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Purpose: Antibodies that target immune checkpoint molecules have demonstrated significant and durable clinical benefit through re-activation and proliferation of pre-existing tumor-infiltrating CD8+ T cells in a broad range of tumor types including lung cancer. Detailed characterization of T cell dynamics at clonal levels using nextgeneration T cell receptor (TCR) sequencing in large cohorts of lung cancer patients is yet reported.Methods: We performed TCR sequencing of peripheral blood samples (and some tumors) obtained from 27 lung cancer patients undergoing single/combined immune checkpoint blockade therapies.Results: In one responder, we found expansion of a single T cell clone (approximately 20% of the all TCR reads) in a metastatic subcutaneous lesion that showed pathologic complete response on day 17 of treatment. The same TCR CDR3 sequence was detected in 6.1% of TCR reads in the peripheral blood prior to treatment initiation and the expansion remained persistent in peripheral blood at 21.0% at week 10 and 24.3% at week 48. In other patients who showed durable response, we also found persistent oligoclonal T cell expansion in their peripheral blood which was not observed in non-responders even while they remained on therapy.Conclusion: We found sustained expansion of oligoclonal T cell clones in lung cancer patients who had durable response to immune checkpoint blockade. This suggests possible use of longitudinal TCR sequencing to assist clinical decision-making, assess synergism with other agents, and most importantly facilitate rational development of alternative treatment strategies for non-responders.

A systematic study has been made of the types and quantities of acid mucopolysaccharides isolated from a variety of tissues and fluids. Identification of the polysaccharides has been based on the precipitation of the polysaccharides as calcium salts, on analysis, on optical rotation and on digestibility by testicular and bacterial hyaluronidases. On this basis, six distinct mucopolysaccharides have been established: hyaluronic acid, chondroitin, the chondroitin sulfates designated as A, B and C, and keratosulfate. Additional mucopolysaccharides have been demonstrated in amyloid tissue and, in aorta, which are resistant to testicular hyaluronidase and contain D-glucosamine, a uronic acid and varying amounts of sulfate. In addition, incompletely sulfated chondroitin sulfates A or C have been obtained by fractional elution from an ion exchange resin. The tissues have been divided into seven groups on the basis of the mucopolysaccharides isolated from them, as follows: 1. (1) Vitreous humor, synovial fluid, filterable fowl tumors, liposarcoma: hyaluronic acid only. 2. (2) Cartilage, adult bone, chondrosarcoma, chordoma: chondroitin sulfate A and/or C. 3. (3) Umbilical cord, loose connective tissue of electric eel, fibroblasts grown in tissue culture: hyaluronic acid as the major with chondroitin sulfate C as a minor component. 4. (4) Pig skin, ligamentum nuchae (the latter contains chondroitin sulfate A in addition): chondroitin sulfate B and hyaluronic acid as major components. 5. (5) Adult connective tissue, tendon, heart valve: chondroitin sulfate B and C with hyaluronate as minor component. 6. (6) Cornea: keratosulfate, chondroitin and chondroitin sulfate A. 7. (7) Incompletely characterized fractions (amyloid, bovine aorta, calf bone). From calf bone, chondroitin sulfate A and C and a large incompletely sulfated fraction were isolated and, in addition, a minor fraction of keratosulfate. From bovine aorta, a major fraction with the properties of chondroitin sulfate A was isolated, accompanied by hyaluronate, chondroitin sulfate B and a variety of glucosamine-containing fractions of varying degrees of sulfation. These latter fractions are believed to be related to heparin. All normal tissues investigated appear to contain two or more mucopolysaccharides in contrast to tumors of mesodermal origin which produce a single type of mucopolysaccharide.

Two types of hyaluronidases have been described: one, found in testicular extracts and in snake venom, randomly cleaves the hexosaminidic bonds and yields a consecutive series from disaccharide through higher homologues of disaccharide units1; the other, found in micro-organisms, yields a Δ4–5 (glucuronido) unsaturated disaccharide2. In this communication, a third type of hyaluronidase is described, found thus far only in extracts of the leech (Hirudo medicinalis), which hydrolyses the endoglucuronidic linkages of hyaluronic acid.

Summary1. From the urine of 4 out of 5 patients with gargoylism, a mixture of mucopolysaccharides was isolated which could be identified as ChS-B and heparitin sulfate—with the former predominating. The urine of one patient, and the liver of another, yielded only heparitin sulfate. 2. The type of polysaccharide excreted by these patients had no obvious correlation with the severity of the disease or the postulated mode of inheritance suggested by the history. 3. It is suggested that gargoylism represents an overproduction of certain mucopolysaccharides due to a genetically determined error of differentiation of fibroblasts.