Establishment of a human polyclonal oral epithelial cell line

Gilchrist, E.P.; Moyer, Mary Pat; Shillitoe, Edward J.; Clare, Nanette; Murrah, Valerie A.

doi:10.1067/moe.2000.107360

Cited by 55 publications

(51 citation statements)

References 26 publications

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“…An SV40 T-antigen mAb was used specifically to detect the GMSM-K cell line. This cell line is Effect of CDT on mixed-cell cultures immortalized by SV40 transfection and expresses the SV40 large T antigen (Gilchrist et al, 2000). It was found that a mAb against the human Thy-1 (CD90) antigen was specific for the fibroblasts (Saalbach et al, 1996).…”

Section: Discussionmentioning

confidence: 99%

“…This cell line was used because attempts to isolate and maintain pure primary cultures of human gingival epithelial cells were inconsistent. Cells were routinely grown in T-75 flasks in keratinocyte serum-free medium (Invitrogen) as reported previously (Kanno et al, 2005;Gilchrist et al, 2000). Cells were passaged in modified Eagle's medium (Medium 199; Gibco), containing 10 % fetal calf serum (FCS), antibiotics (10 000 U penicillin G ml À1 , 25 ìg streptomycin ml À1 ) and an antimycotic (0 .…”

Section: Methodsmentioning

confidence: 99%

“…Fibroblasts express a cell-surface antigen, Thy-1 (CD90), that is a potential candidate for a specific marker (Saalbach et al, 1996(Saalbach et al, , 1999. Since the GMSM-K cell line is virally transformed, the cells express the SV40 large T antigen (Gilchrist et al, 2000). To determine the specificity of these potential cell markers, HPLFs and GMSM-K cells were grown separately in culture and immunolabelled using commercially available primary antibodies against each of these proteins (Fig.…”

Section: Identification and Validation Of Cell-specific Markersmentioning

confidence: 99%

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Differential effect of the cytolethal distending toxin of Actinobacillus actinomycetemcomitans on co-cultures of human oral cells

Kang

Korostoff

Volgina

et al. 2005

Journal of Medical Microbiology

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The periodontal pathogen Actinobacillus actinomycetemcomitans expresses a cytolethal distending toxin (CDT) that typically arrests the growth of eukaryotic cells at either the G 0 /G 1 or G 2 /M phase of the cell cycle. It was previously found that CDT failed to arrest the growth of human periodontal ligament fibroblasts (HPLFs) when grown in pure culture. In contrast, proliferation of an oral epithelial cell line was rapidly inhibited by the toxin. In this study, the feasibility of using mixed-cell cultures and cell-specific markers to evaluate the response of oral cells, when in heterogeneous populations, to CDT was established. Proliferation of epithelial cells was rapidly inhibited and the cells were selectively eliminated in co-culture with HPLFs or cementoblasts by 24-48 h postintoxication. Epithelial cells and HPLFs were detected and counted in co-cultures following cellspecific immunolabelling with antibodies against simian virus 40 large T antigen and the Ab-1 surface antigen, respectively. These results demonstrated that the activities of potential virulence factors, such as CDT, from periodontal pathogens can be successfully examined in mixed-cell cultures. This approach is especially relevant to infectious diseases that affect tissues with a diverse cellular composition, such as the periodontium.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Identification and Validation Of Cell-specific Markersmentioning

confidence: 99%

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Differential effect of the cytolethal distending toxin of Actinobacillus actinomycetemcomitans on co-cultures of human oral cells

Kang

Korostoff

Volgina

et al. 2005

Journal of Medical Microbiology

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“…The human oropharyngeal epidermoid carcinoma KB cell line [43] was obtained from the UCSF Cell Culture Facility. Non-tumor-derived human oral keratinocytes, GMSM-K, [44] were provided by Dr. V.A. Murrah (University of North Carolina at Chapel Hill).…”

Section: Cell Culturementioning

confidence: 99%

Correlation between the levels of survivin and survivin promoter-driven gene expression in cancer and non-cancer cells

Konopka

Spain

Yen

et al. 2009

Cellular and Molecular Biology Letters

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Abbreviations used: DMEM -Dulbecco's modified Eagle's MEM medium; FBS -fetal bovine serum; HNSCC -head and neck squamous cell carcinoma; IAP -inhibitor of apoptosis; OSCC -oral squamous cell carcinoma; RLU -relative light units luciferase driven by the CMV and survivin promoters in murine cells was much higher than that in human cells. The cells displayed very different susceptibilities to transfection; nevertheless, high CMV-driven luciferase activity appeared to correlate with high survivin-promoter driven luciferase expression. The survivin concentration in lysates of cancer cells ranged from 5.8 ± 2.3 to 24.3 ± 2.9 ng/mg protein (mean, 13.7 ng/mg). Surprisingly, elevated survivin protein was determined in lysates of non-tumor-derived cells. Survivin levels for GMSM-K and 184A-1 cells, were 16.7 ± 8.7 and 13.5 ± 6.2 ng/mg protein, respectively. The expression of endogenous survivin did not correlate with the level of survivin promoter-driven transgene activity in the same cells. The expression of survivin by non-tumorigenic, transformed cell lines may be necessary for their proliferative activity. The level of survivin promoter-driven gene expression achieved via liposomal vectors in OSCC cells was too low to be useful in cancer-cell specific gene therapy.

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“…The human oral epithelial cell line GMSM-K (a generous gift from V. A. Murrah and E. P. Gilchrist, University of North Carolina School of Dentistry) was derived from normal oral buccal mucosa of a stillborn fetus and immortalized with simian virus 40 T antigen (14). GMSM-K cells exhibit differentiation markers characteristic of normal keratinocytes, including the expression of lowand high-molecular-weight keratins, squamous cell-specific keratin 14, and epithelial cell-specific antigen Ber-EP4.…”

Section: Cellsmentioning

confidence: 99%

Human Immunodeficiency Virus Type 1 Stimulates the Expression and Production of Secretory Leukocyte Protease Inhibitor (SLPI) in Oral Epithelial Cells: a Role for SLPI in Innate Mucosal Immunity

Jana¹,

Gray

Shugars

2005

J Virol

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The innate immune response is a key barrier against pathogenic microorganisms such as human immunodeficiency virus type 1 (HIV-1). Because HIV-1 is rarely transmitted orally, we hypothesized that oral epithelial cells participate in the innate immune defense against this virus. We further hypothesized that secretory leukocyte protease inhibitor (SLPI), a 12-kDa mucosal antiviral protein, is a component of the host immune response to this virus. Here we demonstrated constitutive expression and production of SLPI in immortalized human oral keratinocytes. Brief exposure of cells to HIV-1 BaL and HXB2 significantly increased SLPI mRNA and protein production compared to that in mock-exposed cells (P < 0.01), as evaluated by real-time quantitative reverse transcription-PCR and enzyme-linked immunosorbent assay. HIV-1-mediated stimulation of SLPI occurred at the transcriptional level, was dose and time dependent, was elicited by heat-inactivated and infectious viruses, and did not depend on cellular infection. Experiments with purified retroviral proteins showed that the stimulatory effect was induced specifically by external envelope glycoproteins from HIV-1 and simian immunodeficiency virus. SLPI responsiveness to HIV-1 was also observed in an unrelated oral epithelial cell line and in normal (nonimmortalized) human oral epithelial cells isolated from healthy uninfected gingival tissues. In this first report of SLPI regulation by HIV-1, we show that the expression and production of the antimicrobial and anti-inflammatory protein can be stimulated in oral epithelial cells by the virus through interactions with gp120 in the absence of direct infection. These findings indicate that SLPI is a component of the oral mucosal response to HIV-1.

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Establishment of a human polyclonal oral epithelial cell line

Cited by 55 publications

References 26 publications

Differential effect of the cytolethal distending toxin of Actinobacillus actinomycetemcomitans on co-cultures of human oral cells

Differential effect of the cytolethal distending toxin of Actinobacillus actinomycetemcomitans on co-cultures of human oral cells

Correlation between the levels of survivin and survivin promoter-driven gene expression in cancer and non-cancer cells

Human Immunodeficiency Virus Type 1 Stimulates the Expression and Production of Secretory Leukocyte Protease Inhibitor (SLPI) in Oral Epithelial Cells: a Role for SLPI in Innate Mucosal Immunity

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