title: Identification of two types of autoreactive T lymphocyte clones cultured from cardiac allograft-infiltrating cells incubated with recombinant mycobacterial heat shock protein 71.
abstract: Recent studies in several laboratories have advanced the concept that during cellular rejection, the allograft undergoes a stress response which regulates the expression of stress proteins (or heat shock proteins, hsp) and triggers the recruitment and activation of hsp-reactive lymphocytes. In a rat model of heterotopic heart transplants we have found that allograft-infiltrating lymphocytes respond to recombinant mycobacterial hsp and irradiated syngeneic spleen cells as a source of self-APC (antigen-presenting cells). This report describes T cell clones generated by culturing ACI into Lewis rat cardiac allograft-derived lymphocytes with mycobacterial hsp71, syngeneic spleen cells and IL-2 (interleukin-2). Two groups of self-APC-reactive T cell clones have been distinguished, all of them are CD3+, CD4+, CD8-. One group is referred to as hsp71-dependent, autoreactive T cells because these clones respond to self-APC but only in the presence of hsp71. No reactivity is seen with mycobacterial hsp65 or when hsp71 is tested with allo-PC from ACI donors or third-party APC from Brown Norway (BN) rats. Treatment of hsp71 with trypsin, polymyxin B or ATP-agarose chromatography abrogates the hsp71 effect thus indicating that structurally intact hsp71 must interact with self-APC which then activate hsp71-dependent, autoreactive T cells. The second group of clones reacts to self-APC and while their response does not require the presence of hsp71, their proliferation is often augmented by hsp71 but not by hsp65. These hsp71-independent, autoreactive clones do not respond to allo-APC from ACI donors or third-party APC from BN rats. Polymyxin or trypsin treatment had no significant effect on their proliferative responses. The data with the anti-TCR-alpha beta monoclonal antibody R73 offer additional evidence for two functionally different types of self-APC reactive CD4 cells infiltrating the allograft. R73 inhibits the proliferation of self-APC induced responses of hsp-71-independent clones as well as the allo-APC induced responses of alloreactive T cell clones. In contrast, this antibody augments the responses of hsp71-dependent T cells. Moreover, these clones can also proliferate in response to self-APC when hsp71 is substituted by R73. The hsp71-dependency of self-APC reactive T cell reactivity represents a previously unrecognized mechanism of cellular immunity to allografts. This mechanism might be related to the peptide binding properties of hsp71 and the ability of stress proteins to function as molecular chaperones in antigen processing.
authors.affiliation: Division of Transplantation Pathology, University of Pittsburgh Medical Center, PA 15261, USA.
authors.forename: K
authors.initials: K
authors.lastname: Liu
authors.forename: R A
authors.initials: RA
authors.lastname: Moliterno
authors.forename: X F
authors.initials: XF
authors.lastname: Fu
authors.forename: R J
authors.initials: RJ
authors.lastname: Duquesnoy
mesh_terms: D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology; D000942:Antigens, Bacterial / Q000276:immunology; D002465:Cell Movement / Q000276:immunology*; D002847:Chromatography, Agarose; D002999:Clone Cells / Q000276:immunology; D016027:Heart Transplantation / Q000276:immunology*; D006360:Heat-Shock Proteins / Q000187:drug effects / Q000276:immunology*; D008213:Lymphocyte Activation; D009161:Mycobacterium / Q000276:immunology; D011112:Polymyxin B / Q000494:pharmacology; D051381:Rats; D011917:Rats, Inbred Lew; D011994:Recombinant Proteins / Q000276:immunology*; D016176:T-Lymphocyte Subsets / Q000276:immunology*; D014184:Transplantation, Homologous; D014357:Trypsin / Q000494:pharmacology
chemical_list: D000942:Antigens, Bacterial; D006360:Heat-Shock Proteins; D011994:Recombinant Proteins; D014357:Trypsin; D011112:Polymyxin B