Abstract
There are no known intestinal cell phenotypic markers characteristic of the transmural idiopathic inflammatory diseases of the small bowel. To address this issue, we developed monoclonal antibodies specific for Crohn's disease tissue by generating a library of hybridomas specific for intestine following murine immunization with intestinal epithelial cells from a patient with Crohn's disease. The epithelial cells were initially harvested from a fresh operative specimen by gentle scraping and digestion with 1% collagenase on ice. Cells were then washed, evaluated for viability, and polytron homogenized. After immunization and subsequent fusion, hybridoma cell lines producing distinct antibody-binding patterns were identified by indirect immunoepifluorescence (IIEF). Wells representing distinct patterns were subcloned and carried to limiting dilution. Of the multiple hybridomas studied, the predominant target antibodies were goblet cell and glycoprotein-like molecules. Patterns of antibody binding identified by IIEF included: brush border, goblet cell, surface glycoprotein, enterocyte membranes, basilar crypt inclusions, circumferential goblet cell, and a heterogeneous goblet cell glycoprotein pattern. Molecular weight determination and cross-reactivity with various human tissues were studied by immunoblotting following sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Several hybridomas were specific for the intestine but were not specific for Crohn's disease.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 935-942 |
| Number of pages | 8 |
| Journal | American Journal of Gastroenterology |
| Volume | 83 |
| Issue number | 9 |
| State | Published - 1988 |
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ASJC Scopus subject areas
- Gastroenterology
Cite this
Generation of monoclonal antibodies to involved ileum of Crohn's disease : Characterization of a panel of antibodies with Goblet cell membrane and brush border-specific reactivity. / Prindiville, Thomas P; Cantrell, M. C.; Gershwin, M. Eric.
In: American Journal of Gastroenterology, Vol. 83, No. 9, 1988, p. 935-942.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Generation of monoclonal antibodies to involved ileum of Crohn's disease
T2 - Characterization of a panel of antibodies with Goblet cell membrane and brush border-specific reactivity
AU - Prindiville, Thomas P
AU - Cantrell, M. C.
AU - Gershwin, M. Eric
PY - 1988
Y1 - 1988
N2 - There are no known intestinal cell phenotypic markers characteristic of the transmural idiopathic inflammatory diseases of the small bowel. To address this issue, we developed monoclonal antibodies specific for Crohn's disease tissue by generating a library of hybridomas specific for intestine following murine immunization with intestinal epithelial cells from a patient with Crohn's disease. The epithelial cells were initially harvested from a fresh operative specimen by gentle scraping and digestion with 1% collagenase on ice. Cells were then washed, evaluated for viability, and polytron homogenized. After immunization and subsequent fusion, hybridoma cell lines producing distinct antibody-binding patterns were identified by indirect immunoepifluorescence (IIEF). Wells representing distinct patterns were subcloned and carried to limiting dilution. Of the multiple hybridomas studied, the predominant target antibodies were goblet cell and glycoprotein-like molecules. Patterns of antibody binding identified by IIEF included: brush border, goblet cell, surface glycoprotein, enterocyte membranes, basilar crypt inclusions, circumferential goblet cell, and a heterogeneous goblet cell glycoprotein pattern. Molecular weight determination and cross-reactivity with various human tissues were studied by immunoblotting following sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Several hybridomas were specific for the intestine but were not specific for Crohn's disease.
AB - There are no known intestinal cell phenotypic markers characteristic of the transmural idiopathic inflammatory diseases of the small bowel. To address this issue, we developed monoclonal antibodies specific for Crohn's disease tissue by generating a library of hybridomas specific for intestine following murine immunization with intestinal epithelial cells from a patient with Crohn's disease. The epithelial cells were initially harvested from a fresh operative specimen by gentle scraping and digestion with 1% collagenase on ice. Cells were then washed, evaluated for viability, and polytron homogenized. After immunization and subsequent fusion, hybridoma cell lines producing distinct antibody-binding patterns were identified by indirect immunoepifluorescence (IIEF). Wells representing distinct patterns were subcloned and carried to limiting dilution. Of the multiple hybridomas studied, the predominant target antibodies were goblet cell and glycoprotein-like molecules. Patterns of antibody binding identified by IIEF included: brush border, goblet cell, surface glycoprotein, enterocyte membranes, basilar crypt inclusions, circumferential goblet cell, and a heterogeneous goblet cell glycoprotein pattern. Molecular weight determination and cross-reactivity with various human tissues were studied by immunoblotting following sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Several hybridomas were specific for the intestine but were not specific for Crohn's disease.
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UR - http://www.scopus.com/inward/citedby.url?scp=0023805301&partnerID=8YFLogxK
M3 - Article
C2 - 3046335
AN - SCOPUS:0023805301
VL - 83
SP - 935
EP - 942
JO - American Journal of Gastroenterology
JF - American Journal of Gastroenterology
SN - 0002-9270
IS - 9
ER -