title: Generation and characterization of cloned T helper cell lines for anti-DNA responses in NZB.H-2bm12 mice.
abstract: We have previously demonstrated that the introduction of the bm12 mutation into NZB mice results in animals that spontaneously produce high titer IgG autoantibodies to dsDNA. The observation that NZB.H-2bm12 develop lupus although NZB.H-2b control mice do not, provides a unique system to study the role of Th cells in the production of antibodies to dsDNA. We have isolated, in the absence of a known stimulating autoantigen, a series of seven autoreactive T cell clones that provide help in vitro for the production of IgG anti-dsDNA antibodies by syngeneic B cells. The data on these seven cloned T cell lines was compared to two cloned T cell lines specific for keyhole limpet hemocyanin. The seven cloned T cell lines, coined clones 19D, 23G, 410F, 410H, C1, C15, and C52 all show significant help in vitro for production of IgM and IgG antibodies to ssDNA and dsDNA; antibody levels increased 7- to 30-fold compared to cultures without T cells. Clones C1, C15, and C52 were furthered studied and were shown to provide help for IgM antihistone and anti-OVA responses but provided significantly less help for IgG antibodies. In contrast, keyhole limpet hemocyanin-specific cloned T cell lines TK2 and TK5 provided help for IgM antibodies to ssDNA, dsDNA, and histone, but failed to significantly increase IgG antibodies to ssDNA, dsDNA, or histone. The cloned T cell lines were restricted to H-2bm12 and proliferated only in response to APC from NZB.H-2bm12 and B6.C-H-2bm12 but not NZB.H-2b or NZB.H-2d mice; their in vitro helper activity was inhibited by antibodies to class II. All cloned T cell lines expressed Thy-1, CD5, and TCR-alpha/beta. Three of the seven clones used TCR-V beta 4. However, the V beta expression of the four remaining autoreactive T cell clones could not be determined. All of the autoreactive cloned T cell lines produce significant IL-4 but no detectable IL-2 or IFN-gamma. We believe that HPLC-purified peptides eluted from I-Abm12 molecules from APC can potentially provide insight on the putative autoantigen.
authors.affiliation: Division of Rheumatology, Allergy and Clinical Immunology, University of California, School of Medicine, Davis 95616.
authors.forename: M
authors.initials: M
authors.lastname: Naiki
authors.forename: B L
authors.initials: BL
authors.lastname: Chiang
authors.forename: D
authors.initials: D
authors.lastname: Cawley
authors.forename: A
authors.initials: A
authors.lastname: Ansari
authors.forename: S J
authors.initials: SJ
authors.lastname: Rozzo
authors.forename: B L
authors.initials: BL
authors.lastname: Kotzin
authors.forename: A
authors.initials: A
authors.lastname: Zlotnik
authors.forename: M E
authors.initials: ME
authors.lastname: Gershwin
mesh_terms: D000818:Animals; D000974:Antibodies, Antinuclear / Q000096:biosynthesis*; D000918:Antibody Specificity; D002999:Clone Cells; D016207:Cytokines / Q000096:biosynthesis; D004247:DNA / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006183:H-2 Antigens / Q000276:immunology*; D007074:Immunoglobulin G / Q000096:biosynthesis; D007075:Immunoglobulin M / Q000096:biosynthesis; D016130:Immunophenotyping; D051379:Mice; D008814:Mice, Inbred NZB; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology*
chemical_list: D000974:Antibodies, Antinuclear; D016207:Cytokines; D006183:H-2 Antigens; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D004247:DNA