This study describes a novel method of inhibiting T-cell function by the use of peptides rationally designed from the T-cell antigen receptor (TCR) alpha-chain transmembrane sequence involved with TCR receptor assembly. The most effective peptide (core peptide, CP) modulating in vitro and in vivo T-cell function contained nine amino acids two of which, lysine and arginine, were hydrophilic and separated by four hydrophobic amino acids. CP without chemical modification or conjugation was able to enter non-T and T cells. Conjugation of CP at the carboxyl terminus with palmitic acid resulted in a greater inhibition of T-cell interleukin-2 (IL-2) production in vitro than peptide alone. When examined for effects in vivo, CP reduced clinical signs of inflammation in three T cell-mediated disease models including adjuvant-induced arthritis, experimental allergic neuritis, and cyclophosphamide-induced diabetes in NOD/Lt(F) mice. This peptide or its analogues has potential as a therapeutic agent in human inflammatory and autoimmune disorders.

Repair of meniscal tears depends in part upon the ability of the resident fibrochondrocytes to produce new extracellular matrix molecules including proteoglycans. Three culture systems have been used to investigate proteoglycan production by meniscal fibrochondrocytes from the inner, middle and outer zones of medial and lateral menisci of the sheep stifle joint. Cultures of meniscal explants, monolayered cells, and cells encapsulated in alginate beads were labeled with 35SO4H2 for 48 h in the absence and presence of transforming growth factor beta (TGF beta) and the proteoglycans were analysed by Sephacryl S-1000 chromatography. In general, the lateral meniscus produced more proteoglycan than the medial. Explants from the inner and middle zones produced predominantly aggrecan-like proteoglycan, together with a smaller proteoglycan population eluting with an average distribution coefficient of around 0.65. The outer meniscal zones synthesized less proteoglycan overall, the majority of which consisted of the smaller proteoglycans. These characteristic proteoglycan size profiles obtained with explant cultures also were preserved when cells isolated from the respective zones were cultured in alginate beads. Monolayer cell cultures, however, produced almost entirely small proteoglycans, regardless of their zone of origin. Chromatography of chondroitinase AC and ABC digested samples indicated that the small proteoglycan population comprised mostly dermatan sulphate-containing proteoglycans. In all meniscal zones and in all culture systems, TGF beta stimulated proteoglycan production by up to 100% and the proteoglycans were slightly larger. TGF beta also stimulated cell division in fibrochondrocyte monolayer cultures. Long term intermittent stimulation of alginate bead cultures with TGF beta resulted in large increases in proteoglycan synthesis, increased aggregation of large proteoglycan monomers, and an increase in the production of the larger of two small proteoglycans, putatively, biglycan.

The thinning of the cornea that occurs in keratoconus has been well described; however, the mechanism of tissue degradation remains unknown. Elevated proteinase activity is one possibility and approximately 20 publications over the last 20 years have addressed this hypothesis. Early studies reported increased collagenase and gelatinase activities in the medium of keratoconus corneal cultures. After the characterization of the matrix metalloproteinase (MMP) enzymes, studies focused on the expression of specific MMPs, in particular the gelatinases, MMP‐2 and MMP‐9. Matrix metalloproteinase‐2 was found to be the major MMP of the cornea and was constitutively produced in normal tissue, whereas MMP‐9 expression was induced by various stimuli, including phorbol esters and even tissue culturing. These studies suggested that there were no differences in the amounts or states of activation of MMP between normal and keratoconus corneas, although the amounts of some proteinase inhibitors, including tissue inhibitors of metalloproteinases, α‐1‐proteinase inhibitor and α‐2‐macroglobulin, were decreased in keratoconus. Most recently, the lysosomal proteinases, cathepsin B and cathepsin G were reported to be elevated in keratoconus corneas, and it is possible that it was cathepsin activity, not MMP activity, that was measured in some early studies. Nevertheless, there are now about 20 human MMPs identified and it is possible that some of these, other than the well known collagenase (MMP‐1) and gelatinases (MMP‐2 and MMP‐9), could be implicated in the pathology of keratoconus. Studies have begun to address more recently described MMPs and it has been reported that the membrane‐bound MT1‐MMP (MMP‐14), which activates latent MMP‐2, was found to have increased expression in keratoconus corneas, whereas the stromelysins, MMP‐3 and MMP‐10, were not.

The observation that MT1-MMP expression may be up-regulated in keratoconus corneas, taken together with the demonstration that human corneal cells can express this enzyme, which in turn can activate latent MMP-2, provide evidence for a possible role for MT1-MMP in the pathogenesis of keratoconus.

Electrostatic amino acid interactions between receptor subunits within the T-cell antigen receptor (TCR) transmembrane domain are critical for the formation of the TCR-CD3 complex. Core peptide, a short peptide corresponding to the TCR-a transmembrane region, containing two positively charged amino acids, is known to inhibit T-cell function in vitro and in vivo. The aim of this study was to examine peptides corresponding to the syntactic transmembrane CD3 region binding to TCR-a for their ability to inhibit T-cell activation in vitro and in vivo. Three peptides matching the transmembrane sequence of CD3-d, -e and -c were synthesized and tested in different biological in vitro and in vivo systems for their effect on T-cell activity. The CD3-peptides had no impact on T-cell function in vitro, but surprisingly, decreased signs of inflammation in the adjuvant arthritis rat model in vivo. Preliminary evidence suggests that peptides with CD3 transmembrane-derived sequences can inhibit an immune response as assessed by adjuvant-induced arthritis. The lack of in vitro activity may lead to a wasteful disregard of active compounds in the process of drug discovery and development.