Skip to main page content
U.S. flag
See More

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Clinical Trial
. 2010 Nov 18;116(20):4099-102.
doi: 10.1182/blood-2010-04-281931. Epub 2010 Jul 28.

Eradication of B-lineage cells and regression of lymphoma in a patient treated with autologous T cells genetically engineered to recognize CD19

James N Kochenderfer  1 , Wyndham H Wilson, John E Janik, Mark E Dudley, Maryalice Stetler-Stevenson, Steven A Feldman, Irina Maric, Mark Raffeld, Debbie-Ann N Nathan, Brock J Lanier, Richard A Morgan, Steven A Rosenberg
Affiliations
Clinical Trial

Eradication of B-lineage cells and regression of lymphoma in a patient treated with autologous T cells genetically engineered to recognize CD19

James N Kochenderfer et al. Blood. .

Abstract

Adoptive transfer of genetically modified T cells is an attractive approach for generating antitumor immune responses. We treated a patient with advanced follicular lymphoma by administering a preparative chemotherapy regimen followed by autologous T cells genetically engineered to express a chimeric antigen receptor (CAR) that recognized the B-cell antigen CD19. The patient's lymphoma underwent a dramatic regression, and B-cell precursors were selectively eliminated from the patient's bone marrow after infusion of anti-CD19-CAR-transduced T cells. Blood B cells were absent for at least 39 weeks after anti-CD19-CAR-transduced T-cell infusion despite prompt recovery of other blood cell counts. Consistent with eradication of B-lineage cells, serum immunoglobulins decreased to very low levels after treatment. The prolonged and selective elimination of B-lineage cells could not be attributed to the chemotherapy that the patient received and indicated antigen-specific eradication of B-lineage cells. Adoptive transfer of anti-CD19-CAR-expressing T cells is a promising new approach for treating B-cell malignancies. This study is registered at www.clinicaltrials.gov as #NCT00924326.

PubMed Disclaimer

Figures

Figure 1
Figure 1
B-lineage cells, including B-cell precursors, were eradicated from the bone marrow after treatment with anti–CD19-CAR-transduced T cells. (A) Representative pretreatment computed tomography scan images and images from 18 weeks after treatment demonstrate regression of lymphoma masses in the chest and abdomen after treatment with chemotherapy followed by anti–CD19-CAR-transduced T cells plus IL-2. (B) Flow cytometric evaluation of a pretreatment bone marrow aspirate was conducted with a forward versus side light scatter analysis gate of lymphoid cells. The left upper quadrant contains CD19+ B-lineage cells (35% of lymphoid cells), and the right lower quadrant contains CD3+ T cells. (C) Flow cytometric evaluation of a pretreatment bone marrow aspirate with a CD19+ analysis gate is shown. κ- and λ-negative, CD19+, mostly immature B-lineage cells that are not part of the malignant lymphoma clone are in the rectangle. The cells outside the rectangle are mostly lymphoma cells. (D) Flow cytometric evaluation of a pretreatment bone marrow aspirate with a forward versus side light scatter analysis gate of lymphoid cells. Immature B-cell precursors in the oval are CD22+ and CD20−. (E) Flow cytometric evaluation of a pretreatment bone marrow aspirate with a forward versus side light scatter analysis gate of lymphoid cells. Immature B-cell precursors in the polyhedral demonstrate decreasing CD10 correlating with increasing CD20 expression. (F) Flow cytometric evaluation of a bone marrow aspirate from 36 weeks after treatment with a forward versus side light scatter analysis gate of lymphoid cells. CD19+ B-lineage cells are absent. (G) Immunohistochemistry staining of a pretreatment bone marrow biopsy reveals a large population of CD19+ cells that includes lymphoma cells as well as nonmalignant B-lineage cells. (H) Immunohistochemistry staining of a bone marrow biopsy from 36 weeks after infusion of anti–CD19-CAR-transduced T cells demonstrates a complete absence of CD19+ cells. (I) High-power view of the same anti-CD19 staining shown in panel H.
Figure 2
Figure 2
Prolonged B-cell depletion after anti–CD19-CAR-transduced T-cell infusion. (A) Immunohistochemistry staining of a pretreatment bone marrow biopsy shows a large population of CD79a+ cells. (B) Thirty-six weeks after anti–CD19-CAR-transduced T-cell infusion, rare CD79a+ cells were detected by immunohistochemisty staining of a bone marrow biopsy. The cells did not appear to be plasma cells morphologically. The number of CD79a+ cells was substantially below normal limits. The arrow indicates one of the rare CD79a+ cells. (C) The blood B-cell count of the patient treated with anti–CD19-CAR-transduced T cells is shown before treatment and at multiple time points after treatment. B cells were measured by flow cytometry for CD19. The dashed line indicates the lower limit of normal. Day 0 is the day of the second anti–CD19-CAR-transduced T-cell infusion. (D) The mean ± SEM blood B-cell count is shown for patients who received infusions of T cells targeted to either the NY-ESO antigen or the gp100 antigen. The patients all received the same chemotherapy and IL-2 regimen as the patient who received anti–CD19-CAR-transduced T cells. NY-ESO and gp100 are not expressed by B cells. Day 0 is the day of T-cell infusion. All available B-cell counts were included for each time point (pretreatment, n = 28; 4-5 weeks after T-cell infusion, n = 29; 8-11 weeks after T-cell infusion, n = 31; 14-19 weeks after T-cell infusion, n = 20). All patients with available samples had a B-cell count in the normal range by 14 to 19 weeks after T-cell infusion. (E) The blood CD3+ T-cell count of the patient treated with anti–CD19-CAR-transduced T cells is shown before treatment and at multiple time points after treatment. (F) The blood NK cell count of the patient treated with anti–CD19-CAR-transduced T cells is shown before treatment and at multiple time points after treatment. NK cells were measured by flow cytometry as CD3−, CD16+, CD56+ cells. (E-F) Day 0 is the day of the second anti–CD19-CAR-transduced T-cell infusion, and the dashed line indicates the lower limit of normal. (G) The serum IgG level of the patient treated with anti–CD19-CAR-transduced T cells is shown before treatment and at multiple time points after treatment. Day 0 is the day of the second anti–CD19-CAR-transduced T-cell infusion. (H) Real-time polymerase chain reaction was performed with a primer and probe set that was specific for the anti-CD19 CAR. Anti–CD19-CAR-transduced T cells were undetectable in pretreatment blood samples. The anti–CD19 CAR transgene was detected in the peripheral blood of the patient who received anti–CD19-CAR-transduced T cells from 1 to 27 weeks after anti–CD19-CAR-transduced T-cell infusion.
See this image and copyright information in PMC

References

    1. Eshhar Z, Waks T, Gross G, Schindler DG. Specific activation and targeting of cytotoxic lymphocytes through chimeric single chains consisting of antibody-binding domains and the gamma or zeta subunits of the immunoglobulin and T-cell receptors. Proc Natl Acad Sci U S A. 1993;90(2):720–724. - PMC - PubMed
    1. Kershaw MH, Teng MWL, Smyth MJ, Darcy PK. Supernatural T cells: genetic modification of T cells for cancer therapy. Nat Rev. 2005;5(12):928–940. - PubMed
    1. Irving BA, Weiss A. The cytoplasmic domain of the T cell receptor zeta chain is sufficient to couple to receptor-associated signal transduction pathways. Cell. 1991;64(5):891–901. - PubMed
    1. Sadelain M, Brentjens R, Riviere I. The promise and potential pitfalls of chimeric antigen receptors. Curr Opin Immunol. 2009;21(2):215–223. - PMC - PubMed
    1. Vera JF, Brenner MK, Dotti G. Immunotherapy of human cancers using gene modified T lymphocytes. Curr Gene Ther. 2009;9(5):396–408. - PMC - PubMed

Publication types

Associated data