The Expression And Significance Of CD48 And CD244 On Peripheral T Cell Of Patients With Rheumatoid Arthritis

Rheumatoid arthritis ( RA) is a whole body autoimmunity disease which character is chronic destructive arthritis. The etiological factor and pathogenesis of RA is uncompletely known. At present, infection, genetic factor, T Lymphocyte, B Lymphocyte and synovial cell are thought to be involved with RA. A-mong these, the imbalance of Th cell and Ts cell plays an importment role. Many reports showed the function of Th cell weakened and the function of Ts cell enhanced relatively, excreted cytokine, growth factor, activated B cells to plas-macyte, then excreted lots of immunoglobulin, meanwhile, joints occurred inflammatory reaction and destruction.The activation of T cells need two independent signals. The two signals are offered by APC when naive T cell recognizes specific antigen. The first signal offered by TCR and CD4 or CD8 binding with MHC - peptid compound presented by APC. The second signal comes from the interaction of T cell and co - stimulation molecu on APC. B7 presents the co - stimulation signal for naive T cell, and its receptor is CD28 on T cells. Once initial activated, T cell can express membran molecus to maintain and modify co - stimulation signal.A number of receptor/ligand pairs have been showen to modulate lymphocyte responses. Among these are members of CD2 subset of the Ig family of re-cepters, including CD2, CD48, CD58, CD84, CD244, signaling lymphocyte activation molecule, and Ly -9. These receptors have been implicated in modulation of T or NK cell responses. One of better characterized interaction is the one occurring between CD2 and CD48 that lowers quantitative thresholds andfine - tunes T cell activation both in vitro and in vivo. The interaction of CD2 -CD48 enhance T cell - APC adhension and trigger T cell activation, optimize T cell reactivity to IL - 12 uniquely. Like CD2, CD244(2B4) interacted with CD48 enhance activation and proliferation of T cell. CD244 binds to CD48 with 4 -9 fold greater affinity than CD2 in rodent. CD2 doesnt bind to CD48, CD2/ CD48 doesn \ modulate lymphocyte responses in human. Investigation show CD244 on activated/memory T cells serves as a ligand for CD48 and by its ability to interact with CD48 provides costimulatory - like function for neighboring T cells;CD244 on NK cell interact with CD48 on T cell can enhance proliferation of CD4+T and CD8 +T cell;CD244 and CD48 on NK cell participate in proliferation of NK cell and enhance secretion of IFN - r.ObjectivesUsing FACS, we explored expression of CD48 and CD244 on peripheral T cell of patients with rheumatoid Arthritis. Analyzed clinical significance of expression of CD48 and CD244 on peripheral T cell of patients with rheumatoid arthritis.Materials and Methods1. Experiment object;30 patients with rheumatoid arthritis in hospital, selected with ARAs standard. 30 health persons were selected as control. The two groups are matched in sex and age.2. Experiment equipment and Reagent:flow cytometer (FACScan, produced by Becton - Dickinson Company, A-merican) , cryogenic centrifugal apparatus ( Produced by Japanese TOSHIBA Company) , thermostatic water circulating instrument ( Produced by Japanese TOSHIBA Company)FITC fluorescence labelling mouse anti human CD48 monoclonal antibody, FITC fluorescence labelling mouse anti human CD244 monoclonal antibody, PE fluorescence labelling mouse anti human CD3 monoclonal antibody, Per - CPfluorescence labelling mouse anti human CD8 monoclonal antibody, erythrocyte bursting liquid, phosptat buffe, above reagents were produced by American Bee-ton - Dickinson Company.3. Experiment method;Added lOOui anti - coagulation blood into two tubes for flow cytommter each. Then added FITC fluorescence labelling mouse anti human CD48 monoclonal antibody lOjxl, PE fluorescence labelling mouse anti human CD3 monoclonal antibody 5 (jlI , Per - CP fluorescence labelling mouse anti human CD8 monoclonal antibody 5jxl to the first tube;added FITC fluorescence labelling mouse anti human CD244 monoclonal antibody IOjjlI , PE fluorescence labelling mouse anti human CD3 monoclonal antibody 5jxl, Per - CP fluorescence labelling mouse anti human CD8 monoclonal antibody 5jxl to the other. Incubated 15 minutes from light, then added 3ml erythrocyte bursting liquid, put the tubes to thermostatic water circulating instrument to pellucidum at 37. Ot. On the condition of 4t lOOOrpm, centrifugated 4 minutes with cryogenic centrifugal apparatus. Taked out of exponents, poured upper schedul. Then added 2ml phosptat buffe, centrifugated 4 minutes. Taked out of exponents, poured upper schedul. Repeated the upper step. Then added 0. 3ml phosptat buffe, detected by flow cytometer. Use " door technique" to mark off lymphocytes, set each standard door 50000 cells. Then analysised the results through " Cell Ques Plot" software. Recorded average fluorescence intensity of lymphocyte surface. Statistics method;use SPSS11.5 software to analysis data.Results(1 ) T cell subgroup;In RA group, the average fluorescence intensity of peripheral CD8 + T cell , CD4 + T cell , CD4/CD8 was 21.15 ±6. 97% ,48.16 ±7. 67% ,2. 60 ± 1. 23 respectively. While, in control group was 26. 57 ±7. 03% ,46. 96 ±7. 30% ,1.91 ±0. 65 respectively. Peripheral CD8 + T cell in RA group was very significantly fewer than control group, P < 0. 01. Peripheral CD4+ T cell in RA.group was higher than control group, P >0. 05. There was no significant difference between two groups. CD4: CD8 rate in RA group was lower than control group,P <0. 05. There was significant difference, P <0. 05.(2) The average fluorescence intensity of CD48 and CD244 on peripheral CD8 + T cell and CD4 + Tcell: The average fluorescence intensity of CD48 and CD244 on peripheral CD8 + T cell in RA group were 31. 63 ±9. 98,6.46 ± 1. 28 respectively,in control group were 38. 76 ± 8. 72,6. 08 ± 1.44 respectively. The average fluorescence intensity of CD48 on peripheral CD8+ T cell in RA group was very significantly fewer than control group, P <0. 01. There was no significant difference between two groups in the average fluorescence intensity of CD244 on peripheral CD8 + T cell, P >0.05. The average fluorescence intensity of CD48 and CD244 on peripheral CD4 + T cell in RA group were 48. 92 ± 10. 18,3.04 ±0. 54 respectively,in control group were 46.48 ± 8.44,3. 23 ±0. 64. The average fluorescence intensity of CD48 on peripheral CD4+ T cell in RA group was higher than control group, P >0. 05. There was on significant difference between two groups. There was no significant difference between two groups in the average fluorescence intensity of CD244 on peripheral CD4+ T cell, P>0.05.Conclusion( 1) There was abnormality in peripheral T cell of patients with rheumatoid arthritis. The reduction of peripheral CD3 + CD8 +T cell make the rate of CD4 + T cell and CD8 + T cell up, then the function of helper T cell stronger relative, the function of suppressor T cell weakened play an important role in the pathogenesis of RA.(2) The expression of CD48 on peripheral CD8 + T cell of patients with rheumatoid arthritis was significantly lower than the control group. That the lower expression of CD48 make the number of suppressive T cell reduce, the function of suppressor T cell weakened play an important role in the pathogenesis of RA.