Quantitative and qualitative analyses of the immune responses induced by a multivalent minigene DNA vaccine

An, Ling; Rodrı́guez, Fernando; Harkins, Stephanie; Zhang, Jie; Whitton, J. Lindsay

doi:10.1016/s0264-410x(99)00546-0

Cited by 45 publications

(31 citation statements)

References 40 publications

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“…The fact that protection against secondary infection was similar in HK or live LM-primed mice suggested that the number of memory cells induced by HKLM priming was sufficient to provide protection before our ability to detect the responding cells. This finding agrees well with previous results using DNA vaccination where small numbers of memory cells are not protective but mount a robust recall response (51). As suggested in that report, memory cell proliferation may not actually be needed for protection if sufficient numbers are generated by optimal priming.…”

Section: Discussionsupporting

confidence: 93%

Sustained Response Initiation Is Required for T Cell Clonal Expansion But Not for Effector or Memory Development In Vivo

Lefrançois

Marzo

Williams

2003

The Journal of Immunology

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The factors determining whether an immune response is productive are poorly understood. To understand the circumstances affecting the early stage of the immune response which determine whether memory is generated, the CD8 T cell response was mapped in detail following immunization with live or heat-killed bacteria. Our results demonstrate that even in response to a weak immunogen, functional memory cell development is linked to effector cell induction in lymphoid and nonlymphoid tissues. The main defect in the response to killed microorganisms is inefficient induction of clonal expansion. This failure is due to a contracted, but costimulation-dependent activation phase in the lymphoid tissues, resulting in rapid but abortive growth. Conversely, the response to live bacteria is characterized by protracted early T cell sequestration in lymphoid tissues. Thus, memory development requires effector induction, while optimal clonal expansion is regulated by the duration of response initiation.

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Section: Discussionsupporting

confidence: 93%

Sustained Response Initiation Is Required for T Cell Clonal Expansion But Not for Effector or Memory Development In Vivo

Lefrançois

Marzo

Williams

2003

The Journal of Immunology

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“…The translational initiation sites in all constructs contained a 'Kozak' consensus sequence, 27 which is important for successful DNA vaccination. 28 The HIV tat PTD was also cloned into the AgeI site of the GFP expression vector peGFP-N1 (Clontech) to generate p-PTD-eGFP. The peptide representing the LCMV CD8…”

Section: Methodsmentioning

confidence: 99%

Full-length proteins attached to the HIV tat protein transduction domain are neither transduced between cells, nor exhibit enhanced immunogenicity

2002

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“…In addition, to facilitate detailed analyses of the epitopespecific CD4 ϩ T-cell responses induced by DNA vaccines, we constructed two plasmids containing minigenes encoding previously characterized LCMV epitopes which are presented by MHC class II I-A b (24). pCMV-NPTh-LII encodes NP 309-328 (SGEGWPYIACRTSIVGRAWE), and pCMV-GPTh-LII encodes GP 61-80 (GLKGPDIYKGVYQFKSVEFD); in each case the minigene ORF is preceded by an ATG start codon, flanked by a good Kozak sequence (2,16). In addition to these four plasmids encoding LIMP-II fusion proteins, four related plasmids were prepared, identical to the above but lacking the LIMP-II tail (pCMV-HBVcore, pCMV-NP, pCMV-NPTh, and pCMV-GPTh).…”

Section: Cell Lines and Viruses The Mc57 Cell Line (H-2mentioning

confidence: 99%

CD4⁺T Cells Induced by a DNA Vaccine: Immunological Consequences of Epitope-Specific Lysosomal Targeting

Rodrı́guez

Harkins

Redwine

et al. 2001

J Virol

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Our previous studies have shown that targeting DNA vaccine-encoded major histocompatibility complex class I epitopes to the proteasome enhanced CD8؉ T-cell induction and protection against lymphocytic choriomeningitis virus (LCMV) challenge. Here, we expand these studies to evaluate CD4 ؉ T-cell responses induced by DNA immunization and describe a system for targeting proteins and minigenes to lysosomes. Full-length proteins can be targeted to the lysosomal compartment by covalent attachment to the 20-aminoacid C-terminal tail of lysosomal integral membrane protein-II (LIMP-II). Using minigenes encoding defined T-helper epitopes from lymphocytic choriomeningitis virus, we show that the CD4 ؉ T-cell response induced by the NP 309-328 epitope of LCMV was greatly enhanced by addition of the LIMP-II tail. However, the immunological consequence of lysosomal targeting is not invariably positive; the CD4 ؉ T-cell response induced by the GP 61-80 epitope was almost abolished when attached to the LIMP-II tail. We identify the mechanism which underlies this marked difference in outcome. The GP 61-80 epitope is highly susceptible to cleavage by cathepsin D, an aspartic endopeptidase found almost exclusively in lysosomes. We show, using mass spectrometry, that the GP 61-80 peptide is cleaved between residues F 74 and K 75 and that this destroys its ability to stimulate virus-specific CD4 ϩ T-cell induction by DNA vaccines. We have previously demonstrated that improving the degradation of endogenously expressed antigens in the proteasome enhanced the induction of CD8 ϩ T-cell responses; covalent linkage of the antigen to the cellular protein ubiquitin marked the fusion protein for rapid hydrolysis in the proteasome, improved class I-antigen presentation and enhanced the protection induced by the DNA vaccines in mice, both against a virus (27, 29) and an invasive melanoma (48). In this report we present a parallel strategy aimed at improving the CD4 ϩ responses induced by DNA immunization. Most CD4 ϩ T-cell responses are induced by proteins endocytosed from the extracellular milieu by specialized antigen-presenting cells (APCs) such as dendritic cells (DC); these proteins are then degraded in the acidic endosomal and lysosomal compartments, where they encounter major histocompatibility complex (MHC) class II molecules, leading to the eventual cell surface presentation of their encoded epitopes. However, although the underlying mechanisms are not fully understood, it has been clearly demonstrated that some proteins synthesized within an APC can be presented by MHC class II molecules and can induce CD4 ϩ T-cell responses (1,4,6,21,25,32). This observation suggested that a DNA vaccine could be designed which should direct endogenously synthesized proteins to the lysosomal compartment of APCs, thus enhancing the induction of CD4 ϩ T cells. To achieve our goal, we have used the lysosomal targeting signal of lysosomal integral membrane protein-II (LIMP-II) (41). Unlike other lysosomal proteins, which usually take an indirect route...

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Quantitative and qualitative analyses of the immune responses induced by a multivalent minigene DNA vaccine

Cited by 45 publications

References 40 publications

Sustained Response Initiation Is Required for T Cell Clonal Expansion But Not for Effector or Memory Development In Vivo

Sustained Response Initiation Is Required for T Cell Clonal Expansion But Not for Effector or Memory Development In Vivo

Full-length proteins attached to the HIV tat protein transduction domain are neither transduced between cells, nor exhibit enhanced immunogenicity

CD4⁺T Cells Induced by a DNA Vaccine: Immunological Consequences of Epitope-Specific Lysosomal Targeting

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Quantitative and qualitative analyses of the immune responses induced by a multivalent minigene DNA vaccine

Cited by 45 publications

References 40 publications

Sustained Response Initiation Is Required for T Cell Clonal Expansion But Not for Effector or Memory Development In Vivo

Sustained Response Initiation Is Required for T Cell Clonal Expansion But Not for Effector or Memory Development In Vivo

Full-length proteins attached to the HIV tat protein transduction domain are neither transduced between cells, nor exhibit enhanced immunogenicity

CD4+T Cells Induced by a DNA Vaccine: Immunological Consequences of Epitope-Specific Lysosomal Targeting

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CD4⁺T Cells Induced by a DNA Vaccine: Immunological Consequences of Epitope-Specific Lysosomal Targeting