2. In ovarian cancer
The researchers analyzed primary and metastatic immune repertoire sequencing (T-cell receptor beta sequencing) in five ovarian cancer patients. From the characteristics of the TIL repertoire of 93 samples, it was found that whether it is the primary site, metastatic tissue of the same patient, or TIL in different parts of the same tissue, or TIL of different patients, the repertoire features are similar. The characteristics of T cell repertoire in peripheral blood are very different, and the TIL CDR3 rearrangement rate of invading tumors is very low. These results indicate that ovarian cancer induces adaptive immunity of TIL, so that the human immune system still maintains a certain response to ovarian cancer. The basic measurement of TIL cells includes the total number, which is expected to be used as a prognostic indicator, which is applicable to various cancers including ovarian cancer.
3. In small lesions
Leukemia minimal residual desease (MRD) refers to a state in which a small amount of leukemia cells remain in the body after complete remission of leukemia induction chemotherapy (or after bone marrow transplantation treatment). These residual leukemia cells have become the source of leukemia recurrence, and the treatment after relapse is very difficult, mainly because the relapsed leukemia cells have developed drug resistance, so the detection of small residual leukemia is of great significance in predicting early recurrence and treatment at the same time. So the examination of minimal residual leukemia cannot be ignored. Patient samples were used to assess minimal residual disease (MRD) after 29 days of treatment; HiSeq sequencing of T cell receptor C and T cell receptor B targeting potential VJ rearrangement combined mutation regions after multiplex PCR; The sample was a cloned T cell receptor (TCR) complementarity determining region 3 (on CDR3) sequence, and the paired prognostic samples were used to evaluate MRD; abnormal T lymphoblasts were found by multi-parameter flow cytometry compared with the above method. High-throughput sequencing of the receptors TCRB and TCRG existed in most of the case groups and subsequent MRDs. Clones were also found (31 T-cell receptors B in 43 and 27 T-cell receptors G in 43). As expected, high-throughput sequencing TCRB and TCRG were able to detect MRDs that were not detectable in flow cytometry (25 / 35HTS vs. 13/35), which highlights the low-throughput sequencing technology in defining a lower detection threshold MRD has the potential to influence clinical diagnosis and treatment decisions. The study found that Immune Repertoire sequencing may become a novel and personalized way to guide clinical diagnosis and treatment of diseases, in which T cell activity is the main determinant. Therefore, the sequencing of lymphoid receptor genes may improve the clinical diagnostic efficacy and the potential of MRD to monitor lymphoproliferative diseases.
4. In hematopoietic stem cell transplantation
The high morbidity and mortality of chronic graft-versus-host disease (GVHD) covers the well-organized different parts of allogeneic hematopoietic stem cell transplantation (HCT) and the use of immune groups in the peripheral blood repertoire. As the allogeneic reaction is mainly a cross-reaction that is randomly distributed throughout the T cell repertoire, the diversity of T cell receptors (T cell receptors) and the allogeneic hematopoietic stem cell transplantation (all -HSCT) is associated with an increased risk of cancer recurrence. Due to technical limitations, the diversity of T cell receptors can be measured by allogeneic hematopoietic stem cell transplantation. By combining 5- RACE PCR and deep sequencing, the T cell receptors of 28 allogeneic hematopoietic stem cell transplant recipients can be quantified using a single oligonucleotide. Repeated blood sample analysis confirms that the frequency of individual temperature coefficients of resistance can be accurately determined. After 6 months, the diversity of T cell receptors in umbilical cord blood recipients and healthy people was similar, while the diversity of CD4 and CD8 T cells in peripheral blood hematopoietic stem cell transplant recipients with T cell depletion was lower 28 times and 14 times. After 12 months these defects have improved in CD4, but not in the CD8 T cell compartment. Overall, this method provides a groundbreaking view of resuscitation in a T cell repertoire, which can be used to identify high-risk groups of infection or relapse after allogeneic hematopoietic stem cell transplantation.
5. In rheumatoid arthritis
The central link in the onset and development of rheumatoid arthritis (RA) has been considered to be antigen-specific T cell activation, using a new next-generation sequencing method to detect multiple cells in a potentially self-reactive cloned T cell repertoire. Recent onset (early) or RA in peripheral blood of each joint and patient. Patients who have recently developed (early) RA (<6 months) (n = 6) or established RA (> 18 months) (n = 6) and screened T cell clones for sequencing over 10,000 T cell receptors (T cell receptors). For each sample, comparative analysis was performed from paired blood samples of T cells. Synovial T cells from two patients were obtained from multiple expanded joints. The degree of each clone is based on the frequency of unique CDR3 sequences within a sample. Cloning more than 0.5% is considered highly cloned (HFC). Results In early RA synovium, the T cell repertoire was mainly 35HEC (median, range 270), accounting for 56% of T cell receptor sequencing. In patients with synovial T cell clones, it was significantly higher than the established RA [11 (in the established RA synovial membrane accounts for 9.8% of T cells 5 to 24), the median, P <0.01]. 34% of highly cloned T cells (range 28% to 40%) are in different joints in the same patient, and only 4% of synovium and blood (P = 0.01) are compared (0% -8%). The results show that RA is a systemic autoimmune disease, and a special T cell clone region is formed in the inflamed and swollen synovium, especially in the early stage of the disease. This indicates that at least RA should address the problem of autoreactive T cells in inflamed and swollen synovial tissue, preferably in the early stages of the disease.