Application of immune repertoire sequencing in solid organ transplantation / 器官移植

Immune repertoire is defined as the sum of T cells and B cells, which possesses high diversity and enables immune system to respond to various antigen stimuli. With the development of sequencing technique, immune repertoire sequencing can be utilized to deeply understand the changes of lymphocyte clones when rejection occurs at the gene level, and also provide the possibility for the emergence of novel non-invasive diagnostic techniques based on immune repertoire sequencing. In recent years, more and more attempts have been made to apply immune repertoire sequencing in solid organ transplantation, especially in the fields of kidney transplantation, liver transplantation, heart transplantation and post-transplantation infection. In this article, research progresses on the application of immune repertoire sequencing in these fields were reviewed, and current status of immune repertoire sequencing in organ transplantation and its potential as a novel technique for early non-invasive diagnosis of rejection were summarized, aiming to provide reference for subsequent development and clinical application of this technique.

Investigação de marcadores moleculares sanguíneos durante a infecção pelo vírus Chikungunya / Investigation of blood molecular markers during Chikungunya virus infection

A febre Chikungunya (CHIKF) é uma infecção viral causada pelo vírus Chikungunya (CHIKV). Os sintomas agudos incluem febre alta de início súbito, erupção cutânea, poliartrite e poliartralgia. Embora a infecção geralmente seja resolvida em menos de duas semanas, muitos pacientes experenciam recorrente dor e inflamação nas articulações, que podem persistir por anos. Esse estudo buscou marcadores moleculares no sangue de infectados pelo CHIKV que estejam associados a dor articular e cronicidade da CHIKF. O sequenciamento de receptores de células B (BCR) e T (TCR) demonstrou que a infecção por CHIKV diminui a diversidade desses receptores. Essa diversidade é ainda menor, durante a fase aguda da infecção, naqueles pacientes que irão desenvolver cronicidade. A menor diversidade de BCR em infectados está associada a um aumento na expressão de genes envolvidos na diferenciação e ativação de osteoclastos pela sinalização RANK/RANKL. Em adição, a cronicidade pode estar relacionada um aumento na expressão do gene ZBTB7A cuja expressão confere maior resistência a apoptose em precursores de osteoclastos naqueles pacientes que vão se tornar crônicos. Caso o envolvimento dos osteoclastos durante a patogênese de CHIKF seja confirmado, os pacientes poderão se beneficiar de abordagens terapêuticas já existentes como alternativas adicionais ao tratamento de CHIKF

Chikungunya fever (CHIKF) is a viral infection caused by the Chikungunya virus (CHIKV). Acute symptoms include sudden-onset high fever, rash, polyarthritis, and polyarthralgia. Although the infection usually resolves within two weeks, many patients experience recurrent joint pain and inflammation, which can persist for years. This study sought molecular markers in the blood of CHIKV-infected individuals that are associated with joint pain and chronicity of CHIKF. Sequencing of B (BCR) and T (TCR) cell receptors demonstrated that CHIKV infection decreases the diversity of these receptors. The diversity is even lower, during the acute phase of the infection, in those patients who will develop chronicity. The lower diversity of BCR in infected individuals is associated with an increase in the expression of genes involved in the differentiation and activation of osteoclasts by RANK/RANKL signaling. In addition, chronicity may be related to an increase in the expression of the ZBTB7A gene whose expression confers greater resistance to apoptosis in osteoclast precursors in those patients who will become chronic. If osteoclast role during CHIKF pathogenesis is confirmed, patients may benefit from existing therapeutic approaches as additional alternatives to CHIKF treatment

Diversity of the T cell receptor β chain complementarity-determining region 3 in peripheral blood of neonates with sepsis: an analysis based on immune repertoire sequencing / 中国当代儿科杂志

OBJECTIVES@#To investigate the diversity of peripheral blood T cell receptor (TCR) β chain complementarity-determining region 3 (CDR3) based on immune repertoire sequencing in neonates with sepsis and the possible pathogenesis of neonatal sepsis.@*METHODS@#A total of 12 neonates with sepsis were enrolled as the case group, and 9 healthy full-term infants, matched for gestational age, birth weight, and age, were enrolled as the control group. Omega nucleic acid purification kit (SQ blood DNA Kit II) was used to extract DNA from peripheral blood samples, TCR β chain CDR3 was amplified by multiplex PCR, and then high-throughput sequencing was performed for the products to analyze the diversity of TCR β chain CDR3 and the difference in expression.@*RESULTS@#The length and type of TCR β chain CDR3 were similar between the case and control groups, and Gaussian distribution was observed in both groups. With D50 and Shannon-Wiener index as the evaluation indices for diversity, the case group had a significantly lower diversity of TCR β chain CDR3 than the control group (@*CONCLUSIONS@#There is a significant change in the diversity of TCR β chain CDR3 in the peripheral blood of neonates with sepsis, suggesting that it might be associated with the immune pathogenesis of neonatal sepsis.

TCR immune repertoire characteristics of patients with drug-induced liver injury due to Polygonum multiflorum preparations / 中国中药杂志

To explore the immune status of patients with drug-induced liver injury caused by Polygonum multiflorum preparations,and analyze their immune characteristics. Case-control design was used to collect the cases of drug-induced liver injury caused by P. multiflorum preparations through key specialized surveillance. Five matching factors,namely type of P. multiflorum preparations,gender,age,basic diseases and concomitant medication were controlled. According to the ratio of 1 ∶ 1,cases of patients who took P. multiflorum preparations but with no liver injury were monitored at prospective hospitals. The demographic information,disease information,medication information and laboratory examination information of the two groups were recorded,and venous blood was collected. The gene sequence was detected by high-throughput sequencing technology,and the characteristics of TCR immune repertoire of the two groups were analyzed. A total of 46 pairs of patients were enrolled in the study. The results showed significant differences in the number of CDR3 and clone species,the length of amino acid sequence in CDR3 region,the abundance of V gene and J gene,the cross-linking of V-J gene and the diversity of immune repertoire between patients with drug-induced liver injury and patients without liver injury. The immunohistochemical diversity and high-frequency V-J cross-linking characteristics of patients with liver injury caused by P. multiflorum preparations were found,which provided a reference for screening out drug users to reduce the occurrence of liver injury caused by P. multiflorum preparations.

Traditional Chinese medicine (tumor) prevention and evaluation model based on ultra-early immune response information amplification index naCTL and multidimensional immunoinformatic index TCR/BCR/HLA effective diversity / 中国中药杂志

The best time of tumor intervention is before the formation of tumor. However,due to the limited number of tumor cells,it is difficult to quantify tumor cells and immunity by the current methods available( such as CTC,ct DNA). This affects the tumor prevention in this period,and the in-depth detection,intervention and evaluation of traditional Chinese medicine( TCM)( tumor) prevention. Due to the limitations of the current detection,the evaluation system turns to detect tumor neoantigen-specific CTL( naCTL) that are directly relating to tumor cells and proliferate to high order of magnitudes after activation,and immune repertoire( TCR/BCR/HLA) effective diversity,introduces immune checkpoints,uses information of " disease" in Western medicine and " syndrome" in TCM( prevention),and sets up a multi-dimensional statistical immunity model using a variety of data analysis and related algorithms. This model can amplify the ultra-early information of tumor,indirectly evaluate the quantity and status of tumor cells,and provide quantitative measurement and new evaluation methods for the normalization of immunity and TCM( tumor) prevention. This model is not only one of important evaluation methods for resisting tumor immunity and treating TCM( tumor) prevention,but also will reveal the scientific connotation of TCM syndrome from the perspective of immunology.

Study of T cell receptor in patients with acute rejection after renal transplantation / 中华肾脏病杂志

Objective To evaluate the immune status of acute rejection recipients,and to improve the short-term and long-term survival rate of renal transplant recipients and grafts,and to investigate dynamically the changes in the immune repertoire of patients with acute rejection.Methods Combined multiplex PCR amplification technique and high throughput sequencing technique,the TCR β chain complementarity determining region 3(CDR3)diversity and repertoire characteristics at different time points during renal transplantation were analyzed,in order to reveal the immunological characteristics of T lymphocytes in patients with acute rejection.Results The diversity of TCR CDR3 in acute rejection patients was reduced to the lowest one day after surgery.The diversity of TCR CDR3 before acute rejection was higher than before.The acute rejection-related upregulated TCR CDR3 amino acid sequences were screened out.In addition,TCR beta chain V and J subfamily showed the phenomenon of advantage usage in pre-acute rejection,which may be due to T cell recognition of transplanted kidney antigens in vivo.Conclusions The immune diversity of patients with acute rejection is significantly lower.In addition,TCR beta chain V and J subfamily show the phenomenon of advantage usage.

Detection of minimal residual disease in children with acute lymphoblastic leukemia and its application / 中华实用儿科临床杂志

Childhood acute lymphoblastic leukemia (ALL) is a kind of abnormal proliferation of malignant tumor diseases originated in lymphocytes,which is the most common malignant tumor in children.Although stratified treatment has significantly improved the efficacy of ALL in children,but 15％-20％ of the patients still have ultimately relapsed due to the minimal residual disease(MRD).MRD refers to the leukemia patients after induction chemotherapy complete remission (or bone marrow transplant),in vivo residual morphology which could not be detected in trace amounts of leukemia cells.The detection methods of MRD mainly include flow cytometry,quantitative real-time polymerase chain reaction and immune repertoire sequencing.MRD level detection is of great important to judge the prognosis,and to guide the risk grouping and individual treatment,and so on.Now,the progress in the clinical application of MRD in children with ALL in recent years was reviewed.

Advances in research and application of immune repertoire analysis in hematological malignancies / 白血病·淋巴瘤

The diversity of immune repertoire (IR) is closely related to immune function and tumors derived from B or T lymphocytes.In recent years,the development of next generation sequencing has greatly promoted the progression and application of IR analysis.The research of IR has become a new hotspot of the American Society of Hematology Annual Meeting in 2016.Relevant reports in this meeting will be reviewed together with advances in research and application of IR analysis in hematological malignancies in recent years.