Impaired anti-mycobacterial T cell functions in HIV-infected patients

The main aim of this research project was to improve our understanding of immune functions in HIV-infected individuals and how we can use this knowledge to develop new treatment strategies. Main achievements in this project: Establishment of HIV virus laboratory work protocols. The project developed procedures and BSL3 laboratory routines to perform HIV virus research work at NTNU/CEMIR. This includes methods to propagate, purify and quantify HIV as well as different HIV infections models for T cell lines and primary T cells. HIV laboratory tools were previously not available at our institute. These methods will also be used in future studies, for example in a recently performed CRISPR/Cas9 whole genome screen for HIV resistance and susceptibility genes (collaboration R. Kandasamy, CEMIR). Such studies are important to identify new targets in order to develop new treatment strategies for HIV. Discovery that HIV infection stimulates receptors of the innate immune system in CD4+ T cells. A surprising discovery in this project was that HIV activates "pattern-recognition receptors" TLR7, 8 and 9 upon uptake in T cells. We found that activation of TLR8 in T cells triggers inflammatory cytokine production and HIV replication. By using HIV patient samples from St. Olavs Hospital, we could demonstrate that TLR8 stimulation of T cells can re-activate latent HIV infection in patients treated by anti-retroviral therapy. This finding represents a previously unknown mechanism by which CD4+ T cells may detect HIV infection and can have the potential to develop new HIV treatment strategies. A manuscript with first authorship from this project has been written and will be submitted to a high impact journal. Development of a novel cancer peptide vaccination method In collaboration with PCI Biotech (Oslo), we could show that the principle of photochemical internalization (PCI) provides a novel vaccination technology to effectively induce cancer-specific cytotoxic T cell responses. Our findings indicate that this method has the potential to improve efficacy of therapeutic cancer peptide vaccination. A manuscript with first authorship from this project has been submitted, clinical trials are under planning. Other project achievements: Methods and resources developed in this research project contributed to several other research projects. In addition, two PhD projects were co-supervised (both to be submitted in spring 2018) and several master projects were successfully performed on the project. Contributions from this research project resulted in the second authorship in a study published in “PNAS” (Awuh et al., 2015), where we investigated the role of the stress-sensor protein KEAP1 in mycobacterial infections. Contributions were also made to investigate role of TLR7/8 in phagososmal escape mechanisms of mycobacteria in macrophages (Gidon et al., PLoS Pathogens, 2017) and to several other publications (Moura Rosa et al., 2016; Gopalakrishnan et al., 2015). The project also contributed with methodology and support to other patient-near research projects at NTNU and St. Olavs Hospital. In collaboration with A. Tøndell (Clinic of Thoracic and Occupational Medicine), methods were established to isolate, purify and characterize T cells and macrophages from non-small cell lung cancer samples. A research project and preliminary results were obtained with biobank material collected at NKSR from pregnant women with SLE and RA (collaboration with the Department of Rheumatology), a PhD project proposal has been written. Findings from this research project will be followed up in a new 5-year project funded by Samarbeidsorganet HMN-NTNU (90176000) with focus on continuation of the HIV work. This project allowed to extend research from the excellent basic research environment on innate immunity at CEMIR/IKM towards more patient-near clinical research by initiating a new research project including HIV patient samples from St. Olavs Hospital. The project contributed to strengthen the field of infection research at the MH Faculty/St. Olavs Hospital and initiated patient-near HIV research at NTNU/CEMIR and St. Olavs Hospital. Funding of this project allowed to establish BSL3 HIV work procedures and protocols and initiated a research project collaboration between NTNU/CEMIR and St. Olavs Hospital on HIV patient samples. These HIV work procedures have now become a valuable resource for several research projects at NTNU/CEMIR and will be implicated in several existing and future research projects. Findings from this project, showing that TLR8 stimulation may affect virus-replication as well as reactivation of latent HIV in patients on ART may be of particular importance. This represents a previously undescribed host defence mechanism against HIV and may have implications for example for development of new treatment strategies or contribution to a better understanding of HIV complications such as “inflamm-aging”. In addition, results from the project could show that PCI peptide vaccination has great potential as novel strategy to realize therapeutic cancer vaccination with peptides derived from the patients’ cancer cells. This method is minimally invasive, well-tolerated and approved for human use. Pre-clinical and clinical phase I cancer peptide vaccination trials with this method are planned.