RESUMO
Vaccination by inactivated vaccine is an effective strategy to prevent the COVID-19 pandemic. However, the detailed molecular immune response at single-cell level is poorly understood. In this study, we systematically delineated the landscape of the pre- and post-vaccination single-cell transcriptome, TCR (T cell antigen receptor) and BCR (B cell antigen receptor) expression profile of vaccinated candidates. The bulk TCR sequencing analysis of COVID-19 patients was also performed. Enrichment of a clonal CD8+ T cell cluster expressing specific TCR was identified in both vaccination candidates and COVID-19 patients. These clonal CD8+ T cells showed high expression of cytotoxicity, phagosome and antigen presentation related genes. The cell-cell interaction analysis revealed that monocytes and dendritic cells could interact with these cells and initiate phagocytosis via ICAM1-ITGAM and ITGB2 signaling. Together, our study systematically deciphered the detailed immunological response during SARS-CoV-2 vaccination and infection. It may facilitate understanding the immune response and the T-cell therapy against COVID-19.
RESUMO
Pyroptosis induced by the N-terminal gasdermin domain (GSDMNT) holds great potential for anti-tumor therapy. However, due to the extreme cytoxicity of GSDMNT, it is challenging to efficiently produce and deliver GSDMNT into tumor cells. Here, we report the development of two strategies to package recombinant adeno-associated virus (rAAV) expressing GSDMNT: 1) drive the expression of GSDMNT by a mammal specific promoter and package the virus in Sf9 insect cells to avoid its expression; 2) co-infect rAAV-Cre to revert and express the double-floxed inverted GSDMNT. We demonstrate that these rAAVs can induce pyroptosis and prolong survival in preclinical cancer models. The oncolytic-viruses induce pyroptosis and evoke a robust immune-response. In a glioblastoma model, rAAVs temporarily open the blood-brain barrier and recruit tumor infiltrating lymphocytes into the brain. The oncolytic effect is further improved in combination with anti-PD-L1. Together, our strategies efficiently produce and deliver GSDMNT into tumor cells and successfully induce pyroptosis, which can be exploited for anti-tumor therapy.
Assuntos
Neoplasias da Mama/terapia , Dependovirus/genética , Glioblastoma/fisiopatologia , Glioblastoma/terapia , Proteínas de Neoplasias/genética , Piroptose , Animais , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Neoplasias da Mama/fisiopatologia , Linhagem Celular Tumoral , Dependovirus/fisiologia , Feminino , Glioblastoma/genética , Glioblastoma/imunologia , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Proteínas de Neoplasias/imunologia , Terapia Viral Oncolítica , Vírus Oncolíticos/genética , Vírus Oncolíticos/fisiologia , Ratos , Ratos Wistar , Células Sf9 , Empacotamento do Genoma ViralRESUMO
The target protein for Xklp2 (TPX2), a microtubule-associated protein, can be used to evaluate more precisely the proliferative behavior of tumor cells. The abnormal expression of TPX2 in various types of malignant tumors has been reported, but less is known for cervical cancer. We studied the relationship between TPX2 expression and the biological behavior of cervical cancer. Immunohistochemistry and RT-PCR were used to detect the expression of TPX2 in cervical cancer tissues. The inhibitory effect of TPX2-siRNA on the growth of HeLa human cervical carcinoma cells was studied in vitro. TPX2 expression was found to be significantly higher in cervical carcinoma compared to normal cervical tissues and CIN. The expression of TPX2 in cervical cancer was correlated with histological grading, FIGO staging and lymph node metastasis. TPX2 RNAi in HeLa cervical cancer cells caused S-phase cell cycle arrest, induced apoptosis and inhibited cell proliferation and invasion. In conclusion, TPX2 shows potential to be used as a new marker for cervical cancer diagnosis and therapy.
