42 research outputs found
Use of Gene Therapy in a Subcutaneous Murine Model of Lung Cancer
OBJECTIVE: To assess the effectiveness of in vivo gene
therapy to treat subcutaneous tumors generated from murine
lung cancer cells.
MATERIAL AND METHODS: C57BL/6 mice received
subcutaneus injections of 5×105 cells from the murine Lewis
lung cancer cell line. By 10 days, subcutaneous tumors of
approximately 5 mm diameter were formed. At that point,
treatment was provided by intratumor injection of a
replication-defective recombinant adenovirus carrying the
gene for thymidine kinase (AdCMV-Tk) or interleukin (IL)
12 (AdCMV-IL12), or by injection of syngeneic dendritic
cells previously transduced with adenovirus containing the
IL-12 gene (DC-IL12). Control groups were treated with
saline or adenovirus containing the gene for β-galactosidase
(AdCMV-LacZ), which functions as a reporter gene and
does not have a therapeutic effect. The number of animals in
each group ranged from 14 to 25 in experiments using
adenovirus and from 10 to 12 in experiments using dendritic
cells. Tumor size was followed for 3 weeks in the case of
treatment with adenovirus and 4 weeks for treatment with
dendritic cells.
RESULTS: A significant reduction in subcutaneous tumor
growth was observed in the groups treated with AdCMVTk,
AdCMV-IL12, and DC-IL12 compared with control
groups treated with saline or AdCMV-LacZ. The difference
was statistically significant from day 7 of treatment in the
AdCMV-Tk group, from day 9 in the AdCMV-IL12 group,
and from day 10 in the DC-IL12 group, and in all cases it
was maintained until the end of the follow-up period.
CONCLUSIONS: Gene therapy with AdCMV-Tk, AdCMVIL12,
or DC-IL12 is effective in our model of subcutaneous
tumors arising from cells of the Lewis lung cancer cell line.
The treatment leads to a significant reduction in tumor
growth compared with control groups
Effective tumor immunotherapy: start the engine, release the brakes, step on the gas pedal,...and get ready to face autoimmunity
Cellular immune responses can destroy cancer cells, achieving the cure of experimental malignancies. An expanding wealth of knowledge on the molecular basis of how to prime and amplify a T cell response has fueled a number of strategies successful at treating established tumors (rather than merely preventing tumor grafting). The most efficacious approaches operate at different stages, including: 1) priming the immune response using tumor antigen-expressing dendritic cells or tumor cells transfected with genes that render them immunogenic, 2) sustaining and amplifying immunity using agonistic monoclonal antibodies against costimulatory molecules or immune-potentiating cytokines, and 3) eliminating mechanisms that self-regulate the strength of the immune response, such as inhibitory receptors or regulatory T cells. A rational combination of such approaches holds great hope for cumulative and synergistic effects, but there is also evidence that they can open the flood-gates for unwanted inflammatory reactions. The next decade can be envisioned as the time when the first reproducibly efficacious combination regimes for cancer immunotherapy will become available and widely used in the clinic, as clinicians learn the best strategies and try to harness their potentially damaging effects
Clinical implications of antigen transfer mechanisms from malignant to dendritic cells: Exploiting cross-priming
Expansion and activation of cytolytic T lymphocytes bearing high-affinity T-cell receptors specific for tumor antigens is a major goal of active cancer immunotherapy. Physiologically, T cells receive promitotic and activating signals from endogenous professional antigen-presenting cells (APC) rather than directly from malignant cells. This phenomenon fits with the broader concept of cross-presentation that earlier was demonstrated for minor histocompatibility and viral antigens. Many mechanisms have been found to be capable of transferring antigenic material from malignant cells to APC so that it can be processed and subsequently presented by MHC class I molecules expressed on APC. Dendritic cells (DC) are believed to be the most relevant APC mediating cross-presentation because they can take up antigens from apoptotic, necrotic, and even intact tumor cells. There exist specific molecular mechanisms that ensure this transfer of antigenic material: 1) opsonization of apoptotic bodies; 2) receptors for released heat shock proteins carrying peptides processed intracellularly; 3) Fc receptors that uptake immunocomplexes and immunoglobulins; and 4) pinocytosis. DC have the peculiar capability of reentering the exogenously captured material into the MHC class I pathway. Exploitation of these pieces of knowledge is achieved by providing DC with complex mixtures of tumor antigens ex vivo and by agents and procedures that promote infiltration of malignant tissue by DC. The final outcome of DC cross-presentation could be T-cell activation (cross-priming) but also, and importantly, T-cell tolerance contingent upon the activation/maturation status of DC. Artificial enhancement of tumor antigen cross-presentation and control of the immune-promoting status of the antigen-presenting DC will have important therapeutic implications in the near future
Potentiation of therapeutic immune responses against malignancies with monoclonal antibodies
Immunotherapeutic monoclonal antibodies (mAbs) can be
defined as those that exert their functions by tampering with
immune system cell molecules, causing an enhancement of
antitumor immune responses. Some of these antibodies are
agonistic ligands for surface receptors involved in the activation
of lymphocytes and/or antigen-presenting cells, whereas
others are antagonists of mechanisms that normally limit the
intensity of immune reactions. Several mAbs of this category
have been described to display in vivo antitumor activity in
mouse models. Only anti–CTLA-4 (CD152) mAb has entered
clinical trials, but the preclinical effects described for anti-
CD40, anti-CD137 (4-1BB), anti-CD102 (intercellular adhesion
molecule-2), and regulatory T cell-depleting mAbs should lead
to their prompt clinical development. Their use in combination
with immunizations against tumor antigens has been reported
to be endowed with synergistic properties. This new group of
antitumor agents holds promise for at least additive effects with
conventional therapies of cancer and deserves intensive translational
research
Improving efficacy of interleukin-12-transfected dendritic cells injected into murine colon cancer with anti-CD137 monoclonal antibodies and alloantigens
Intralesional administration of cultured dendritic cells (DCs) engineered to produce IL-12 by in vitro infection with recombinant adenovirus frequently displays eradicating efficacy against established subcutaneous tumors derived from the CT26 murine colon carcinoma cell line. The elicited response is mainly mediated by cytolytic T lymphocytes. In order to search for strategies that would enhance the efficacy of the therapeutic procedure against less immunogenic tumors, we moved onto malignancies derived from the inoculation of MC38 colon cancer cells that are less prone to undergo complete regression upon a single intratumoral injection of IL-12-secreting DCs. In this model, we found that repeated injections of such DCs, as opposed to a single injection, achieved better efficacy against both the injected and a distantly implanted tumor; that the use of semiallogeneic DCs that are mismatched in one MHC haplotype with the tumor host showed slightly better efficacy; and that the combination of this treatment with systemic injections of immunostimulatory anti-CD137 (4-1BB) monoclonal antibody achieved potent combined effects that correlated with the antitumor immune response measured in IFN-gamma ELISPOT assays. The elicited systemic immune response eradicates concomitant untreated lesions in most cases. Curative efficacy was also found against some tumors established for 2 weeks when these strategies were used in combination. These are preclinical pieces of evidence to be considered in order to enhance the therapeutic benefit of a strategy that is currently being tested in clinical trials. Supplementary Material for this article can be found on the International Journal of Cancer website at http://www.interscience.wiley.com/jpages/0020-7136/suppmat/index.html
Anti-ICAM-2 monoclonal antibody synergizes with intratumor gene transfer of interleukin-12 inhibiting activation-induced T-cell death
PURPOSE:
Systemic treatment with an anti-ICAM-2 monoclonal antibody (mAb; EOL4G8) eradicates certain established mouse tumors through a mechanism dependent on the potentiation of a CTL-mediated response. However, well-established tumors derived from the MC38 colon carcinoma cell line were largely refractory to this treatment as well as to intratumor injection of a recombinant adenovirus encoding interleukin-12 (IL-12; AdCMVIL-12). We sought to design combined therapy strategies with AdCMVIL-12 plus anti-ICAM-2 mAbs and to identify their mechanism of action.
EXPERIMENTAL DESIGN:
Analysis of antitumor and toxic effects were performed with C57BL/6 mice bearing established MC38 tumors. Anti-ovalbumin T-cell receptor transgenic mice and tumors transfected with this antigen were used for in vitro and in vivo studies on activation-induced cell death (AICD) of CD8(+) T cells.
RESULTS:
Combined treatment with various systemic doses of EOL4G8 mAb plus intratumor injection of AdCMVIL-12 induced complete regression of MC38 tumors treated 7 days after implantation. Unfortunately, most of such mice succumbed to a systemic inflammatory syndrome that could be prevented if IFN-gamma activity were neutralized once tumors had been rejected. Importantly, dose reduction of EOL4G8 mAb opened a therapeutic window (complete cure of 9 of 18 cases without toxicity). We also show that ICAM-2 ligation by EOL4G8 mAb on activated CTLs prevents AICD, thus extending IFN-gamma production.
CONCLUSIONS:
Combination of intratumor gene transfer of IL-12and systemic anti-ICAM-2 mAb display synergistic therapeutic and toxic effects. CTL life extension resulting from AICD inhibition by anti-ICAM-2 mAbs is the plausible mechanism of action
Enhancement of CD4 and CD8 immunity by anti-CD137 (4-1BB) monoclonal antibodies during hepatitis C vaccination with recombinant adenovirus
The induction of protective or therapeutic cellular immunity against hepatitis C virus (HCV) is a difficult goal. In a previous work we showed that immunization with a recombinant adenovirus encoding HCV-NS3 (RAdNS3) could partially protect mice from challenge with a vaccinia virus encoding HCV antigens. We sought to investigate whether systemic administration of an immunostimulatory monoclonal antibody directed against the lymphocyte surface molecule CD137 could enhance the immunity elicited by RAdNS3. It was found that treatment with anti-CD137 mAb after the administration of a suboptimal dose of RAdNS3 enhanced cytotoxic and T helper cell responses against HCV NS3. Importantly, the ability of RAdNS3 to induce protective immunity against challenge with a recombinant vaccinia virus expressing HCV proteins was markedly augmented. Thus, combination of immunostimulatory anti-CD137 mAb with recombinant adenoviruses expressing HCV proteins might be useful in strategies of immunization against HCV
Pancreatic cancer escape variants that evade immunogene therapy through loss of sensitivity to IFNgamma-induced apoptosis
Combined injections into experimental tumor nodules of adenovirus encoding IL-12 and certain chemokines are capable to induce immune-mediated complete regressions. In this study, we found that the combination of two adenoviruses, one encoding IL-12 and other MIP3alpha (AdCMVIL-12+AdCMVMIP3alpha) was very successful in treating CT-26-derived colon carcinomas. However, in experimental tumors generated from the pancreatic carcinoma cell line Panc02 such combined treatment induces 50% of macroscopic complete regressions, although local relapses within 1 week are almost constant. We derived cell lines from such relapsing tumors and found that experimental malignancies derived from their inoculum were not amenable to treatment in any case with AdCMVIL-12+AdCMVMIP-3alpha. Importantly, relapsing cell lines were insensitive to in vitro induction of apoptosis by IFNgamma, in clear contrast with the original Panc02 cells. Comparative analyses by cDNA arrays of relapsing cell lines versus wild-type Panc02 were performed revealing an important number of genes (383) whose expression levels were modified more than two-fold. These changes grouped in certain gene ontology categories should harbor the mechanistic explanations of the acquired selective resistance to IFNgamma
The immunotherapy potential of agonistic anti-CD137 (4-1BB) monoclonal antibodies for malignancies and chronic viral diseases
Pharmacological intervention on the immune system to
achieve more intense lymphocyte responses has potential application
in tumour immunology and in the treatment of chronic
viral diseases. Immunostimulating monoclonal antibodies are
defined as a new family of drugs that augment cellular immune
responses. They interact as artificial ligands with functional proteins
of the immune system, either activating or inhibiting their
functions. There are humanized monoclonal antibodies directed
to the inhibitory receptor CD152 (CTLA-4) that are being tested
in clinical trials with evidence of antitumoural activity. As a
drawback, anti-CTLA-4 monoclonal antibodies induce severe
autoimmunity reactions in a fraction of the patients. Anti-CD137
monoclonal antibodies have the ability to induce potent
immune responses mainly mediated by cytotoxic lymphocytes
with the result of frequent complete tumour eradications in
mice. Comparative studies in experimental models indicate that
the antitumour activity of anti-CD137 monoclonal antibodies is
superior to that of anti-CD152. CD137 (4-1BB) is a leukocyte differentiation
antigen selectively expressed on the surface of activated
T and NK lymphocytes, as well as on dendritic cells. Monoclonal
antibodies acting as artificial stimulatory ligands of this
receptor (anti-CD137 agonist antibodies) enhance cellular antitumoural
and antiviral immunity in a variety of mouse models.
Paradoxically, anti-CD137 monoclonal antibodies are therapeutic
or preventive in the course of model autoimmune diseases in
mice. In light of these experimental results, a number of
research groups have humanized antibodies against human
CD137 and early clinical trials are about to start
Low surface expression of B7-1 (CD80) is an immunoescape mechanism of colon carcinoma
Artificially enforced expression of CD80 (B7-1) and CD86 (B7-2) on tumor cells renders them more immunogenic by triggering the CD28 receptor on T cells. The enigma is that such B7s interact with much higher affinity with CTLA-4 (CD152), an inhibitory receptor expressed by activated T cells. We show that unmutated CD80 is spontaneously expressed at low levels by mouse colon carcinoma cell lines and other transplantable tumor cell lines of various tissue origins. Silencing of CD80 by interfering RNA led to loss of tumorigenicity of CT26 colon carcinoma in immunocompetent mice, but not in immunodeficient Rag-/- mice. CT26 tumor cells bind CTLA-4Ig, but much more faintly with a similar CD28Ig chimeric protein, thus providing an explanation for the dominant inhibitory effects on tumor immunity displayed by CD80 at that expression level. Interestingly, CD80-negative tumor cell lines such as MC38 colon carcinoma and B16 melanoma express CD80 at dim levels during in vivo growth in syngeneic mice. Therefore, low CD80 surface expression seems to give an advantage to cancer cells against the immune system. Our findings are similar with the inhibitory role described for the dim CD80 expression on immature dendritic cells, providing an explanation for the low levels of CD80 expression described in various human malignancies