Gene replacement therapy for non-small cell lung cancer: a review.

Current therapy such as radiation and chemotherapy controls less than 50% of lung cancers, summoning the development of novel therapeutic strategies that can directly target the underlying mechanisms of tumorigenesis. The clinical trials summarized in this article clearly demonstrate that contrary to initial predictions that gene therapy would not be suitable for cancer, gene replacement therapy is a viable potential addition to the arsenal for cancer. Gene expression has been documented and occurs even in the presence of an antiadenovirus immune response.

Induction of p53-regulated genes and tumor regression in lung cancer patients after intratumoral delivery of adenoviral p53 (INGN 201) and radiation therapy.

Purpose: We designed a prospective single arm Phase II study to evaluate the feasibility and mechanisms of apoptosis induction after Ad-p53 (INGN 201) gene transfer and radiation therapy in patients with non-small cell lung cancer.

Experimental Design: Nineteen patients with nonmetastatic non-small cell lung cancer who were not eligible for chemoradiation or surgery were treated as outpatients with radiation therapy to 60 Gy over 6 weeks in conjunction with three intratumoral injections of Ad-p53 (INGN 201) on days 1, 18, and 32.

Results: Seventeen of 19 patients completed all planned radiation and Ad-p53 (INGN 201) gene therapy as outpatients.

P53 gene replacement for cancer--interactions with DNA damaging agents.

Clinical trials of p53 gene replacement have provided information that will be useful in the design of future gene therapy strategies. Direct intratumor injection has low toxicity and thus can be readily combined with existing treatments. Post-injection gene expression can be documented and occurs in the presence of an anti-adenovirus immune response.

Gene therapy approaches for the management of non-small cell lung cancer.

Targeting the specific genetic lesions responsible for carcinogenesis and cancer progression is an attractive strategy for developing more effective anticancer therapies and reducing treatment-related toxicity. The restoration of defective tumor suppressor gene pathways by replacement of tumor suppressor genes in cancer cells has been studied in lung cancer. The most extensively studied agent is the wild-type p53 tumor suppressor gene delivered by an adenoviral vector.

Gene replacement strategies for treating non-small cell lung cancer.

Fewer than 15% of the 170,000 patients who develop lung cancer each year will survive their disease, which shows the need for novel, more specific, and less toxic therapeutic strategies. Recent advances in molecular biology have made it possible to ascertain which genetic alterations contribute to the etiology of cancer. For example, the tumor-suppressor gene, p53, responsible for directing repair of damaged DNA or committing a cell to apoptosis, is mutated or otherwise altered in more than 50% of cancers, including 40% to 70% of non-small cell lung cancers.

Difficult bile duct stone recurrence after endoscopy and extracorporeal shockwave lithotripsy.

Background/aims: In a prospective study, we investigated stone recurrence in high risk patients with difficult common bile duct stones treated with extracorporeal shockwave lithotripsy (ESWL) after futile endoscopic attempts at stone extraction with sphincterotomy.

Methodology: Endoscopic stone extraction proved unsuccessful in 35 of 659 patients presenting with common bile duct stones (5.5%, 11 males and 24 females: mean age 71.

Role of F4/80+ cells during induction of hapten-specific contact hypersensitivity.

F4/80, a monoclonal antibody that binds to a surface molecule on mature macrophages and certain dendritic cells, has been used to explore the role of epidermal and dermal cells as antigen-presenting cells (APC) during the induction of contact hypersensitivity (CH) in mice. Systemic administration of the antibody appeared to have little or no physical or functional effect on intraepidermal Langerhans' cells, even though a subpopulation of these cells expressed the F4/80 ligand. None the less, systematically administered F4/80 antibodies were able to impair CH induction when dinitrofluorobenzene (DNFB) was painted on normal body wall skin of BALB/c mice [an ultraviolet B (UVB)-resistant strain].

Fresh and cultured Langerhans cells display differential capacities to activate hapten-specific T cells.

Langerhans cells (LC) that have been cultured for 3 days acquire potent T cell-activating properties when compared to freshly prepared, uncultured LC. By contrast, fresh LC are superior to cultured LC in the ability to process native protein Ag. To define further the disparate functional properties of these epidermally derived APC, freshly isolated and cultured epidermal cells (EC) enriched for LC were prepared from BALB/c mice.

pH-dependent intracellular quenching of the indicator carboxy-SNARF-1.

Carboxy-SNARF-1 is an emission-changing, pH-sensitive probe for measurements of intracellular pH. However, the protonated and deprotonated forms of the dye interact differently with intracellular constituents, and this imposes new requirements on the calibration of the system. Whole spectra of intracellular and extracellular C.

Effects of chloroquine on antigen-presenting functions of epidermal cells from normal and psoriatic skin.

The lysosomotropic drug chloroquine has been added to cultures containing peripheral blood mononuclear cells (PBMC) and allogeneic antigen-presenting cells obtained from the epidermis of normal human skin or from skin of patients with psoriasis. We found that in the presence of chloroquine, the allostimulatory properties of freshly obtained, normal epidermal antigen-presenting cells (EAPC) were profoundly impaired. By contrast, normal EAPC (cultured for 72 h prior to exposure to alloreactive T cells), as well as fresh EAPC from psoriatic skin were not impaired by chloroquine.

T-lymphocyte-activating properties of epidermal antigen-presenting cells from normal and psoriatic skin: evidence that psoriatic epidermal antigen-presenting cells resemble cultured normal Langerhans cells.

Fresh and cultured human Langerhans cells display disparate functional programs, based on their capacities to activate autologous and allogeneic T cells, and with respect to their susceptibility to inhibition by transforming growth factor-beta (TGF beta). We have compared the functional properties of epidermal antigen-presenting cells (APC) procured from uninvolved and involved skin of patients with psoriasis with fresh and cultured normal epidermal cells. Freshly obtained psoriatic epidermal APC resembled cultured normal epidermal cells in their superior capacity to activate syngeneic and allogeneic T cells; fresh normal epidermal cells failed to activate syngeneic T cells, and induced only modest proliferation among allogeneic T cells.

Comparison of effects of transforming growth factor-beta and cyclosporin A on antigen-presenting cells of blood and epidermis.

The antigen-processing and -presenting functions of freshly obtained epidermal Langerhans cells (fresh LC) and 72-h cultured Langerhans cells (cultured LC) differ remarkably. It has been proposed that the disparate functional programs revealed in vitro may correspond directly with distinct in vivo physiologic functions--fresh LC are the in vitro equivalent of intraepidermal LC and cultured LC are equivalent to LC that have migrated from skin to the draining lymph node. As an approach to studying this proposal, we have compared the effects of two immunosuppressive agents, cyclosporin A (CsA) and transforming growth factor-beta (TGF beta), on the alloantigen-presenting capabilities of fresh LC, cultured LC, and peripheral blood mononuclear cells (PBMC).

Functional dichotomy between Langerhans cells that present antigen to naive and to memory/effector T lymphocytes.

The general thrust of this volume is to review the roles of accessory cells in regulating T and B lymphocytes. To that end, we have summarized the evidence that indicates the crucial role that Langerhans cells play in the induction and expression of immunity to antigens that gain access to, or arise within, skin. Langerhans cells accomplish this important goal by their abilities to (a) activate naive T cells to antigens not previously encountered by the host, and (b) activate memory/effector T cells specific for previously encountered antigens.

Identification of an HSV-1/HSV-2 cross-reactive T cell determinant.

The results from a number of studies have documented that the HSV glycoprotein gD is an important target for neutralizing antibodies. In contrast, little is known about the Th cell determinants present on HSV that are required for anti HSV gD antibody production. In our study we have immunized BALB/c mice with a recombinant source of HSV-1 gD lacking the carboxyl-terminal 93 amino acids.

Influence of microenvironmental factors on human Langerhans cell function in vitro.

When murine epidermal cells are placed in tissue culture for 48-72 hr, the Langerhans cell subpopulation acquires a markedly enhanced capacity to activate autologous as well as allogeneic T cells. This change is mediated largely, if not exclusively, by GM-CSF derived from keratinocytes present in the same cultures. When human epidermal cells are cultured under similar conditions, Langerhans cells display much more limited functional changes.

In vitro evidence that Langerhans cells can adopt two functionally distinct forms capable of antigen presentation to T lymphocytes.

Monodisperse suspensions of epidermal Langerhans cells (LC) have been examined for their capacity to process and present Ag immediately upon extraction from mouse epidermis (fresh LC) and after 72 h in tissue culture (cultured LC). Cultured, but not fresh, LC stimulated proliferation among autologous T cells, whereas fresh, but not cultured, LC proved to be superior at processing native OVA for presentation to an OVA peptide-specific, MHC-restricted T cell hybridoma. Cultured LC were also more effective at stimulating proliferation among allogeneic T cells.

Studies on the capacity of B cells as well as T cells to serve as accessory cells for the activation of herpes simplex virus-specific cytolytic T cells.

Experiments were conducted in an effort to determine the ability of B and T lymphocytes to serve as APC for the activation of HSV-primed splenic T cells to become class I-restricted, HSV-specific CTL. The results showed that both freshly isolated splenic B cells as well as LPS and dextran sulfate (L/D)-activated B cells were effective at stimulating the generation of CTL during a 5-day in vitro culture. There was no requirement for the addition of exogenous IL-2 to the culture and, since murine B cells do not appear to express either membrane or secreted IL-1, this lymphokine appears to either not be required for the activation of virus-specific CTL or to be provided by the T cells themselves.

Radiation safety and protection in U.S. dental hygiene programs.

A survey of radiation safety and protection measures used by programs teaching dental hygiene indicated some areas for concern. No barriers or radiation shieldings were used between operator and patient in four programs. Radiation monitoring devices were not worn by faculty operators in 16% of the programs.

Ia-specific mixed leukocyte reactive T cell hybridomas: analysis of their specificity by using purified class II MHC molecules in synthetic membrane system.

This study was undertaken to determine the nature of the antigens recognized in allogeneic and syngeneic mixed leukocyte reactions (MLR). Specifically, we wished to determine whether Ia antigens alone were recognized by MLR-reactive T cells, or whether the specificity was determined by the corecognition of non-MHC antigens together with syngeneic or allogeneic Ia. To do this we used 11 T cell hybrids that were characterized as being specific for Iad and were tested their capacity to respond to isolated I-Ad or I-Ed that had been incorporated into liposomes and had bound to the surface of glass beads.

Antigen presentation by splenic B cells: resting B cells are ineffective, whereas activated B cells are effective accessory cells for T cell responses.

In this study, we have investigated the ability of splenic B cells to act as antigen-presenting cells. Previous data had established that lipopolysaccharide (LPS)-activated B cells were effective antigen-presenting cells; however, the relative capacity of resting B cells to carry out this function remains controversial. Splenic B cells from naive BALB/c mice were depleted of macrophages, dendritic cells, and T cells, and were fractionated on the basis of cell density by using Percoll gradient centrifugation.

Radiology requirements in United States dental hygiene programs.

A survey of accredited dental hygiene programs in the United States revealed little standardization of requirements for dental radiology. However, most programs satisfied suggested minimum guidelines for didactic instruction in radiology. Six programs had no preclinical laboratory requirements, and seven had no clinical requirements.