Expansion of HIV-specific CD4+ and CD8+ T cells by dendritic cells transfected with mRNA encoding cytoplasm- or lysosome-targeted Nef

Kavanagh, Daniel G.; Kaufmann, Daniel E.; Sunderji, Sherzana; Frahm, Nicole; Gall, Sylvie Le; Boczkowski, David; Rosenberg, Eric S.; Stone, David R.; Johnston, Mary N.; Wagner, Bradford S.; Zaman, M. Tauheed; Berthou, Christian; Gilboa, Eli; Walker, Bruce D.; Bhardwaj, Nina

doi:10.1182/blood-2005-04-1513

Cited by 56 publications

(49 citation statements)

References 31 publications

(45 reference statements)

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“…Immunization against native simian HIV Ags with DNA or vaccinia virus preparations has been shown to increase the breadth of the CTL response after simian HIV challenge (68), and it is possible that endosomal/lysosomal targeting, such as that provided by the LAMP/gag or DC-LAMP/gag chimeras, could further enhance the diversity of the epitope response. Finally, Kavanagh et al (69) recently showed that DC transfected with HIV Nef targeted to the lysosome by LAMP was able to expand a greater repertoire of T cells from HIV patients than the DC transfected with HIV Nef. These results imply that LAMP-targeted Ag generated a greater or distinct repertoire of peptides than the native Nef, and it was considered as a promising approach to HIV immunotherapy.…”

Section: Discussionmentioning

confidence: 99%

Dendritic Cell-Lysosomal-Associated Membrane Protein (LAMP) and LAMP-1-HIV-1 Gag Chimeras Have Distinct Cellular Trafficking Pathways and Prime T and B Cell Responses to a Diverse Repertoire of Epitopes

Arruda

Sim²,

Chikhlikar

et al. 2006

The Journal of Immunology

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Ag processing is a critical step in defining the repertoire of epitope-specific immune responses. In the present study, HIV-1 p55Gag Ag was synthesized as a DNA plasmid with either lysosomal-associated membrane protein-1 (LAMP/gag) or human dendritic cell-LAMP (DC-LAMP/gag) and used to immunize mice. Analysis of the cellular trafficking of these two chimeras demonstrated that both molecules colocalized with MHC class II molecules but differed in their overall trafficking to endosomal/lysosomal compartments. Following DNA immunization, both chimeras elicited potent Gag-specific T and B cell immune responses in mice but differ markedly in their IL-4 and IgG1/IgG2a responses. The DC-LAMP chimera induced a stronger Th type 1 response. ELISPOT analysis of T cell responses to 122 individual peptides encompassing the entire p55gag sequence (15-aa peptides overlapping by 11 residues) showed that DNA immunization with native gag, LAMP/gag, or DC-LAMP/gag induced responses to identical immunodominant CD4+ and CD8+ peptides. However, LAMP/gag and DC-LAMP/gag plasmids also elicited significant responses to 23 additional cryptic epitopes that were not recognized after immunization with native gag DNA. The three plasmids induced T cell responses to a total of 39 distinct peptide sequences, 13 of which were induced by all three DNA constructs. Individually, DC-LAMP/gag elicited the most diverse response, with a specific T cell response against 35 peptides. In addition, immunization with LAMP/gag and DC-LAMP/gag chimeras also promoted Ab secretion to an increased number of epitopes. These data indicate that LAMP-1 and DC-LAMP Ag chimeras follow different trafficking pathways, induce distinct modulatory immune responses, and are able to present cryptic epitopes.

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Section: Discussionmentioning

confidence: 99%

Dendritic Cell-Lysosomal-Associated Membrane Protein (LAMP) and LAMP-1-HIV-1 Gag Chimeras Have Distinct Cellular Trafficking Pathways and Prime T and B Cell Responses to a Diverse Repertoire of Epitopes

Arruda

Sim²,

Chikhlikar

et al. 2006

The Journal of Immunology

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“…Recently, we (47,48,61) and others (9,15,22,28,40,54,57) have shown that transfection with mRNA is more effective than mRNA lipofection, peptide pulsing, or viral transduction to generate primary (65) and memory (57) responses. Furthermore, we demonstrated that DC from treatment-naïve HIV-1-seropositive subjects can efficiently be transfected with HIV gag and env mRNA, derived either from consensus subtype B or autologous viral or proviral HIV, and that these DC readily trigger autologous CD4 ϩ and CD8 ϩ T cells to release IFN-␥ and IL-2 in a short-term ex vivo enzyme-linked immunospot (ELISPOT) assay (60).…”

mentioning

confidence: 99%

Efficient In Vitro Expansion of Human Immunodeficiency Virus (HIV)-Specific T-Cell Responses bygagmRNA-Electroporated Dendritic Cells from Treated and Untreated HIV Type 1-Infected Individuals

Gulck

Vanham

Heyndríckx

et al. 2008

J Virol

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“…DNA transfection of DC has been used successfully in some studies but is limited by poor expression levels and toxicity (2,29,41,45,49), although methods to enhance expression based on DC maturation have been proposed (28). mRNA transfection of DC is being increasingly utilized for cancer immunotherapy and in vitro stimulation of virus-specific T cells, but approaches to deliver mRNA into primary DC cultures have been limited (16,18,25,39,40,43,48,50). Transfection of DC with mRNA offers the potential for highlevel expression of wild-type viral transgenes that have poor expression in mammalian cells when traditional methods of gene delivery are used (8,26).…”

mentioning

confidence: 99%

High-Level Antigen Expression and Sustained Antigen Presentation in Dendritic Cells Nucleofected with Wild-Type Viral mRNA but Not DNA

Melhem

Gleason

Liu

et al. 2008

Clin Vaccine Immunol

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Dendritic cells (DC) are potent antigen-presenting cells that hold promise as cell-based therapeutic vaccines for infectious diseases and cancer. Ideally, DC would be engineered to express autologous viral or tumor antigens to ensure the presentation of relevant antigens to host T cells in vivo; however, expression of wild-type viral genes in primary cell lines can be problematic. Nucleofection is an effective means of delivering transgenes to primary cell lines, but its use in transfecting DNA or mRNA into DC has not been widely investigated. We show that nucleofection is a superior means of transfecting human and monkey monocyte-derived DC with DNA and mRNA compared to lipofection and conventional electroporation. However, the delivery of DNA and mRNA had significantly different outcomes in transfected DC. DC nucleofected with DNA encoding green fluorescent protein (GFP) had poor antigen expression and viability and were refractory to maturation with CD40 ligand. In contrast, >90% of DC expressed uniform and high levels of GFP from 3 h to 96 h postnucleofection with mRNA while maintaining a normal maturation response to CD40 ligation. Monkey DC nucleofected with wild-type, non-codon-optimized mRNA encoding simian immunodeficiency virus Gag stimulated robust antigen-specific effector T-cell responses at 24 h and 48 h postnucleofection, reflecting sustained antigen presentation in transfected DC, whereas no detectable T-cell response was noted when DC were nucleofected with DNA encoding the same Gag sequence. These data indicate that mRNA nucleofection may be an optimal means of transfecting DC with autologous tumor or viral antigen for DC-based immunotherapy.

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Expansion of HIV-specific CD4+ and CD8+ T cells by dendritic cells transfected with mRNA encoding cytoplasm- or lysosome-targeted Nef

Cited by 56 publications

References 31 publications

Dendritic Cell-Lysosomal-Associated Membrane Protein (LAMP) and LAMP-1-HIV-1 Gag Chimeras Have Distinct Cellular Trafficking Pathways and Prime T and B Cell Responses to a Diverse Repertoire of Epitopes

Dendritic Cell-Lysosomal-Associated Membrane Protein (LAMP) and LAMP-1-HIV-1 Gag Chimeras Have Distinct Cellular Trafficking Pathways and Prime T and B Cell Responses to a Diverse Repertoire of Epitopes

Efficient In Vitro Expansion of Human Immunodeficiency Virus (HIV)-Specific T-Cell Responses bygagmRNA-Electroporated Dendritic Cells from Treated and Untreated HIV Type 1-Infected Individuals

High-Level Antigen Expression and Sustained Antigen Presentation in Dendritic Cells Nucleofected with Wild-Type Viral mRNA but Not DNA

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