Relative contribution of "determinant selection" and "holes in the T-cell repertoire" to T-cell responses.

Using BALB/c and CBA/J mice, the I-region associated (Ia) binding capacity and T-cell immunogenicity of a panel of 14 overlapping peptides that span the entire sequence of the protein staphylococcal nuclease (Nase) was examined to evaluate major histocompatibility gene complex (MHC) control of T-cell responses. Ia binding and Ia-restricted T-cell immunogenicity could be determined for a total of 54 peptide-MHC combinations. Only 30% of the 54 instances examined involved detectable Ia binding, but they represented almost all (12 of 13) of the immune responses found. However, binding to Ia was not sufficient to ensure T-cell immunogenicity, since only 70% of the binding events were productive--i.e., were associated with an immune response. Thus, Ia molecules have the expected characteristics of a highly permissive capacity for antigen interaction that allows them to function as restriction elements for a large universe of antigens. On the other hand, since the Ia molecules cannot distinguish between self and non-self, not all antigen-Ia interactions would be permitted to elicit a T-cell response. It appears that both Ia binding ("determinant selection") and T-cell repertoire act in concert to define the immune response status of an individual toward any particular T-cell epitope.

Studies on the mechanism of stimulation of T cells by the Mycoplasma arthritidis-derived mitogen. Role of class II IE molecules.

A mitogen derived from the supernatant of broth cultures of Mycoplasma arthritidis (MAS-P) stimulates a proliferative response by normal, unprimed T cells and interleukin 2 production by some, but not all, T cell hybridomas. The response requires an IE-positive accessory cell (AC). The direct participation of IE, and not IA, in this system was confirmed by two sets of experiments. First, L cells transfected with IE, but not IA, provided effective AC function for both normal T cells and the T cell hybridoma DO-11.10. Second, we have taken a more direct approach by showing that purified IE incorporated in liposomes and used to coat glass beads can support the MAS-P response of the DO-11.10 T cell hybridoma in the absence of intact AC or other AC molecules. Although the receptor for IE-MAS-P has not been identified, we have eliminated from consideration two potential T cell recognition structures. Monoclonal antibody to the antigen-major histocompatibility complex specific receptor failed to inhibit the MAS-P response of DO-11.10 or the T cell line LBRM-33. Furthermore, the L3T4 molecule did not appear to be involved since an L3T4-negative variant of DO-11.10 responded well to the mitogen. In addition, we show that both Lyt-2-positive and L3T4-positive T cells respond to this class II-restricted stimulus. Thus, we postulate the existence of a non-T cell receptor, non-L3T4 receptor that recognizes MAS-P in association with a presumed nonpolymorphic region of IE.

T cell and IR gene regulation of expression of a cross-reactive idiotype.

The immune response to the chemically defined DNP-oligo-L-lysine antigens is under Ir gene control in the guinea pig. Strain 2 (responder) guinea pigs mount a highly specific T cell-mediated and humoral immune response capable of discriminating closely related DNP-peptides. In contrast, strain 13 (nonresponder) animals fail to mount a cell-mediated response, and produce antibody that is only DNP-specific. In these studies a common idiotypic determinant is defined on highly specific anti-epsilon,DNP-Lys10 antibody produced by strain 2 guinea pigs. Ir gene nonresponder antibody is idiotype-negative. The presence of the idiotypic determinant distinguishes responder anti-epsilon,DNP-Lys10 antibody from responder antibodies elicited by structurally related antigens such as epsilon,DNP-Lys9 and alpha,DNP-Lys10-. Investigation of the regulation of idiotype expression demonstrated that production of idiotype-positive antibody requires the presence of viable, antigen-responsive T cells. Moreover, genetic analysis revealed that expression of the shared idiotype correlates directly with the phenotypic expression of I region genes (DTH responsiveness, Ia antigens) in (2 x 13)F1 x 13 backcross and randomly bred Hartley guinea pigs. Thus, Ir gene regulation of the immune response may be reflected in the v region specificities expressed by antigen-specific B cell clones.

Characterization of growth factors in human cartilage.

Growth factor activity has been identified in the chondrocytes and extracellular matrix (ECM) fractions of human costal cartilage. There was about five times more growth factor activity in the ECM than was found to be associated with the chondrocytes. The growth factor activity in chondrocytes was found to be associated with chromatin. Both the chromatin-associated growth factor (CAGF) activity and extracellular matrix growth factor (EMGF) activity were characterized for molecular weight, charge, and the effect of reduction by sulfhydryl reducing reagents. Biorex cation exchange chromatography showed that both CAGF and EMGF were cationic. CAGF and EMGF have molecular weights between 15,000 and 18,000 as determined by size exclusion chromatography on HPLC TSK 3000 columns equilibrated with guanidine-HCl and dithiothreitol.