Download 542. Photothermal Therapy in Combination with Immunotherapy to

Photothermal Therapy in Combination with Immunotherapy to Inhibit Cancer Growth and Metastasis
Jun Chen
CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, Multi-disciplinary Research Division,
Institute of High Energy Physics, Chinese Academy of Sciences, Beijing, 100049, China.
*Email: [email protected].
orthotopic MB49 bladder cancer in mice. Therefore, the
Statement of Purpose: In this study, we report the design
present study explored the feasibility and efficacy of
of nanocarbon-based systems that combine photothermal
CS@GO nano-delivery as a feasible system for sustained
treatment with immunotherapy. We have developed
delivery of the immune cytokine SA-GM-CSF by the
chitosan-coated carbon nanotubes that assemble the GMintravesical route. A variety of thermosensitive
CSF for photothermal immunotherapy in a orthotopic
nanoparticles have been studied as vehicles for sustained
mouse bladder cancer model. Post laser excitation, the
drug delivery. Chitosan(CS)/graphene(GO) as SA-GMcarbon nanotubes-mediated photothermal
CSF delivery has great advantanges, such as nontoxic,
immunotherapy, compared with immunotherapy or
degradation, storage effect and its thermal effect. To
photothermal therapy alone, exhibits more potent innate
validate the effectiveness of nano-delivery system for
and adaptive immune responses, resulting in combined
intravesical SA-GM-CSF, we explored its antitumor
anticancer effects against primary tumors and metastatic
effect, local and system immune therapeutic effects. First,
tumors.
the temperature of mice can be heated up to 42~45 ℃ or
even
higher, the bladder tumor cells are hyperthermia or
Methods: Preparation and characterization of NPs
directly
killed. And our studies showed the intravesical
(CS, CS@NGO): The CS@NGO NPs were prepared by
CS@GO-SA-GM-CSF was better than SA-GM-CSF
a nonsolvent counterion complexation method established
alone in reducing the weight of tumors and prolonging the
in the reference. Bioactivity assays of free and cell
survival time. Moreover, the DC(CD11c+CD11b+ and
membrane-bound SA-GM-CSF bifunctional fusion
CD11c+CD86+), T cells subpopulation(CD4+and
proteins: Flow cytometric analysis and bone marrow cell
CD8+T) and natural killer cell(NK cells ) in CS@GOproliferation assays were performed to test the
SA-GM-CSF group level have significantly increased.
bifunctionality of SA-GM-CSF fusion protein.
Therefore, mGM–CSF, DCs and T lymphocytes have a
Intravesical immobilization of SA-GM-CSF/CS and
close relationship with the antitumor response. MB49
SA-GM-CSF/CS@NGO fusion protein for
cells can be identified and killed by CD4+and CD8+T
immunotherapy of superficial bladder cancer: The
lymphocytes.
mouse orthotopic model of MB49 bladder cancer was
used to evaluate the feasibility and efficacy of the novel
immunotherapy through intravesical immobilization of
SA-GM-CSF@CS and SA-GM-CSF/CS@NGO
bifunctional fusion protein on the biotinylated mucosal
surface of bladder wall. Cytokine detection: Serum
samples were isolated from mice after various treatments
and diluted for analysis. IL-1β (Dakewe biotech), tumor
necrosis factor (TNF-α, Dakewe biotech), IL-12 (Dakewe
biotech), and IL-6 (Dakewe biotech) were analyzed with
ELISA kits according to vendors’ instructions. All
measurements were carried out in triplicate. FCM
analysis of immune effector cells in each treatment
group tumor-bearing mice: The percentages of T
lymphocytes (in blood and bladder) were investigated by
FCM. The bladder were harvested from tumor-bearing
mice and teased apart into single cell suspension by
Figure 1. The different nano@SA-GM-CSF stimulate the
pressing with plunger of a syringe. After washing by PBS
recruitment
and activation of innate and adaptive immune
for twice, cells were stained with anti-mouse-CD4 APC,
cells.(A)
Flow cytometry plot of CD4+,
anti-mouse-CD8aPE, anti-mouse-CD80 FITC, antiCD8+,CD11b+CD11c
+,CD11c+CD86 +cells isolated
mouse-CD86 PE, anti-mouse-CD11c FITC and antifrom
bladder
mouse-CD11b APC for 30 min at 4 ℃ according to the
Conclusions:
In
conclusion,
the immune adjuvant loaded
manufacturer instructions. After washing by FACS (2%
by
multifunctional
nanoparticles
offers a new way for
FBS) and filtering by a nylon membrane, cell samples
bladder
cancer
treatment
through
intravesical therapy.
were analyzed by FCM through FL 1, FL 2, FL 3 and FL
The
photothermal
effect
can
boost
the immuno-response
4 channel.
and achieve better therapeutic efficacy.
Results: Intravesical SA-GM-CSF delivery involves the
direct administration of the SA-GM-CSF into the urinary
bladder through a catheter and has been investigated for