The purpose of this proposal is to investigate the role of band 3 and senescent cell antigen (SCA), an aging antigen that is a degradation product of band 3, in neurological aging and disease. Since SCA is intimately involved in cellular aging and removal, we will elucidate its role in neural aging and disease. We will begin by investigating band 3 structure and function in adult and aging brain. We will determine whether it undergoes the same age related changes as the anion transporter, """"""""band 3,"""""""" in other cells. The following anion transporter, band 3, changes are observed with cellular aging: increased degradation detected by immunoblotting with band 3 antibodies, decreased anion transport activity (increased Km; decreased Vmax), decreased number of high affinity ankyrin binding sites, and binding of physiologic IgG autoantibodies in situ (1- 27). In addition, the anion transporter, band 3, undergoes an as yet undefined change that results in binding of """"""""980"""""""" antibodies to aged band 3. We will quantitate the amount of SCA IgG and amount of SCA on neural tissue of various ages. Kinetic studies will be performed on pieces of perfused brains obtained immediately and at various time intervals following death to determine whether there is an increase in SCA following cell death. Titers of antibodies to band 3, synthetic peptides of band 3 and SCA will be determined with ELISA in both serum and cerebral spinal fluid (CSF) to determine if antibodies are present to promote phagocytosis of neurons by glial cells. Mice will be used for the initial aging studies. However, results will be tested in parallel experiments on human tissue. We will compare the changes in neurological diseases such as Alzheimer's Disease (AD) to those occurring during normal brain aging. We continue working on and characterizing 5 antibodies we have that distinguish between normal human brain tissue and tissue from individuals with AD.
The specific aims of the proposed research are: 1. Determine band 3 structural and functional changes during brain aging. 2. Quantitate brain band 3, altered band 3, and aging antigen content during aging. 3. Mechanism of band 3 alterations in aging. 4. Determine whether differences in serum and CSF antibodies to brain band 3 synthetic peptides and SCA are detected between young and old normal individuals (both mice and humans). 5. Determine whether differences in serum and CSF antibodies to brain band 3 peptides and SCA are detected between normal individuals and those with diseases such as Alzheimer's disease, multiple sclerosis, Huntington's chorea, and age-matched controls. 6. Develop monospecific and monoclonal antibodies to SCA, aged band 3, and breakdown products of band 3 for immunoblots, immunohistochemistry, immunoelectron microscopy, and affinity chromatography. 7. Identify structural and functional alterations in band 3 in neurological diseases such as AD. 8. Quantitate brain band 3, altered band 3, and aging antigen content in AD and other neurological diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Project (R01)
Project #
5R01AG008444-07
Application #
2050214
Study Section
Neurological Sciences Subcommittee 1 (NLS)
Project Start
1989-07-12
Project End
1996-06-30
Budget Start
1994-07-01
Budget End
1996-06-30
Support Year
7
Fiscal Year
1994
Total Cost
Indirect Cost
Name
University of Arizona
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
City
Tucson
State
AZ
Country
United States
Zip Code
85721
Poulin, J E; Cover, C; Gustafson, M R et al. (1996) Vitamin E prevents oxidative modification of brain and lymphocyte band 3 proteins during aging. Proc Natl Acad Sci U S A 93:5600-3
Kay, M M; Lake, D; Cover, C (1995) Band 3 and its peptides during aging, radiation exposure, and Alzheimer's disease: alterations and self-recognition. Adv Exp Med Biol 383:167-93
Salhany, J M; Schopfer, L M; Kay, M M et al. (1995) Differential sensitivity of stilbenedisulfonates in their reaction with band 3 HT (Pro-868-->Leu). Proc Natl Acad Sci U S A 92:11844-8
Kay, M M; Wyant, T; Goodman, J (1994) Autoantibodies to band 3 during aging and disease and aging interventions. Ann N Y Acad Sci 719:419-47
Goodman, J R; Gamble, D; Kay, M M (1994) Distribution and function of multiple anion transporter proteins in brain tumor cell lines in relation to glucose transport. Brain Res Bull 33:411-7
Kay, M M (1994) Regulatory autoantibody and cellular aging and removal. Adv Exp Med Biol 347:161-92
Kay, M M; Bosman, G; Johnson, R C et al. (1994) Molecular basis of human band-3 mutation associated with increased anion transport. Exp Clin Immunogenet 11:209-21
Kay, M M (1993) Generation of senescent cell antigen on old cells initiates IgG binding to a neoantigen. Cell Mol Biol (Noisy-le-grand) 39:131-53
Bruce, L J; Kay, M M; Lawrence, C et al. (1993) Band 3 HT, a human red-cell variant associated with acanthocytosis and increased anion transport, carries the mutation Pro-868-->Leu in the membrane domain of band 3. Biochem J 293 ( Pt 2):317-20
Marchalonis, J J; Schluter, S F; Wilson, L et al. (1993) Natural human antibodies to synthetic peptide autoantigens: correlations with age and autoimmune disease. Gerontology 39:65-79

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