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Evolutionary approaches are regularly used to isolate single molecules with desired activities from large populations of nucleic acids (approximately 10(15)). Several methods have also been developed to generate libraries of mRNA-encoded peptides and proteins for the in vitro selection of functional polypeptides. In principal, such mRNA encoding systems could be used with libraries of nonbiological polymers if the ribosome can be directed to polymerize tRNAs carrying unnatural amino acids. The fundamental problem is that current chemical aminoacylation systems cannot easily produce sufficient amounts of the numerous misacylated tRNAs required to synthesize a complex library of encoded polymers. Here, we show that bulk-aminoacylated tRNA can be transformed into N-monomethylated aminoacyl tRNA and translated. Because poly-N-methyl peptide backbones are refractory to proteases and are membrane permeable, our method provides an uncomplicated means of evolving novel drug candidates.

S-antigen is a highly pathogenic retinal autoantigen for the induction of experimental autoimmune uveitis (EAU). EAU is predominantly a T cell mediated autoimmune disease of the uveal tract and retina of the eye and the pinealocytes of the pineal gland. Using synthetic peptides it has been possible to identify several B cell and T cell epitopes in the molecule. In addition, synthetic peptides derived from proteins of diverse origin with amino acid sequence homology to pathogenic regions of S-antigen induce an EAU which is indistinguishable from the disease induced by native S-antigen. These studies aid in the understanding of immune mechanisms in EAU and provide a basis for the pathogenesis of uveitis in humans.

The immune responses against the sequential polypeptides; (T-G-A-Gly)n, (T-A-G-Gly)n, (Phe-G-A-Gly)n and (Phe-A-G-Gly)n were studied in inbred guinea pigs and mice. Strain 13 guinea pigs responded to (Phe-G-A-Gly)n and (T-G-A-Gly)n whereas strain 2 guinea pigs responded to (T-A-G-GLY)n and (Phe-A-G-Gly)n. These responses which are linked to MHC, are only against the helical form of the polymers which have conformational determinants. Significant cross reactions at the humoral and T cell levels (PELS) are exhibited with the following reciprocal combinations: (Phe-G-A-Gly)n and (T-G-A-Gly)n; (T-A-G-Gly)n and (Phe-A-G-Gly)n. With mice, the polymers were shown to be T dependent with the following response patterns: mice of H-2b haplotype respond against (T-G-A-Gly)n; those of H-2b, f and r haplotypes respond against (T-A-G-Gly)n. There are no responders against (Phe-G-A-Gly)n and only mice of H-2f respond against (Phe-A-G-Gly)n. "Nonresponders" respond against the MBSA aggregates of all of these polymers. The Ir gene(s) controlling these T cell dependent H-linked responses mapped to the IA subregion. Antibody responses against (T-G-A-Gly) and (T-A-G-Gly) were quite variable, and were most marked in, F1 mice of (responder and nonresponder) and in backcross populations of (F1 x R) and (F1 x NR). However, the T cell proliferative responses performed with nylon wool purified T cells gave clear cut and predictable distinctions between "responders" and nonresponders and linkage with responding haplotype. Hypotheses advanced to explain these findings relate to the poor immunogenicity (antibody) of these polymers, which have a restricted number of repeating determinants, the B cell mitogenic properties of these polymers and the possible involvement of suppressor cells. The specificities of the humoral responses, i.e. cross reactions, were similar to those found in guinea pigs. However, in contrast to the guinea pig studies cross stimulation with structurally related polymers occurred only in those situations where the immunizing and "crossreacting" polymers were both immunogenic in mice of the same haplotype, i.e., (T-A-G-Gly)n and (Phe-A-G-Gly)n in mice of H-2f haplotypes.

A plaque-forming cell (PFC) assay to measure the immune response of mice to the synthetic random copolymer of glutamic acid and phenylalanine (GPhe) is described here. GPhe can be coupled to sheep red blood cells (SRBC) using "aged" CrCl3. Both IgM and IgG plaques are detected in the murine GPhe response. Mice of H-2 haploytpes p and q are high responders, a and k are medium or low responders and b, d and s are nonresponders as detected by the PFC assay.

Synthetic polyaminoacid antigens were used to examine the specificity of transfer of delayed-type hypersensitivity with spleen cell dialysates in mice. Dialysates from GAT10-sensitized donors sensitized recipients to GAT10, but not GLA5 or cytochrome c. Dialysates from GLA5-sensitized donors sensitized recipients to GLA5, but not GAT10 or cytochrome c. We interpret these findings as consistent with the concept that passive transfer of delayed hypersensitivity with dialyzable materials is an immunologically specific event.

The genetic control of the immune response of inbred strains of mice to certain antigens has been demonstrated to be governed by a set of Ir genes linked to the major histocompatibility complex (H-2) of mice (1,2). Until recently, the control was thought to be governed by single, dominant genes, located within the I region of the H-2 complex. Merryman et al. (3) originally demonstrated that the immune response to the synthetic terpolymer L-glutamic acid, L-lysine, L-phenylaline (GLphi) is under dominant, H-2-linked Ir gene control (4-7). This was shown both by crossing two nonresponder parental strains to produce responder offspring in the F(1) generation, and by the analysis of appropriate recombinant stains of mice. The two complementing genes have been mapped in the IA and IC regions of the H-2 complex, and have been termed beta and alpha, respectively (5,6). Thus, any strain of mouse may contain neither, one, or both genes. Only mice containing both genes are capable of responding to GLphi. It has been shown using F(1) hybrid and recombinant strains of mice, that the alpha- and beta-genes can complement each other in either the cis (on the same chromosome) or in the trans (on different chromosomes) position (8). In this paper we report the results of studies aimed at answering the question of whether or not the alpha- and beta- genes can complement each other when they are present in different lymphoid cells. To this end we have constructed allophenic mice composed of two nonresponder strains (A and C57BL/6), which show gene complementation in the F(1) generation. Allophenic mice are chimeras containing two cell types coexisting in a "normal" environment. The mice were tested for the specific cellular composition of the two parental cell types and were found to possess a complete range in the relative proportion of the two cell types. This report demonstrates that regardless of the mixture of cell types present in the allophenic mice, none of them were responders to GLphi. Thus no complementation of the alpha- and beta-genes is seen when the two genes are present in different cells.