Andre Radensky's Sigma Xi Showcase slideshare on T-cell immunotherapy and the chimeric antigen receptor

1. The Artificial Chimeric Antigen Receptor
Provides the Prospect for Efficacious
Treatment of B-Cell Malignancies
Through Immunotherapy
A Literary Analysis by André Radensky

2. Research Objectives
I. Create an artificial T-Cell Receptor that will recognize
the CD19 glycoprotein on B-cells
II. Amplify the patient’s immune response and have
altered T-Cells proliferate in vivo
III. Effectively rid patients of Acute Lymphoblastic
Leukemia or other B-cell cancers
http://naked-
science.ru/sites/default/files/imagecache/node_image_full/isto
ck_000019908954medium-purchased.jpg
Modified T-cells destroying
cancerous B-cell

3. Background
❖ adaptive immune system
➢ T-cell (or T lymphocyte) - specific subtype
of white blood cell that is involved in cell
mediated immunity, which binds directly to
the cell and induces apoptosis (or
programmed cell death) or the release of
cytokines
■ have a T-cell receptor (TCR) that
allows the cell to recognize and bind
to antigens that are marked by the
major histocompatibility complex
http://digitalcommons.wayne.edu/cgi/viewcontent.cgi?ar
ticle=1259&context=oa_theses
➢ B-cell (or B lymphocyte) - a different subtype of white blood cell (or
leukocyte) that is involved in humoral immunity, which is the production of
antibodies
■ have a distinct glycoprotein called CD19 found on the surface of all B-

4. ❖ standard cancer treatment
➢ includes: surgery, chemotherapy regimen, radiation, and stem cell
transplantation
➢ chemotherapy targets all rapidly replicating cells - harmful
implications from bone marrow suppression as target blood cells
❖ activation immunotherapy alternative - a targeted therapy for a specific
type of cell that uses the amplification of an immune response as the
remedy
❖ this therapy has been proven effective in hematological cancers - in
this experiment, the focus was placed on B-cell malignancies (and
particularly B-cell Acute Lymphoblastic Leukemia or ALL)
Background (continued)

5. Methodologies
I. Retrieval of autologous blood cells
II. Leukapheresis of blood cells to obtain
only white blood cells or leukocytes,
which are the cells to be genetically
altered
III. Genetic modification of T-cell’s
genome by addition of CD19-BB-z
transgene by disabled HIV vector
A. this transgene encodes the
artificial
T-cell Receptor that will recognize the
CD19 protein
http://www.wikidoc.org/index.php/Apheresis
HIV vector
T-Cell
CD19-BB-z
transgene

6. Methodologies
IV. Chimeric Antigen Receptor T-cells propagate ex
vivo by paramagnetic beads, which help induce
an immune response that causes the replication
V. Patient may be given chemotherapy to reduce
the number of T-cells in vivo to have a greater
chance of successful transplantation
VI. Altered T-cells are then transplanted back into
patient where they proliferate in vivo
http://www.sciencedaily.com/releases/2011/08/11081
0141248.htm
CAR T-cell
paramagnetic beads

7. Results
❖ Successful introduction of transgene by lentiviral vector and creation
of chimeric antigen receptor in T-cells
❖ Effective ex vivo proliferation of modified T-cells and notably in vivo
replication
❖ use of varying number of cells in transplantation had similar
implication in terms of cancer treatment - variation of 0.76×10^6 to
20.6×10^6 cells per kilogram of weight of patient
❖ 19/30 patients sustained remission following treatment with high
chances (68%) of persistence of altered T-cells for continued cancer
prevention
❖ negative effects - cytokine release syndrome and loss of
heterozygosity

8. Discussionl❖ Other studies performed on patients with chronic lymphocytic
leukemia and multiple myeloma corroborate the compelling
evidence seen in this one on acute lymphoblastic leukemia
❖ The CTL019 cells that expressed the CAR that destroyed all B-cells
because it recognized the glycoprotein on all B-cells was efficacious
in ridding the patient of the B-cell cancer
❖ The lentiviral (HIV) vector’s ability to add the CD19-BB-z gene to the
genome of T-cells has great utility as all subsequently replicated
cells will also express the artificial T-cell receptor
❖ Cytokine release syndrome was rarely fatal, but this issue as well as
loss of heterozygosity removing the entire gene that encodes for the
receptor need to be addressed before widespread adoption
❖ Application can be spread to other B-cell malignancies and even
further to other non-hematological cancers - only requisite is a
distinct protein on the cancerous cells and the ability to program the

9. Acknowledgements
(1) Maude, Shannon L., et al. "Chimeric Antigen Receptor T Cells for Sustained Remissions in Leukemia." New
England Journal of Medicine 371.16 (2014): 1507-17. Print.
(2) "T Cell Therapy (CTL019)." The Children's Hospital of Philadelphia. Children's Hospital of Philadelphia, n.d.
Web. 10 Dec. 2014. <http://www.chop.edu/centers-programs/cancer-center/t-cell-therapy-ctl019#.VIiEJ6TF-
Qy>.
(3) Jarvis, Lisa M. "The Immune System Fights Back." Chemical & Engineering News. American Chemical Society,
6 Oct. 2014. Web. 10 Dec. 2014. <http://cen.acs.org/articles/92/i40/Immune-System-Fights-Back.html>.
(4) Kudchodkar, Sagar B., and Marcela V. Maus. "Chimeric Antigen Receptor (CAR) T-Cell Immunotherapy for
Leukemia and Beyond." OncLive. Intellisphere LLC, 29 Aug. 2014. Web. 10 Dec. 2014.
<http://www.onclive.com/publications/contemporary-oncology/2014/august-2014/chimeric-antigen-receptor-car-
t-cell-immunotherapy-for-leukemia-and-beyond/ 4>.
(5) "University of Pennsylvania's Personalized Cellular Therapy for Leukemia Receives FDA's Breakthrough
Therapy Designation." Penn Medicine. The Trustees of the University of Pennsylvania, 7 July 2014. Web. 10
Dec. 2014. <http://www.uphs.upenn.edu/news/ News_Releases/2014/07/ctl019/>.
(6) Alfred L. Garfall, Joseph A Fraietta, and Marcela V Maus. "Immunotherapy with Chimeric Antigen Receptors for
Multiple Myeloma." Discovery Medicine 18.91 (2014): n. pag. Print.
(7) http://www.fredhutch.org/en/news/spotlight/imports/evasion-of-adoptive-t-cell-therapy-through-loss-of-mhc.html
(8) Lee, DW, R. Gardner, and DL Porter. "Current concepts in the diagnosis and management of cytokine release
syndrome." National Center for Biotechnology Information. U.S. National Library of Medicine, 10 July 2014.
Web. 10 Dec. 2014. <http://www.ncbi.nlm.nih.gov/ pubmed/24876563>.
(9) "T-Cell Therapy Eradicates an Aggressive Leukemia in Two Children." Penn Medicine. The Trustees of the
University of Pennsylvania, 25 Mar. 2013. Web. 10 Dec. 2014. <http://
www.uphs.upenn.edu/news/News_Releases/2013/03/grupp/>.