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Induction of pluripotent protective immunity following immunisation with a chimeric vaccine against human cytomegalovirus.

Zhong J, Rist M, Cooper L, Smith C, Khanna R - PLoS ONE (2008)

Bottom Line: In spite of numerous attempts successful licensure of a HCMV vaccine formulation remains elusive.More importantly, immunization with adenoviral chimeric vaccine also afforded protection against challenge with recombinant vaccinia virus encoding HCMV antigens and this protection was associated with the induction of a pluripotent antigen-specific cellular and antibody response.These studies demonstrate that the adenovirus chimeric HCMV vaccine provides an excellent platform for reconstituting protective immunity to prevent HCMV diseases in different clinical settings.

View Article: PubMed Central - PubMed

Affiliation: Australian Centre for Vaccine Development, Tumour Immunology Laboratory, Division of Immunology, Queensland Institute of Medical Research, Brisbane, Australia.

ABSTRACT
Based on the life-time cost to the health care system, the Institute of Medicine has assigned the highest priority for a vaccine to control human cytomegalovirus (HCMV) disease in transplant patients and new born babies. In spite of numerous attempts successful licensure of a HCMV vaccine formulation remains elusive. Here we have developed a novel chimeric vaccine strategy based on a replication-deficient adenovirus which encodes the extracellular domain of gB protein and multiple HLA class I & II-restricted CTL epitopes from HCMV as a contiguous polypeptide. Immunisation with this chimeric vaccine consistently generated strong HCMV-specific CD8(+) and CD4(+) T-cells which co-expressed IFN-gamma and TNF-alpha, while the humoral response induced by this vaccine showed strong virus neutralizing capacity. More importantly, immunization with adenoviral chimeric vaccine also afforded protection against challenge with recombinant vaccinia virus encoding HCMV antigens and this protection was associated with the induction of a pluripotent antigen-specific cellular and antibody response. Furthermore, in vitro stimulation with this adenoviral chimeric vaccine rapidly expanded multiple antigen-specific human CD8(+) and CD4(+) T-cells from healthy virus carriers. These studies demonstrate that the adenovirus chimeric HCMV vaccine provides an excellent platform for reconstituting protective immunity to prevent HCMV diseases in different clinical settings.

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Related in: MedlinePlus

Expansion of gB-specific T cells following in vitro stimulation of human PBMC with Ad-gBCMVpoly.PBMC from a panel of healthy virus carriers (referred to as D1–D17) were co-cultured with autologous PBMC infected with Ad-gBCMVpoly (MOI: 5∶1 or 1∶1) at a responder to stimulator ratio of 2∶1. These cultures were supplemented with rIL-2 (10 U/ml) on day 3 and every 3–4 days thereafter. On day 14, these T cell cultures were tested against a panel of pooled overlapping gB peptides (20 aa long, overlapping by 10 aa) using intracellular cytokine assays. The data presented in the figure shows the percentage of gB-specific CD8+ and CD4+ T cell recovered from each donor following stimulation with Ad-gBCMVpoly.
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pone-0003256-g007: Expansion of gB-specific T cells following in vitro stimulation of human PBMC with Ad-gBCMVpoly.PBMC from a panel of healthy virus carriers (referred to as D1–D17) were co-cultured with autologous PBMC infected with Ad-gBCMVpoly (MOI: 5∶1 or 1∶1) at a responder to stimulator ratio of 2∶1. These cultures were supplemented with rIL-2 (10 U/ml) on day 3 and every 3–4 days thereafter. On day 14, these T cell cultures were tested against a panel of pooled overlapping gB peptides (20 aa long, overlapping by 10 aa) using intracellular cytokine assays. The data presented in the figure shows the percentage of gB-specific CD8+ and CD4+ T cell recovered from each donor following stimulation with Ad-gBCMVpoly.

Mentions: Another important aspect of the current study was aimed at exploring the potential efficacy of Ad-gBCMVpoly to recall memory T cell responses from healthy seropositive individuals. PBMC from healthy donors were stimulated with irradiated autologous PBMC-infected with Ad-gBCMVpoly. Following stimulation these T cells were assessed for antigen specificity using intracellular cytokine assays. Data for the gB-specific CD4+ and CD8+ T cell responses are summarised in Figure 7, while the T cell responses towards the epitopes within the polyepitope sequence are presented in Tables 2 & 3. To identify the gB-specific T cell responses we used an overlapping set of peptides based on the gB sequence from Ad169 strain of HCMV. This analysis showed that following stimulation with Ad-gBCMVpoly, more than 88% of the individuals showed expansion of gB-specific CD4+ T cells. These T cell expansions raged from 2–36% of the total CD3+ CD4+ T cells (Figure 7). CD8+ T cell responses directed towards gB epitopes were detected in 70.5% donors which ranged from 2–15% of the total CD3+ CD8+ T cells. T cells from each donor recognized multiple gB epitopes and most of the donors demonstrated a selective expansion of gB-specific CD4+ or CD8+ T cells.


Induction of pluripotent protective immunity following immunisation with a chimeric vaccine against human cytomegalovirus.

Zhong J, Rist M, Cooper L, Smith C, Khanna R - PLoS ONE (2008)

Expansion of gB-specific T cells following in vitro stimulation of human PBMC with Ad-gBCMVpoly.PBMC from a panel of healthy virus carriers (referred to as D1–D17) were co-cultured with autologous PBMC infected with Ad-gBCMVpoly (MOI: 5∶1 or 1∶1) at a responder to stimulator ratio of 2∶1. These cultures were supplemented with rIL-2 (10 U/ml) on day 3 and every 3–4 days thereafter. On day 14, these T cell cultures were tested against a panel of pooled overlapping gB peptides (20 aa long, overlapping by 10 aa) using intracellular cytokine assays. The data presented in the figure shows the percentage of gB-specific CD8+ and CD4+ T cell recovered from each donor following stimulation with Ad-gBCMVpoly.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2533118&req=5

pone-0003256-g007: Expansion of gB-specific T cells following in vitro stimulation of human PBMC with Ad-gBCMVpoly.PBMC from a panel of healthy virus carriers (referred to as D1–D17) were co-cultured with autologous PBMC infected with Ad-gBCMVpoly (MOI: 5∶1 or 1∶1) at a responder to stimulator ratio of 2∶1. These cultures were supplemented with rIL-2 (10 U/ml) on day 3 and every 3–4 days thereafter. On day 14, these T cell cultures were tested against a panel of pooled overlapping gB peptides (20 aa long, overlapping by 10 aa) using intracellular cytokine assays. The data presented in the figure shows the percentage of gB-specific CD8+ and CD4+ T cell recovered from each donor following stimulation with Ad-gBCMVpoly.
Mentions: Another important aspect of the current study was aimed at exploring the potential efficacy of Ad-gBCMVpoly to recall memory T cell responses from healthy seropositive individuals. PBMC from healthy donors were stimulated with irradiated autologous PBMC-infected with Ad-gBCMVpoly. Following stimulation these T cells were assessed for antigen specificity using intracellular cytokine assays. Data for the gB-specific CD4+ and CD8+ T cell responses are summarised in Figure 7, while the T cell responses towards the epitopes within the polyepitope sequence are presented in Tables 2 & 3. To identify the gB-specific T cell responses we used an overlapping set of peptides based on the gB sequence from Ad169 strain of HCMV. This analysis showed that following stimulation with Ad-gBCMVpoly, more than 88% of the individuals showed expansion of gB-specific CD4+ T cells. These T cell expansions raged from 2–36% of the total CD3+ CD4+ T cells (Figure 7). CD8+ T cell responses directed towards gB epitopes were detected in 70.5% donors which ranged from 2–15% of the total CD3+ CD8+ T cells. T cells from each donor recognized multiple gB epitopes and most of the donors demonstrated a selective expansion of gB-specific CD4+ or CD8+ T cells.

Bottom Line: In spite of numerous attempts successful licensure of a HCMV vaccine formulation remains elusive.More importantly, immunization with adenoviral chimeric vaccine also afforded protection against challenge with recombinant vaccinia virus encoding HCMV antigens and this protection was associated with the induction of a pluripotent antigen-specific cellular and antibody response.These studies demonstrate that the adenovirus chimeric HCMV vaccine provides an excellent platform for reconstituting protective immunity to prevent HCMV diseases in different clinical settings.

View Article: PubMed Central - PubMed

Affiliation: Australian Centre for Vaccine Development, Tumour Immunology Laboratory, Division of Immunology, Queensland Institute of Medical Research, Brisbane, Australia.

ABSTRACT
Based on the life-time cost to the health care system, the Institute of Medicine has assigned the highest priority for a vaccine to control human cytomegalovirus (HCMV) disease in transplant patients and new born babies. In spite of numerous attempts successful licensure of a HCMV vaccine formulation remains elusive. Here we have developed a novel chimeric vaccine strategy based on a replication-deficient adenovirus which encodes the extracellular domain of gB protein and multiple HLA class I & II-restricted CTL epitopes from HCMV as a contiguous polypeptide. Immunisation with this chimeric vaccine consistently generated strong HCMV-specific CD8(+) and CD4(+) T-cells which co-expressed IFN-gamma and TNF-alpha, while the humoral response induced by this vaccine showed strong virus neutralizing capacity. More importantly, immunization with adenoviral chimeric vaccine also afforded protection against challenge with recombinant vaccinia virus encoding HCMV antigens and this protection was associated with the induction of a pluripotent antigen-specific cellular and antibody response. Furthermore, in vitro stimulation with this adenoviral chimeric vaccine rapidly expanded multiple antigen-specific human CD8(+) and CD4(+) T-cells from healthy virus carriers. These studies demonstrate that the adenovirus chimeric HCMV vaccine provides an excellent platform for reconstituting protective immunity to prevent HCMV diseases in different clinical settings.

Show MeSH
Related in: MedlinePlus