Efficient stimulation of HIV-1-specific T cells using dendritic cells electroporated with mRNA encoding autologous HIV-1 Gag and Env proteins.

Infection with human immunodeficiency virus type 1 (HIV-1) is characterized by dysfunction of HIV-1-specific T cells. To control the virus, antigen-loaded dendritic cells (DCs) might be useful to boost and broaden HIV-specific T-cell responses. In the present study, monocyte-derived DCs from nontreated HIV-1-seropositive patients were electroporated with codon-optimized ("humanized") mRNA encoding consensus HxB-2 (hHXB-2) Gag protein. These DCs elicited a strong HIV-1 Gag-specific interferon-gamma (IFN-gamma) response by an HLA-A2-restricted CD8+ T-cell line. Moreover, hHXB-2 gag mRNA-electroporated DCs also triggered IFN-gamma secretion by autologous peripheral blood mononuclear cells (PBMCs), CD4+ T cells, and CD8+ T cells from all patients tested. Next, a novel strategy was developed using autologous virus sequences. Significant specific IFN-gamma T-cell responses were induced in all patients tested by DCs electroporated with patients' autologous polymerase chain reaction (PCR)-amplified and in vitro-transcribed proviral and plasma viral mRNA encoding either Gag or Env. The stimulatory effect was seen on PBMCs, CD8+ T cells, and CD4+ T cells, demonstrating both major histocompatibility complex (MHC) class I and MHC class II antigen presentation. Moreover, a significant interleukin-2 (IL-2) T-cell response was induced by DCs electroporated with hHxB-2 or proviral gag mRNA. These findings open a major perspective for the development of patient-specific immunotherapy for HIV-1 disease.

[1]  R. Steinman,et al.  Proliferating dendritic cell progenitors in human blood , 1994, The Journal of experimental medicine.

[2]  Simon C Watkins,et al.  Adenovirus-Transduced Dendritic Cells Injected into Skin or Lymph Node Prime Potent Simian Immunodeficiency Virus-Specific T Cell Immunity in Monkeys 1 , 2003, The Journal of Immunology.

[3]  C. van Broeckhoven,et al.  Messenger RNA Electroporation of Human Monocytes, Followed by Rapid In Vitro Differentiation, Leads to Highly Stimulatory Antigen-Loaded Mature Dendritic Cells1 , 2002, The Journal of Immunology.

[4]  R. Siliciano,et al.  Latency in Human Immunodeficiency Virus Type 1 Infection: No Easy Answers , 2003, Journal of Virology.

[5]  M. Pope.  Dendritic cells as a conduit to improve HIV vaccines. , 2003, Current molecular medicine.

[6]  G. Pantaleo,et al.  HIV-1-specific IFN-gamma/IL-2-secreting CD8 T cells support CD4-independent proliferation of HIV-1-specific CD8 T cells. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[7]  S. Coppens,et al.  Design and evaluation of an in-house HIV-1 (group M and O), SIVmnd and SIVcpz antigen capture assay. , 1998, Journal of virological methods.

[8]  G. Pantaleo,et al.  Functional Heterogeneity of Memory CD4 T Cell Responses in Different Conditions of Antigen Exposure and Persistence1 , 2005, The Journal of Immunology.

[9]  R. Steinman,et al.  Dendritic cells and the control of immunity , 1998, Nature.

[10]  T. Chun,et al.  Latent reservoirs of HIV: obstacles to the eradication of virus. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[11]  Wei Lu,et al.  Therapeutic dendritic-cell vaccine for chronic HIV-1 infection , 2004, Nature Medicine.

[12]  R. Koup,et al.  Correlates of immune protection in HIV-1 infection: what we know, what we don't know, what we should know , 2004, Nature Medicine.

[13]  D. Richman,et al.  Recovery of replication-competent HIV despite prolonged suppression of plasma viremia. , 1997, Science.

[14]  T. Di Pucchio,et al.  Potent Immune Response against HIV-1 and Protection from Virus Challenge in hu-PBL-SCID Mice Immunized with Inactivated Virus-pulsed Dendritic Cells Generated in the Presence of IFN-α , 2003, The Journal of experimental medicine.

[15]  S. Saebøe-Larssen,et al.  mRNA-based electrotransfection of human dendritic cells and induction of cytotoxic T lymphocyte responses against the telomerase catalytic subunit (hTERT). , 2002, Journal of immunological methods.

[16]  R. Koup,et al.  Optimal antigens for HIV vaccines based on CD8+ T response, protein length, and sequence variability. , 2002, DNA and cell biology.

[17]  G. Boiteau,et al.  J. Virol. Methods , 1996, Journal of Virological Methods.

[18]  J. Lisziewicz,et al.  Latent infection of CD4+ T cells provides a mechanism for lifelong persistence of HIV-1, even in patients on effective combination therapy , 1999, Nature Medicine.

[19]  Adeeba Kamarulzaman,et al.  AIDS Res Hum Retroviruses , 2006 .

[20]  O. Majdic,et al.  Cultured human Langerhans cells resemble lymphoid dendritic cells in phenotype and function. , 1989, The Journal of investigative dermatology.

[21]  C. van Broeckhoven,et al.  Highly efficient gene delivery by mRNA electroporation in human hematopoietic cells: superiority to lipofection and passive pulsing of mRNA and to electroporation of plasmid cDNA for tumor antigen loading of dendritic cells. , 2001, Blood.

[22]  R. Wagner,et al.  Impact of Codon Usage Modification on T Cell Immunogenicity and Longevity of HIV-1 Gag-Specific DNA Vaccines , 2003, Intervirology.

[23]  R. Phillips,et al.  Escape of human immunodeficiency virus from immune control. , 1997, Annual review of immunology.

[24]  Thomas C. Quinn,et al.  A Novel Assay Allows Genotyping of the Latent Reservoir for Human Immunodeficiency Virus Type 1 in the Resting CD4+ T Cells of Viremic Patients , 2005, Journal of Virology.

[25]  J. Mascola,et al.  Macaque dendritic cells infected with SIV-recombinant canarypox ex vivo induce SIV-specific immune responses in vivo. , 2004, AIDS research and human retroviruses.

[26]  Mamoru Ito,et al.  Induction of Protective Immune Responses against R5 Human Immunodeficiency Virus Type 1 (HIV-1) Infection in hu-PBL-SCID Mice by Intrasplenic Immunization with HIV-1-Pulsed Dendritic Cells: Possible Involvement of a Novel Factor of Human CD4+ T-Cell Origin , 2003, Journal of Virology.

[27]  C. Wilson,et al.  HIV-1-specific CTL responses primed in vitro by blood-derived dendritic cells and Th1-biasing cytokines. , 1999, Journal of immunology.

[28]  S. Rowland-Jones,et al.  Role of cellular immunity in protection against HIV infection. , 1997, Advances in immunology.

[29]  C. Broeckhoven,et al.  mRNA-electroporated mature dendritic cells retain transgene expression, phenotypical properties and stimulatory capacity after cryopreservation , 2002, Leukemia.

[30]  Jerome A. Zack,et al.  HIV-1 entry into quiescent primary lymphocytes: Molecular analysis reveals a labile, latent viral structure , 1990, Cell.

[31]  D. Hanau,et al.  Dendritic cells cross-present HIV antigens from live as well as apoptotic infected CD4+ T lymphocytes. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[32]  Alison Johnson,et al.  Diminished Proliferation of Human Immunodeficiency Virus-Specific CD4+ T Cells Is Associated with Diminished Interleukin-2 (IL-2) Production and Is Recovered by Exogenous IL-2 , 2003, Journal of Virology.

[33]  E. Gilboa,et al.  Dendritic cells pulsed with RNA are potent antigen-presenting cells in vitro and in vivo , 1996, The Journal of experimental medicine.

[34]  D. Nixon,et al.  Amplification of low-frequency antiviral CD8 T cell responses using autologous dendritic cells , 2002, AIDS.

[35]  D. Nixon,et al.  Requirement of Mature Dendritic Cells for Efficient Activation of Influenza A-Specific Memory CD8+ T Cells1 , 2000, The Journal of Immunology.

[36]  C. Broeckhoven,et al.  Gene-based cancer vaccines: an ex vivo approach , 2001, Leukemia.

[37]  M. Altfeld,et al.  Cytotoxic T-lymphocyte (CTL) responses directed against regulatory and accessory proteins in HIV-1 infection. , 2002, DNA and cell biology.

[38]  M A Nowak,et al.  Presence of an inducible HIV-1 latent reservoir during highly active antiretroviral therapy. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[39]  R. Lempicki,et al.  Gene expression and viral prodution in latently infected, resting CD4+ T cells in viremic versus aviremic HIV-infected individuals , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[40]  B. Autran,et al.  Yeast-Derived Human Immunodeficiency Virus Type 1 p55gag Virus-Like Particles Activate Dendritic Cells (DCs) and Induce Perforin Expression in Gag-Specific CD8+ T Cells by Cross-Presentation of DCs , 2003, Journal of Virology.

[41]  R Brookmeyer,et al.  Identification of a reservoir for HIV-1 in patients on highly active antiretroviral therapy. , 1997, Science.

[42]  C. Rouzioux,et al.  Dendritic cells generated in the presence of granulocyte-macrophage colony-stimulating factor and IFN-α are potent inducers of HIV-specific CD8 T cells , 2003, AIDS.

[43]  J. Andrieu,et al.  In Vitro Human Immunodeficiency Virus Eradication by Autologous CD8+ T Cells Expanded with Inactivated-Virus-Pulsed Dendritic Cells , 2001, Journal of Virology.

[44]  B. Sullenger,et al.  Emerging clinical applications of RNA , 2002, Nature.

[45]  R. Steinman,et al.  Mature Dendritic Cells Infected with Canarypox Virus Elicit Strong Anti-Human Immunodeficiency Virus CD8+and CD4+ T-Cell Responses from Chronically Infected Individuals , 2001, Journal of Virology.

[46]  E. Gilboa,et al.  Cancer immunotherapy with mRNA‐transfected dendritic cells , 2004, Immunological reviews.

[47]  G. Schuler,et al.  Human dendritic cells transfected with either RNA or DNA encoding influenza matrix protein M1 differ in their ability to stimulate cytotoxic T lymphocytes , 2000, Gene Therapy.

[48]  Simon C Watkins,et al.  Induction of Anti-Human Immunodeficiency Virus Type 1 (HIV-1) CD8+ and CD4+ T-Cell Reactivity by Dendritic Cells Loaded with HIV-1 X4-Infected Apoptotic Cells , 2002, Journal of Virology.

[49]  P. Ponsaerts,et al.  Cancer immunotherapy using RNA‐loaded dendritic cells , 2003, Clinical and experimental immunology.

[50]  S. Coppens,et al.  Simplified strategy for detection of recombinant human immunodeficiency virus type 1 group M isolates by gag/env heteroduplex mobility assay. Study Group on Heterogeneity of HIV Epidemics in African Cities. , 2000, Journal of virology.

[51]  G. Pantaleo,et al.  Skewed representation of functionally distinct populations of virus-specific CD4 T cells in HIV-1-infected subjects with progressive disease: changes after antiretroviral therapy. , 2004, Blood.

[52]  Z. Grossman,et al.  HIV-1 Viremia Prevents the Establishment of Interleukin 2–producing HIV-specific Memory CD4+ T Cells Endowed with Proliferative Capacity , 2003, The Journal of experimental medicine.

[53]  J. Sleasman,et al.  HIV-1 infection , 2003 .

[54]  M P Busch,et al.  Rapid molecular epidemiology of human immunodeficiency virus transmission. , 1995, AIDS research and human retroviruses.