The extended objective of this proposal is to examine the molecular and genetic mechanisms governing lymphocyte circulation and their relationship to murine lymphocyte and lymphoid tumor endothelial cell recognition. Lymphocyte circulation through the peripheral lymph nodes and Peyer's patches is dependent upon the expression of two distinct developmentally regulated receptor systems (one specific for peripheral lymph node, one specific for Peyer's patches) responsible for the adhesion of lymphocytes to the high endothelial (HE) cells of the lymph organ postcapillary venules. The monoclonal antibody MEL-14, specific for the lymphocyte cell surface high endothelial venule receptor structure, has been successfully used to isolate the gene sequences encoding a portion of the peripheral lymph node receptor. These sequences encode ubiquitin; we have also collected amino acid sequence data that supports this finding, as well as evidence for a second polypeptide chain within the receptor structure. The isolation of the DNA sequences encoding this portion of the receptor structure will be achieved by either immunological or nucleic acid hybridization techniques. Monoclonal antibodies will be prepared to the native cell surface molecule (and/or the cloned DNA sequences expressed as bacterial fusion proteins). The genomic structure and the expression of these DNA sequences as both transcripts and cell surface proteins will be examined. The functional consequences of the expression (and misexpression) of these gene sequences in normal tissues, lymphoid tumors and DNA transfectants will be examined. The expression of HEV receptors may contribute to the metastatic potential of lymphoid tumors; an understanding of the molecular basis of this potential may allow more reliable prognosis and treatment of these tumors. This model cell recognition system is one of the best characterized, most thoroughly validated (in vivo and in vitro) examples of specific cell interactions between lymphoid and nonlymphoid cells. The mechanisms responsible for this interaction may provide general models applicable to many aspects of tissue development.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA042571-03
Application #
3184012
Study Section
Experimental Immunology Study Section (EI)
Project Start
1986-05-01
Project End
1989-04-30
Budget Start
1988-05-01
Budget End
1989-04-30
Support Year
3
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
075524595
City
Seattle
State
WA
Country
United States
Zip Code
98109
St John, T; Meyer, J; Idzerda, R et al. (1990) Expression of CD44 confers a new adhesive phenotype on transfected cells. Cell 60:45-52
Nottenburg, C; Gallatin, W M; St John, T (1990) Lymphocyte HEV adhesion variants differ in the expression of multiple gene sequences. Gene 95:279-84
Gallatin, W M; Wayner, E A; Hoffman, P A et al. (1989) Structural homology between lymphocyte receptors for high endothelium and class III extracellular matrix receptor. Proc Natl Acad Sci U S A 86:4654-8
Nottenburg, C; Rees, G; St John, T (1989) Isolation of mouse CD44 cDNA: structural features are distinct from the primate cDNA. Proc Natl Acad Sci U S A 86:8521-5
Idzerda, R L; Carter, W G; Nottenburg, C et al. (1989) Isolation and DNA sequence of a cDNA clone encoding a lymphocyte adhesion receptor for high endothelium. Proc Natl Acad Sci U S A 86:4659-63
Sive, H L; St John, T (1988) A simple subtractive hybridization technique employing photoactivatable biotin and phenol extraction. Nucleic Acids Res 16:10937