1. CpG oligodeoxynucleotides allow for effective adoptive T-cell therapy in chronic retroviral infection
by Anke R. M. Kraft, Frank Krux, Simone Schimmer, Claes Ohlen, Philip D. Greenberg, and Ulf Dittmer
Blood
Volume 109(7):
April 1, 2007
©2007 by American Society of Hematology

2. Virus load after adoptive transfer of virus-specific CD8+ T cells.
Virus load after adoptive transfer of virus-specific CD8+ T cells. Chronically FV-infected (B10 × A.BY) F1 mice were treated with 4 × 106 naive, virus-specific TCRtg CD8+ T cells and 15 nmol of the B-type CpG-ODN (CpG-1668: 5′-tcc atg acg ttc ctg atg ct-3′) (CpG) or control ODN without the CpG motif (5′-tcc atg agc ttc ctg atg ct-3′)12 (Control) were injected twice, once on the day of T-cell transfer and again 4 days later. As controls, 1 group of animals received CpG-ODNs without T cells and 1 group of chronically infected mice did not receive any treatment (none). At 1 week (closed symbols) or 3 weeks (open symbols) after transfer, the spleen cells of the recipients were harvested and virus loads were determined in an infectious center assay.9 Mean values for each group are indicated by bars. The differences in viral loads between the groups of mice receiving CpG-ODNs or control ODNs together with T cells were statistically significant (unpaired t test).
Anke R. M. Kraft et al. Blood 2007;109:
©2007 by American Society of Hematology

3. Production of IFNγ and granzyme B and detection of exocytosis in adoptively transferred CD8+ T cells.
Production of IFNγ and granzyme B and detection of exocytosis in adoptively transferred CD8+ T cells. Chronically FV-infected (B10 × A.BY) F1 mice were treated with 4 × 106 naive, virus-specific CD8+ T cells from B6 mice carrying a TCRtg specific for the H-2Db–restricted gagL epitope of FV. The transferred CD8+ T cells were genetically labeled (CD45.1) to distinguish the host cell population from the donor cell population. B-type CpG-ODN 1668 (CpG) or control ODN without CpG motif (Control) (15 nmol) were injected twice. At 1 week after transfer, spleen cells of the recipient were harvested, and host (CD45.1−) and donor (CD45.1+) CD8+ T cells were stained for granzyme B (gzmB), IFNγ, and CD107a.9 Accumulated results are shown in the left column, with representative flow data in the right column. (A) The difference in the percentages of CD8+ donor T cells (CD45.1+) was statistically significant between the mice receiving CpG-ODNs or control ODNs. (B) Harvested cells were restimulated for 5 hours in vitro with α-CD3 and α-CD28 (closed symbols) or with 5 μg/mL FV DbGagL peptide and α-CD28 (open symbols) to detect intracellular IFNγ. Nonstimulated CD45.1+ CD8+ cells showed a background of less than 0.05% IFNγ-positive cells. The difference in absolute numbers of CD8+ donor (CD45+) T cells (CD45.1–) producing IFNγ was statistically significant between the mice receiving CpG-ODNs or control ODNs. (C) Harvested cells were analyzed directly ex vivo to detect intracellular granzyme B. The difference in absolute numbers of CD8+ donor and host T cells (CD45.1+) producing granzyme B was statistically significant between the mice receiving CpG-ODNs or control ODNs. Similar results were obtained for granzyme A production by CD8+ T cells (data not shown). (D) Harvested cells were analyzed directly ex vivo to detect CD107a expression. The difference in absolute numbers of CD8+ donor T cells (CD45.1+) expressing CD107a was statistically significant between the mice receiving CpG-ODNs or control ODNs. Percentages of positive donor (top right) and host (bottom right) cells are given in the respective quadrants. Mean values for each group are indicated by bars. Statistical analyses were done by unpaired t test.
Anke R. M. Kraft et al. Blood 2007;109:
©2007 by American Society of Hematology