Author: Zahra Rashidijahanabad

Publisher:

Published: 2021

Total Pages: 143

ISBN-13:

Chimeric antigen receptor (CAR) T cells and bispecific antibodies (BsAbs) are exciting directions to harness the power of the immune system to fight cancer. Chapter 1 is focused on GD2 ganglioside and the mucin-1 (MUC1) protein, two important tumor associated carbohydrate antigens, and latest advances in CAR T cells and bispecific antibodies targeting these two antigens are presented. The roles of co-stimulatory molecules, structures of the sequences for antigen binding, methods for CAR and antibody construction, as well as strategies to enhance solid tumor penetration and reduce T cell exhaustion and death are discussed. Furthermore, approaches to reduce "on target, off tumor" side effects are introduced. Besides CAR T cells and bispecific antibodies, carbohydrate-based vaccines hold great promise for a number of diseases, which will be the focus of the rest of this dissertation. Several challenges are associated with carbohydrate antigens in regard to inducing specific and protective antibodies as they are poorly immunogenic and the resulting antibodies induced by immunizing with carbohydrates only, typically have low affinity. Currently, developing carbohydrate-based vaccines requires covalent conjugation of the carbohydrate antigen with a protein carrier for optimal antibody response. Thus, generated antibodies have higher affinity against glycan structures. In chapter 2, a potential conjugate vaccine was developed by linking O-specific polysaccharide (OSP) antigen purified from Vibrio cholerae Inaba with QÎø virus like particles (VLPs) efficiently via squarate chemistry as one of the first examples of polysaccharide conjugation to VLPs. The QÎø-OSP conjugate was characterized with mass photometry on the whole particle level. Pertinent immunologic display of OSP was confirmed by immunoreactivity of the conjugate with convalescent phase samples from humans with cholera. Mouse immunization with the QÎø-OSP conjugate showed that the construct generated prominent and long-lasting IgG antibody responses against OSP, and the resulting antibodies could recognize the native lipopolysaccharide from Vibrio cholerae Inaba. This was the first time that QÎø was conjugated with a bacterial polysaccharide for vaccine development, broadening the scope of this powerful carrier. Tumor associated carbohydrate antigens (TACAs) are another class of attractive carbohydrate antigens for the development of anti-cancer immunotherapy with respect to monoclonal antibodies and vaccines. Tetrasaccharide sialyl-Lewisa is an attractive therapeutic target for cancer therapy since it is widely expressed on epithelial tumors of the gastrointestinal tract. The overexpression of sLea appears to be a key event in invasion and metastasis of many tumors and results in susceptibility to antibody-mediated lysis. In chapter 3, sialyl-Lewisa conjugate vaccine with QÎø was developed. The resulting construct, QÎø-sLea, induced antibody production in vivo and the resulting antibodies showed high selectivity for sLea antigen in in vitro studies and effectively reduced tumor growth in mice.