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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.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.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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HCMV-specific effector and memory humoral and cellular immune responses following immunisation with Ad-gBCMVpoly vaccine.A, HCMV-specific CD8+ T cell responses following immunisation with Ad-gBCMVpoly. These T cell responses were assessed using ELISPOT assays on day 10, 25 and 75 post immunisation. The results are expressed as Mean±SE of spot forming cells (SFC) per 106 splenocytes. B, gB-specific antibody responses in serum samples from immunised mice on days 10, 25 and 75. Serum samples on day 0 were collected before the immunisation. C, Virus neutralizing capacity of antibody responses induced following immunisation with Ad-gBCMVpoly. Serum samples from these mice were pre-incubated with HCMV virus Ad169 and then these virus preps were used to infect MRC-5. Following overnight incubation virus infectivity was assessed using IE-1/IE-2 expression as outlined in the “Material and Methods” section. D, Avidity maturation of gB-specific antibody responses in Ad-gBCMVpoly immunised mice. E, Immunoglobulin subclass analysis of gB-specific antibody responses in HHD-2 vaccinated mice. Serum samples were collected from three different groups of mice on days 10, 25 and 75 post-immunisation. A minimum of five mice from each group were assessed for HCMV epitope-specific T cell reactivity and humoral immune responses. All statistical analyses were conducted using GraphPad Prism 4 software.
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pone-0003256-g004: HCMV-specific effector and memory humoral and cellular immune responses following immunisation with Ad-gBCMVpoly vaccine.A, HCMV-specific CD8+ T cell responses following immunisation with Ad-gBCMVpoly. These T cell responses were assessed using ELISPOT assays on day 10, 25 and 75 post immunisation. The results are expressed as Mean±SE of spot forming cells (SFC) per 106 splenocytes. B, gB-specific antibody responses in serum samples from immunised mice on days 10, 25 and 75. Serum samples on day 0 were collected before the immunisation. C, Virus neutralizing capacity of antibody responses induced following immunisation with Ad-gBCMVpoly. Serum samples from these mice were pre-incubated with HCMV virus Ad169 and then these virus preps were used to infect MRC-5. Following overnight incubation virus infectivity was assessed using IE-1/IE-2 expression as outlined in the “Material and Methods” section. D, Avidity maturation of gB-specific antibody responses in Ad-gBCMVpoly immunised mice. E, Immunoglobulin subclass analysis of gB-specific antibody responses in HHD-2 vaccinated mice. Serum samples were collected from three different groups of mice on days 10, 25 and 75 post-immunisation. A minimum of five mice from each group were assessed for HCMV epitope-specific T cell reactivity and humoral immune responses. All statistical analyses were conducted using GraphPad Prism 4 software.

Mentions: Although co-immunisation with Ad-gB and Ad-CMVpoly induced both humoral and cellular immune responses against HCMV, delivery of this formulation in a human setting may face significant regulatory constraints. To overcome this potential limitation, we constructed another recombinant adenovirus expressing the extracellular domain of gB and HCMV polyepitope as a single polypeptide (referred as Ad-gBCMVpoly). HHD-2 mice were immunised with the Ad-gBCMVpoly vaccine (7.5×108 pfu/mouse) and both humoral and cellular immune responses were evaluated at the indicated time points. Data presented in Figure 4A shows that immunisation with Ad-gBCMVpoly vaccine induced a long-tem memory CD8+ T cell response towards the HLA A2-restricted epitopes from HCMV. Furthermore these animals also showed strong gB-specific antibody response and similar to the data presented in Figure 3B, the levels of gB-specific antibody dropped by day 75 post immunisation (Figure 4B). A significant increase in the neutralizing activity of the antibody response was observed (Figure 4C), which was co-incident with avidity maturation (Figure 4D). On the other hand, there was no antibody isotype switching at different time point after immunisation (Figure 4E). These observations clearly demonstrated that covalent linking of the gB with the polyepitope sequence does not impair the immunogenicity of each of the components of the vaccine.


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)

HCMV-specific effector and memory humoral and cellular immune responses following immunisation with Ad-gBCMVpoly vaccine.A, HCMV-specific CD8+ T cell responses following immunisation with Ad-gBCMVpoly. These T cell responses were assessed using ELISPOT assays on day 10, 25 and 75 post immunisation. The results are expressed as Mean±SE of spot forming cells (SFC) per 106 splenocytes. B, gB-specific antibody responses in serum samples from immunised mice on days 10, 25 and 75. Serum samples on day 0 were collected before the immunisation. C, Virus neutralizing capacity of antibody responses induced following immunisation with Ad-gBCMVpoly. Serum samples from these mice were pre-incubated with HCMV virus Ad169 and then these virus preps were used to infect MRC-5. Following overnight incubation virus infectivity was assessed using IE-1/IE-2 expression as outlined in the “Material and Methods” section. D, Avidity maturation of gB-specific antibody responses in Ad-gBCMVpoly immunised mice. E, Immunoglobulin subclass analysis of gB-specific antibody responses in HHD-2 vaccinated mice. Serum samples were collected from three different groups of mice on days 10, 25 and 75 post-immunisation. A minimum of five mice from each group were assessed for HCMV epitope-specific T cell reactivity and humoral immune responses. All statistical analyses were conducted using GraphPad Prism 4 software.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2533118&req=5

pone-0003256-g004: HCMV-specific effector and memory humoral and cellular immune responses following immunisation with Ad-gBCMVpoly vaccine.A, HCMV-specific CD8+ T cell responses following immunisation with Ad-gBCMVpoly. These T cell responses were assessed using ELISPOT assays on day 10, 25 and 75 post immunisation. The results are expressed as Mean±SE of spot forming cells (SFC) per 106 splenocytes. B, gB-specific antibody responses in serum samples from immunised mice on days 10, 25 and 75. Serum samples on day 0 were collected before the immunisation. C, Virus neutralizing capacity of antibody responses induced following immunisation with Ad-gBCMVpoly. Serum samples from these mice were pre-incubated with HCMV virus Ad169 and then these virus preps were used to infect MRC-5. Following overnight incubation virus infectivity was assessed using IE-1/IE-2 expression as outlined in the “Material and Methods” section. D, Avidity maturation of gB-specific antibody responses in Ad-gBCMVpoly immunised mice. E, Immunoglobulin subclass analysis of gB-specific antibody responses in HHD-2 vaccinated mice. Serum samples were collected from three different groups of mice on days 10, 25 and 75 post-immunisation. A minimum of five mice from each group were assessed for HCMV epitope-specific T cell reactivity and humoral immune responses. All statistical analyses were conducted using GraphPad Prism 4 software.
Mentions: Although co-immunisation with Ad-gB and Ad-CMVpoly induced both humoral and cellular immune responses against HCMV, delivery of this formulation in a human setting may face significant regulatory constraints. To overcome this potential limitation, we constructed another recombinant adenovirus expressing the extracellular domain of gB and HCMV polyepitope as a single polypeptide (referred as Ad-gBCMVpoly). HHD-2 mice were immunised with the Ad-gBCMVpoly vaccine (7.5×108 pfu/mouse) and both humoral and cellular immune responses were evaluated at the indicated time points. Data presented in Figure 4A shows that immunisation with Ad-gBCMVpoly vaccine induced a long-tem memory CD8+ T cell response towards the HLA A2-restricted epitopes from HCMV. Furthermore these animals also showed strong gB-specific antibody response and similar to the data presented in Figure 3B, the levels of gB-specific antibody dropped by day 75 post immunisation (Figure 4B). A significant increase in the neutralizing activity of the antibody response was observed (Figure 4C), which was co-incident with avidity maturation (Figure 4D). On the other hand, there was no antibody isotype switching at different time point after immunisation (Figure 4E). These observations clearly demonstrated that covalent linking of the gB with the polyepitope sequence does not impair the immunogenicity of each of the components of the vaccine.

Bottom Line: 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.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