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Characterization and transduction of a retroviral vector encoding human interleukin-4 and herpes simplex virus-thymidine kinase for glioma tumor vaccine therapy.

Vaccination with cytokine-transduced tumor cells represents a potentially important approach to the treatment of central nervous system tumors. We have recently demonstrated the therapeutic efficacy of tumor cell vaccines expressing the murine interleukin 4 (IL-4) and the herpes simplex virus-thymidine kinase in a rat brain tumor model in which nonirradiated vaccine cells can be eliminated by the subsequent administration of ganciclovir. In this report, we demonstrate the construction and characterization of a retroviral vector that encodes human IL-4, neomycin phosphotransferase, and herpes simplex virus-thymidine kinase genes for use in human clinical trials. An MFG-based retroviral vector was used to generate the recombinant retrovirus, TFG-hIL4-Neo-Tk, in which a long terminal repeat-driven polycistronic transcript encodes three cDNAs that are linked and coexpressed using two intervening internal ribosome entry site fragments from the encephalomyocarditis virus. The amphotropic retroviral vector TFG-hIL4-Neo-Tk was then used to infect human primary glioma cultures and skin-derived fibroblasts. After infection and G418 selection, cells produced 89-131 ng/10(6) cells/48 hours of human IL-4, which was determined to be biologically active. Transduced glioma cells were highly sensitive to the cytotoxic effect of ganciclovir. These data demonstrate the suitability of the TFG-hIL4-Neo-Tk vector for therapeutic studies of cytokine-transduced autologous tumor vaccination in patients with malignant gliomas.

Animals↗

[Effect of antibiotic therapy and vaccine therapy on the phagocytic reaction in mice infected with Staphylococcus].

The time course of changes in the activity, intensity and completeness of phagocytosis with leukocytes of the peritoneal exudate was studied on mice with experimental staphylococcal infection treated with rifampicin, lincomycin and inactivated staphylococcal vaccine used alone or in combination. It was shown that immunization of the animals with inactivated staphylococcal vaccine promoted stimulation of the phagocytic defense. Rifampicin and lincomycin applied therapeutically induced a decrease in the activity, intensity and completeness of phagocytosis. It should be noted that rifampicin had a less pronounced inhibitory effect than lincomycin. The combined use of vaccine and antibiotics with therapeutic purposes promoted an increase in phagocytosis as compared to the use of the antibiotics alone. The combined therapy sometimes resulted in completeness of phagocytosis making it reach the control values (the 10th and 15th days, rifampicin and vaccine). It should be noted that a more pronounced stimulation of the activity, intensity and completeness of the phagocytosis was observed with the use of the combination of rifampicin and the vaccine.

Animals↗

Mechanism of action of vaccine therapy in murine hepatitis B virus carriers: vaccine-induced activation of antigen presenting dendritic cells.

BACKGROUND/AIMS: Vaccine therapy in which vaccine containing hepatitis B surface antigen (HBsAg) is injected has shown therapeutic activity (vaccine therapy) in some human and murine chronic hepatitis B virus (HBV)-carriers. Using HBV-transgenic mice (HBV-Tg), an animal model of the HBV-carrier state, the mechanism underlying the antiviral and immune modulatory capacity of vaccine therapy was studied. METHODS: Placebo-controlled, double-blind trials of vaccine therapy were conducted in HBV-Tg; some HBV-Tg responded to the therapy, whereas others were non-responders. The titers of HBV-markers, the functions of lymphocytes and antigen presenting dendritic cells were compared between vaccine responders and vaccine non-responders. RESULTS: The prevaccinated titers of HBsAg, hepatitis B e-antigen (HBeAg), HBV DNA and the responses of lymphocytes to polyclonal mitogens were almost unchanged between responders and non-responders, but the levels of proliferation of HBsAg-specific lymphocytes from non-responders was significantly lower than responders (p<0.05). The capacity of dendritic cells to induce proliferation of T cells and production of antibody to HBsAg (anti-HBs) was significantly higher in responders compared with non-responders (p<0.05). Injection with HBsAg resulted in upregulation of MHC class II and CD86 antigens (p<0.05) on dendritic cells and increased production of IL-12, IL-2 and TNF-alpha in cultures (p<0.05) in vaccine responders but not in non-responders. CONCLUSIONS: The activation of dendritic cells following injection with vaccine containing HBsAg is the vital factor underlying the therapeutic potentiality of vaccine therapy in HBV carriers.

Animals↗

[Vaccine therapy of malignant tumors].

Vaccine therapy of neoplasms increases anti-tumor immune responses by enhancing the immunogenicity of tumor cells. One possible way of vaccine therapy is treatment with live apathogenic virus vaccines, in particular with Newcastle disease virus and other apathogenic avian viruses. Frequent regression of tumors and subjective improvement by vaccine therapy of patients may be explained by: a) infection of tumor cells by the virus and so enhancing their immunogenicity, b) activation of cells of the immune system, increased production of interferons, tumor necrosis factor-alpha and other cytokines, c) increased production of ACTH and beta endorphin by monocytes and lymphocytes resulting in an improvement in the general well-being. According to the previous results and the experimental data, extended trials with avian virus vaccines seem to be promising.

Humans↗

Prospects for vaccine therapy for pancreatic cancer.

Vaccine therapy is being tested for many forms of cancer. The identification of immunogenic target molecules in pancreatic cancer is providing candidates for new vaccines targeting this cancer. Early clinical trials have demonstrated safety for all vaccines tested. Immunogenicity has been variable, but some vaccines have induced responses in a high proportion of vaccinated patients. Most trials have to some extent been able to document increased survival associated with immune responses. Based on this, several vaccines are now entering controlled trials. Recent feasibility studies have demonstrated that vaccination can be combined with standard chemotherapy and indicate that some synergy effects are to be expected. This has paved the way for larger clinical studies combining gemcitabin with cancer vaccination. Characterization of regulatory pathways involved in negative control of the immune system and development of new drugs to intercept such pathways has opened for ways to manipulate the immune response in the clinical setting. Vaccination therapy for pancreatic cancer is moving into an exiting area with opportunities to attack not only the tumour as such, but also to deal with important regulatory mechanisms that have negatively influenced clinical efficacy so far.

Antigens, Neoplasm↗

Vaccine therapy for prostate cancer.

Vaccine therapy may provide an alternative for prostate cancer patients whose disease no longer responds to hormone therapy. Administration of dendritic cells pulsed with prostate-specific membrane antigen (PSMA) induces cellular immune responses against the tumor with virtually no adverse effects. About 30% of the evaluable patients were identified as partial responders, based on the National Prostate Cancer Project (NPCP) criteria. In addition, there was a 50% decrease of serum prostate-specific antigen or resolution of previously measurable lesions on imaging. Dendritic cell vaccine therapy may have a synergistic effect, when combined with other therapies.

Cancer Vaccines↗

Prognostic importance of antigen-presenting dendritic cells during vaccine therapy in a murine hepatitis B virus carrier.

As, the outcome of vaccine therapy was extremely heterogeneous in both human and murine hepatitis B virus (HBV)-carriers, the experiments presented here were performed to find out a prognostic marker of vaccine therapy using an animal model of HBV-carrier state, HBV-transgenic mice (Tg). Neither the prevaccinated titres of viral markers, such as hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg) or HBV DNA, nor the function of lymphocytes prior to vaccination, had significant influence on the outcome of vaccine therapy. Two independent, placebo-controlled, trials of vaccine therapy for 12 months, one in 17 HBV-Tg and the other in 26 HBV-Tg (total, n=43) showed that the eight of 17 and 15 of 26 HBV-Tg that had potent dendritic cell (DC) function at the start of vaccine therapy became completely negative for HBsAg, HBeAg and reduced HBV DNA, whereas all 19 HBV-Tg that had poor DC function at the start of vaccine therapy became complete non-responders, although, the prevaccinated titres of HBsAg, and HBeAg were similar in all 43 HBV-Tg. Further study to find the mechanism underlying this revealed that there was up-regulation of major histocompatibility complex (MHC) class II, CD86 antigens on DC and increased production of interleukin-12 (IL-12) by DC and of IL-2, and tumour necrosis factor-alpha (TNF-alpha) in DC/T-cell cultures when vaccine containing HBsAg was injected in HBV-Tg with potent DC function but not in HBV-Tg with poor DC function. This is the first report on the prognostic importance of DC during an immune therapy. Degree of activation of DC following vaccination would possibly help to predict the outcome of vaccine therapy in human HBV-carriers. These data also provide the scientific and logical basis to up-regulate the function of the DC before an immune therapy.

Adjuvants, Immunologic↗

Vaccine therapy for renal cell carcinoma.

Several potential vaccines have been evaluated for the treatment of patients with renal cell carcinoma (RCC). They include dendritic cells pulsed with tumor lysate, a dendritic cell-tumor cell hybrid, irradiated tumor cells admixed with adjuvants, and a heat shock protein-peptide complex. Promising results have been obtained in several early clinical trials, but issues of tumor immunosuppression and lack of identified tumor-associated antigens must be addressed before vaccine therapy can be applied successfully in advanced RCC. In this patient population, vaccine therapy will likely be required in combination with other forms of immunotherapy, such as interleukin-2 and thalidomide. In contrast, vaccine therapy alone may be sufficient for high-risk patients in the adjuvant setting.

Journal Article↗

Helper virus-free herpes simplex virus type 1 amplicon vectors for granulocyte-macrophage colony-stimulating factor-enhanced vaccination therapy for experimental glioma.

Subcutaneous vaccination therapy with glioma cells, which are retrovirally transduced to secrete granulocyte-macrophage colony-stimulating factor (GM-CSF), has previously proven effective in C57BL/6 mice harboring intracerebral GL261 gliomas. However, clinical ex vivo gene therapy for human gliomas would be difficult, as transgene delivery via retroviral vectors occurs only in dividing cells and ex vivo glioma cells have a low growth fraction. To circumvent this problem, a helper virus-free herpes simplex virus type 1 (HSV-1) amplicon vector was used. When primary cultures of human glioblastoma cells were infected with HSV-1 amplicon vectors at an MOI of 1, more than 90% of both dividing and nondividing cells were transduced. When cells were infected with an amplicon vector, HSVGM, bearing the GM-CSF cDNA in the presence of Polybrene, GM-CSF secretion into the medium during the first 24 hr after infection was 1026 ng/10(6) cells, whereas mock-infected cells did not secrete detectable GM-CSF. Subcutaneous vaccination of C57BL/6 mice with 5 x 10(5) irradiated HSVGM-transduced GL261 cells 7 days prior to intracerebral implantation of 10(6) wild-type GL261 cells yielded 60% long-term survivors (>80 days), similar to the 50% long-term survivors obtained by vaccination with retrovirally GM-CSF-transduced GL261 cells. In contrast, animals vaccinated with the same number of nontranduced GL261 cells or with GL261 cells infected with helper virus-free packaged HSV-1 amplicon vectors carrying no transgene showed only 10% long-term survivors. In conclusion, helper virus-free HSV-1 amplicon vectors appear to be effective for cytokine-enhanced vaccination therapy of glioma, with the advantages that both dividing and nondividing tumor cells can be infected, no viral proteins are expressed, and these vectors are safe and compatible with clinical use.

Animals↗

[Vaccine therapy of prostate cancer].

Vaccine therapy of prostate cancer has been increasingly studied in trials over the past few years. The different vaccine techniques are quite variable and are predominantly used in patients with advanced hormone-refractory prostate cancer. In this review, vaccine techniques using tumor cells, dendritic cells or poxvirus are analyzed. For theses approaches phase-III trials are being planned, have been initiated or are already completed. Many trials demonstrate the efficacy with regard to endpoints such as stimulation of the immune system and/or biochemical response and/or clinical response. Recently, one vaccine approach using autologous dendritic cells demonstrated a statistically significant prolongation of overall survival compared to placebo in patients with hormone-refractory prostate cancer. Side effects of vaccination are generally mild. At present, there are trials are being planned or are already ongoing that combine vaccine with other treatment strategies or enroll patients with earlier disease stages.

Aged↗

[On vaccine therapy of chronic urogenital chlamydia infection].

The efficiency of vaccine therapy of a small group of patients with chronic urogenital chlamydial infection can be regarded as preliminary results. The group included only patients treated with antibiotics many times without success; vaccine therapy led to stable cure and elimination of Chlamydia from the organism. Therefore, vaccine therapy of the above mentioned patient population is justified.

Bacterial Vaccines↗

Activation and maturation of antigen-presenting dendritic cells during vaccine therapy in patients with chronic hepatitis due to hepatitis B virus.

BACKGROUND/AIMS: The function and phenotype of antigen-presenting dendritic cells (DC) have been checked in patients with chronic hepatitis (CH) undergoing vaccine therapy to have insight about the mechanism of this immune therapy. SUBJECTS AND METHODS: Eleven patients with CH due to hepatitis B virus (HBV) were injected with vaccine containing 20 &mgr;g of hepatitis B surface antigen (HBsAg) once at every 2 weeks for 12 consecutive times. The serum interluekin-12 levels were estimated and the function and phenotype of peripheral blood DC were checked before, during and after the vaccine therapy. RESULTS: Sustained normalization of DNA-polymerase activity and alanine aminotransferase were seen in six patients (complete responders [CR]) due to vaccine therapy. The levels of serum interleukin-12 (P<0.01), the stimulatory capacity of peripheral blood DC (P<0.05) and the numbers of CD83-positive mature DC and CD86-positive activated DC (P<0.05) were significantly increased due to vaccination in CH-B patients, especially in younger patients. CONCLUSIONS: A role of DC during vaccine therapy is shown. This study also indicates that more effective vaccine therapy can be developed by upregulating the function of DC in situ in younger patients.

Journal Article↗

Technology Insight: vaccine therapy for prostate cancer.

The lack of effective therapies for advanced prostate cancer mandates continued development of alternative treatment strategies. Insights into the regulation of immune responses and the malignant process have facilitated the emergence of new immune-based strategies, currently under investigation in clinical trials. Like other forms of targeted therapy, cancer vaccines hold the promise of achieving cancer control without inducing overt toxicity. Many prostate cancer vaccines at different phases of development have been tested in clinical trials. Vaccination strategies under consideration include: immunization with defined antigenic preparations such as synthetic peptides, proteins or plasmid DNA; antigen-loaded dendritic cells; manipulated tumor cells; or with viral vectors engineered to express immunogenic genes. Although the underlying mechanisms of immunization may vary, all strategies share the common goal of eliciting immune responses against prostate tumor-associated antigens or of enhancing an otherwise weak antitumor response in the cancer patient. Unlocking the therapeutic potential of cancer vaccines will require a thorough understanding of cellular and molecular mechanisms that modulate the immune response. In this review, we provide an overview of vaccine-based strategies for prostate cancer therapy, discuss their mechanisms of action, and provide relevant clinical trial data.

Cancer Vaccines↗

[Probability of vaccine therapy for pancreas cancer].

To explore the possibility of vaccine therapy for pancreatic cancer using ras oncogenic protein related antigen we studied whether cytotoxic T lymphocyte for tumor rejection can be induced by vaccination using the peptide relating mutated ras protein. Splenocytes from mice immunized by peptide (9-15 mer) relating mutated ras protein with normal splenocytes specifically increased in the culture with the peptide. The harvested splenic lymphocytes from the mice had peptide specific lysis against B 6 fibroblast incubated with the peptide. These results suggested that immunization using the peptide relating mutated ras peptide with autologous antigen presenting cell can be a vaccine therapy for pancreatic cancer.

Animals↗

[Immunity indices during the vaccinal therapy of protracted forms of dysentery in children].

The results obtained in the study of the dynamics of systemic and local immunity characteristics in children with prolonged and chronic dysentery under the influence of vaccinal therapy are presented. The vaccine, containing soluble antigenic complexes isolated from Shigella sonnei cells by disintegration with hydroxylamine, was introduced intrarectally in doses of 2-4 mg. The course of treatment consisted of 5-6 administrations. The vaccinal therapy resulted in an increase in the level of immunoglobulins and the titer of specific antibodies, particularly IgA, in sera and fecal filtrates. These data coincided with an increase in the number of IgA-producing cells in bioptic samples of the mucous membrane of the large intestine. The vaccinal therapy contributed to the cessation of the release of bacteria in 82.5% of the patients.

Administration, Rectal↗

[Indices of alpha-interferon and of lymphocyte natural killer activity in genital herpes and the effect on them of specific vaccinal therapy and interferon therapy].

Fifty-six patients with recurrent genital herpes (RGH) and 53 normal subjects were examined. No interferon was found in the blood sera of the patients either in the period of the disease relapse or during remission. The leukocyte capacity to produce interferon in vitro (leukocyte interferon reaction, LIR) in the patients was found to be 4-5 times lower than in normal subjects. Study of the normal killer activity of lymphocytes in patients with RGH as compared with that in normal subjects revealed its decrease in 37.5% of the patients and only in 5.1% of normal subjects. Interferon therapy with purified human leukocyte interferon given to 36 patients with RGH resulted in clinical improvement in 88.9% of the patients that was accompanied by increasing normal killer activity of lymphocytes. Vaccine therapy given to 35 patients resulted in a stable clinical effect in 91.4% of the patients, however, without activation of LIR in them. It is concluded that the therapeutic effects of interferon therapy and vaccine therapy have different mechanisms.

Adult↗

Specific hepatitis B vaccine therapy in inactive HBsAg carriers: a randomized controlled trial.

BACKGROUND: Hepatitis B virus (HBV) vaccine therapy has two major areas of application: for preventive purposes and for treating patients with chronic hepatitis B. This study aimed to investigate the effect of therapeutic vaccination of inactive hepatitis B surface antigen (HbsAg) carriers using a recombinant hepatitis B vaccine in a randomized-controlled study. PATIENTS AND METHODS: The 71 studied patients had never received prior antiviral therapies, were anti-HBe positive, had undetectable HBV-DNA and persistently normal alanine transaminase levels. 31 patients were given three 20 mg intramuscular injections of a preS2/S vaccine (GenHevac-B) on days 0, 30 and 60 and the remaining 40 patients were included in the control group. The efficacy of vaccination was evaluated by testing for HBsAg seroconversion to anti-HBs. Post-vaccination follow-up was for 12 months after the first dose. RESULTS: At the end of the follow-up, three out of 31 patients (10%) who received vaccine therapy were able to clear HBsAg from their sera and concomitantly develop anti-HBs antibodies. In contrast, none of the 40 control patients who did not received vaccine therapy had decreased their levels of HBsAg or elicited anti-HBs antibodies (p = 0.079). In three vaccinated patients serum HBsAg became undetectable approximately by the 3rd month of vaccine therapy and HBsAg seroconversion was seen to be durable in all patients in the follow-up period. CONCLUSION: This study offers the first direct evidence, based on a controlled study, that the recombinant HBV vaccine has no great effect in enhancing the rate of HBsAg seroconversion in inactive HBsAg carriers. More efficient strategies, such as an increase in the dose and number of immunizations, should be evaluated further in large controlled trials.

Adolescent↗