Simultaneous activation of viral antigen-specific memory CD4+ and CD8+ T-cells using mRNA-electroporated CD40-activated autologous B-cells.
Recently, it has become obvious that not only CD8 T-cells, but also CD4 T-helper cells are required for the induction of an effective, long-lasting cellular immune response. In view of the clinical importance of cytomegalovirus (CMV) and human immunodeficiency virus (HIV) infection, we developed 2 strategies to simultaneously reactivate viral antigen-specific memory CD4 and CD8 T-cells of CMV-seropositive and HIV-seropositive subjects using mRNA-electroporated autologous CD40-activated B cells. In the setting of HIV, we provide evidence that CD40-activated B cells can be cultured from HAART-naive HIV-1 seropositive patients. These cells not only express and secrete the HIV p24 antigen after electroporation with codon-optimized HIV-1 gag mRNA, but can also be used to in vitro reactivate Gag antigen-specific interferon-gamma-producing CD4 and CD8 autologous T-cells. For the CMV-specific approach, we applied mRNA coding for the pp65 protein coupled to the lysosomal-associated membrane protein-1 to transfect CD40-activated B cells to induce CMV antigen-specific CD4 and CD8 T-cells. More detailed analysis of the activated interferon-gamma-producing CMV pp65 tetramer positive CD8 T-cells revealed an effector memory phenotype with the capacity to produce interleukin-2. Our findings clearly show that the concomitant activation of both CD4 and CD8 (memory) T-cells using mRNA-electroporated CD40-B cells is feasible in CMV and HIV-1-seropositive persons, which indicates the potential value of this approach for application in cellular immunotherapy of infectious diseases.
Van den Bosch
Glenn A
GA
Laboratory of Experimental Hematology, Antwerp University (UA), Antwerp University Hospital (UZA), Wilrijkstraat 10, 2650 Edegem, Belgium.
Van Gulck
Ellen
E
Ponsaerts
Peter
P
Nijs
Griet
G
Lenjou
Marc
M
Apers
Ludwig
L
Kint
Ilse
I
Heyndrickx
Leo
L
Vanham
Guido
G
Van Bockstaele
Dirk R
DR
Berneman
Zwi N
ZN
Van Tendeloo
Viggo F I
VF
D000328:Adult; D000956:Antigens, Viral / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D019013:CD40 Antigens / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002478:Cells, Cultured; D003586:Cytomegalovirus Infections / Q000276:immunology*; D018274:Electroporation; D005240:Feasibility Studies; D005260:Female; D015683:Gene Products, gag / Q000276:immunology; D015658:HIV Infections / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology; D006801:Humans; D007376:Interleukin-2 / Q000378:metabolism; D008213:Lymphocyte Activation; D052118:Lysosomal-Associated Membrane Protein 1 / Q000276:immunology; D051907:Lysosomal Membrane Proteins / Q000276:immunology; D008297:Male; D008875:Middle Aged; D010641:Phenotype; D010750:Phosphoproteins / Q000276:immunology; D012333:RNA, Messenger / Q000276:immunology*; D012367:RNA, Viral / Q000276:immunology; D014763:Viral Matrix Proteins / Q000276:immunology
D000956:Antigens, Viral; D019013:CD40 Antigens; D015683:Gene Products, gag; D007376:Interleukin-2; C489116:LAMP1 protein, human; D052118:Lysosomal-Associated Membrane Protein 1; D051907:Lysosomal Membrane Proteins; D010750:Phosphoproteins; D012333:RNA, Messenger; D012367:RNA, Viral; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa