Cui Kong

The Skewness of Alpha Beta T Cell Receptors in Peripheral Blood of the Patients with Type 1 Diabetes.

AIM To detect the skewness of TCR Vα and TCR Vβ of patients with type 1 diabetes (T1D). METHODS The heparinized venous blood was collected from ten patients with T1D. The peripheral blood lymphocytes (PBL) were isolated and used to extract mRNA. Reverse amplyfication was performed for cDNA synthesis. The skewness of TCR Vα and Vβ was detected with real-time florescence quantitative polymerase chain reaction (RQ-PCR) and analyzed by DNA melting-curve analysis technique, respectively. RESULTS Among the TCR Vα genes, the skewness frequency rate (SFR) of Vα22 was 30%; both of Vα5 and Vα24 were 20%; Vα28 was the only restricted-clone gene with the SFR of 10%. In all the Vβ genes, Vβ7 and Vβ17 were the the highest expression genes, and their SFRs were both 60%. Vβ11 was near them with the SFR of 40%; the restricted clonal genes were Vβ18 and Vβ20, their SFRs were 10% and 20%, respectivley. CONCLUSION There are skewd genes in TCR Vα and TCR Vβ, which are probably relative to the onset of T1D.

The short review on the studies of T cells receptors relate to type 1 diabetes

Although type 1 diabetes (T1D) is a common autoimmune disease, and there have been many experimental and clinical researches on it, yet the exact mechanisms still remain unclear. What is the fact without doubt that T cells play an important role in the progress of T1D. Because the identification of T cells depends on the identification of MHC which binds the peptides of auto-antigens, the responses of T cells specific to this combination might make the T cell receptor (TCR) genes changed, especially the complementarity determining region 3 (CDR3) genes. According to this theory, it is possible to unclose the immune mechanisms of T1D from the changes of the specific TCR. This paper focuses on the current studies of TCR relative to T1D.

In silico analysis of tcr vβ7 of two patients with type 1 diabetes mellitus

Objective: To compare the sequences and crystal structures of variable region of beta chain 7 (Vβ7) of T cell receptor (TCR) of two patients with type 1 diabetes mellitus (T1DM). Patients and Methods: The skewness of TCR Vβ7 of two T1DM patients were detected with real-time florescence quantitative polymerase chain reaction (FQ-PCR) and deoxyribonucleic acid (DNA) melting curve analysis technique followed by being sequenced, the crystal structures of them were simulated according to CPH models 2.0 Server, IMGT database, and RasMol 2 software. Results: The whole sequences of TCR Vβ7 of T1DM patient-1 were “CASRTAGQYEQYFGPGTR”, that of patient-2 were “CASRTAGQYEQFFGPGTR”; the only difference between them lied on the 12th amino acid. The crystal structures of Vβ7 of the two patients simulated with backbone model were rather similar, while that with sphere model were obviously different. Conclusion: Although the TCR Vβ7 of the T1DM patients share the similar gene sequences, their crystal structures simulated with sphere model are different, and the mechanism needs further study.

SKEWNESS OF TCR Vβ OF PERIPHERAL BLOOD AND SYNOVIAL FLUID OF PATIENTS WITH RHEUMATOID ARTHRITIS

To date, the complete mechanism of rheumatoid arthritis (RA) remain unclear, T cells have been proposed to play an important role in the disease initiation and progression. Presently, some researchers have reported that there were skewed TCR Vβ in different samples of experimental animals or RA patients, such as in the peripheral blood, joints or synovial fluid, however, most of the results were not coincident or even conflict with each other. In this article, with real-time fluorescence quantitative PCR with DNA melting curving technique, we detected the bias of TCR Vβ of RA patients, and found that although most of TCR Vβ usage were different between peripheral blood and synovial fluid, the overview of all the Vβ skewness was similar between the two samples.

TCR Vβ Usage of Peripheral Blood and Liver Infiltrating Lymphocytes in Patients with Chronic Hepatitis B

INTRODUCTION Chronic hepatitis B (CHB) is still a public health problem and its mechanism remains unclear. In this study, we detect the skewness of T cell receptor beta chain variable gene (TCR Vβ) in peripheral blood lymphocytes (PBL) and the liver infiltrating lymphocytes (LIL) of patients with CHB; and hope to provide information for further research on the pathogenic mechanism of CHB. MATERIAL AND METHODS Fifteen patients with CHB, ten healthy volunteers and three patients with liver cysts were recruited as the subjects. The usage of TCR Vβ of PBL and LIL were measured and compared; the associations of the TCR Vβ usage of PBL with some hematological indices, including human leukocyte antigen (HLA) alleles, percents of CD4+ and CD8+ T cells, sera levels of HBV-DNA and IFN-γ, were analyzed. RESULTS In PBL, Vβ12 and Vβ13.1 were the highest predominant usage genes which usage frequencies were all 46.7%; Vβ23 was the key limited usage gene (40.0%). In LIL, the mainly predominant and limited usage gene was Vβ13.1 (73.3%) and Vβ23 (46.7%), respectively. About half of the patients with CHB with HLA-DR9 or HLA-DR12 showed the predominant usage of Vβ5.2 or Vβ13.2. In patients with CHB, the percentage of CD4+ T cells was 33.41 ± 5.39 %, that of CD8+ T cells was 28.67 ± 6.77 %; the concentration of IFN-γ was 182.52 ± 44.16 pg/mL. Compared to the healthy controls, there were significant differences for these data (P < 0.05). Neither ALT nor HBV-DNA was relative to the usage of TCR Vβ. CONCLUSIONS PBL and LIL share the common sknewness of TCR Vβ genes, which probably relates to some hematological indices. However, the roles of such similarities and associations in the development of CHB need further study.INTRODUCTION Chronic hepatitis B (CHB) is still a public health problem and its mechanism remains unclear. In this study, we detect the skewness of T cell receptor beta chain variable gene (TCR Vβ) in peripheral blood lymphocytes (PBL) and the liver infiltrating lymphocytes (LIL) of patients with CHB; and hope to provide information for further research on the pathogenic mechanism of CHB. MATERIAL AND METHODS Fifteen patients with CHB, ten healthy volunteers and three patients with liver cysts were recruited as the subjects. The usage of TCR Vβ of PBL and LIL were measured and compared; the associations of the TCR Vβ usage of PBL with some hematological indices, including human leukocyte antigen (HLA) alleles, percents of CD4+ and CD8+ T cells, sera levels of HBV-DNA and IFN-γ, were analyzed. RESULTS In PBL, Vβ12 and Vβ13.1 were the highest predominant usage genes which usage frequencies were all 46.7%; Vβ23 was the key limited usage gene (40.0%). In LIL, the mainly predominant and limited usage gene was Vβ13.1 (73.3%) and Vβ23 (46.7%), respectively. About half of the patients with CHB with HLA-DR9 or HLA-DR12 showed the predominant usage of Vβ5.2 or Vβ13.2. In patients with CHB, the percentage of CD4+ T cells was 33.41 ± 5.39 %, that of CD8+ T cells was 28.67 ± 6.77 %; the concentration of IFN-γ was 182.52 ± 44.16 pg/mL. Compared to the healthy controls, there were significant differences for these data (P < 0.05). Neither ALT nor HBV-DNA was relative to the usage of TCR Vβ. CONCLUSIONS PBL and LIL share the common sknewness of TCR Vβ genes which probably relates to some hematological indices. However, the roles of such similarities and associations in the development of CHB need further study.

Preparation and identification of transfer factor specific to Staphylococcus aureus in vitro

The objective of the study was to explore the methods for preparing transfer factor specific to Staphylococcus aureus (SA‐STF) in vitro. Under the optimum conditions, the spleen cells of rabbits were immunized with SA in vitro to prepare SA‐STF, and the immune activities were identified with the phagocytosis and sterilization, skin delayed‐type hypersensitivity, and immune protection tests. The concentration of polypeptide was 2.26 ± 0.27 mg/mL, and ribose was 0.684 ± 0.094 mg/mL. The phagocytosis and sterilization rates of the STF group were 70.9 ± 12.4% and 62.1 ± 12.2%, respectively, and compared with the non‐specific transfer factor (NTF) group, there were no significant differences (P = 0.074 and 0.069, respectively). The skin was inflamed and marked nodules formed at the injection site in the mice of the STF group rather than the NTF and control groups. The survival rate of the STF‐1 group was significantly higher than the survival rates of the STF‐2 (P = 0.024) and NTF groups (P = 0.016). SA‐STF was prepared and characterized successfully in vitro, and it probably is a biological candidate for therapy or adjuvant therapy for diseases caused by Staphylococcus aureus.

Comparison of Beta Variable Gene Usage of T Cell Receptor in Peripheral Blood and Synovial Fluid of Rheumatoid Arthritis Patients

Objective To compare the beta variable (Vβ) gene usage of T cell receptor (TCR) in peripheral blood (PB) and synovial fluid (SF) of the patients with rheumatoid arthritis (RA). Methods The total RNAs were extracted from PB and SF of 12 RA patients and reverse-transcribed and amplified by polymerase chain reaction (PCR), then the TCR Vβ gene usages were detected with real-time fluorescence quantitative PCR with DNA melting curving technique. Results For the four patients with acute RA, Vβ1 and Vβ13.1 were the key advantage usage genes in PB, while in SF that were Vβ10 and Vβ13.1. Except for Vβ11, there were no other genes found to be restrictively used both in PB and SF. For the eight patients with chronic RA, in PB, the main advantage usage genes were Vβ1, Vβ5.1, Vβ7 and Vβ13.1, the limited usage genes were Vβ2, Vβ11, Vβ21 and Vβ22. In SF, the predominant usage genes were Vβ1, Vβ6, Vβ7, Vβ13.1 and Vβ18, the limited usage genes were Vβ2, Vβ10, Vβ11, Vβ12, Vβ17, Vβ22, Vβ23 and Vβ24. Conclusions TCR Vβ gene usages in PB are similar to that in SF of RA patients, and this is probably an indication for the further studies of the pathogenesis and therapy of RA in future.