Gene modification of primary tumor cells for active immunotherapy of human breast and ovarian cancer

Philip, Ramila; Brunette, Elisa; Clary, Bryan M.; Kilinski, Lydia; Murugesh, Deepa K.; Sorich, M; Yau, Josephine; Lebkowski, Jane; Lyerly, H. Kim; Philip, Mohan

doi:10.1016/0959-8049(95)99937-u

Cited by 3 publications

(5 citation statements)

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“…The empty HD-Ad vector (C4HSU-HD-Ad) used as a control in the in vitro experiments testing IL-10 expression from SMCs was described by Koehler et al (13). The HD-Ad vector for human IL-10 expression was constructed similarly to HD-Ad-CMVlacZ, but the CMV promoter and poly A signal were taken from the pMP6a plasmid (27). All vectors were produced and purified as described (13,36).…”

Section: Methodsmentioning

confidence: 99%

Modulation of Alloimmune Responses by Interleukin-10 Prevents Rejection of Implanted Allogeneic Smooth Muscle Cells and Restores Postinfarction Ventricular Function

Dhingra

et al. 2015

Cell Transplant

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Interleukin-10 (IL-10) gene transduction into allogeneic smooth muscle cells (SMCs) was evaluated to improve the long-term benefits of allogeneic cell transplantation into infarcted myocardium. Allogeneic cells, including SMCs, have been demonstrated to restore cardiac function and repair the infarcted myocardium, but late rejection of the transplanted cells by the host immune system may reverse the benefits of cell therapy. In a rat myocardial infarction model, three groups of rats were injected with either unmodified autologous, unmodified allogeneic, or allogeneic + IL-10 SMCs into the infarct region. Three weeks later, most of the allogeneic cells were rejected, whereas autologous cells were engrafted in the myocardium. IL-10 gene transduction of the allogeneic SMCs significantly improved the cell survival. To understand the mechanism of this improved survival, we evaluated the host immune responses against the SMCs. Allogeneic SMCs expressing IL-10 decreased leukocyte-mediated cytotoxicity in coculture, decreased the number of cytotoxic CD8(+) T-cells, and increased the number of CD4(+)CD25(+) regulatory T-cells in vitro and in vivo. Furthermore, IL-10 prevented the production of antidonor antibodies by the recipients against the allogeneic SMCs. Transplantation of unmodified autologous SMCs, but not unmodified allogeneic SMCs, significantly improved fractional shortening and left ventricular dimensions compared to the media-injected control group. However, IL-10 gene-enhanced allogeneic SMCs improved ventricular function, increased wall thickness, and decreased scar length in association with their enhanced survival. We conclude that IL-10 gene-enhanced cell therapy with allogeneic SMCs prevents detrimental alloimmune responses in the recipient, thereby increasing the survival of transplanted allogeneic SMCs and more effectively restoring cardiac function.

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Section: Methodsmentioning

confidence: 99%

Modulation of Alloimmune Responses by Interleukin-10 Prevents Rejection of Implanted Allogeneic Smooth Muscle Cells and Restores Postinfarction Ventricular Function

Dhingra

et al. 2015

Cell Transplant

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“…The bacterial ␤-galactosidase gene included a 5Ј nuclear localization signal (NLS) from the SV40 large T protein. pMP6aTElacZ was created by inserting the TE and ␤-galactosidase gene of pK18mTElacZ into the multicloning site of pMP6a (27).…”

Section: Methodsmentioning

confidence: 99%

“…After the SEAP gene, both these vectors include the SV40 t intron and SV40 minimal (237 nt) early poly(A) signal. These were replaced with the SV40 late poly(A) signal from pMP6a (27) (originally from pREP4, Invitrogen, Carlsbad, CA, U.S.A.) to create pK18SEAP-LpA and pCMVSEAP-LpA.…”

Section: Methodsmentioning

confidence: 99%

“…This is due to the fact that the parental vector pK18mTElacZ does not contain the SV40 late poly(A) signal but the weaker SV40 medium-length early poly(A) signal. Compared with the strong expression vector pMP6aTElacZ [based on pMP6a (27)], which contains a CMV promoter/ enhancer, hybrid intron, and SV40 late poly(A) signal, our new pK18mTElacZ-K18i6x7pA vector demonstrated excellent expression in lung epithelial cells with substantially reduced fibroblast expression. In contrast with our work with the SEAP reporter gene, we consistently observed higher levels of lacZ expression in fibroblasts versus epithelial cells with all expression vectors.…”

Section: Optimized Lung Gene Therapy Vectormentioning

confidence: 99%

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A Human Epithelium-Specific Vector Optimized in Rat Pneumocytes for Lung Gene Therapy

Koehler

Chow²,

Plumb³

et al. 2000

Pediatr Res

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Gene therapy vectors based on mammalian promoters offer the potential for increased cell specificity and may be less susceptible than viral promoters to transcription attenuation by host cytokines. The human cytokeratin 18 (K18) gene is naturally expressed in the lung epithelia, a target site for gene therapies to treat certain genetic pediatric lung diseases. Our original vector based on the promoter and 5Ј control elements of K18 offered excellent epithelial cell specificity but relatively low expression levels compared with viral promoters. In the present study, we found that adding a stronger SV40 poly(A) signal boosted primary rat lung epithelial cell expression but greatly reduced cell specificity. Addition of a 3Ј portion of the K18 gene to our vector as a 3Ј untranslated region (UTR) improved epithelial cellspecific expression by reducing expression in lung fibroblasts. The effect of the 3Ј UTR was not related to gross differences in cell-specific splicing. A deletion variant of this UTR further increased lung epithelial cell expression while retaining some cell specificity. These data illustrate the possibilities for using 3Ј UTR to regulate cell-specific transgene expression. Our improved K18 vector should prove useful for pediatric lung gene therapy applications. The epithelial surface of the airways presents an attractive opportunity for the topical delivery of gene-based treatments for pediatric lung diseases. CF and ␣ 1 -antitrypsin deficiency, the two most common fatal monogenic lung disorders in the Caucasian population, have garnered much attention as possible targets for lung gene therapy (1, 2). Other lung disorders that may someday benefit from gene therapies include genetic surfactant protein B deficiency (3) and inflammatory lung diseases (2). CF is caused by a defect in the gene encoding CFTR, a chloride channel residing in the epithelium of the lungs and other organs (4, 5). CF affects multiple organ systems, but it is the lung disease, characterized by thick mucous, persistent bacterial infection, and inflammation, that causes the majority of morbidity and mortality associated with CF (5, 6). In theory, early intervention to deliver a functional CFTR gene to just 5 to 10% of the defective lung cells of CF patients could lead to an improvement in the lung airway surface fluid and an improved prognosis (7,8). Attempts at human gene therapy proved the feasibility of this approach (9), but challenges remain before CF gene therapy becomes a reality.Viral vectors such as those derived from human adenovirus can efficiently deliver DNA to cells, but host immune responses to both the virus and transgene are problematic (10, 11). Complexes of DNA and cationic liposome have been widely tested and have the advantages of relatively low toxicity, low immunogenicity, and ease of preparation (12) but are currently inefficient as DNA delivery vehicles (13,14). It may be possible to counter this problem by choosing a strong expression vector to drive the production of CFTR. It may also be desirable to hav...

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“…Liposomes may be used for this purpose [58], but genes may also be delivered 'naked'. The feasibility of such an approach was first recognized when DNA injected intramuscularly was found to be taken up by a small proportion of host cells, in which it may be expressed locally for extended periods of time [59].…”

Section: Vectors For Gene Transfermentioning

confidence: 99%

Gene therapy for carcinoma of the breast: Genetic immunotherapy

Strong

1999

Breast Cancer Res

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Advances in gene transfer technology have greatly expanded the opportunities for developing immunotherapy strategies for breast carcinoma. Genetic immunotherapy approaches include the transfer of genes encoding cytokines and costimulatory molecules to modulate immune function, as well as genetic immunization strategies which rely on the delivery of cloned tumor antigens. Improved gene transfer vectors, coupled with a better understanding of the processes that are necessary to elicit an immune response and an expanding number of target breast tumor antigens, have led to renewed enthusiasm that effective immunotherapy may be achieved. It is likely that immunotherapeutic interventions will find their greatest clinical application as adjuvants to traditional first-line therapies, targeting micrometastatic disease and thereby reducing the risk of cancer recurrence.

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Gene modification of primary tumor cells for active immunotherapy of human breast and ovarian cancer

Cited by 3 publications

References 0 publications

Modulation of Alloimmune Responses by Interleukin-10 Prevents Rejection of Implanted Allogeneic Smooth Muscle Cells and Restores Postinfarction Ventricular Function

Modulation of Alloimmune Responses by Interleukin-10 Prevents Rejection of Implanted Allogeneic Smooth Muscle Cells and Restores Postinfarction Ventricular Function

A Human Epithelium-Specific Vector Optimized in Rat Pneumocytes for Lung Gene Therapy

Gene therapy for carcinoma of the breast: Genetic immunotherapy

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