Pe Cy7, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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pe cy7 anti cd8  (Cytek Biosciences)
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Cytek Biosciences pe cy7 anti cd8
Schematic representation showing the relative location within HSV-1 VP11/12 of the potential <t> CD8 </t> + T cell epitopes studied.
Pe Cy7 Anti Cd8, supplied by Cytek Biosciences, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti-cd8 pe-cy7 pra-t8  (Becton Dickinson)
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anti-cd8-pe-cy7 mab sk1  (Becton Dickinson)
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Becton Dickinson anti-cd8-pe-cy7 mab sk1
Human lymphocytes are susceptible to DV infection and replication. (A) Representative flow cytometry density plots of purified cells showing the frequencies of CD4+ and <t>CD8+</t> T cells infected by DV. The number within each box refers to the frequency of cells within each gate. (B and C) Average frequencies of CD4+ and CD8+ T cell infection by the four DV serotypes. Mock infection (circles) and DV1 BR90 (squares), DV2 265 (triangles), DV3 98 (inverted triangles), and DV4 360 (diamonds) infection. Bars show the mean values for samples from six healthy donors. *, P ≤ 0.05 compared to the results for infection by four DV serotypes. (D and E) Confocal fluorescence microscopy of CD4+ and CD8+ T cells labeled for DNA (DAPI; blue), DV (anti-E protein MAb; red), and immunophenotyping markers (anti-CD4 <t>or</t> <t>anti-CD8</t> MAb; green). Scale bar = 10 μm. (A, D, E) Data are representative of six independent experiments. (F and G) Flow cytometry data indicating the average frequencies of DV-infected CD4+ (F) and CD8+ (G) T lymphocytes (CD14+-depleted cells from six healthy donors) after virus treatment with different concentrations of heparin (2 to 20 μg). (H and I) Average frequencies, using a flow cytometry assay, of CD4+ (H) and CD8+ (I) T lymphocytes infected with DV1 to DV4 after cell treatment with different concentrations of heparinase III (0.5 to 5.0 IU) and infection with the four DV serotypes. Data represent the mean values ± standard errors of the means (SEM) for cells from six healthy donors. *, P ≤ 0.05 compared to the results for mock-infected controls. (J and M) DV progeny in cell culture supernatants from purified CD4+ (J) and CD8+ (M) T lymphocytes infected with the four DV serotypes were quantified using a focus-forming assay in C6/36 cells. (K and N) DV replication on CD4+ (K) and CD8+ (N) T lymphocytes (purified via cell sorting) was determined by real-time PCR assay. Quantification of DV nonstructural protein 3 (NS3) gene mRNA, normalized to the housekeeping gene 18S, for comparison of the levels in infected cells and mock-infected cells. (L and O) Qualitative determination of the levels of DV nonstructural protein 1 (NS1) in cell culture supernatants of purified CD4+ (L) and CD8+ (O) T lymphocytes infected by DV by using a capture ELISA from PanBio (values >11 indicate NS1 secretion). Mock infected (circles) and DV1 BR90 (squares), DV2 265 (triangles), DV3 98 (inverted triangles), and DV4 360 (diamonds) infected. Bars show the mean values for cells from six healthy donors. *, P ≤ 0.05 compared to the results for infection by four DV serotypes.
Anti Cd8 Pe Cy7 Mab Sk1, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cd56-pe-cy5.5  (Becton Dickinson)
90
Becton Dickinson cd56-pe-cy5.5
Human lymphocytes are susceptible to DV infection and replication. (A) Representative flow cytometry density plots of purified cells showing the frequencies of CD4+ and <t>CD8+</t> T cells infected by DV. The number within each box refers to the frequency of cells within each gate. (B and C) Average frequencies of CD4+ and CD8+ T cell infection by the four DV serotypes. Mock infection (circles) and DV1 BR90 (squares), DV2 265 (triangles), DV3 98 (inverted triangles), and DV4 360 (diamonds) infection. Bars show the mean values for samples from six healthy donors. *, P ≤ 0.05 compared to the results for infection by four DV serotypes. (D and E) Confocal fluorescence microscopy of CD4+ and CD8+ T cells labeled for DNA (DAPI; blue), DV (anti-E protein MAb; red), and immunophenotyping markers (anti-CD4 <t>or</t> <t>anti-CD8</t> MAb; green). Scale bar = 10 μm. (A, D, E) Data are representative of six independent experiments. (F and G) Flow cytometry data indicating the average frequencies of DV-infected CD4+ (F) and CD8+ (G) T lymphocytes (CD14+-depleted cells from six healthy donors) after virus treatment with different concentrations of heparin (2 to 20 μg). (H and I) Average frequencies, using a flow cytometry assay, of CD4+ (H) and CD8+ (I) T lymphocytes infected with DV1 to DV4 after cell treatment with different concentrations of heparinase III (0.5 to 5.0 IU) and infection with the four DV serotypes. Data represent the mean values ± standard errors of the means (SEM) for cells from six healthy donors. *, P ≤ 0.05 compared to the results for mock-infected controls. (J and M) DV progeny in cell culture supernatants from purified CD4+ (J) and CD8+ (M) T lymphocytes infected with the four DV serotypes were quantified using a focus-forming assay in C6/36 cells. (K and N) DV replication on CD4+ (K) and CD8+ (N) T lymphocytes (purified via cell sorting) was determined by real-time PCR assay. Quantification of DV nonstructural protein 3 (NS3) gene mRNA, normalized to the housekeeping gene 18S, for comparison of the levels in infected cells and mock-infected cells. (L and O) Qualitative determination of the levels of DV nonstructural protein 1 (NS1) in cell culture supernatants of purified CD4+ (L) and CD8+ (O) T lymphocytes infected by DV by using a capture ELISA from PanBio (values >11 indicate NS1 secretion). Mock infected (circles) and DV1 BR90 (squares), DV2 265 (triangles), DV3 98 (inverted triangles), and DV4 360 (diamonds) infected. Bars show the mean values for cells from six healthy donors. *, P ≤ 0.05 compared to the results for infection by four DV serotypes.
Cd56 Pe Cy5.5, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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pe cy7 conjugated anti cd8 antibody  (Cytek Biosciences)
94
Cytek Biosciences pe cy7 conjugated anti cd8 antibody
Human lymphocytes are susceptible to DV infection and replication. (A) Representative flow cytometry density plots of purified cells showing the frequencies of CD4+ and <t>CD8+</t> T cells infected by DV. The number within each box refers to the frequency of cells within each gate. (B and C) Average frequencies of CD4+ and CD8+ T cell infection by the four DV serotypes. Mock infection (circles) and DV1 BR90 (squares), DV2 265 (triangles), DV3 98 (inverted triangles), and DV4 360 (diamonds) infection. Bars show the mean values for samples from six healthy donors. *, P ≤ 0.05 compared to the results for infection by four DV serotypes. (D and E) Confocal fluorescence microscopy of CD4+ and CD8+ T cells labeled for DNA (DAPI; blue), DV (anti-E protein MAb; red), and immunophenotyping markers (anti-CD4 <t>or</t> <t>anti-CD8</t> MAb; green). Scale bar = 10 μm. (A, D, E) Data are representative of six independent experiments. (F and G) Flow cytometry data indicating the average frequencies of DV-infected CD4+ (F) and CD8+ (G) T lymphocytes (CD14+-depleted cells from six healthy donors) after virus treatment with different concentrations of heparin (2 to 20 μg). (H and I) Average frequencies, using a flow cytometry assay, of CD4+ (H) and CD8+ (I) T lymphocytes infected with DV1 to DV4 after cell treatment with different concentrations of heparinase III (0.5 to 5.0 IU) and infection with the four DV serotypes. Data represent the mean values ± standard errors of the means (SEM) for cells from six healthy donors. *, P ≤ 0.05 compared to the results for mock-infected controls. (J and M) DV progeny in cell culture supernatants from purified CD4+ (J) and CD8+ (M) T lymphocytes infected with the four DV serotypes were quantified using a focus-forming assay in C6/36 cells. (K and N) DV replication on CD4+ (K) and CD8+ (N) T lymphocytes (purified via cell sorting) was determined by real-time PCR assay. Quantification of DV nonstructural protein 3 (NS3) gene mRNA, normalized to the housekeeping gene 18S, for comparison of the levels in infected cells and mock-infected cells. (L and O) Qualitative determination of the levels of DV nonstructural protein 1 (NS1) in cell culture supernatants of purified CD4+ (L) and CD8+ (O) T lymphocytes infected by DV by using a capture ELISA from PanBio (values >11 indicate NS1 secretion). Mock infected (circles) and DV1 BR90 (squares), DV2 265 (triangles), DV3 98 (inverted triangles), and DV4 360 (diamonds) infected. Bars show the mean values for cells from six healthy donors. *, P ≤ 0.05 compared to the results for infection by four DV serotypes.
Pe Cy7 Conjugated Anti Cd8 Antibody, supplied by Cytek Biosciences, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Schematic representation showing the relative location within HSV-1 VP11/12 of the potential CD8 + T cell epitopes studied.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: HLA-A02:01-Restricted Epitopes Identified from the Herpes Simplex Virus Tegument Protein VP11/12 Preferentially Recall Polyfunctional Effector Memory CD8 + T Cells from Seropositive Asymptomatic Individuals and Protect “Humanized” HLA-A*02:01 Transgenic Mice Against Ocular Herpes

doi: 10.4049/jimmunol.1402606

Figure Lengend Snippet: Schematic representation showing the relative location within HSV-1 VP11/12 of the potential CD8 + T cell epitopes studied.

Article Snippet: Anti-mouse antibodies were: CD8 (clone 53-6.7) PE-Cy7, CD44 (clone IM7) APC-Cy7, CD62L (MEL-14) A700, CD107a (clone 1D4B) FITC, CD107b (clone Ha1/29) FITC, IFN-γ (clone XMG1.2) A700 (BD Pharmingen); Granzyme B (clone GB11) A647, (BioLegend).

Techniques: Sequencing

PBMCs (~10 × 106) derived from ten HLA-A*02:01 positive HSV-1 seropositive ASYMP individuals and from ten HLA-A*02:01 positive HSV-1 seropositive SYMP individuals were analyzed ex vivo by FACS for the frequency of CD8+ T cells specific to three VP11/12 epitopes using HLA-A*0201 VP11/12 peptide-tetramer complexes representing each of the three high to medium peptide binders (VP11/1266–74, VP11/12220–228 and VP11/12702–710 epitopes), as shown in Fig. 1. (A) Representative FACS data of the frequencies of CD8+ T cells, specific to VP11/1266–74, VP11/12220–228 and VP11/12702–710 epitopes, detected in PBMCs from one HLA-A*02:01 positive HSV-1 seropositive ASYMP individual (top panels) and one HLA-A*02:01 positive HSV-1 seropositive SYMP individual (lower panels). (B) Average frequencies of PBMC-derived CD8+ T cells, specific to VP11/1266–74, VP11/12220–228 and VP11/12702–710 epitopes, detected from ten HLA-A*02:01 positive HSV-1 seropositive ASYMP individuals compared to ten HLA-A*02:01 positive HSV-seropositive SYMP individuals. The results are representative of 2 independent experiments in each individual. The indicated P values, calculated using one-way ANOVA Test, show statistical significance between SYMP and ASYMP individuals.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: HLA-A02:01-Restricted Epitopes Identified from the Herpes Simplex Virus Tegument Protein VP11/12 Preferentially Recall Polyfunctional Effector Memory CD8 + T Cells from Seropositive Asymptomatic Individuals and Protect “Humanized” HLA-A*02:01 Transgenic Mice Against Ocular Herpes

doi: 10.4049/jimmunol.1402606

Figure Lengend Snippet: PBMCs (~10 × 106) derived from ten HLA-A*02:01 positive HSV-1 seropositive ASYMP individuals and from ten HLA-A*02:01 positive HSV-1 seropositive SYMP individuals were analyzed ex vivo by FACS for the frequency of CD8+ T cells specific to three VP11/12 epitopes using HLA-A*0201 VP11/12 peptide-tetramer complexes representing each of the three high to medium peptide binders (VP11/1266–74, VP11/12220–228 and VP11/12702–710 epitopes), as shown in Fig. 1. (A) Representative FACS data of the frequencies of CD8+ T cells, specific to VP11/1266–74, VP11/12220–228 and VP11/12702–710 epitopes, detected in PBMCs from one HLA-A*02:01 positive HSV-1 seropositive ASYMP individual (top panels) and one HLA-A*02:01 positive HSV-1 seropositive SYMP individual (lower panels). (B) Average frequencies of PBMC-derived CD8+ T cells, specific to VP11/1266–74, VP11/12220–228 and VP11/12702–710 epitopes, detected from ten HLA-A*02:01 positive HSV-1 seropositive ASYMP individuals compared to ten HLA-A*02:01 positive HSV-seropositive SYMP individuals. The results are representative of 2 independent experiments in each individual. The indicated P values, calculated using one-way ANOVA Test, show statistical significance between SYMP and ASYMP individuals.

Article Snippet: Anti-mouse antibodies were: CD8 (clone 53-6.7) PE-Cy7, CD44 (clone IM7) APC-Cy7, CD62L (MEL-14) A700, CD107a (clone 1D4B) FITC, CD107b (clone Ha1/29) FITC, IFN-γ (clone XMG1.2) A700 (BD Pharmingen); Granzyme B (clone GB11) A647, (BioLegend).

Techniques: Derivative Assay, Ex Vivo

The phenotype of CD8+ T cells specific to VP11/12 peptide/Tetramer complexes representing each of the two immunodominant VP11/12220–228 and VP11/12702–710 epitopes were analyzed in term of TEM and TCM cells. (A) Representative FACS data of the frequencies of CD44highCD62LlowCD8+ TEM cells and CD44highCD62LhighCD8+ TCM cells specific to VP11/12220–228 peptide/Tetramer complexes from one HLA-A*02:01 positive HSV-1 seropositive ASYMP individual and from one HLA-A*02:01 positive HSV-1 seropositive SYMP individual. (B) Average frequencies of blood-derived CD8+ T cells specific to VP11/12220–228 peptide/Tetramer complexes with either TEM or TCM phenotype analyzed from 10 ASYMP and 10 SYMP individuals. (C) Representative data of the frequencies of CD44highCD62LlowCD8+ TEM and CD44highCD62LhighCD8+ TCM cells specific to VP11/12702–710 epitope from one ASYMP individual and one SYMP individual. (D) Average frequencies of blood-derived CD8+ T cells specific to VP11/12702–710 epitope with either TEM or TCM phenotype analyzed from 10 ASYMP and 10 SYMP individuals. Representative FACS data of the frequencies of CD45RAlowCCR7lowCD8+ TEM cells and CD45RAlowCCR7highCD8+ TCM cells, specific to (E) VP11/12220–228 epitope and (F) VP11/12702–710 epitope, analyzed in one ASYMP individual and one SYMP individual. Average frequencies of PBMC-derived CD8+ T cells with either TEM or TCM phenotype, specific to (G) VP11/12220–228 epitope and (H) VP11/12702–710 epitope, analyzed from 10 ASYMP and 10 SYMP individuals. The results are representative of 2 independent experiments in each individual. The indicated P values, calculated using one-way ANOVA Test, show statistical significance between SYMP and ASYMP individuals.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: HLA-A02:01-Restricted Epitopes Identified from the Herpes Simplex Virus Tegument Protein VP11/12 Preferentially Recall Polyfunctional Effector Memory CD8 + T Cells from Seropositive Asymptomatic Individuals and Protect “Humanized” HLA-A*02:01 Transgenic Mice Against Ocular Herpes

doi: 10.4049/jimmunol.1402606

Figure Lengend Snippet: The phenotype of CD8+ T cells specific to VP11/12 peptide/Tetramer complexes representing each of the two immunodominant VP11/12220–228 and VP11/12702–710 epitopes were analyzed in term of TEM and TCM cells. (A) Representative FACS data of the frequencies of CD44highCD62LlowCD8+ TEM cells and CD44highCD62LhighCD8+ TCM cells specific to VP11/12220–228 peptide/Tetramer complexes from one HLA-A*02:01 positive HSV-1 seropositive ASYMP individual and from one HLA-A*02:01 positive HSV-1 seropositive SYMP individual. (B) Average frequencies of blood-derived CD8+ T cells specific to VP11/12220–228 peptide/Tetramer complexes with either TEM or TCM phenotype analyzed from 10 ASYMP and 10 SYMP individuals. (C) Representative data of the frequencies of CD44highCD62LlowCD8+ TEM and CD44highCD62LhighCD8+ TCM cells specific to VP11/12702–710 epitope from one ASYMP individual and one SYMP individual. (D) Average frequencies of blood-derived CD8+ T cells specific to VP11/12702–710 epitope with either TEM or TCM phenotype analyzed from 10 ASYMP and 10 SYMP individuals. Representative FACS data of the frequencies of CD45RAlowCCR7lowCD8+ TEM cells and CD45RAlowCCR7highCD8+ TCM cells, specific to (E) VP11/12220–228 epitope and (F) VP11/12702–710 epitope, analyzed in one ASYMP individual and one SYMP individual. Average frequencies of PBMC-derived CD8+ T cells with either TEM or TCM phenotype, specific to (G) VP11/12220–228 epitope and (H) VP11/12702–710 epitope, analyzed from 10 ASYMP and 10 SYMP individuals. The results are representative of 2 independent experiments in each individual. The indicated P values, calculated using one-way ANOVA Test, show statistical significance between SYMP and ASYMP individuals.

Article Snippet: Anti-mouse antibodies were: CD8 (clone 53-6.7) PE-Cy7, CD44 (clone IM7) APC-Cy7, CD62L (MEL-14) A700, CD107a (clone 1D4B) FITC, CD107b (clone Ha1/29) FITC, IFN-γ (clone XMG1.2) A700 (BD Pharmingen); Granzyme B (clone GB11) A647, (BioLegend).

Techniques: Derivative Assay

(A) VP11/12 epitope-primed CD8+ T cells from ASYMP individuals express high level of lytic granules compared to CD8+ T cells from SYMP individuals. FACS was used to determine the level of expression of GzmB, GzmK and PFN on tetramer gated CD8+ T cells specific to VP11/1266–74, VP11/12220–228 and VP11/12702–710 epitopes, as described in Materials & Methods. Samples were acquired on BD LSR II and data analysis was performed using FlowJo. The numbers on the top of each histogram represent mean fluorescent intensity (MFI) depicting the level of expression of each cytotoxic molecule. Numbers in bold represent mean fluorescent intensity (MFI) of ASYMP individual. (B) Representative FACS data of VP11/1266–74, VP11/12220–228 and VP11/12702–710 epitope-specific CD107a/bhighCD8+ T cells from one ASYMP vs. one SYMP individual. Average percentage (C) and average absolute number (D) of VP11/12220–228 epitope-specific CD107a/bhighCD8+ T cells from 10 ASYMP and 10 SYMP individuals. (E) Representative FACS data of VP11/1266–74, VP11/12220–228 and VP11/12702–710 epitope-specific IFN-γhighCD8+ T cells from one ASYMP vs. one SYMP individual. Average percentage (F) and average absolute number (G) of VP11/12220–228 epitope-specific IFN-γhighCD8+ T cells from 10 ASYMP and 10 SYMP individuals. (H) Summary pie charts showing the average amount of each cytokine produced by CD8+ T-cells from ASYMP patients (n = 10, black) and SYMP patients (n = 8, white), as detected by Luminex assay. The average frequencies of cytokine-producing CD8+ T cells from SYMP and ASYMP individuals are shown inside each pie chart. (I) Each pie chart represents the overall mean of CD8+ T cell functions from 10 HLA-A*02:01-positive ASYMP and 10 SYMP individuals in responses to stimulation with either SYMP or ASYMP VP11/12 peptides. Each sector of the pie chart represents the number of CD8+ T cell functions produced.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: HLA-A02:01-Restricted Epitopes Identified from the Herpes Simplex Virus Tegument Protein VP11/12 Preferentially Recall Polyfunctional Effector Memory CD8 + T Cells from Seropositive Asymptomatic Individuals and Protect “Humanized” HLA-A*02:01 Transgenic Mice Against Ocular Herpes

doi: 10.4049/jimmunol.1402606

Figure Lengend Snippet: (A) VP11/12 epitope-primed CD8+ T cells from ASYMP individuals express high level of lytic granules compared to CD8+ T cells from SYMP individuals. FACS was used to determine the level of expression of GzmB, GzmK and PFN on tetramer gated CD8+ T cells specific to VP11/1266–74, VP11/12220–228 and VP11/12702–710 epitopes, as described in Materials & Methods. Samples were acquired on BD LSR II and data analysis was performed using FlowJo. The numbers on the top of each histogram represent mean fluorescent intensity (MFI) depicting the level of expression of each cytotoxic molecule. Numbers in bold represent mean fluorescent intensity (MFI) of ASYMP individual. (B) Representative FACS data of VP11/1266–74, VP11/12220–228 and VP11/12702–710 epitope-specific CD107a/bhighCD8+ T cells from one ASYMP vs. one SYMP individual. Average percentage (C) and average absolute number (D) of VP11/12220–228 epitope-specific CD107a/bhighCD8+ T cells from 10 ASYMP and 10 SYMP individuals. (E) Representative FACS data of VP11/1266–74, VP11/12220–228 and VP11/12702–710 epitope-specific IFN-γhighCD8+ T cells from one ASYMP vs. one SYMP individual. Average percentage (F) and average absolute number (G) of VP11/12220–228 epitope-specific IFN-γhighCD8+ T cells from 10 ASYMP and 10 SYMP individuals. (H) Summary pie charts showing the average amount of each cytokine produced by CD8+ T-cells from ASYMP patients (n = 10, black) and SYMP patients (n = 8, white), as detected by Luminex assay. The average frequencies of cytokine-producing CD8+ T cells from SYMP and ASYMP individuals are shown inside each pie chart. (I) Each pie chart represents the overall mean of CD8+ T cell functions from 10 HLA-A*02:01-positive ASYMP and 10 SYMP individuals in responses to stimulation with either SYMP or ASYMP VP11/12 peptides. Each sector of the pie chart represents the number of CD8+ T cell functions produced.

Article Snippet: Anti-mouse antibodies were: CD8 (clone 53-6.7) PE-Cy7, CD44 (clone IM7) APC-Cy7, CD62L (MEL-14) A700, CD107a (clone 1D4B) FITC, CD107b (clone Ha1/29) FITC, IFN-γ (clone XMG1.2) A700 (BD Pharmingen); Granzyme B (clone GB11) A647, (BioLegend).

Techniques: Expressing, Produced, Luminex

Representative (A) and average frequencies (B) of VP220–228 epitope-specific CD8+ T cells from 10 ASYMP and vs. 10 SYMP individuals that are CD27highCD28high. Representative (C) and average frequencies (D) of VP220–228 epitope-specific CD8+ T cells from 10 ASYMP and vs. 10 SYMP individuals that are PD-1high. (E and H) The expression patterns of T-bet and Eomes transcription factors was analyzed, at RNA level using RT-PCR, from total CD8+ T cells derived from either SYMP (open square) or ASYMP individuals (closed square). Representative FACS data of the percentage of Eomes (F) and T-bet (I) positive HSV-1 VP11/12220–228 epitope-specific CD8+ T cells, derived from one SYMP individual (right) and one ASYMP individual (left). Average frequencies of the expression patterns of Eomes (G) and T-bet (J) transcription factors analyzed by FACS at the protein level from gated VP11/12220–228-specific CD8+ TEM cells derived from 10 SYMP (open circles) and 10 ASYMP individuals (closed circles). The results are representative of 3 independent experiments in each individual. The indicated P values, calculated using one-way ANOVA Test, show statistical significance between SYMP and ASYMP individuals.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: HLA-A02:01-Restricted Epitopes Identified from the Herpes Simplex Virus Tegument Protein VP11/12 Preferentially Recall Polyfunctional Effector Memory CD8 + T Cells from Seropositive Asymptomatic Individuals and Protect “Humanized” HLA-A*02:01 Transgenic Mice Against Ocular Herpes

doi: 10.4049/jimmunol.1402606

Figure Lengend Snippet: Representative (A) and average frequencies (B) of VP220–228 epitope-specific CD8+ T cells from 10 ASYMP and vs. 10 SYMP individuals that are CD27highCD28high. Representative (C) and average frequencies (D) of VP220–228 epitope-specific CD8+ T cells from 10 ASYMP and vs. 10 SYMP individuals that are PD-1high. (E and H) The expression patterns of T-bet and Eomes transcription factors was analyzed, at RNA level using RT-PCR, from total CD8+ T cells derived from either SYMP (open square) or ASYMP individuals (closed square). Representative FACS data of the percentage of Eomes (F) and T-bet (I) positive HSV-1 VP11/12220–228 epitope-specific CD8+ T cells, derived from one SYMP individual (right) and one ASYMP individual (left). Average frequencies of the expression patterns of Eomes (G) and T-bet (J) transcription factors analyzed by FACS at the protein level from gated VP11/12220–228-specific CD8+ TEM cells derived from 10 SYMP (open circles) and 10 ASYMP individuals (closed circles). The results are representative of 3 independent experiments in each individual. The indicated P values, calculated using one-way ANOVA Test, show statistical significance between SYMP and ASYMP individuals.

Article Snippet: Anti-mouse antibodies were: CD8 (clone 53-6.7) PE-Cy7, CD44 (clone IM7) APC-Cy7, CD62L (MEL-14) A700, CD107a (clone 1D4B) FITC, CD107b (clone Ha1/29) FITC, IFN-γ (clone XMG1.2) A700 (BD Pharmingen); Granzyme B (clone GB11) A647, (BioLegend).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Derivative Assay

Three groups of age-matched male HLA Tg mice (n = 10 each) were immunized subcutaneously, on days 0 and 21, with a mixture of three human CD8+ TEM cell epitope peptides (VP11/66–74, VP11/12220–228 and VP11/12702–710) delivered either with a novel CD4+ T cell epitope (VP11/12129–143) emulsified in CpG1826 adjuvant (Group 1) or with the promiscuous CD4+ T-cell PADRE epitope peptide emulsified in CpG1826 adjuvant (Group 2). CpG1826 adjuvant alone was used as control (Group 3, mock-vaccinated). Two weeks after the final immunization, all animals were challenged ocularly with 2 × 105 pfu of HSV-1 (strain McKrae). The eye disease was detected and scored two weeks after immunization as described in Material & Methods (A and B). Virus titrations were determined from eye swabs on day 7 post-infection, as described in Material & Methods (C). Survival was determined in a window of 30 days post-challenge, as described in Material & Methods (D). The results are representative of 2 independent experiments. The indicated P values, calculated using one-way ANOVA Test, show statistical significance between protected (ASYMP) and non-protected (SYMP) mice.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: HLA-A02:01-Restricted Epitopes Identified from the Herpes Simplex Virus Tegument Protein VP11/12 Preferentially Recall Polyfunctional Effector Memory CD8 + T Cells from Seropositive Asymptomatic Individuals and Protect “Humanized” HLA-A*02:01 Transgenic Mice Against Ocular Herpes

doi: 10.4049/jimmunol.1402606

Figure Lengend Snippet: Three groups of age-matched male HLA Tg mice (n = 10 each) were immunized subcutaneously, on days 0 and 21, with a mixture of three human CD8+ TEM cell epitope peptides (VP11/66–74, VP11/12220–228 and VP11/12702–710) delivered either with a novel CD4+ T cell epitope (VP11/12129–143) emulsified in CpG1826 adjuvant (Group 1) or with the promiscuous CD4+ T-cell PADRE epitope peptide emulsified in CpG1826 adjuvant (Group 2). CpG1826 adjuvant alone was used as control (Group 3, mock-vaccinated). Two weeks after the final immunization, all animals were challenged ocularly with 2 × 105 pfu of HSV-1 (strain McKrae). The eye disease was detected and scored two weeks after immunization as described in Material & Methods (A and B). Virus titrations were determined from eye swabs on day 7 post-infection, as described in Material & Methods (C). Survival was determined in a window of 30 days post-challenge, as described in Material & Methods (D). The results are representative of 2 independent experiments. The indicated P values, calculated using one-way ANOVA Test, show statistical significance between protected (ASYMP) and non-protected (SYMP) mice.

Article Snippet: Anti-mouse antibodies were: CD8 (clone 53-6.7) PE-Cy7, CD44 (clone IM7) APC-Cy7, CD62L (MEL-14) A700, CD107a (clone 1D4B) FITC, CD107b (clone Ha1/29) FITC, IFN-γ (clone XMG1.2) A700 (BD Pharmingen); Granzyme B (clone GB11) A647, (BioLegend).

Techniques: Infection

(A) Representative FACS data of the frequencies of CD44highCD62LlowCD8+ TEM and CD44highCD62LlowCD8+ TCM cells specific to VP11/12220–228 peptide/Tetramer complexes detected the corneas of one protected “ASYMP” vs. one non-protected “SYMP” HLA Tg mice. The spleen, cornea, and trigeminal ganglia were assayed for CD8+ T cell responses on day 9 post-infection. (B) Average frequencies of VP11/12220–228 epitope-specific TCM and TEM CD8+ cells detected in the corneas of five protected ASYMP and vs. five non-protected SYMP HLA Tg mice. ASYMP mice had a clinical score of 1.0 in Fig. 6B. SYMP mice had a score of 5 or 4 in Fig. 6B). Representative (C) and average frequencies (D) of VP11/1266–74 epitope-specific TCM and TEM CD8+ cells detected in the cornea five protected ASYMP and vs. five non-protected SYMP HLA Tg mice. (E) Representative data of level of expression of GzmB on cornea-derived CD8+ T cells specific to VP11/66–74, VP11/12220–228 and VP11/12702–710 detected from protected “ASYMP” mice (black histogram) and non-protected “SYMP” mice (white histogram). (F) Average frequencies of GzmB(+)CD8+ T cells specific to VP11/66–74, VP11/12220–228 and VP11/12702–710 detected from the corneas of protected “ASYMP” mice (black circles) and non-protected “SYMP” mice (white circles). (G) Representative data of the % IFN-γ(+)CD8+ T cells specific to VP11/66–74, VP11/12220–228 and VP11/12702–710 detected from the corneas of protected “ASYMP” mice (right) and non-protected “SYMP” mice (left). (H) Average frequencies of IFN-γ(+)CD8+ T cells specific to VP11/66–74, VP11/12220–228 and VP11/12702–710 detected from the corneas of protected “ASYMP” mice (black circles) and non-protected “SYMP” mice (white circles). 1×106 cells for each assay and the results are representative of 2 independent experiments. The indicated P values, calculated using one-way ANOVA Test, show statistical significance between SYMP and ASYMP mice.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: HLA-A02:01-Restricted Epitopes Identified from the Herpes Simplex Virus Tegument Protein VP11/12 Preferentially Recall Polyfunctional Effector Memory CD8 + T Cells from Seropositive Asymptomatic Individuals and Protect “Humanized” HLA-A*02:01 Transgenic Mice Against Ocular Herpes

doi: 10.4049/jimmunol.1402606

Figure Lengend Snippet: (A) Representative FACS data of the frequencies of CD44highCD62LlowCD8+ TEM and CD44highCD62LlowCD8+ TCM cells specific to VP11/12220–228 peptide/Tetramer complexes detected the corneas of one protected “ASYMP” vs. one non-protected “SYMP” HLA Tg mice. The spleen, cornea, and trigeminal ganglia were assayed for CD8+ T cell responses on day 9 post-infection. (B) Average frequencies of VP11/12220–228 epitope-specific TCM and TEM CD8+ cells detected in the corneas of five protected ASYMP and vs. five non-protected SYMP HLA Tg mice. ASYMP mice had a clinical score of 1.0 in Fig. 6B. SYMP mice had a score of 5 or 4 in Fig. 6B). Representative (C) and average frequencies (D) of VP11/1266–74 epitope-specific TCM and TEM CD8+ cells detected in the cornea five protected ASYMP and vs. five non-protected SYMP HLA Tg mice. (E) Representative data of level of expression of GzmB on cornea-derived CD8+ T cells specific to VP11/66–74, VP11/12220–228 and VP11/12702–710 detected from protected “ASYMP” mice (black histogram) and non-protected “SYMP” mice (white histogram). (F) Average frequencies of GzmB(+)CD8+ T cells specific to VP11/66–74, VP11/12220–228 and VP11/12702–710 detected from the corneas of protected “ASYMP” mice (black circles) and non-protected “SYMP” mice (white circles). (G) Representative data of the % IFN-γ(+)CD8+ T cells specific to VP11/66–74, VP11/12220–228 and VP11/12702–710 detected from the corneas of protected “ASYMP” mice (right) and non-protected “SYMP” mice (left). (H) Average frequencies of IFN-γ(+)CD8+ T cells specific to VP11/66–74, VP11/12220–228 and VP11/12702–710 detected from the corneas of protected “ASYMP” mice (black circles) and non-protected “SYMP” mice (white circles). 1×106 cells for each assay and the results are representative of 2 independent experiments. The indicated P values, calculated using one-way ANOVA Test, show statistical significance between SYMP and ASYMP mice.

Article Snippet: Anti-mouse antibodies were: CD8 (clone 53-6.7) PE-Cy7, CD44 (clone IM7) APC-Cy7, CD62L (MEL-14) A700, CD107a (clone 1D4B) FITC, CD107b (clone Ha1/29) FITC, IFN-γ (clone XMG1.2) A700 (BD Pharmingen); Granzyme B (clone GB11) A647, (BioLegend).

Techniques: Infection, Expressing, Derivative Assay

Expression and dynamic changes in CTLA-4 in T cells of SIV-infected ChRMs. Twelve Chinese rhesus macaques were infected with SIVmac239 as described by Tian et al. ( <xref ref-type=Tian et al., 2018 ). PBMCs from monkeys before infecti on (Normal, n = 12), at acute infection stage (Acute, n = 12), at early chronic infection stage (Early chronic, n = 6), at chronic infection (Chronic, n = 5) and AIDS stages (AIDS, n = 6, time point=3) were used for CTLA-4 detecting. Intracellular CTLA-4 was stained with fluorescent antibodies and detected by flow cytometry. Frequencies of CTLA-4+ CD4+ T cell subsets (A) and CTLA-4+ CD8+ T cell subsets (B), and dynamic changes in CTLA-4 in CD4+ T cells (C) and CD8+ T cells (D) are shown. Single independent experiment was including for each animal and each time point. * P < 0.05, ** P < 0.01, *** P < 0.001. " width="100%" height="100%">

Journal: Virus Research

Article Title: Increased cAMP-PKA signaling pathway activation is involved in up-regulation of CTLA-4 expression in CD4+ T cells in acute SIVmac239-infected Chinese rhesus macaques

doi: 10.1016/j.virusres.2024.199313

Figure Lengend Snippet: Expression and dynamic changes in CTLA-4 in T cells of SIV-infected ChRMs. Twelve Chinese rhesus macaques were infected with SIVmac239 as described by Tian et al. ( Tian et al., 2018 ). PBMCs from monkeys before infecti on (Normal, n = 12), at acute infection stage (Acute, n = 12), at early chronic infection stage (Early chronic, n = 6), at chronic infection (Chronic, n = 5) and AIDS stages (AIDS, n = 6, time point=3) were used for CTLA-4 detecting. Intracellular CTLA-4 was stained with fluorescent antibodies and detected by flow cytometry. Frequencies of CTLA-4+ CD4+ T cell subsets (A) and CTLA-4+ CD8+ T cell subsets (B), and dynamic changes in CTLA-4 in CD4+ T cells (C) and CD8+ T cells (D) are shown. Single independent experiment was including for each animal and each time point. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The following anti-human flow cytometry antibodies cross-reactive with macaques were used: anti-CD3 APC—Cy7 (clone SP34–2), anti-CD8 PE-Cy7 (clone PRA-T8), anti-CD28 APC (clone CD28.8), anti-CD95 FITC (clone DX2) and anti-Ki67 PE (clone B56) were purchased from BD; anti-CD4 PerCP-Cy5.5 (clone OKT4), anti-CTLA-4 PE (clone BNI3), anti-HLA-DR APC (clone L243), and anti-PD-1 PE (clone EH12.2H7).

Techniques: Expressing, Infection, Staining, Flow Cytometry

Correlations between CTLA-4 levels and disease progression markers. Data corresponding to the time points of CTLA-4 detecting were extracted. Correlations between frequency of CTLA-4+CD4+ T cells and viral load (A), CD4+ T cell number (B), CD4/CD8 ratio (C), HLA-DR+CD38+CD4+ T cells (D), HLA-DR+CD38+CD8+ T cells (E), Ki67+CD4+ T cells (F), and Ki67+CD8+ T cells (G) were assessed using Pearson and Spearman rank correlation analyses.

Journal: Virus Research

Article Title: Increased cAMP-PKA signaling pathway activation is involved in up-regulation of CTLA-4 expression in CD4+ T cells in acute SIVmac239-infected Chinese rhesus macaques

doi: 10.1016/j.virusres.2024.199313

Figure Lengend Snippet: Correlations between CTLA-4 levels and disease progression markers. Data corresponding to the time points of CTLA-4 detecting were extracted. Correlations between frequency of CTLA-4+CD4+ T cells and viral load (A), CD4+ T cell number (B), CD4/CD8 ratio (C), HLA-DR+CD38+CD4+ T cells (D), HLA-DR+CD38+CD8+ T cells (E), Ki67+CD4+ T cells (F), and Ki67+CD8+ T cells (G) were assessed using Pearson and Spearman rank correlation analyses.

Article Snippet: The following anti-human flow cytometry antibodies cross-reactive with macaques were used: anti-CD3 APC—Cy7 (clone SP34–2), anti-CD8 PE-Cy7 (clone PRA-T8), anti-CD28 APC (clone CD28.8), anti-CD95 FITC (clone DX2) and anti-Ki67 PE (clone B56) were purchased from BD; anti-CD4 PerCP-Cy5.5 (clone OKT4), anti-CTLA-4 PE (clone BNI3), anti-HLA-DR APC (clone L243), and anti-PD-1 PE (clone EH12.2H7).

Techniques:

Effects of selective cAMP-phosphodiesterase inhibitor on CTLA-4 expression. Freshly separated PBMCs from four healthy monkeys were rested overnight and then incubated with cAMP-phosphodiesterase inhibitor for 48 h. Intracellular CTLA-4 was then stained with fluorescent antibodies and detected by flow cytometry. CTLA-4 expression levels in CD4+ T cells (A) and CD8+ T cells (B) are shown ( n = 4). Each study contained three independent replicates. And single independent measurement was included for each sample. One representative result was displayed. All data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001.

Journal: Virus Research

Article Title: Increased cAMP-PKA signaling pathway activation is involved in up-regulation of CTLA-4 expression in CD4+ T cells in acute SIVmac239-infected Chinese rhesus macaques

doi: 10.1016/j.virusres.2024.199313

Figure Lengend Snippet: Effects of selective cAMP-phosphodiesterase inhibitor on CTLA-4 expression. Freshly separated PBMCs from four healthy monkeys were rested overnight and then incubated with cAMP-phosphodiesterase inhibitor for 48 h. Intracellular CTLA-4 was then stained with fluorescent antibodies and detected by flow cytometry. CTLA-4 expression levels in CD4+ T cells (A) and CD8+ T cells (B) are shown ( n = 4). Each study contained three independent replicates. And single independent measurement was included for each sample. One representative result was displayed. All data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The following anti-human flow cytometry antibodies cross-reactive with macaques were used: anti-CD3 APC—Cy7 (clone SP34–2), anti-CD8 PE-Cy7 (clone PRA-T8), anti-CD28 APC (clone CD28.8), anti-CD95 FITC (clone DX2) and anti-Ki67 PE (clone B56) were purchased from BD; anti-CD4 PerCP-Cy5.5 (clone OKT4), anti-CTLA-4 PE (clone BNI3), anti-HLA-DR APC (clone L243), and anti-PD-1 PE (clone EH12.2H7).

Techniques: Expressing, Incubation, Staining, Flow Cytometry

Human lymphocytes are susceptible to DV infection and replication. (A) Representative flow cytometry density plots of purified cells showing the frequencies of CD4+ and CD8+ T cells infected by DV. The number within each box refers to the frequency of cells within each gate. (B and C) Average frequencies of CD4+ and CD8+ T cell infection by the four DV serotypes. Mock infection (circles) and DV1 BR90 (squares), DV2 265 (triangles), DV3 98 (inverted triangles), and DV4 360 (diamonds) infection. Bars show the mean values for samples from six healthy donors. *, P ≤ 0.05 compared to the results for infection by four DV serotypes. (D and E) Confocal fluorescence microscopy of CD4+ and CD8+ T cells labeled for DNA (DAPI; blue), DV (anti-E protein MAb; red), and immunophenotyping markers (anti-CD4 or anti-CD8 MAb; green). Scale bar = 10 μm. (A, D, E) Data are representative of six independent experiments. (F and G) Flow cytometry data indicating the average frequencies of DV-infected CD4+ (F) and CD8+ (G) T lymphocytes (CD14+-depleted cells from six healthy donors) after virus treatment with different concentrations of heparin (2 to 20 μg). (H and I) Average frequencies, using a flow cytometry assay, of CD4+ (H) and CD8+ (I) T lymphocytes infected with DV1 to DV4 after cell treatment with different concentrations of heparinase III (0.5 to 5.0 IU) and infection with the four DV serotypes. Data represent the mean values ± standard errors of the means (SEM) for cells from six healthy donors. *, P ≤ 0.05 compared to the results for mock-infected controls. (J and M) DV progeny in cell culture supernatants from purified CD4+ (J) and CD8+ (M) T lymphocytes infected with the four DV serotypes were quantified using a focus-forming assay in C6/36 cells. (K and N) DV replication on CD4+ (K) and CD8+ (N) T lymphocytes (purified via cell sorting) was determined by real-time PCR assay. Quantification of DV nonstructural protein 3 (NS3) gene mRNA, normalized to the housekeeping gene 18S, for comparison of the levels in infected cells and mock-infected cells. (L and O) Qualitative determination of the levels of DV nonstructural protein 1 (NS1) in cell culture supernatants of purified CD4+ (L) and CD8+ (O) T lymphocytes infected by DV by using a capture ELISA from PanBio (values >11 indicate NS1 secretion). Mock infected (circles) and DV1 BR90 (squares), DV2 265 (triangles), DV3 98 (inverted triangles), and DV4 360 (diamonds) infected. Bars show the mean values for cells from six healthy donors. *, P ≤ 0.05 compared to the results for infection by four DV serotypes.

Journal: Journal of Virology

Article Title: Human T Lymphocytes Are Permissive for Dengue Virus Replication

doi: 10.1128/JVI.02181-17

Figure Lengend Snippet: Human lymphocytes are susceptible to DV infection and replication. (A) Representative flow cytometry density plots of purified cells showing the frequencies of CD4+ and CD8+ T cells infected by DV. The number within each box refers to the frequency of cells within each gate. (B and C) Average frequencies of CD4+ and CD8+ T cell infection by the four DV serotypes. Mock infection (circles) and DV1 BR90 (squares), DV2 265 (triangles), DV3 98 (inverted triangles), and DV4 360 (diamonds) infection. Bars show the mean values for samples from six healthy donors. *, P ≤ 0.05 compared to the results for infection by four DV serotypes. (D and E) Confocal fluorescence microscopy of CD4+ and CD8+ T cells labeled for DNA (DAPI; blue), DV (anti-E protein MAb; red), and immunophenotyping markers (anti-CD4 or anti-CD8 MAb; green). Scale bar = 10 μm. (A, D, E) Data are representative of six independent experiments. (F and G) Flow cytometry data indicating the average frequencies of DV-infected CD4+ (F) and CD8+ (G) T lymphocytes (CD14+-depleted cells from six healthy donors) after virus treatment with different concentrations of heparin (2 to 20 μg). (H and I) Average frequencies, using a flow cytometry assay, of CD4+ (H) and CD8+ (I) T lymphocytes infected with DV1 to DV4 after cell treatment with different concentrations of heparinase III (0.5 to 5.0 IU) and infection with the four DV serotypes. Data represent the mean values ± standard errors of the means (SEM) for cells from six healthy donors. *, P ≤ 0.05 compared to the results for mock-infected controls. (J and M) DV progeny in cell culture supernatants from purified CD4+ (J) and CD8+ (M) T lymphocytes infected with the four DV serotypes were quantified using a focus-forming assay in C6/36 cells. (K and N) DV replication on CD4+ (K) and CD8+ (N) T lymphocytes (purified via cell sorting) was determined by real-time PCR assay. Quantification of DV nonstructural protein 3 (NS3) gene mRNA, normalized to the housekeeping gene 18S, for comparison of the levels in infected cells and mock-infected cells. (L and O) Qualitative determination of the levels of DV nonstructural protein 1 (NS1) in cell culture supernatants of purified CD4+ (L) and CD8+ (O) T lymphocytes infected by DV by using a capture ELISA from PanBio (values >11 indicate NS1 secretion). Mock infected (circles) and DV1 BR90 (squares), DV2 265 (triangles), DV3 98 (inverted triangles), and DV4 360 (diamonds) infected. Bars show the mean values for cells from six healthy donors. *, P ≤ 0.05 compared to the results for infection by four DV serotypes.

Article Snippet: Additionally, DV infection of lymphocytes was measured using the 4G2 antibody together with the phenotypic markers anti-CD3-APC-H7 MAb (clone SK7), anti-CD4-AmCyan MAb (clone RPA-T4), and anti-CD8-PE-Cy7 MAb (clone SK1) (all antibodies from BD Biosciences).

Techniques: Infection, Flow Cytometry, Purification, Fluorescence, Microscopy, Labeling, Cell Culture, Focus Forming Assay, FACS, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

CD4+ and CD8+ T lymphocytes are naturally infected by DV in acute-phase patients. (A, C, E, and G) Representative flow cytometry density data showing CD4+ (A) and CD8+ (C) T lymphocyte, B cell (CD19+) (E), and monocyte (CD14+) (G) infection by DV1 in PBMCs from an acute-phase patient. Lymphocyte infection was observed by labeling the viral proteins with anti-flavivirus E protein MAb (4G2), anti-DV NS3 MAb (1722-1B), or an isotype control (anti-GFP MAb). The number within each box refers to the frequency of cells within each gate. Phenotype staining was performed using anti-CD3, anti-CD4, anti-CD8, anti-CD19, and anti-CD14 MAbs. Cell infection was shown using PBMCs from a patient negative for DV infection (PBMC#13) and a patient infected with DV1 (PBMC#3). (B, D, F, and H) Average frequencies of CD4+ (B) and CD8+ (D) T lymphocytes, B cells (F), and monocytes (H) infected by DV (anti-DV NS3 and anti-DV E MAbs) in PBMCs from seven negative (circles) and six DV-infected patients (squares). α, P ≤ 0.05 comparing the results for anti-DV MAb (4G2 or 1722-1B) staining in PBMCs from DV-infected and noninfected patients; β, P ≤ 0.05 comparing the results for staining with 4G2 or 1722-1B and the isotype control (anti-GFP MAb).

Journal: Journal of Virology

Article Title: Human T Lymphocytes Are Permissive for Dengue Virus Replication

doi: 10.1128/JVI.02181-17

Figure Lengend Snippet: CD4+ and CD8+ T lymphocytes are naturally infected by DV in acute-phase patients. (A, C, E, and G) Representative flow cytometry density data showing CD4+ (A) and CD8+ (C) T lymphocyte, B cell (CD19+) (E), and monocyte (CD14+) (G) infection by DV1 in PBMCs from an acute-phase patient. Lymphocyte infection was observed by labeling the viral proteins with anti-flavivirus E protein MAb (4G2), anti-DV NS3 MAb (1722-1B), or an isotype control (anti-GFP MAb). The number within each box refers to the frequency of cells within each gate. Phenotype staining was performed using anti-CD3, anti-CD4, anti-CD8, anti-CD19, and anti-CD14 MAbs. Cell infection was shown using PBMCs from a patient negative for DV infection (PBMC#13) and a patient infected with DV1 (PBMC#3). (B, D, F, and H) Average frequencies of CD4+ (B) and CD8+ (D) T lymphocytes, B cells (F), and monocytes (H) infected by DV (anti-DV NS3 and anti-DV E MAbs) in PBMCs from seven negative (circles) and six DV-infected patients (squares). α, P ≤ 0.05 comparing the results for anti-DV MAb (4G2 or 1722-1B) staining in PBMCs from DV-infected and noninfected patients; β, P ≤ 0.05 comparing the results for staining with 4G2 or 1722-1B and the isotype control (anti-GFP MAb).

Article Snippet: Additionally, DV infection of lymphocytes was measured using the 4G2 antibody together with the phenotypic markers anti-CD3-APC-H7 MAb (clone SK7), anti-CD4-AmCyan MAb (clone RPA-T4), and anti-CD8-PE-Cy7 MAb (clone SK1) (all antibodies from BD Biosciences).

Techniques: Infection, Flow Cytometry, Labeling, Staining