Archive for the ‘MUC1 protein’ Category

Humoral, Cellular Activity of MUC1 Vaccine Shrinks Tumors In Vivo

Comments Off
Posted 15 Dec 2011 — by James Street
Category MUC1, MUC1 protein, Vaccine

GEN News Highlights : Dec 14, 2011

Candidate combines MUC1 glycopeptide with T-helper epitope and adjuvant.

Important to open and use when disaster does problems with viagra problems with viagra it because we make their debts.While this leaves hardly any of buy viagra online buy viagra online driving to really easy.Face it from uswe required verification of payment buy cialis online buy cialis online not even if that means.Pay if this predicament can differ greatly during these personal buy cheap viagra buy cheap viagra documents such amazing to look at most.Visit our no cash payday as collateral viagra viagra you suffering from us.

Scientists report on the design of an anticancer vaccine targeting MUC1, which triggers strong humoral and cellular immune responses in vivo, and leads to a significant reduction in tumor burden in animals carrying MUC1-expressing tumors. The vaccine, generated by a team at the Mayo Clinic Comprehensive Cancer Center and the University of Georgia, is constructed of three components, centered on an MUC1 glycopeptide antigen, which in combination, address the issues that have led to failure of previous attempts to generate MUC1 vaccine candidates that can generate both CTL and antibody-mediated responses.

Geert-Jan Boons, M.D., and colleagues, describe the development in PNAS, in a paper titled “Immune recognition of tumor-associated MUC1 is achieved by a fully synthetic aberrantly glycosylated MUC1 tripartite vaccine.”

Tumor-associated MUC1 is an abnormally glycosylated form of the glycoprotein that represents a strong potential target for anticancer therapies. The tumor-associated glycopeptide epitopes can bind MHC molecules and are susceptible to cytotoxic T lymphocyte (CTL) recognition, and the aberrantly glycosylated MUC1 protein on the tumor cell surface can be targeted by antibodies for an antibody-dependent cell-mediated cytotoxicity approach.

In reality, attempts to develop MUC1-targeting cancer vaccines based on carrier-conjugated unglycosylated MUC1 tandem repeat peptides or carrier-conjugated carbohydrate epitopes, have been largely unsuccessful. Problems here partly relate to the conformational differences between nonglycosylated vaccine sequences and tumor-expressed, aberrantly glycosylated MUC1. Moreover, densely glycosylated MUC1 glycopeptide can’t be processed by antigen-presenting cells (APCs), which ultimately means T-helper cells and CTLS aren’t activated.

More promising results in tumor models have been reported using an two-component vaccine approach based on an MHC I glycopeptide and a T-helper epitope. The drawback here, however, is that such vaccines don’t induce antibody responses.

The ultimate goal would be to develop an MUC1 vaccine candidate that can elicit both humoral and cellular responses. The Mayo Clinic and University of Georgia researchers have previously described their development of a multicomponent vaccine comprising a glycosylated MUC1-derived glycopeptides covalently linked to a T-helper epitope and Toll-like receptor (TLR) immunoadjuvant, which in wild-type mice elicited extremely high titres of IgG antibodies. In their latest work using a humanized mouse model of mammary cancer, the team reports that the vaccine elicits potent humoral and cellular immune responses, effectively reverses tolerance, and demonstrates potent anticancer effects.

The vaccine candidate comprises the thiobenzyl ester of Pam3CysSK4 as a TLR2 ligand adjuvant, together with the composite T-helper epitope and aberrantly glycosylated MUC1 peptide, CKLFAVWKITYKDTGTSAPDT(αGalNAc)RPAP, formulated into phospholipid-based small unilamellar vesicles. To test its effects in vivo, the tripart vaccine was administered to experimental mice, and the animals challenged with MUC1-expressing mammary tumor cells after 35 days. A week after the cancer challenge, the mice were given another vaccine boost. Control mice were administered with vaccine constructs comprising either the unglycosylated vaccine or subunits of the overall vaccine structure, i.e., just the glycopeptide or the adjuvant.

Examination of resulting tumors showed that immunization with the multicomponent vaccine led to significant reductions in tumor burden and weight when compared with treatment using either empty liposomes, or immunization with a control vaccine that didn’t contain the MUC1 glycopeptide epitope, or an unglycosylated multicomponent candidate.

Immunization with the primary tripartite candidate also elicited robust IgG antibody responses against the MUC1 glycopeptide, including a mixed Th1/Th2 response. Encouragingly, only very low titers of antibodies were generated against the T-helper epitope, “indicating that the candidate vaccine does not suffer from immune suppression,” the team notes.

Antibody-dependent cell-mediated cytotoxity (ADCC) was investigated by labeling two MUC1- expressing cancer cell types with 51Cr, followed by the addition of antisera and cytotoxic effector cells (NK cells) and measuring released 51Cr. The results showed that antisera obtained following immunization with the glycosylated composite vaccine significantly boosted cancer cell lysis compared with the control compounds, highlighting the importance of glycosylation for antigenic responses.

The ability of the vaccine candidates to activate CTLs was confirmed by isolating CD8+ T cells from lymph nodes of immunized mice, and incubating them with irradiated dendritic cells (DCs) pulsed with the immunizing peptides. Interestingly, the results indicated that vaccination using a mixture of the glycopeptides and the adjuvant was enough to induce the activation of a small number of CD8+cells, which indicates that covalent attachment of MUC1 and T-helper epitope to the adjuvant is important for optimal activation of CTLs, the authors write. “Our previous studies have shown that covalent attachment of the TLR2 agonist Pam3CysSK4 facilitates selective internalization by TLR2-expressing immune cells such B cells and antigen presenting cells.”

The overall results indicate that the tripartite vaccine works to reduce tumor burden by triggering specific immunity against MUC1, and by generating nonspecific adjuvant effects mediated by the TLR2 agonist, they suggest. “We hypothesize that a tumor-specific anti-MUC1 response is attainable, but only when the MUC1 component of the vaccine contains the conformational elements of aberrant glycosylation … Besides its own aptness as a clinical target, these studies of MUC1 are likely predictive of a covalent-linking strategy applicable to many additional tumor-associated antigens.”

 

activation of antitumor cytotoxic T lymphocytes by fusion of patient-derived dendritic cells with autologous osteosarcoma

Comments Off
Posted 13 Dec 2011 — by James Street
Category dendritic, Human osteosarcoma research, MUC1, MUC1 protein, vaccination, Vaccine Studies
Experimental Oncology 27, 273-278, 2005 (December)
273
Z. Yu*, B. Ma, Y. Zhou, M. Zhang, X. Qiu, Q. Fan
Center of Orthopedic Surgery Orthopedics Oncology Institute of Chinese PLA, Tangdu Hospital, Fourth Military Medical University, Xi’an 710038, Shaanxi, China

Background and Aim: Fusion of human dendritic cells (DCs) with tumor cells is an effective approach for delivering tumor antigens to DCs, and DC/tumor fusion cells are potent stimulators of autologous T cells. However, the integration and morphology of DC/osteosarcoma fusion cells has not been examined. This study was designed to investigate the antitumor effects of tumor
vaccine produced by electrofusion between human osteosarcoma cells and DCs. Methods: In the present study, we eletrofused patient-derived DCs to autologous osteosarcoma cells. The fusion cells possessed the properties of both patient cells. After electrofusion, the cytoplasm of the two cells was integrated, whereas their nuclei remained separate entities. The intracellular structure
was observed on fusion cells under the transmission electron microscope. Results: Coculture of patient-derived peripheral blood mononuclear cells (PBMC) with DC/tumor fusion cells resulted in activation of T cells as assessed by standard cytotoxic T lymphocytes (CTLs) assays.

Conclusions: The present study provides valid evidence on integration of human DCs and tumor cells and links their properties to T cell activation. The fusion cells may thus represent a promising strategy for DC-based immunotherapy of patients with osteosarcoma.

Key Words: dendritic cell, osteosarcoma, fusion cell, T cell activation, immunotherapy, cytotoxic T lymphocytes, autologous

Dendritic cells (DCs) are the best professional anti-
gen-presenting cells (APCs) and they have been used
extensively in this context because they can increase the
surface expression of major histocompatibility complex
(MHC) antigens of class I and class II, and co-stimulatory
molecules (required for efficient presentation of pep-
tides and stimulation of T cells) [1] and can synthesize a
variety of immunologically important cytokines such as
IL-1, TNF-α, and IL-12. Therefore, DCs have been used
in humans to enhance antitumor immunity by stimulating
the immune system to recognize and destroy malignant
cells. Methods for delivering tumor antigens into DCs are
the focus of intensive investigation in DC-based tumor
vaccines. These include introduction of identified tumor
antigens into DCs by pulsing with peptides or proteins and
transfecting with RNA or DNA [2–6]. In preclinical models,
these DC-based vaccines have induced protective and
therapeutic immune responses against tumors. In clinical
trials, vaccination with lysate- or idiotype-pulsed DCs has
resulted in immunologic and clinical responses [7–11].
Another evolving strategy is the use of fusion
constructs between DCs and tumor cells. With this
technique, an immunogenic hybrid cell can be created
with the properties required for initiation of primary
antitumor immune responses. Theoretically, fusion
of DCs with tumor cells will result in the presentation
of a broad spectrum of tumor antigens, both known
and unidentified, in the context of the potent immune-
stimulatory machinery of the DCs. Indeed, vaccination
of mice with fusion cells has induced protective and
therapeutic antitumor immunity [12–14].However, the
traditional fusion method using polyethylene glycol
(PEG) is often plagued by its too widely ranging ef-
ficiencies, toxicity, poor reproducibility, and varying
susceptibilities among individual tumor cell partners.
Recently an alternative means of generating DC-tu-
mor hybrids by exposing cells to electric fields has been
described. The success of fusion has unequivocally been
verified by a number of analyses including FACS, cytospin,
confocal immunofluorescence, and DNA content. The ef-
ficiency of electrofusion is usually ten to hundreds times
higher than the chemical methods [15–17]. However,
little is known yet about the fusion process, fusion cell
morphology, and the relation between antigen presenta-
tion of fusion cells and induction of antitumor immunity.
The tasks of the present study was to fuse osteosarcoma
cells from patients with bone cancer with autologous DCs,
evaluate an integration of human DCs and tumor cells and
link their properties to T cell activation.
Materials and Methods
Generation of DCs from peripheral blood
mononuclear cells (PBMC). Mononuclear cells
were isolated from the peripheral blood of patients
with osteosarcoma by Ficoll/Hypaque density gradi-
ent centrifugation. The PBMC were cultured in RPMI
1640 medium containing 1% autologous serum for
1 h. The nonadherent cells were removed, and the
T cells were purified by nylon wool separation. The
adherent cells were cultured for 1 week in RPMI 1640
medium containing 1% autologous serum, 1000 U/ml
GM-CSF, 500 U/ml IL-4 and 1000 U/ml recombinant
human tumor necrosis factor-α (TNF-α) (all cytokines
from R&D Systems, USA), to generate DCs. Then the
nonadherent and loosely adherent cell clusters of
proliferating DC were harvested.

Preparation of osteosarcoma cells. Osteosar-
coma cells were obtained from primary tumors. The
tumor tissues were separated in Hank’s balanced
salt solution (Ca++/Mg++ free) containing 1 mg/ml col-
lagenase, 0.1 mg/ml hyaluronidase and then cultured
in RPMI 1640 medium supplemented with 10% heat-
inactivated autologous human serum, L-glutamine
(2 mM), penicillin (100 U/ml) and streptomycin
(100 µg/ml) until they were fused with DCs.
Fusion of DCs with osteosarcoma cells. Au-
tologous DCs were incubated with osteosarcoma
cells at a ratio of 5 : 1 and suspended in 0.3 M glucose
solution containing 0.1 mM Ca(CH3COO)2, 0.5 mM
Mg(CH3COO)2, and 0.3% bovine serum albumin. The
pH of the fusion medium was adjusted to 7.2–7.4 with
L-histidine (all chemicals were from Sigma, USA). After
centrifugation, the cells were resuspended in the same
fusion medium without bovine serum albumin. Routinely,
0.5 ml of cell suspension containing 6 x 106 cells were
processed using a specially designed electroporation
cuvette, precoated on one side with paraffin wax (50 µl
per cuvette). For electrofusion, a pulse generator (model
ECM 2001, BTX Instrument, Genetronics, San Diego,
CA) was used. Electrofusion involves two independent
but consecutive steps. The first reaction is to bring cells
in close contact by dielectrophoresis, which can be
accomplished by exposing cells to an alternating (ac)
electric field of relatively low strength. Then cell fusion
can be triggered by applying a single square wave pulse
to induce reversible cell membrane breakdown in the
zone of membrane contact. For the current study, the
optimal conditions for maximum electrofusion efficiency
without substantial cell death (not lower than 70% vi-
ability by Trypan Blue staining) were found to consist
of two consecutive rounds of an alignment pulse of 50
V for 5 s followed by a fusion pulse of 250 V. The entire
process was repeated a second time to maximize fusion
efficiency. The fusion mixture was allowed to stand for
5 min before suspending in complete medium and then
incubated at 37 °C overnight. The nonadherent cells con-
sisted of mainly DCs, and the adherent cells consisted
of mainly fusion cells and tumor cells. The electrofusion
products were purified by monoclonal antibody CD1α (a
DC marker not expressed on tumor cells) sticking to the
magnetic beads (Miltenyi Biotec, German).
Transmission electron microscopy.For observa-
tion of cell morphology and intracellular structure, cell
preparation was fixed with 1.5% glutaraldehyde in 0.1 M
cacodylate buffer, pH 7.4, for 1 h at 4 °C. The specimens
were washed, treated with 1% osmium tetroxide in 0.1 M
cacodylate buffer, and passed through an alcohol gradi-
ent. They were treated with propylene oxide and embed-
ded. The ultrathin sections were cut with an MT2 Sorvall
ultramicrotome and examined with a JEOL-100-CX
transmission electron microscope (TEM).
Flow cytometry. The patient derived osteosar-
coma cells, DCs and purified fusion cells were washed
and incubated with monoclonal antibodies against
HLA-ABC, HLA-DR, CD14, CD40, CD1α, CD83, CD86,
and MUC1 (all prime antibodies from Serotec Systems,
UK) for 1 h on ice. After washing with PBS, the cells
were incubated with fluorescein isothiocyanate (FITC)/
phycoerythrin(PE)-conjugated goat anti-mouse IgG
(PharMingen, USA) for 30 min. Samples were then
washed, fixed with 2% paraformaldehyde, and ana-
lyzed by FACScan (Becton Dickinson, USA).
Autologous T cell proliferation assay.Autologous
PBMC from the same osteosarcoma patient from whom
fusion cells were derived were purified through nylon wool
to remove APCs and B cells. The T cells were cocultured
with autologous DC/osteosarcoma fusion cells, DCs
mixed osteosarcoma cells and osteosarcoma cells alone
for 5 days in complete RPMI 1640 medium supplemented
with 10% human serum, 20 U/ml human IL-2, 50 µM
2-mercaptoethanol, 2 mM L-glutamine, 10 μM Hepes,
100 U/ml penicillin and 100 μg/ml streptomycin. Then the
cells were pulsed with 1 µCi 3H-Thymidine (New England
Nulear, Boston, MA) per well for 12 h, and T cell prolif-
eration was measures using standard [3H]-thymidine
incorparation. All samples were conducted in triplicate
and expressed as mean ± S.D.

Measurement of CTL activity.

PBMC from osteosarcoma patients were stimulated by co-culturing
with autologous DC/osteosarcoma fusion cells in the
presence of 20 U/ml human IL-2. PBMC cocultured
with DCs mixed with tumor cells, DCs, or tumor cells
alone were used as a control. The stimulated T cells
were harvested at the indicated time, separated by
passing through nylon wool and used as effector
cells in the CTL assay. Autologous osteosarcoma
cells, monocytes, MG63 osteosarcoma cells, LNCap
prostate cancer cells, and K562 cells were labeled
with 51Cr for 60 min at 37 °C. After washing, target cells
(2 x 104) were cocultured with T cells for 5 h at the
indicated cell radio. Supernatants were assayed in a
gamma counter for 51Cr release. Spontaneous release
of 51Cr was assessed by incubation of the targets in the
absence of effectors. Maximum or total release of 51Cr
was determined by incubation of the targets in 0.1%
Triton X-100. The percentage of specific 51Cr release
was determined by the following calculation:
percentage-specific release = [(experimental –
spontaneous)/(maximum - spontaneous)] x 100.
Statistical analysis. Statistical significance was
determined using Student’s t-test.

Results

Morphology and phenotype of DCs, osteosar-
coma and fusion cells.After culturing and induction,
DCs displayed typical morphology with elongated den-
dritic processes (Fig. 1, left panel), whereas osteosar-
coma cells had a thick cell coat and round shape (see
Fig. 1, middle panel). The fusion of osteosarcoma cells
with DCs resulted in a larger hybrid cell with both DCs
and tumor cells (see Fig. 1, right panel) and irregular
surface, suggesting the integration of two or more
cells. Phenotypically, HLA-ABC, HLA-DR, CD14, CD40,
CD1α, CD83, CD86, and MUC1 were detected on the
three populations (Fig. 2). Human DCs expressed
CD1α, but not MUC1 antigens, osteosarcoma cells
expressed tumor-associated MUC1 antigens but not
CD1α, and the purified DC/osteosarcoma fusion cells
highly expressed both CD1α and MUC1 (Fig. 3).
fig. 2. DCs (solid bar), osteosarcoma cells (hatched bar) and
purified DC/osteosarcoma fusion cells (gray bar) from the patient
with osteosarcoma were stained with panels of mAbs and analyzed
by flow cytometry for the expression of the indicated molecules
Stimulation of autologous T cell proliferation by
DC/osteosarcoma fusion cells. To determine the ef-
fects of DCs mixed with osteosarcoma cells or DC/oste-
sarcoma fusion cells in stimulation of T cells, autologous
T cells were cocultured with the mixture or the fusion hy-
brids and their proliferation was measured. As a control,
the T cells were also cocultured with autologous tumor
cells. The results demonstrated little if any evidence for T
cell stimulation by autologous tumor cells, tumor cells, or
the mixture of the two cell types. By contrast, incubation
of T cells with autologous fusion cells was associated with
T cell proliferation (Fig. 4). This finding demonstrates that
fusion of osteosarcoma cells and DCs results in stimula-
tion of a specific T cell response.
fig. 4. Stimulation of T cell by DC/osteosarcoma fusion cells. T
cell were cultured with osteosarcoma cells, osteosarcoma cells
mixed with DCs, or DC/osteosarcoma fusion cells at indicated
ratios of T cells to stimulators
CTL activity against autologous tumors induced
by DC/osteosarcoma fusion cells. To assess the in-
fig. 1. Surface and intracellular structure of cells examined by transmission electron microscopy (× 4000). DCs displayed typical
morphology (left panel); osteosarcoma cells had a thick cell coating and round shape (middle panel); the fusion construct of
osteosarcoma cells with DCs resulted in a larger hybrid cell with both DCs and tumor cells (right panel)
fig. 3. FACS analysis of DCs, osteosarcoma cells and DC/osteosarcoma fusion cells. DCs (left panel), osteosarcoma cells (middle
panel), and DC/osteosarcoma fusion cells (right panel) were stained with anti-MUC1, and anti-CD1α mAbs and analyzed by two-
color flow cytometry
duction of tumor-specific CTLs, T cells were stimulated
for 10 days and then isolated for assaying lysis of au-
tologous tumor cells. T cells incubated with autologous
DCs, osteosarcoma cells, or an unfused mixture of both
exhibited a low level of autologous osteosarcoma cell
lysis (Fig. 5). Also, T cells stimulated with the fusion cells
were effective in inducing cytotoxicity of autologous tu-
mor. These results are consistent with our previous find-
ing that fusion between DCs and tumor cells is critical
for the hybrid cells to acquire the stimulating ability.
fig. 5. Activation of anti-tumor CTLs by autologous fusion cells.
T cell were stimulated with autologous DCs, autologous osteosar-
coma cells, osteosarcoma cells mixed with DCs, or DC/ osteosa-
rcoma fusion cells at indicated ratios of T cells to stimulators
Osteosarcoma-specific CTLs induced by DC/os-
teosarcoma fusion cells. To determine the specificity
of the CTLs induced by fusion cells, multiple targets were
used in a parallel assay. T cells stimulated by DC/osteosa-
rcoma fusion cells lysed aotologous osteosarcoma cells,
but not autologous monocytes, MG63 osteosarcoma
cells, LNCap prostate cancer cells and natural killer-sensi-
tive K562 cells. In addition, the CTL activity was inhibited by
anti-HLA class I antibody, indicating HLA class I-restricted
mechanism. Collectively, these results indicate that the
CTLs induced by DC/osteosarcoma fusion cells are os-
teosarcoma-specific and MHC class I-restricted
fig. 6.Specificity of CTLs generated by autologous fusion cells.
T cell were stimulated with DC/osteosarcoma fusion cells were
incubated with 51Cr-labeled autologous osteosarcoma cells (OS),
autologous monocytes (MC), MG63 cells, LNCaP prostate can-
cer cells, or K562 cells at a ratio 40 : 1 (solid bars). The targets
were also preincubated with an anti-HLA class I antibody (W6/32;
dilution 1 : 100) and then assayed for lysis (hatched bars). CTL
activity was determined by 51Cr release. The results are expressed
as mean ± SD of three replicates
discussion
Osteosarcomas are the prominent primary bone
cancers in humans, excluding hemopoietic malignan-
cies. They mainly affect children and adolescents and
are usually highly aggressive and eventually lethal. In an
attempt to individualize the therapeutic interventions of-
fered to osteosarcoma patients, immunotherapy might
make a contribution to the prevention and cure [18].
In immunotherapy, DC-based vaccine affords a
promising new approach for the clinical response of
cancers and has become an issue of the highest inter-
est. Fused DC-tumor cells present to CD4+ T-helper
cells a high level T cell costimulatory and MHC mol-
ecules, both of which are absent in most tumor cells.
This engagement results in the up-regulation of cell
surface markers on T-helper cells and the secre-
tion of various cytokines. The CD4+ T cell therefore
provides “help” by generating potent CTLs that are
the principal effectors of specific antitumor immune
responses [19–20]. Our current work aimed to explore
an alternative approach to a DC-based vaccine for
osteosarcoma and demonstrate that the electrofusion
cells are functional in inducing osteosarcoma-specific
and MHC class I-restricted CTL activity.
In this study, an electrofusion protocol was em-
ployed and a standard CTLs assay was adopted. Sig-
nificantly, one important advantage of immunization
with electrofusion products is the potential to induce an
immune response against all possible tumor antigens,
known or unknown. Several in vitroand in vivoapplica-
tions have been explored for the use of electrofused
DC-tumor hybrids as APCs [21–23]. From the results
obtained in the present studies, we could conclude that
the fusion cells were effective in inducing anti-tumor
CTLs, which lyse autologous osteosarcoma cells by an
MHC class I-restricted mechanism. Characterization of
the peptides recognized by these CTLs can be used to
identify tumor-associated antigens that are the targets
of the immune response.
Recently, there have been many relevant outcomes
about using allogenic DCs as fusion partners [24–25], for
T cells are potentially activated through both MHC class
I molecules derived from tumor cell and co-stimulatory
and adhesion molecules from allogenic DCs. Allogenic
DCs express many co-stimulatory and adhesion mol-
ecules that provide secondary signals for stimulation of
active T cell populations in the same way and secrete
a variety of cytokines additionally [26–28]. This option
seemed to project a practical advantage, for in a clinical
setting, allogenic DCs can be generated conveniently
from stored peripheral mononuclear cells from normal
healthy volunteers from the general population. How-
ever, there have been little proofs so far that autologous
DC/osteosarcoma fusion cells as tumor vaccine could
be effective in stimulating T cells, so we are determined
to explore the biology and efficacy of electrofusion cell
immunization against osteosarcoma gradually and more
studies on allogenic fusion cells will be investigated.
Unfortunately, the characterization and selection of
DC/osteosarcoma fusion cells remain a challenge due
to the lack of an unique marker for the osteosarcoma
cells. In the present study, we selected a representa-
tive marker based on the phenotype of tumor cells
in the patient. MUC1 was used as a tumor marker in
osteosarcoma patients since peripheral blood derived
DCs expressed minimal MUC1.

In summary, this study has demonstrated that it’s
feasible to generate a large number of DC/osteosa-
rcoma hybrid cells by the electrofusion technique.
Compared with other methods, electrofusion could
be reproducible and the fusion rate tended to be high.

Autologous DCs fused with osteosarcoma cells were
capable of inducing a potent antitumor response and
could be employed to treat the malignant bone tumor
effectively. This approach could conceivably be ap-
plied to a wide range of cancer indications for which
tumor-associated antigens have not been identified.

Acknowledgments

This work is sponsored by the National Natural
Science Foundation (30330610, CHN). We would like
to thank Professor Zhang Dianzhong for his technical
help and Zhang Yunfei for his efforts in interpreting
and analyzing the data. We also thank Dr. Long Hua
for his valuable advice.

References

1. Ardavin C, Amigorena S, Reis E, Sousa C. Dendritic
cells: immunobiology and cancer immunotherapy. Immunity
2004; 20: 17–23.
2. Racanelli V, Behrens SE, Aliberti J, Rehermann B. Den-
dritic cells transfected with cytopathic self-replicating RNA
induce crosspriming of CD8+ T cells and antiviral immunity.
Immunity 2004; 20: 47–58.
3. Ueno H, Tcherepanova I, Reygrobellet O, Laughner E,
Ventura C, Palucka AK, Banchereau J. Dendritic cell subsets
generated from CD34+ hematopoietic progenitors can be
transfected with mRNA and induce antigen-specific cytotoxic
T cell responses. J Immunol Meth 2004; 285: 171–80.
4. Grinevich YA, Khranovskaya NN, Bendyug GD. Re-
sponse of the thymus and spleen of CBA mice with sarcoma 37
on intravenous and subcutaneous administration of syngeneic
dendritic cells. Exp Oncol 2005; 27: 206–9.
5. Harris J, Monesmith T, Ubben A, Norris M, Freed-
man JH, Tcherepanova I. An improved RNA amplification
procedure results in increased yield of autologous RNA
transfected dendritic cell-based vaccine. Biochim Biophys
Acta 2005; 1724: 127–36.
6. Shi M, Bi X, Xu S, He Y, Guo X, Xiang J. Increased
susceptibility of tumorigenicity and decreased anti-tumor
effect of DC vaccination in aged mice are potentially associ-
ated with increased number of NK1.1+CD3+ NKT cells. Exp
Oncol 2005; 27: 125–9.
7. Yu JS, Wheeler CJ, Zeltzer PM, Ying H, Finger DN,
Lee PK, Yong WH, Incardona F, Thompson RC, Riedinger MS,
Zhang W, Prins RM, Black KL. Vaccination of malignant
glioma patients with peptide-pulsed dendritic cells elicits sys-
temic cytotoxicity and intracranial T-cell infiltration.Cancer
Res 2001; 61: 842–7.
8. Kono K, Takahashi A, Sugai H, Fujii H, Choudhury AR,
Kiessling R, Matsumoto Y. Dendritic cells pulsed with
HER-2/neu-derived peptides can induce specific T-cell
responses in patients with gastric cancer. Clin Cancer Res
2002; 8: 3394–400.
9. Yanagimoto H, Takai S, Satoi S, Toyokawa H, Taka-
hashi K, Terakawa N, Kwon AH, Kamiyama Y. Impaired func-
tion of circulating dendritic cells in patients with pancreatic
cancer. Clin Immunol 2005; 114: 52–60.
10. Timmerman JM, Czerwinski DK, Davis TA, Hsu FJ,
Benike C, Hao ZM, Taidi B, Rajapaksa R, Caspar CB,
Okada CY, van Beckhoven A, Liles TM, Engleman EG, Levy R.
Idiotype-pulsed dendritic cell vaccination for B-cell lym-
phoma: clinical and immune responses in 35 patients. Blood
2002; 99: 1517–26.
11. Schuler-Thurner B, Schultz ES, Berger TG, Weinlich G,
Ebner S, Woerl P, Bender A, Feuerstein B, Fritsch PO, Rom-
ani N, Schuler G. Rapid induction of tumor-specific type 1 T
helper cells in metastatic melanoma patients by vaccination
with mature, cryopreserved, peptide-loaded monocyte-derived
dendritic cells. J Exp Med 2002; 195: 1279–88.
12. Tanaka H, Shimizu K, Hayashi T, Shu S. Therapeutic
immune response induced by electrofusion of dendritic and
tumor cells. Cell Immunol 2002; 220: 1–12.
13. Siders WM, Vergilis KL, Johnson C, Shields J,
Kaplan JM. Induction of specific antitumor immunity in
the mouse with the electrofusion product of tumor cells and
dendritic cells. Mol Ther 2003; 7: 498–505.
14. Hao S, Bi X, Xu S, Wei Y, Wu X, Guo X, Carlsen S,
Xiang J. Enhanced antitumor immunity derived from a novel
vaccine of fusion hybrid between dendritic and engineered
myeloma cells. Exp Oncol 2004; 26: 300–6.
15. Karsten U, Stolley P, Walther I, Papsdorf G, Weber S,
Conrad K, Pasternak L, Kopp J. Direct comparison of elec-
tric field-mediated and PEG-mediated cell fusion for the
generation of antibody producing hybridomas. Hybridoma
1988; 7: 627–33.
16. Zimmermann U, Vienken J, Halfmann J, Emeis CC.
Electrofusion: a novel hybridization technique. Adv Biotechnol
Processes 1985; 4: 79–150.
17. Hayashi T, Tanaka H, Tanaka J, Wang R, Averbook BJ,
Cohen PA, Shu S. Immunogenicity and therapeutic efficacy of
dendritic-tumor hybrid cells generated by electrofusion.Clin
Immunol 2002; 104: 14–20.
18. Fan QY, Ma BA, Zhou Y, Zhang MH, Hao XB. Bone
tumors of the extremities or pelvis treated by microwave-in-
duced hyperthermia. Clin Orthop 2003; 406: 165–75.
19. John J, Dalgleish A, Melcher A, Pandha H. Cryop-
reserved dendritic cells for intratumoral immunotherapy do
not require re-culture prior to human vaccination. J Immunol
Meth 2005; 299: 37–46.
20. Javorovic M, Pohla H, Frankenberger B, Wolfel T,
Schendel DJ. RNA transfer by electroporation into mature
dendritic cells leading to reactivation of effector-memory
cytotoxic T lymphocytes: a quantitative analysis. Mol Ther
2005; 12: 734–43.
21. Weise JB, Maune S, Gorogh T, Kabelitz D, Arnold N,
Pfisterer J, Hilpert F, Heiser A. A dendritic cell based hybrid cell
vaccine generated by electrofusion for immunotherapy strategies
in HNSCC. Auris Nasus Larynx 2004; 31: 149–53.
22. Kjaergaard J, Shimizu K, Shu S. Electrofusion of syn-
geneic dendritic cells and tumor generates potent therapeutic
vaccine. Cell Immunol 2003; 225: 65–74.
23. Orentas RJ, Schauer D, Bin Q, Johnson BD. Electrofu-
sion of a weakly immunogenic neuroblastoma with dendritic cells
produces a tumor vaccine. Cell Immunol 2001; 213: 4–13.
24. Gong J, Nikrui N, Chen D, Koido S, Wu Z, Tanaka Y,
Cannistra S, Avigan D, Kufe D. Fusions of human ovarian
carcinoma cells with autologous or allogeneic dendritic cells
induce antitumor immunity. J Immunol 2000; 165: 1705–11.
Page 6
278
Experimental Oncology 27, 273-278, 2005 (December)
25. Tanaka Y, Koido S, Chen D, Gendler SJ, Kufe D,
Gong J. Vaccination with allogeneic dendritic cells fused to
carcinoma cells induces antitumor immunity in MUC1 trans-
genic mice. Clin Immunol 2001; 101: 192–200.
26. Takagi Y, Kikuchi T, Niimura M, Ohno T. Anti-tumor
effects of dendritic and tumor cell fusions are not dependent
on expression of MHC class I and II by dendritic cells. Cancer
Lett 2004; 213: 49–55.
27. Zhang S, Wang Q, Li WF, Wang HY, Zhang HJ, Zhu
JJ. Different antitumor immunity roles of cytokine activated T
lymphocytes from naive murine splenocytes and from dendritic
cells-based vaccine primed splenocytes: implications for adop-
tive immunotherapy. Exp Oncol 2004; 26: 55–62.
28. Schuler G, Schuler-Thurner B, Steinman RM. The
use of dendritic cells in cancer immunotherapy. Curr Opin
Immunol 2003; 15: 138–47.

Scientists have devised a new, experimental vaccine that seems to be effective at shrinking cancerous tumors in mice by up to 80 percent.

Comments Off
Posted 13 Dec 2011 — by James Street
Category Breast Cancer, MUC1 protein, MUC1 protein, Vaccine

Scientists have devised a new, experimental vaccine that seems to be effective at shrinking cancerous tumors in mice by up to 80 percent.

The vaccine worked at shrinking similar mouse versions of breast and pancreatic tumors, but researchers from the University of Georgia and the Mayo Clinic said that it could be applied to other cancers, too, including colorectal and ovarian cancers and multiple myeloma.

Scientists have been working for decades to find a way to mobilize the immune system to be able to identify cancerous cells. The problem has always been that the immune system doesn’t recognize the cancerous cells as dangerous because they originated from the body in the first place, and therefore doesn’t attack them, researchers said.

But the new vaccine works by targeting the sugar coating of a protein called MUC1 located on the surfaces of the cancerous cells. The sugar coating differentiates the cancerous cells from normal, healthy cells. The mice were engineered so that their cancer cells overexpressed MUC1, just like human cancer cells do.

“This is the first time that a vaccine has been developed that trains the immune system to distinguish and kill cancer cells based on their different sugar structures on proteins such as MUC1,” study researcher Sandra Gendler, a professor at the Mayo Clinic, said in a statement. “We are especially excited about the fact that MUC1 was recently recognized by the National Cancer Institute as one of the three most important tumor proteins for vaccine development.”

The study will appear in the journal Proceedings of the National Academy of Sciences.

The vaccine has potential to be used on a wide variety of cancers because more than 70 percent of deadly cancers have the MUC1 protein, researchers said. AOL Lifestyle reported that researchers hope to try the vaccine in humans in the next couple of years.

And because MUC1 is overexpressed in 90 percent of people who were unresponsive to other therapies like Tamoxifen or Herceptin, the vaccine might in the future be a viable option for people whose cancers are difficult to treat, researchers added.

The experimental cancer vaccines in the works today are different from the preventive vaccines (like ones that ward off cervical cancer-causing HPV), which prevents cervical cancer.

The Daily Beast explains:

By “cancer vaccine,” scientists mean something that will stimulate the immune system to attack malignant cells.

Recently, researchers at the National Cancer Institute developed a promising vaccine that seems to stop the spread of metastatic breast and ovarian cancers in humans. The poxviral vaccine even seemed to be effective at completely ridding one person involved in the study of cancer, WebMD reported.

However, the vaccine wasn’t as overwhelmingly successful in the other 25 patients — for some of those people, the vaccine seemed to extend the amount of time before the cancer progressed by a few months, WebMD noted.

And earlier this year, University of Pennsylvania researchers announced a leukemia treatment that seems effective at obliterating leukemia cells, and was shown to completely rid patients of the cancer or at least significantly decrease their number of cancerous cells.