Development of autoreactive L3T4⁺ T cells from double‐negative (L3T4⁻/Ly‐2⁻) Thy‐1⁺ spleen cells of normal mice

Abstract

Thy-1+/L3T4-/Ly-2- spleen cells were purified from normal C57BL/6 (B6) and C,B-17 mice. Cells within this subset expressed the T cell receptor (TcR) for antigen: the majority of cells in this subset were CD3+; a fraction of the cells was stained with the monoclonal antibody (mAb) F23.1; and the TcR molecule was immunoprecipitable with mAb F23.1 from cells within this subset. In limiting dilution analyses, about 1/30 cells within this subset were growth inducible in vitro by stimulation with phorbol myristate acetate (PMA) plus ionomycin; conditioned media containing interleukin (IL) 1, IL2, IL3 or IL4 activity neither triggered nor promoted in vitro growth of these cells. The in vitro generated T cells displayed the Thy-1+/L3T4+/Ly-2- surface phenotype and were self-reactive, i.e., proliferated preferentially in response to syngeneic stimulator cells, and secreted IL2 and IL3 only in response to syngeneic but not allogeneic stimulator cells. The proliferative response of these cells to syngeneic stimulator cells was blocked by anti-self Ia mAb. This autoreactive helper T cell subset was not inducible in purified Thy-1+ spleen cell subsets from athymic nude mice or scid mice. Autoreactive helper T cells did not express detectable levels of the IL2 receptor (IL2R), and their proliferative response was not blocked by anti-IL2R mAb. From PMA plus ionomycin-stimulated double-negative Thy-1+ spleen cells, 14 T cell clones were established in long-term culture which displayed the CD3+CD4+CD8- surface phenotype and were self-reactive.

Decreases in αβ T cell receptor negative T cells and CD8 cells, and an increase in CD4 +CD8+ cells in active Hashimoto's disease and subacute thyroiditis

“…Another possibility is that peripheral xji TCR T eells may differentiate into autoreactive CD4' T helper cells, which in turn increase the extent of thyroid-specific autoimmune eytolysis. since peripheral CD4 CD8 T eells are precursors of autoreaetive CD4* T helper cells [31]. Evidence for this explanation was the relative inerease of CD4cells in destructive thyrotoxieosis seen in this study.…”

Multiple Effects of Codon Usage Optimization on Expression and Immunogenicity of DNA Candidate Vaccines Encoding the Human Immunodeficiency Virus Type 1 Gag Protein

“…Cells were suspended in ␣-MEM medium (Gibco-BRL) supplemented with 10 mM HEPES buffer, 50 M ␤-mercaptoethanol, and 10% FCS. Then, 10% of a selected batch of concanavalin A-stimulated rat spleen cell supernatants (30) were added to the culture medium as a source of growth factors. Responder cells (3 ϫ 10 7 ) were cocultured with 1.5 ϫ 10 6 syngeneic, 9-mer p24(CA) peptide-pulsed A20 cells (gamma irradiated with 20,000 rads) in 10 ml of tissue culture medium in upright 25-cm 3 tissue culture flasks in a humidified atmosphere with 7% CO 2 at 37°C.…”

Hepatitis B virus small surface antigen particles are processed in a novel endosomal pathway for major histocompatibility complex class I‐restricted epitope presentation

“…Single, cell suspensions were aseptically prepared from lymph nodes of these mice 7 days post immunization. Cells were suspended in Click/FPMI 1640 tissue culture medium supplemented with 10 mM Hepes, 5 X M 2-ME, antibiotics, 5% v/v FCS (Pan Systems, Aidenbach, FRG), and 5% v/v of a selected batch of Con A-stimulated rat spleen cell supernatant [17]. Responder cells (3 x lo7) were co-cultured with 1 x lo6 syngeneic, S-antigen-expressing transfectants (irradiated with 20000 rad) in 10 ml medium in upright 25 cm2 tissue culture flasks in a humidified atmosphere of 7% COz at 37 "C. Blast cells harvested from these cultures were separated on a discontinuous density gradient and restimulated weekly at 3 x lo4 celldm1 with irradiated P815/S transfectants in conditioned medium.…”