In the United States and throughout the world, cancer incidence and mortality has increased dramatically in both developed and developing nations. Cancer causes ~13% of human deaths with 7.6 million people dying from cancer in 2007. More people in the US die of lung cancer than breast, colon, kidney, and prostate cancers combined. Recent studies show that veterans are 25 to 75 percent more likely to develop lung cancer than people who did not serve in the military. Advances in cancer immunotherapy are leading to breakthroughs in treatment. Adoptive transfer of T cells expressing chimeric antigen receptors (CAR-T) results in durable remis- sions for B cell malignancies. Checkpoint blockade with antibodies against PD-1, PD-L1, and CTLA-4 results in partial and complete responses in patients with a variety of malignancies. Yet, significant limitations exist. With the exception of patients with melanoma, only a minority of patients respond to checkpoint blockage. Common cancers such as prostate and colorectal cancer generally do not respond. Thus, additional approaches are needed to realize the full potential of cancer immunotherapy. Treatment with ?? T cells expressing V?2V?2 TCRs is one such approach. Unlike ?? T cells, the response of V?2V?2 T cells is not MHC restricted but instead requires the Ig superfamily protein, butyrophilin 3A1, to sense the foreign-microbial isoprenoid metabolite, HMBPP, and the self-metabolite, IPP. This sensing allows tumor cells to be recognized and killed by V?2V?2 T cells independent of their mutational burden. V?2V?2 T cells safely expand to very high numbers during many infections (up to 1 in 2 circulating T cells) where they kill infected cells and secrete inflammatory Th1 cytokines, chemokines, and growth factors. Two approaches are being used to treat cancer with V?2V?2 T cells. The first is to immunize with stimula- tors such as the bromohydrin analog of HMBPP or the aminobisphosphonate zoledronic acid with low-dose IL- 2. Although treatment has resulted in partial remissions, these vaccines eventually cause anergy and deletion of the V?2V?2 T cells. The second is to adoptively transfer V?2V?2 T cells. This approach is safe and has in- duced complete remissions in three patients with solid tumors, and induced partial remissions or stable dis- ease in others. However, for widespread adoption, V?2V?2 T cell therapy needs to be more effective. Live bacterial vaccines have been used to prevent tuberculosis, typhoid fever, and tularemia. The bacteria produce compounds that activate innate immunity and antigens that stimulate ?? T cells to provide help to the V?2V?2 T cells as they expand. We have now identified an attenuated Listeria strain that consistently ex- pands V?2V?2 T cells. Listeria preferentially accumulate in tumors which should allow the specific tar- geting of adoptively-transferred V?2V?2 T cells to tumors as well as TCR stimulation at the tumor site by HMBPP. We also find that V?2V?2 T cells rapidly express checkpoint receptors such as CTLA-4, PD- 1, TIM-3, and LAG-3 upon stimulation and that adding PD-1 checkpoint blockade markedly enhances prostate tumor immunity by V?2V?2 T cells in a mouse model. We have also identified a novel bisphosphonate prodrug. To accomplish these goals, we will: delete inlB in ?actA prfA* (G155S) Listeria vaccine and assess dosing and timing of immunization, assess the ability of Listeria bacteria to target and acti- vate adoptively transferred V?2V?2 T cells to control tumors, and assess effectiveness of combining checkpoint blockade and a new PTA bisphosphonate prodrug with adoptively transferred V?2V?2 T cells. We have an outstanding team with an excellent track record and have extensive experience working with ?? T cells and isoprenoid metabolism. We have established in vivo models and techniques. An effective Liste- ria vaccine have been identified. The molecular methods to create more vaccines are well developed. In con- clusion, immunotherapy using bacterial vaccines for V?2V?2 T cells has the potential to be broadly applicable for the treatment of many different tumors both by direct activation and through potentiating adoptive transfer.

Public Health Relevance

Many studies have found higher rates of cancer incidence and mortality among veterans. In the United States, cancer accounts for 25% of all deaths with 30% of these from lung cancer. U.S. veterans who served in the Vietnam, Korean, or the 1991 Persian Gulf war are at high-risk for lung cancer since they are 25 to 75 percent more likely to develop lung cancer than people who did not serve. Also, veterans were exposed to various environmental hazards and microbial infections. Despite advances in our understanding of the causes of cancer, progress in treatment is still limited for many types of cancers especially solid tumors. Our vaccine will stimulate a type of blood cells called ?gamma delta? T cells for treatment of a number of different tumor types by immunotherapy.

National Institute of Health (NIH)
Veterans Affairs (VA)
Non-HHS Research Projects (I01)
Project #
Application #
Study Section
Special Emphasis Panel (ZRD1)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Iowa City VA Medical Center
Iowa City
United States
Zip Code
Tanaka, Yoshimasa; Murata-Hirai, Kaoru; Iwasaki, Masashi et al. (2018) Expansion of human ?? T cells for adoptive immunotherapy using a bisphosphonate prodrug. Cancer Sci 109:587-599
Nada, Mohanad H; Wang, Hong; Workalemahu, Grefachew et al. (2017) Enhancing adoptive cancer immunotherapy with V?2V?2 T cells through pulse zoledronate stimulation. J Immunother Cancer 5:9
Tanaka, Yoshimasa; Iwasaki, Masashi; Murata-Hirai, Kaoru et al. (2017) Anti-Tumor Activity and Immunotherapeutic Potential of a Bisphosphonate Prodrug. Sci Rep 7:5987
Sakai, Yuki; Mizuta, Satoshi; Kumagai, Asuka et al. (2017) Live Cell Labeling with Terpyridine Derivative Proligands to Measure Cytotoxicity Mediated by Immune Cells. ChemMedChem 12:2006-2013
Collins, Cheryl C; Bashant, Kathleen; Erikson, Cuixia et al. (2016) Necroptosis of Dendritic Cells Promotes Activation of ?? T Cells. J Innate Immun 8:479-92
Matsumoto, Kenji; Hayashi, Kosuke; Murata-Hirai, Kaoru et al. (2016) Targeting Cancer Cells with a Bisphosphonate Prodrug. ChemMedChem 11:2656-2663
Wang, Hong; Morita, Craig T (2015) Sensor Function for Butyrophilin 3A1 in Prenyl Pyrophosphate Stimulation of Human V?2V?2 T Cells. J Immunol 195:4583-94
Workalemahu, Grefachew; Wang, Hong; Puan, Kia-Joo et al. (2014) Metabolic engineering of Salmonella vaccine bacteria to boost human V?2V?2 T cell immunity. J Immunol 193:708-21
Sugie, Tomoharu; Murata-Hirai, Kaoru; Iwasaki, Masashi et al. (2013) Zoledronic acid-induced expansion of ?? T cells from early-stage breast cancer patients: effect of IL-18 on helper NK cells. Cancer Immunol Immunother 62:677-87
Wang, Hong; Henry, Olivier; Distefano, Mark D et al. (2013) Butyrophilin 3A1 plays an essential role in prenyl pyrophosphate stimulation of human V?2V?2 T cells. J Immunol 191:1029-42

Showing the most recent 10 out of 13 publications