Novel Chemotactic-Antigen DNA Vaccine against Cancer

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Novel Chemotactic-Antigen DNA Vaccine against
Cancer ????????????
Shuren Zhang ???
Cancer Institute, Hospital Chinese Academy of
Medical Sciences Peking Union Medical Collage
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Regulation of Cytokines secretion by secondary
lymphoid tissue chemokine (SLC)
Intratumoral SLC administration promotes Th1
cytokine and antiangiogenic chemokine release and
a decline in immunosuppressive mediators
Th1 cytokine IFN-g, GM-CSF, IL-12, MIG, and
IP-10 Immunosuppressive mediators PGE2, IL-10,
and TGF-b angiogenic factor VEGF
The Journal of Immunology, 2000, 164 45584563.
Steven M. Dubinett
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Chemotactic-Antigen DNA Vaccine (CADV)
????????
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Effect Mechanisms of CADV
Gene transfer and expression
Recruiting DC and T cell catching Ag by
ligands Ag presentation
T and B cell activation, anti-Tumor Effects
Injection or Gene gun
EP
Antigen presentation by DCs through
ligand-mediated internalization can be
enhanced up to 10,000-fold over fluid-phase
antigen pinocytosis
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Mouse Model of CADV
HPV16E7
Her2/P53
mSLC
Fc
CMV
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Mouse Model of CADV (pSLC-E7-Fc)
E7
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RT-PCR and Western Blotting Analysis
(A) RT-PCR detection of B16F10 cells transfected
with various recombinant plasmids. DNA marker
(M),Lane 1 (vector control), Lane 2 (SLC
fragment), Lane 3 (Sig-E7 fusion fragment), Lane
4 (SLC-E7 fusion fragment),Lane 5 (Sig-Fc fusion
fragment), Lane 6 (SLC-Fc fusion fragment), Lane
7 (Sig-E7-Fc fusion fragment), Lane 8 (SLC-E7-Fc
fusion fragment). (B) Western blotting analysis
of B16F10 transfectants. SLC-E7-Fc fusion protein
(50kD), E7-Fc fusion protein (38kD),SLC-E7 fusion
protein (22kD).
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Chemotaxis Assay by Chemotaxis Chamber
pSLC-E7-Fc pcDNA

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Therapeutic immunization with pSLC-E7-Fc induces
C3 tumor regression
HPV16-E7 as tumor antigen. E7C3 tumor cell
subcutaneous inoculated in C57BL/6 mice on day0.
The tumor bearing mouse immunized by gene gun
with different gene vaccine on day 6, day 13 and
day 20.
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Therapeutic immunization with pSLC-E7-Fc induces
TC-1 tumorregression
HPV16-E7 as tumor antigen. E7TC-1 tumor cell
subcutaneous inoculated in C57BL/6 mice on day0.
The tumor bearing mouse immunized by gene gun
with different gene vaccine on day 15, day 22 and
day 28.
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Tumor incidence after TC-1 tumor recharlenged

Recharlenge with 1.8 x 105 TC-1, after 90days
later of first TC-1 inoculation.
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Prostate Cancer Chemotactic-Antigen DNA Vaccine
????????????
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human SLC with Chemotactic Activity for mouse
immune cells
Human PBMC
Mouse spleen cell
A and B Human PBMC C and D mouse
spleen cell A and C Negative control (200) B
and D SLC-3P-Fc (200)
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Transfection with pSLC-3P-Fc chemoattracts
lymphocytes to the immunization site
A B C
pathology HE staining of dermis tissues sections
from C57BL/6 mice, non-treated mice (A), treated
with pC (B) or pSLC-3P-Fc (C). Images are
representative of multiple microscopic fields
observed in at least three mice per group.
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Immunotherapy effects of human pSLC-3P-Fc gene
vaccine on mouse model
B16-3P tumor cell subcutaneous inoculated in
C57BL/6 mice on day0. The tumor bearing mouse
immunized by gene gun with different gene vaccine
on day 4, day 9 and day 14.
3P PSA-PAP-PSM Target B16-3P
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Induction of human CTL against cancer by CADV in
vitro
Day 0 Day 1 Day 6 Day 13
MLC Mixed Lymphocyte Culture
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SLC-3P-Fc DNA Vaccine induced CTL by MLC
EffectorTarget601
A0201 -
hPSA -
- -
- - hPSM -
- - -
- hPAP -
- - -
-
A Stimulating cell tansfected by pSLC-3P-Fc
B Stimulating cell tansfected by pcDNA
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Universal Chemo-Antigen DNA Vaccine
(CADV) ??????????
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SLC-TERT-Fc DNA Vaccine induced human CTL by MLC
The CTLs induced by pSLC-T-Fc-transfected
PBMC, pC-transfected PBMC, and non-transfected
PBMC were tested for cytolytic activity against
MCF-7, SK-BR3, U2OS/T, U2OS, LNCap, PC-3M,
SK-OV-3, and T2 .
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Assessment of involvement of NK cells, CD4 and
CD8T cells in antitumor effect
CD8 CD4 NK
FIGURE 8 Assessment of involvement of NK cells,
CD4 and CD8T cells in antitumor effect. Animals
were s.c. challenged with 5 x 104 B16/Te cells on
day 0, then treated with B16/SLC-Te-Fc or B16/C
plus anti-4-1BB (n 4) as Figure 6a described.
Cell subsets were depleted by the injection of
GK1.5 (for CD4 T cells), 2.43 (for CD8 T cells)
ascites fluids (100 ml/mouse) on days 2, 4 and 6,
then twice a week before killing. For NK cells
depletion, asialo-GM1 antibodies (200 mg/mouse)
were injected for 5 consecutive days beginning on
day 2, then twice a week before killing. The
control Ig was injected in the same way. (a)
Abrogation of CTL-mediated cytotoxicity. (b) The
s.c. tumor size was measured twice per week.
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Evaluation of therapeutic effect on established
pulmonary metastases by CADV-tumor anti-41BB
B16/SLC-Te-Fc anti-4-1BB B16/SLC-Te-Fc
Ig B16/C anti-4-1BB B16/C Ig
C57BL/6 mice were challenged via tail vein with 1
x 105 B16/Te tumor cells on day 0 (n 5), and
and were treated with CADV modified tumor cells
immunization on days 7 and 11 followed by either
rat Ig or anti-4-1BB MAbs administration (i.p.)
on days 8, 11 and 14. The lung metastatic nodules
were counted on day 23. p lt 0.001 versus B16/C
Ig and B16/C anti-4-1BB by Student paired
t-test p lt 0.05 versus B16/CCL21-Te-Fc 1 Ig by
Student paired t-test.
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The Advantageous Features of Chemotactic-Antigen
DNA Vaccine
1. The CADV could facilitate the co-localization
of DC, T, B and NK cells by SLC and induce
effective cell-mediated immune response and
humoral immune response . 2. CADV fusion protein
can be forcefully captured by DC (Fc/FcR). This
approach can efficiently increase antigen
presentation by DCs, to induce both
antigen-specific Th and CTL. 3. Tumor antigen
specific memory T cells can be highly induced by
CADV to prevent tumor recurrence and
metastasis. 4. CADV is a recombinant plasmid DNA.
It is suitable for industrialisation. 5. CADV is
a vaccine platform. Tumor or non-tumor vaccines
can be easily made by this system.
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Publications
1?Specific antitumor immune response induced by a
novel DNA vaccine composed of multiple CTL and T
helper cell epitopes of prostate cancer
associated antigen, Immunology Letters
9985-93,2005 2?Enhancement of Antitumor Immunity
by a Novel Chemotactic Antigen DNA Vaccine
Encoding Chemokine and ultiepitope of Prostate
Tumor Associate Antigens, Immunology, 117
419-430, 2006 3?Enhancement of DNA vaccine
potency by sandwiching antigen-coding gene
between secondary lymphoid tissue chemokine (SLC)
and IgG Fc fragment genes, Cancer Biology and
Therapy,Apr 205(4), 2006 4?Enhanced anti-tumor
effect against human telomerase reverse
transcriptase (hTERT) by vaccination with
chemotactic-hTERT gene-modified tumor cell and
its combination with anti-4-1BB monoclonal
antibodies, International Journal of Cancer 119,
2006 5?Induction of protective and therapeutic
antitumour immunity using a novel
tumour-associated antigen-specific DNA
vaccine,Immunology and Cell Biology 84, 18 ,
2006 6?Potent Systemic Antitumor Immunity
Induced by Vaccination with Chemotactic-Prostate
Tumor Associated Antigen Gene-Modified Tumor Cell
and Blockade of B7-H1,Journal of Clinical
Immunology, 27(1)117-130, 2007 7?Synergistic
antitumor effect of chemotactic-prostate tumor
associated antigen gene-modified tumor cell
vaccine and anti-CTLA-4 mAb in murine tumor
model,Immunology Letters, 1139098, 2007 8?A
novel chemotactic-antigen DNA vaccine against
cancer, Future Oncology, Apr4(2)299-303,2008
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Thank you !
Lin Chen
Sun Wenxin
Qin Hanjun
Lin Xiaoyan
Liu Rongzhi