Further studies on a hybrid cell-surface antigen associated with human chromosome 11 using a monoclonal antibody

Jones, Carol; Kimmel, Kathryn A.; Carey, Thomas E.; Miller, York E.; Lehman, David W.; Mackenzie, D. H.

doi:10.1007/bf01543049

Cited by 13 publications

(7 citation statements)

References 22 publications

(9 reference statements)

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“…Complement, as normal rabbit serum, was obtained from Dutchland Laboratory (Denver, PA). Lethal polyclonal and monoclonal antibodies against the a1 marker and polyclonal antibodies against a2 and a3 were prepared as described (19,21 survival curve ofthe AL hybrid, an initial shoulder is followed by an exponential fall in the number of survivors. A certain fraction of the killed cells will also have incurred mutation among the markers of the human chromosome.…”

Section: Methodsmentioning

confidence: 99%

Measurement of low levels of x-ray mutagenesis in relation to human disease.

Waldren¹,

Correll²,

Sognier³

et al. 1986

Proc. Natl. Acad. Sci. U.S.A.

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We previously demonstrated that conventional methods for measurement of mutagenesis in mammalian cells are subject to serious error that causes underestimation of environmental contributions to cancer and genetic disease. This error has been corrected by use of somatic cell hybrids containing a single human chromosome on which the marker genes are carried and by using doses of mutagenic agents so low that little cell killing occurs. This method permits direct measurement of the effects of low doses of radiation and other mutagens without resort to the controversial extrapolation procedure customarily used to estimate effects of doses in the neighborhood of actual human exposures. The new data demonstrate that the true mutagenesis efficiency at the low doses of ionizing radiation that approximate human exposures is more than 200 times greater than those obtained with conventional methods. This methodology also permits evaluation of localized mutations, large and small chromosomal deletions, and nondisjunctional processes and can be used for mutagens that need metabolic activation as well as for cooperatively acting agents. The two opposing classical views that in mammalian cells extrapolation to low doses of x-radiation is linear, on the one hand, or involves a threshold, on the other, are both demonstrated to be incorrect at least for the conditions here considered. The actual curve exhibits a downward concavity so that the mutational efficiency is maximal at low doses. These data may have important implications for human health.

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Section: Methodsmentioning

confidence: 99%

Measurement of low levels of x-ray mutagenesis in relation to human disease.

Waldren¹,

Correll²,

Sognier³

et al. 1986

Proc. Natl. Acad. Sci. U.S.A.

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“…Thus loss of tumor suppressor genes at a variety of loci may be important in colorectal, bladder, and other epithelial cancers. T h e localization of the gene controlling the expression of the E7 cell surface antigen to a site near the Wilms' tumor locus on llp13 (at the MICl locus) (Jones et al, 1983;Fisher et al, 1984;Scoggin et al, 1985) suggests that reduced E7 expression in human SCC cell lines relative to normal cultured keratinocytes may indicate the presence of l l p abnormalities. A statistically significant correlation of 1 l p deletions and breakpoints (P < 0.02) with poor E7 expression supports this hypothesis.…”

Section: Discussionmentioning

confidence: 99%

“…T h e E7 antigen is strongly expressed on the membrane of most cell types in vitro, as detected by hemadsorption assays . In the presence of complement, UM-E7 antibody is cytotoxic to interspecific hybrid cells that contain an intact human 1 l p (Jones et al, 1983). In immunoperoxidase assays on tissue specimens from skin, mucosa, endometrium, cervix, esophagus, and bowel, the cells stained by this antibody include epithelial cells, fibroblasts, and cells in blood vessels and neural elements.…”

Section: Materials and Methods Monoclonal Antibody E7mentioning

confidence: 99%

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11p Deletions and breakpoints in squamous cell carcinoma: Association with altered reactivity with the UM‐E7 antibody

Bradford

Kimmel

Dyke

et al. 1991

Genes Chromosomes & Cancer

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The UM-E7 monoclonal antibody raised against the UM-SCC-I human squamous cell carcinoma (SCC) cell line identifies a cell surface antigen that is strongly expressed in normal tissues. The locus (MICI) controlling the expression of E7 and related cell surface antigens has been mapped to chromosome band 11p13. This band has been identified as a region of cancer-associated aberrations and as the probable locus of a tumor suppressor gene. Although E7 antigen expression is strong in normal keratinocytes, it varies among squamous carcinoma cell lines. Some SCC lines (12/26) exhibit weak expression of the E7 antigen, whereas other SCC cell lines (14/26) and 21 cell lines from other tumor types express the antigen strongly. On the basis of these observations and of mapping data, we postulated that low E7 antigen expression in a subset of SCC cell lines might be associated with chromosomal rearrangement or deletion involving the E7 locus on 11p. Fully evaluable karyotypes were prepared from 19 SCC cell lines, including 11 with weak and eight with strong E7 expression. Eight of the 11 lines with weak E7 expression had 11p abnormalities. Four of these contained 11p deletions, and four others had a breakpoint in 11p. In contrast, none of the cell lines in the group with strong E7 expression had an 11p deletion, although one had a rearrangement with an 11p breakpoint. In the four tumors with visible 11p deletions, the smallest region of overlap corresponded to the 11p13-p14 region. The mean log10 50% endpoint E7 titer in the group with 11p deletions or breakpoints was nearly two orders of magnitude lower than that of the lines with no 11p abnormality (1.95 +/- 0.53) (P less than 0.02). Our results indicate that the UM-E7 antibody identifies tumors with 11p13-p14 deletions and other 11p rearrangements and that the 11p region is a site of nonrandom chromosome rearrangement in a subset of human squamous cancers. The strong association of loss of antigen expression with visible 11p deletion or rearrangement in some tumors suggests that other tumors with this phenotype may contain submicroscopic lesions of 11p13-p14.

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“…The E7.1 mouse IgM mAb was raised against a human squamous cell carcinoma line UM-SCC-1 [12]. We have recently demonstrated its identity as a CD59 antibody by ELISA and Western blot analyses using purified human CD59, and by FACS analyses of cells transfected with or deficient in CD59 [19].…”

Section: Methodsmentioning

confidence: 99%

An Aberrant Form of CD59 Derived from HeLa Cells

Davies

Vannais

Fernie

et al. 2001

Exp Clin Immunogenet

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We isolated a CD59 cDNA from a HeLa cell library which encoded a mutated form of CD59, having a single base substitution (G to T) that changed Arg55 to Met. Since this mutation occurred in the vicinity of the putative active site of CD59, we expressed the aberrant form of the protein in Chinese hamster ovary cells in order to test for effects upon function. We found that the mutation did not influence complement inhibitory activity of CD59. However, the epitopes recognised by the function-blocking CD59 monoclonal antibodies BRIC229 and YTH 53.1 were significantly affected. The G to T substitution caused loss of an Mnl I restriction site which permitted PCR-RFLP analysis. All of 52 human subjects studied, and our in-house HeLa cells, were homozygous for the normal CD59 sequence, indicating that the altered sequence was not due to normal variation in the general population. Therefore this mutation probably arose spontaneously in the HeLa cell line used to generate the commercially obtained cDNA library.

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Further studies on a hybrid cell-surface antigen associated with human chromosome 11 using a monoclonal antibody

Cited by 13 publications

References 22 publications

Measurement of low levels of x-ray mutagenesis in relation to human disease.

Measurement of low levels of x-ray mutagenesis in relation to human disease.

11p Deletions and breakpoints in squamous cell carcinoma: Association with altered reactivity with the UM‐E7 antibody

An Aberrant Form of CD59 Derived from HeLa Cells

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