Skip to Main Content
 

Global Search Box

 
 
 
 

Files

File List


case1055268413.pdf (6.2 MB)

ETD Abstract Container

Abstract Header

Disulfide bond formation between dimeric immunoglobulin A and the polymeric immunoglobulin receptor in cultured epithelial cells and rat liver

Chintalacharuvu, Koteswara Rao
http://rave.ohiolink.edu/etdc/view?acc_num=case1055268413

Abstract Details

1991, Doctor of Philosophy, Case Western Reserve University, Pathology.
Antibodies of the IgA isotype predominate in external secretions and provide the first line of immunological defense at mucosal surfaces. Dimeric IgA (dIgA) is transported across mucosal and exocrine glandular epithelium by the polymeric immunoglobulin receptor (pIgR). The initial association between dIgA and pIgR on the basolateral surface of epithelial cells is noncovalent. During transcellular routing the pIgR-dIgA complex is stabilized through the formation of two disulfide bonds between dIgA and pIgR. We have studied disulfide bond formation between dIgA and pIgR in cultured epithelial cells and rat liver. When human dIgA was added to the basolateral surface of a polarized monolayer of Madin-Darby canine kidney cells transfected with cDNA for rabbit pIgR (pWE cells) grown on a permeable support, dIgA was specifically transported and released into the apical medium. Analysis by SDS-PAGE under non-reducing conditions demonstrated that transported dIgA was non-covalently associated with the ectoplasmic domain of pIgR (secretory component, SC), suggesting that disulfide bond formation is not required for the transport of dIgA into e xternal secretions. In rats, the primary route of transport of dIgA into intestinal secretions is by hepatic transcytosis into bile. To assess the relative efficiency of transcytosis of covalently bound and noncovalently bound dIgA, rats were injected intravenously with either native dIgA or dIgA in which free sulfhydryl groups had been blocked with iodoacetamide. There was no difference in efficiency of transport between covalently bound dIgA and noncovalently bound dIgA, suggesting that disulfide bond formation does not facilitate transport of dIgA into bile. We have studied the kinetics of transport of dIgA across hepatocytes and the time at which the disulfide bond between dIgA and pIgR is formed, and conclude that disulfide bonding between dIgA and pIgR is a late event during hepatic transcytosis. We hypothesize that the enzyme protein disulfide isomerase (PDI), a ubiquitous enzyme which plays a major role in post-translational modifications of proteins, may catalyze the formation of disulfide bonds between dIgA and pIgR in vivo. As a first step in testing this hypothesis, we have purified PDI from human placenta and rat liver and have generated a series of anti-PDI monoclonal antibodies. These antibodies should prove useful for blocking PDI activity in the transcytotic pathway and testing subsequent effects on disulfide bond formation between dIgA and pIgR.
Charlotte Kaetzel (Advisor)
227 p.
Disulfide bond formation; dimeric immunoglobulin; polymeric immunoglobulin receptor epithelial cells; rat liver

Recommended Citations

Citations

  • Chintalacharuvu, K. R. (1991). Disulfide bond formation between dimeric immunoglobulin A and the polymeric immunoglobulin receptor in cultured epithelial cells and rat liver [Doctoral dissertation, Case Western Reserve University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=case1055268413

    APA Style (7th edition)

  • Chintalacharuvu, Koteswara. Disulfide bond formation between dimeric immunoglobulin A and the polymeric immunoglobulin receptor in cultured epithelial cells and rat liver. 1991. Case Western Reserve University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=case1055268413.

    MLA Style (8th edition)

  • Chintalacharuvu, Koteswara. "Disulfide bond formation between dimeric immunoglobulin A and the polymeric immunoglobulin receptor in cultured epithelial cells and rat liver." Doctoral dissertation, Case Western Reserve University, 1991. http://rave.ohiolink.edu/etdc/view?acc_num=case1055268413

    Chicago Manual of Style (17th edition)

case1055268413
448
© 1991, all rights reserved.
This open access ETD is published by Case Western Reserve University School of Graduate Studies and OhioLINK.