title: CD4-CD8- thymocytes from MRL-lpr/lpr mice exhibit abnormal proportions of alpha beta- and gamma delta-TCR+ cells and demonstrate defective responsiveness when activated through the TCR.
abstract: MRL-lpr/lpr (lpr) mice develop profound lymphadenopathy resulting from the accumulation of CD4-CD8- (double-negative, DN) cells in the peripheral lymphoid organs. Earlier studies from our laboratory demonstrated an increased proportion of DN cells in the thymus of lpr mice with age. Inasmuch as the DN thymocytes constitute a heterogenous population of cells, in the present study, we investigated the TCR phenotype of DN thymocytes and their responsiveness to activation through the TCR. The DN thymocytes of young (1 month of age) lpr mice contained approximately 65% CD3+ cells of which approximately 60% were alpha beta-TCR+ and approximately 39% were gamma delta-TCR+ as detected by using pan anti-TCR mAbs. In old (4-6 months of age) or young MRL-(+/+) mice, similar proportions of CD3+, alpha beta- or gamma delta-TCR+ DN thymocytes were detected. Interestingly, however, in old (4-6 months of age) lpr mice, the CD3+ T cells increased to approximately 86% and the majority of these (approximately 81%) were alpha beta-TCR+ and only approximately 3% were gamma delta-TCR+. Also, in old lpr mice, there was a 10-fold increase in the absolute number of alpha beta-TCR+ DN cells in the thymus, whereas, the absolute number of gamma delta-TCR+ DN cells in the thymus did not alter significantly. Furthermore, a majority (approximately 84%) of the old lpr DN thymocytes expressed CD45R, similar to the peripheral DN T cells. In contrast, only a small number (approximately 1%) of DN thymocytes from young lpr or MRL-(+/+) mice expressed CD45R. The DN thymocytes from young lpr or MRL-(+/+) mice demonstrated strong and similar proliferative responsiveness to stimulation with PMA + calcium ionophore or PMA + IL-2, or to immobilized mAb directed against the TCRs (CD3, alpha beta and gamma delta). In contrast, the DN thymocytes and the DN peripheral T cells from old lpr mice demonstrated marked defect in responding to the above stimuli. The present study suggests that with the onset of lymphadenopathy, the DN cells in the thymus of old lpr mice are increasingly skewed toward the alpha beta-TCR repertoire, the majority of which express CD45R and respond poorly to mitogenic stimuli or when activated through the TCR. It is suggested that migration of such cells continuously to the periphery may result in severe lymphadenopathy seen in old MRL-lpr/lpr mice.
authors.affiliation: Department of Biology, Virginia Polytechnic Institute and State University, Blacksburg 24061.
authors.forename: V N
authors.initials: VN
authors.lastname: Kakkanaiah
authors.forename: M
authors.initials: M
authors.lastname: Nagarkatti
authors.forename: J A
authors.initials: JA
authors.lastname: Bluestone
authors.forename: P S
authors.initials: PS
authors.lastname: Nagarkatti
mesh_terms: D000367:Age Factors; D000818:Animals; D015703:Antigens, CD / Q000032:analysis; D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis; D001327:Autoimmune Diseases / Q000276:immunology*; D017252:CD3 Complex; D002469:Cell Separation; D006649:Histocompatibility Antigens / Q000032:analysis; D007119:Immunoblastic Lymphadenopathy / Q000276:immunology*; D007376:Interleukin-2 / Q000494:pharmacology; D015847:Interleukin-4 / Q000494:pharmacology; D017493:Leukocyte Common Antigens; D008213:Lymphocyte Activation; D051379:Mice; D008817:Mice, Mutant Strains; D011948:Receptors, Antigen, T-Cell / Q000032:analysis / Q000145:classification / Q000502:physiology*; D011994:Recombinant Proteins; D016176:T-Lymphocyte Subsets / Q000276:immunology*; D013950:Thymus Gland / Q000166:cytology
chemical_list: D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D017252:CD3 Complex; D006649:Histocompatibility Antigens; D007376:Interleukin-2; D011948:Receptors, Antigen, T-Cell; D011994:Recombinant Proteins; D015847:Interleukin-4; D017493:Leukocyte Common Antigens