Tumor Antigen and Human Chorionic Gonadotropin in CaSki Cells: A New Epidermoid Cervical Cancer Cell Line

Pattillo, Roland A.; Hussa, Robert O.; Story, Michael T.; Rückert, A; Shalaby, M. R.; Mattingly, Richard F.

doi:10.1126/science.867042

Cited by 225 publications

(134 citation statements)

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“…CaSki cells were grown in RPMI medium with 10% FCS (Gibco) (1,10,12,23). CaSki cells contain approximately 500 integrated copies of HPV-16 in their genome, whereas HeLa cells have 20-50 copies of integrated HPV-18 and SiHa cells have 1-2 copies of integrated HPV 16 (18).…”

Section: Cell Culturementioning

confidence: 99%

Detection of human papillomavirus type 16/18 DNA in cervicovaginal cells by fluorescence based in situ hybridization and automated image cytometry

et al. 1994

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Automatic fluorescence image cytometry (AFIC) is a fast, sensitive, and reliable approach for screening slide-based clinical specimens. In this study, we applied AFIC to identify cancer-associated human papillomavirus (HPV) genotypes 16 and 18 in individual cells of cervical smears using a sensitive fluorescence based in situ hybridization (FISH) assay. HPV sequences were labeled by FISH and the cells imaged using an epi-fluorescence microscope coupled to a low-light color CCD camera. Before application to clinical specimens, AFIC was assessed using fluorescent calibration beads and cervical cancer cell lines containing known numbers of integrated HPV genomes per nucleus. Assessment showed that our AFIC had a linear response, was quantitatively accurate, and had the sensitivity to detect one HPV genome per nucleus. After acquisition of images, computer algorithms identified every cell nucleus (via a fluorescent DNA counterstain) and quantified the FISH signal per nucleus. AFIC was employed to screen 27 patient specimens for HPV 16/18, of which 12 were positive. The HPV status of the specimens positively correlated with the pathological diagnosis, and since AFIC automatically and correctly located every cell, it was possible to directly compare morphology and HPV status in the same cell. In conclusion, the combination of FISH and AFIC is a sensitive and quantitative method to detect high risk HPV sequences in cervical Smears. Q 1994 Wiley-Liss, Inc.Key terms: Fluorescent in situ hybridization, cervical smears, human papillomavirus, automated fluorescence image cytometry Cervical dysplasia and carcinoma are common among women throughout the world. Approximately 500,000 women worldwide are diagnosed annually as having cervical cancer (24) and the mortality rate associated with this malignancy is second to breast cancer (20). In the United States, it is estimated that 13,000 women develop invasive cervical cancer annually (25).Substantial evidence has accumulated associating specific human papillomaviruses (HPVs) with human anogenital disorders, most notably cervical cancer (3,11,13,35). To date, over 60 different types of HPV have been described, of which about 20 have been associated with anogenital lesions (8). These 20 HPVs have been divided into "high risk" and "low risk" groups based on their association with benign vs. ma-'Supported by grants from NIH (CA51323 and the Office of Research on Womens Health), the Whitaker Foundation, and a contract with American Innovision, Inc.

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Section: Cell Culturementioning

confidence: 99%

Detection of human papillomavirus type 16/18 DNA in cervicovaginal cells by fluorescence based in situ hybridization and automated image cytometry

et al. 1994

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“…The identity of the sequences (Table I) for each peptide was confirmed by mass spectrometry. [11][12][13][14][15][16][17][18][19][20] monospecific polyclonal CTL line D4 D4, a monospecific polyclonal CTL line was generated as described previously. 9 Briefly, this HPV16 E7 11 -20 specific CTL line was generated from a patient with stage IIB cervical cancer.…”

Section: Peptidesmentioning

confidence: 99%

“…9 Briefly, this HPV16 E7 11 -20 specific CTL line was generated from a patient with stage IIB cervical cancer. 8 Single cell sorting based on CD8 (tricolor-anti CD8, Caltag, Burlingame, CA)/HPV16 E7 [11][12][13][14][15][16][17][18][19][20] tetramer (PE labeled) double staining was then carried out using a FACSVantage (Becton-Dickinson, Mountain View, CA).…”

Section: Peptidesmentioning

confidence: 99%

“…7 We have previously shown that it is possible to detect CTL specific for HPV16 E7 [11][12][13][14][15][16][17][18][19][20] in the blood of patients with both CaCx 8 and CIN3 9 and in the blood of healthy donors. 9 These CTL can mediate the lysis of B cells endogenously expressing the E7 protein 8 and the HPV 16 transformed cervical cancer cell line CaSki.…”

mentioning

confidence: 99%

“…9 These CTL can mediate the lysis of B cells endogenously expressing the E7 protein 8 and the HPV 16 transformed cervical cancer cell line CaSki. 9 Since the E7 [11][12][13][14][15][16][17][18][19][20] peptide from HPV16 has homology to the E7 peptide of the phylogenetically related types 31 and 52, we used these CTL to investigate in vitro the potential for crossrecognition between these HPV types.…”

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confidence: 99%

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Cross‐typic specificity and immunotherapeutic potential of a human HPV16 E7‐specific CTL line

Youde

McCarthy

Thomas

et al. 2004

Intl Journal of Cancer

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Cervical cancer (CaCx) is strongly associated with human papillomavirus (HPV) infection, particularly HPV types 16 and 18. The constitutive expression of HPV E6 and E7 proteins in CaCx makes them attractive targets for CTL based immunotherapy. However cervical carcinomas may have features, e.g., antigen processing defects, that limit the effectiveness of HPV specific CTL. Furthermore most vaccine development has concentrated on HPV type 16, and it is not clear whether such vaccines could induce CTL able to cross-react on related oncogenic HPV types, e.g., HPV31 and 52. To investigate these potentially important parameters in vitro, we used a CTL (D4) specific for HPV16 E7 11-20 . D4 was able to kill a variety of HPV16؉ CaCx cell lines including those with suspected (CaSki) or known antigen processing defects (C33A), and with low HPV DNA copy number (SiHa). D4 was also able to cross react on a related peptide from HPV52 E7 but not HPV31 E7. Further analysis suggested that D4 cross reactivity against related peptides was influenced both by TCR contact residues and a certain threshold for peptide binding. The HPV cross-reactivity was confirmed at the whole protein level as D4 was also able to recognize the endogenously processed forms of HPV16 and 52 E7 but not 31 E7. These results suggest that HPV16 E7 11-20 would be a useful epitope for immunotherapy in both HPV 16 and 52 tumours. Despite this, it is difficult to generate these CTL in response to vaccination, emphasizing the need for definition of novel epitopes and more efficient vaccination strategies.Key words: HPV; CTL; cervical cancer; immunotherapy; cross-reactivity Cervical cancer (CaCx) is the second most common cause of cancer in women world wide and with premalignant cervical intraepithelial neoplasia (CIN3) is associated with HPV infection. In developing countries where 80% of cases occur, this is the principal female cancer. 1 The DNA of HPVs, particularly 16 and 18, are found in Ͼ99% of CaCx patients. 2 It is the E6 and E7 proteins, which are consistently retained and expressed in cervical tumor cells, that give the virus its transforming properties. 3,4 HPVs are defined as low and high risk, high-risk types being associated with invasive cervical cancer, while low risk types are associated with warts. There are 11 HPV types that are consistently classified as high-risk types: 16,18,31,33,35, 39, 45, 51, 52, 56 and 58. These types are further divided into classes: type A (16, 31, 33, 35, 52 and 58), C (18, 39 and 45) and D (51 and 56).HPV type-specific prevalence varies geographically. Worldwide, types 16 and 18 dominate but in other non-Caucasian populations, 31, 52 and 58 dominate. The infection rate with each HPV type varies from country to country, i.e., HPV16 is found in 43.9% of cases in the Philippines and 72.4% in Morocco. According to Mûnoz et al., 95% of all infections were caused by 8 types: 16, 18, 45, 31, 33, 52, 58 and 35. They suggest that vaccination against the 5 most common types could prevent 90% of cases of CaCx. 5 The geographi...

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Interferon-? and -?, but not tumor necrosis factor-?, demonstrate immunoregulatory effects on carcinoma cell lines infected with human papillomavirus

Bornstein

Lahat

Kinarty

et al. 1997

Cancer

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Tumor Antigen and Human Chorionic Gonadotropin in CaSki Cells: A New Epidermoid Cervical Cancer Cell Line

Cited by 225 publications

References 10 publications

Detection of human papillomavirus type 16/18 DNA in cervicovaginal cells by fluorescence based in situ hybridization and automated image cytometry

Detection of human papillomavirus type 16/18 DNA in cervicovaginal cells by fluorescence based in situ hybridization and automated image cytometry

Cross‐typic specificity and immunotherapeutic potential of a human HPV16 E7‐specific CTL line

Interferon-? and -?, but not tumor necrosis factor-?, demonstrate immunoregulatory effects on carcinoma cell lines infected with human papillomavirus

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