The Monoclonal Antibody SM5-1 Recognizes a Fibronectin Variant Which is Widely Expressed in Melanoma

Overview

Journal BMC Cancer

Publisher Biomed Central

Specialty Oncology

Date 2006 Jan 13

PMID 16405722

Citations 3

Authors

Uwe Trefzer

Yingwen Chen

Gunda Herberth

Maja Ann Hofmann

Felix Kiecker

Yajun Guo

Wolfram Sterry

Affiliations

Soon will be listed here.

Abstract

Background: Previously we have generated the monoclonal antibody SM5-1 by using a subtractive immunization protocol of human melanoma. This antibody exhibits a high sensitivity for primary melanomas of 99% (248/250 tested) and for metastatic melanoma of 96% (146/151 tested) in paraffin embedded sections. This reactivity is superior to the one obtained by HMB-45, anti-MelanA or anti-Tyrosinase and is comparable to anti-S100. However, as compared to anti-S100, the antibody SM5-1 is highly specific for melanocytic lesions since 40 different neoplasms were found to be negative for SM5-1 by immunohistochemistry. The antigen recognized by SM5-1 is unknown.

Methods: In order to characterize the antigen recognized by mAb SM5-1, a cDNA library was constructed from the metastatic human melanoma cell line SMMUpos in the Uni-ZAP lambda phage and screened by mAb SM5-1. The cDNA clones identified by this approach were then sequenced and subsequently analyzed.

Results: Sequence analysis of nine independent overlapping clones (length 3100-5600 bp) represent fibronectin cDNA including the ED-A, but not the ED-B region which are produced by alternative splicing. The 89aa splicing variant of the IIICS region was found in 8/9 clones and the 120aa splicing variant in 1/9 clones, both of which are included in the CS1 region of fibronectin being involved in melanoma cell adhesion and spreading.

Conclusion: The molecule recognized by SM5-1 is a melanoma associated FN variant expressed by virtually all primary and metastatic melanomas and may play an important role in melanoma formation and progression. This antibody is therefore not only of value in immunohistochemistry, but potentially also for diagnostic imaging and immunotherapy.

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References

Pierschbacher M, Ruoslahti E . Cell attachment activity of fibronectin can be duplicated by small synthetic fragments of the molecule. Nature. 1984; 309(5963):30-3. DOI: 10.1038/309030a0. View

Mosher D . Binding of plasma fibronectin to cell layers of human skin fibroblasts. J Cell Biol. 1983; 97(2):466-72. PMC: 2112508. DOI: 10.1083/jcb.97.2.466. View

Humphries M, Olden K, Yamada K . A synthetic peptide from fibronectin inhibits experimental metastasis of murine melanoma cells. Science. 1986; 233(4762):467-70. DOI: 10.1126/science.3726541. View

Paul J, Schwarzbauer J, Tamkun J, Hynes R . Cell-type-specific fibronectin subunits generated by alternative splicing. J Biol Chem. 1986; 261(26):12258-65. View

Sekiguchi K, Klos A, Hirohashi S, Hakomori S . Human tissue fibronectin: expression of different isotypes in the adult and fetal tissues. Biochem Biophys Res Commun. 1986; 141(3):1012-7. DOI: 10.1016/s0006-291x(86)80145-0. View

Borsi L, Carnemolla B, Castellani P, Rosellini C, Vecchio D, Allemanni G . Monoclonal antibodies in the analysis of fibronectin isoforms generated by alternative splicing of mRNA precursors in normal and transformed human cells. J Cell Biol. 1987; 104(3):595-600. PMC: 2114534. DOI: 10.1083/jcb.104.3.595. View

Humphries M, Komoriya A, Akiyama S, Olden K, Yamada K . Identification of two distinct regions of the type III connecting segment of human plasma fibronectin that promote cell type-specific adhesion. J Biol Chem. 1987; 262(14):6886-92. View

Garcia-Pardo A, Rostagno A, Frangione B . Primary structure of human plasma fibronectin. Characterization of a 38 kDa domain containing the C-terminal heparin-binding site (Hep III site) and a region of molecular heterogeneity. Biochem J. 1987; 241(3):923-8. PMC: 1147649. DOI: 10.1042/bj2410923. View

Ruoslahti E, Pierschbacher M . New perspectives in cell adhesion: RGD and integrins. Science. 1987; 238(4826):491-7. DOI: 10.1126/science.2821619. View

10.

Patel R, Odermatt E, Schwarzbauer J, Hynes R . Organization of the fibronectin gene provides evidence for exon shuffling during evolution. EMBO J. 1987; 6(9):2565-72. PMC: 553675. DOI: 10.1002/j.1460-2075.1987.tb02545.x. View

11.

Schwarzbauer J, Patel R, Fonda D, Hynes R . Multiple sites of alternative splicing of the rat fibronectin gene transcript. EMBO J. 1987; 6(9):2573-80. PMC: 553677. DOI: 10.1002/j.1460-2075.1987.tb02547.x. View

12.

Norton P, Hynes R . Alternative splicing of chicken fibronectin in embryos and in normal and transformed cells. Mol Cell Biol. 1987; 7(12):4297-307. PMC: 368112. DOI: 10.1128/mcb.7.12.4297-4307.1987. View

13.

Hynes R . Fibronectins: a family of complex and versatile adhesive glycoproteins derived from a single gene. Harvey Lect. 1985; 81:133-52. View

14.

Kalebic T, Williams J, Talmadge J, Kravitz B, Locklear K, Siegal G . A novel method for selection of invasive tumor cells: derivation and characterization of highly metastatic K1735 melanoma cell lines based on in vitro and in vivo invasive capacity. Clin Exp Metastasis. 1988; 6(4):301-18. DOI: 10.1007/BF01753577. View

15.

Short J, Fernandez J, Sorge J, Huse W . Lambda ZAP: a bacteriophage lambda expression vector with in vivo excision properties. Nucleic Acids Res. 1988; 16(15):7583-600. PMC: 338428. DOI: 10.1093/nar/16.15.7583. View

16.

Pierschbacher M, Dedhar S, Ruoslahti E, Argraves S, Suzuki S . An adhesion variant of the MG-63 osteosarcoma cell line displays an osteoblast-like phenotype. Ciba Found Symp. 1988; 136:131-41. DOI: 10.1002/9780470513637.ch9. View

17.

Carnemolla B, Balza E, Siri A, Zardi L, Nicotra M, Bigotti A . A tumor-associated fibronectin isoform generated by alternative splicing of messenger RNA precursors. J Cell Biol. 1989; 108(3):1139-48. PMC: 2115391. DOI: 10.1083/jcb.108.3.1139. View

18.

Kornblihtt A, Baralle F . Human fibronectin: molecular cloning evidence for two mRNA species differing by an internal segment coding for a structural domain. EMBO J. 1984; 3(1):221-6. PMC: 557323. DOI: 10.1002/j.1460-2075.1984.tb01787.x. View

19.

Ebbinghaus C, Scheuermann J, Neri D, Elia G . Diagnostic and therapeutic applications of recombinant antibodies: targeting the extra-domain B of fibronectin, a marker of tumor angiogenesis. Curr Pharm Des. 2004; 10(13):1537-49. DOI: 10.2174/1381612043384808. View

20.

Ruoslahti E . Fibronectin in cell adhesion and invasion. Cancer Metastasis Rev. 1984; 3(1):43-51. DOI: 10.1007/BF00047692. View