Adoptively transferred effector cells derived from naive rather than central memory CD8+ T cells mediate superior antitumor immunity - PubMed (original) (raw)

. 2009 Oct 13;106(41):17469-74.

doi: 10.1073/pnas.0907448106. Epub 2009 Sep 24.

Zachary A Borman, Lydie Cassard, Luca Gattinoni, Rosanne Spolski, Zhiya Yu, Luis Sanchez-Perez, Pawel Muranski, Steven J Kern, Carol Logun, Douglas C Palmer, Yun Ji, Robert N Reger, Warren J Leonard, Robert L Danner, Steven A Rosenberg, Nicholas P Restifo

Affiliations

Adoptively transferred effector cells derived from naive rather than central memory CD8+ T cells mediate superior antitumor immunity

Christian S Hinrichs et al. Proc Natl Acad Sci U S A. 2009.

Abstract

Effector cells derived from central memory CD8(+) T cells were reported to engraft and survive better than those derived from effector memory populations, suggesting that they are superior for use in adoptive immunotherapy studies. However, previous studies did not evaluate the relative efficacy of effector cells derived from naïve T cells. We sought to investigate the efficacy of tumor-specific effector cells derived from naïve or central memory T-cell subsets using transgenic or retrovirally transduced T cells engineered to express a tumor-specific T-cell receptor. We found that naïve, rather than central memory T cells, gave rise to an effector population that mediated superior antitumor immunity upon adoptive transfer. Effector cells developed from naïve T cells lost the expression of CD62L more rapidly than those derived from central memory T cells, but did not acquire the expression of KLRG-1, a marker for terminal differentiation and replicative senescence. Consistent with this KLRG-1(-) phenotype, naïve-derived cells were capable of a greater proliferative burst and had enhanced cytokine production after adoptive transfer. These results indicate that insertion of genes that confer antitumor specificity into naïve rather than central memory CD8(+) T cells may allow superior efficacy upon adoptive transfer.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Fig. 1.

Fig. 1.

Effector CD8+ T cells derived from naïve or central memory cells acquire cytolytic phenotype and function. (A) Production of IFN-γ by freshly isolated cells in an overnight coculture assay. The error bars represent the standard error of the mean. (B) The number of cell divisions 2 days after peptide stimulation, as determined by CFSE dilution from a FACS histogram. (C) Schematic delineating generation of primary and secondary effector cells from naïve and central memory CD8+ T cells. (D) Cytolytic function of TEFFN and TEFFCM as determined by 51Cr release assay. The target peptide, gp10025–33 or NP366–374 is indicated in parenthesis. (E) Flow cytometric analysis indicating expression of granzyme B and perforin and (F) L-selectin and CD44. The open histograms indicate isotype antibody controls. All figures shown are representative of at least two independent experiments.

Fig. 2.

Fig. 2.

Naïve and central memory cells confer distinct genetic signatures and developmental programs to their effector cell progeny. Microarray analysis of the gene expression of (A) rested and (B) stimulated TEFFN and TEFFCM. Probe sets with FDR ≤2% and ≥2-fold differences in expression are shown. Red and green colors indicate increased and decreased expression respectively. The scales are log10. (C) Relative expression of Eomes by TEFFN or TEFFCM as determined by real time RT-PCR. The x axis indicates the number of times the cells were stimulated with antigen and IL-2. Error bars represent the standard error of the mean. (D) Flow cytometric determination of the expression of CD62L and CD44 by TEFFN and TEFFCM through serial stimulations. The numbers on the dot plots indicate the frequency of cells with central memory (CD62L+ CD44high) phenotype. (E) Expression of CD62L and KLRG1. Quadrant frequencies are shown. (F) Graph derived from panel E. Frequencies of cells from the CD62L+ or CD62L− fraction of TEFFN or TEFFCM with KLRG1high phenotype. (G) IFN-γ and IL-2 production, in coculture assays, by effector cells generated by primary or secondary stimulation of naïve or central memory cells. (H) Intracellular IFN-γ expression following peptide stimulation of TEFFN or TEFFCM. The mean fluorescence intensities are indicated. All figures shown are representative of at least two independent experiments.

Fig. 3.

Fig. 3.

Use of effector cells derived from naïve rather than central memory cells improves adoptive cell transfer therapy. (A) Tumor responses following treatment with 4 × 106 effector cells generated from naïve or central memory subsets. All groups except “No treatment” received adjuvant vaccine and IL-2. TEFFN versus TEFFCM, P < 0.01. (_B_) Production of IFN-γ and IL-2 by adoptively transferred cells reisolated from spleens 6 days following infusion. Cell numbers were equalized for the assay. (_C_) Tumor treatment with 7.65 × 105 wild-type or IFN-γ−/− effector cells. TEFFN versus IFN-γ−/− TEFFN, _P_ < 0.05. (_D_) Tumor treatment with 1.05 × 106 wild-type or IL-2−/− effector cells. TEFFN versus IL-2−/− TEFFN, _P_ > 0.05. All figures shown are representative of at least two independent experiments.

Fig. 4.

Fig. 4.

Effector cells derived directly from naïve cells possess greater potential for clonal expansion following infusion. (A and B) The number of cells present in spleens and lymph nodes, respectively, following adoptive transfer of TEFFN or TEFFCM. Error bars indicate the standard error of the mean. (n = 4 for each group and each time point). (C) The frequency of transferred cells in recipient spleens displaying KLRG1high phenotype 5 days following infusion. The transverse lines represent the means. P = 0.0024. All figures shown are representative of at least two independent experiments.

Fig. 5.

Fig. 5.

Genetically engineered open repertoire cells reproduce the central findings of the transgenic mouse model. (A) Elaboration of IFN-γ and IL-2 by TCR transduced naïve- or central memory-derived effector cells. (B) Tumor growth following treatment with transduced effector cells generated by 4 × 106 primary or (C) 2 × 107 secondary stimulation of naïve or central memory cells. All figures shown are representative of at least two independent experiments.

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