Roles Of Platelets And Red Blood Cells In Modulating T Lymphocytes And Their Relationships In This Process

Immune system has three basic function including immune defence,immune homeostasis and immune surveillance.The major immunologic cells are T lymphocytes whose subsets play an important role in removing the pathogen and outcome of many diseases,including tumor,autoallergic disease,inflammatory and so on.Immune response is a complicated process,whose completion depends on the coordination of lymphocytes and many other immunological cells.Normal immune response is regulated by an immunological network,which is formed by T lymphocytes and many kinds of immunological cells.Because of the complicated immune regulatory mechanism,which can be affected by cells,cytokine microenvironment and other factors,previous studies failed to inform how immune balance is maintained by hematological system.The different statement of immune imbalance and clinical manifestation of many diseases make it difficult to treat etiologically.Therefore,it is important to shed some light on the immune regulatory mechanism.Blood is composed by plasma and blood cells,which include red blood cells, white blood cells and platelet.As the studies goes on,more and more files informed that thrombocytes and erythrocytes played an important role in innate immunity and adaptive immunity.There are still many controversies on how RBC and platelet affect lymphcytes.From 1981 when Siegel gave the new concept of "RBC immune system",many researchers studied the relationship between blood cells and immune response and drew a theory of "the main way to transmit through erythrocyte".The point was most of the antigen would be immune adhered and disposed by erythrocytes before presenting to leucocytes and thrombocytes.Those antigens would induce a series of immune response of leucocytes.However,many studies showed that thrombcytes not only played an important role in the process of haemostasis and inflammatory response,but also in immune response.For instance,erythrocytes can bridge the innate immunity and adaptive immunity and regulate the degree of immune response,as well as promote the maturity of dendritic cells and affect the generation and dividatur of T and B leucocytes.Many researchers informed that platelet could modulate the degree of immune response through connecting innate immunity and adaptive immunity.Since both erythrocytes and thrombcytes play important roles in immune response,we do not know the immune imbalance of thrombocytes is the reason or the result of other blood cells immune imbalance.And the relationship among blood cells in the immunological network is still unknown.To explore the immunity relationships of RBC and platelets,we activated haematogenic immunity used inactivated tumor cell S₁₈₀(5×10⁹/L) However,when we carried out the experiment,we observed the activation of thrombocytes,but no aggregation.Studies have showed that thrombocytes could easily be activated to adhere,aggregate and release immunological active materials,and played important roles in inflammatory disease,tumor and so on,Furthermore,as a complicated process, immune response can be induced by different activated thrombocytes.Therefore,all the activaties of thrombocytes and the immune response which is induced by thrombocytes can not be showed in the blood immunological system only activated by S₁₈₀(5×10⁹/L).In order to reveal the roles that thrombocytes and erythrocytes play in blood immunological system,we build a new blood immunological system,which can impersonate all the activaties of thrombocytes and can be used as a way to study how thrombocytes and erythrocytes regulate T lymphocytes in immune response.In this new system we proved that platelet is the first reason to cause the imbalance of T lymphocyte subset,and RBC can maintain the balance of T lymphocyte subset;the imbalance of platelet is an important turning point of imbalance of immune system,and the imbalance of RBC is the result of the imbalance of immune system.Part 1.Empirical study on the effects of platelets and red blood cells in modulating T lymphocytesObjectiveInvestigate the roles of thrombocytes and erythrocytes in the process of regulating T lymphocytes and their relationship in this process.MethodsHematological immune systems was divided into four groups according to different conditions,including whole blood cell group,non-PL group,non-RBC and the group without RBC or PL.Each group was stimulated by the antigen which is inactivated S180(5×10⁹/L).And set control group which was stimulated by normal sodium(NS).FCM was utilized to determine the expressions of immune moleculars on some cells. ResultsComparison of the lvel of CD₃⁺CD₄⁺,CD3⁺CD4⁺/CD3⁺ and CD3⁺CD8⁺/CD3⁺ in whole blood cell group and which in the non-PL group:experimental group and control group had no significant differences(p＞0.05).Comparison of which in non-RBC group,the level of CD₃⁺CD₄⁺ and CD3⁺CD4⁺/CD3⁺ +in whole blood cell group increased,but CD3⁺CD8⁺/ CD3⁺ decresed:experimental group and control group had significant differences(p＜0.01).Comparison of which in the group without RBC or PL,the level of CD₃⁺CD₄⁺ and CD3⁺CD4⁺/ CD3⁺ in non- RBC group increased,but CD3⁺CD8⁺/ CD3⁺ decresed:experimental group(p＜0.05),control group had no significant differences(p＞0.05).Comparison of which in the group without RBC or PL,the level of CD₃⁺CD₄⁺ and CD3⁺CD4⁺/CD3^{+ +}in non- PL group increased,but CD3⁺CD8⁺/CD3⁺ decresed:experimental group and control group had significant differences(p＜0.001).CD₃⁺CD₄⁺ had a negative correlation wi(?)h CD₃⁺CD₈⁺ in whole blood cell group,the group non-PL and the group without RBC or PL: experimental group and control group had significant differences(p＜0.05),but they had no significant correlation in non-RBC group:experimental group and control group had no significant differences(p＞0.05).The level of CD₅₉ on platelets in whole blood cell group was less than that in non-RBC group:experimental group(p＞0.05), control group(p＜0.05);The level of CD_62P on platelets in whole blood cell group was higher than that in non-RBC group:experimental group and control group had significant differences(p＜0.05).The level of CD35 on RBC in whole blood cell group was higher than that in non-PL group:experimental group and control group had significant differences(p＜0.05).The level of the expressions of CD_62P on platelets was higher in whole blood cell experimental group than that in whole blood cell control group(p＜0.05);but the level of the expressions of CD35 on RBC was lower (p＜0.05).ConclusionPlatelet is the first reason to cause the imbalance of T lymphocyte subset,and RBC can maintain the balance of T lymphocyte subset.Part 2.Construction of a new blood immunological system which can be induced the imbalance Of platelets and study on the roles of thrombocytes and erythroeytes in regulatting the expressions of CD4+CD25+ on T lymphocytes Objective1.To select a proper antigen and build a new haematogenic immunological system which can be induced the imbalance Of platelets.2.To explore the roles of thrombocytes and erythrocytes in affecting the expressions of CD4+CD25+ on T lymphocytes with the new haematogenic immunological system.Methods1.To select a proper antigen:to selet a antigen which can induce all acticities of thrombocytes.(1) To compare the changes of platelet aggregations induced by yeast fungus and S180 of different doses by means of Wright's staining,obsering with light microscope..(2) To observe the changes of platelet aggregations induced by yeast fungus and S180 by means of Wright's staining when different complement activation pathways were blocked.,observing with light microscope.(3)To measure the curve of platelet aggregations induced by yeast fungus,S180,yeast fungus coated with activated complements and S180 coated with activated complements using platelet aggregometer2.Construction of a new haematogenic immunological system which can induce all acticities of thrombocytes:haematogenic immunological system was activated with the selected proper antigen.3.FCM was used to detecet the expressions of CD4⁺CD25⁺ on lymphercytes.Results1.Yeast fungus and S180 both can cause all activities of pletlet through the way of complement activation correlating with the changes of their doses:S₁₈₀(5×10⁹/L,1×10¹⁰/L)could not cause platelet aggregation,S₁₈₀(2×10¹⁰/L,4×10¹⁰/L) could;and yeast fungus(2×10⁹L,4×10⁹/L) couldnot,yeast fungus(8×10⁹/L,16×10⁹/L) could.2.In five minutes platelet aggregometer showed:yeast fungus(16×10⁹/L) and S180(4×10¹⁰/L) could not cause the curve of platelet aggregations,but yeast fungus coated with actived complements(16×10⁹/L) and S180 coated with actived complements(4×10¹⁰/L) could.3.Construction of a new haematogenic immunological system and which can be induced the imbalace of thrombocytes:S₁₈₀ of hypo-dose(5×10⁹/L) and S₁₈₀ of hypso-dose(3×10¹⁰/L) were both acted as the selected proper antigens,to activate haematogenic immunological system;as well as setting up a control haematogenic immunological system stimulated by NS;which means a control haematogenic immunological system stimulated by NS,a experimental haematogenic immunological system activated by L_S180(5×10⁹/L S₁₈₀) and a experimental haematogenic immunological system activated by H_S180(3×10¹⁰/L S₁₈₀) were set up at the same time.4.Comparison of which in whole blood cell group and which in non- PL group: control group and L_S180 experimental group had no significant differences(p＞0.05), H_S180experimental group increased(p＜0.05);Comparison of which in non-RBC group, The level of the expressions of CD4⁺CD25⁺ on lymphercytes in whole blood cell group:control group and L_S180 experimental group both decresed(p＞0.05),but H_S180 experimental group incresed(p＜0.05);Comparison of which in non- RBC group and which in the group without RBC or PL:control group and L_S180 expernnental group had no significant differences(p＞0.05),,but H_S180 experimental group decreased (p＜0.05);Comparison of which in non- PL group and which in the group without RBC or PL:control group and L_S180 experimental group and H_S180experimental group all decreased(p＜0.05).Conclusion1.A new haematogenic immunological system was build,and which can be induced the imbalace of thrombocytes.2.Erythrocytes can dual-regulate the level of the expressins of CD4⁺CD25⁺ on lymphercytes,and thrombocyte aggregation is the key reason of the up-regulation roles of erythrocytesPart 3.Study on the roles of thrombocytes and erythrocytes in causing the direaction that Th differentiate to Th1 or Th2 misregistratedObjectiveStudy on the roles of thrombocytes and erythrocytes in causing the direaction that Th differentiate to Th1 or Th2 misregistrated.MethodsELASE was used to determine the levelof IFN-rand IL-10Results 1.The level of IFN-r/1L-10 in whole blood cell group was less than that in non-PL group:control group and L_S180 experimental group had no significant differences(p＞0.05),H_S180experimental group(p＜0.05);Comparison of which in whole blood cell group and which in non- RBC group:all of control group,L_S180 experimental group and H_S180experimental group had no significant differences(p＞0.05);Comparison of which in non- RBC group and which in the group without RBC or PL:control group and L_S180 experimental group increased (p＜0.05),but H_S180 experimental group decreased(p＜0.05);Comparison of which in non- PL group and which in the group without RBC or PL:control group and L_S180 experimental group increased(p＜0.05),H_S180experimental group had no significant differences(p＞0.05).2.The level of IFN-r in whole blood cell group was higher than that in non- PL group:control group and L_S180 experimental group(p＜0.05),H_S180experimental group had no significant differences(p＞0.05);Comparison of which in whole blood cell group and which in non- RBC group:control group and H_S180experimental group increased(p＜0.05),L_S180experimental group had no significant differences(p＞0.05).; Comparison of which in non- RBC group and which in the group without RBC or PL:Control group increased(p＜0.05),L_S180experimental group had no significant differences(p＞0.05),H_S180experimental group decresed(p＜0.05);Comparison of which in non- PL group and which in the group without RBC or PL:control group increased(p＜0.05),L_S180 experimental group and H_S180experimental group had no significant differences(p＞0.05).3.The level of IL-10in whole blood cell group was higher than that in non- PL group:control group had no significant differences(p＞0.05),L_S180 experimental group and H_S180experimental group incresed(p＜0.05);Comparison of which in whole blood cell group and which in non- RBC group:control group and L_S180experimental group had no significant differences(p＞0.05),H_S180 experimental group increased(P＜0.05).; Comparison of which in non- RBC group and which in the group without RBC or PL: control group and H_S180experimental group had no significant differences(p＞0.05), L_S180experimental group decresed(P＜0.05);Comparison of which in non- PL group and which in the group without RBC or PL:control group had no significant differences(p＞0.05).,L_S180 experimental group decreased(p＜0.05),H_S180experimental group increased(p＜0.05). ConclusionPlatelet is a turing point to cause the direaction that Th differentiate to Th1 or Th2 changed;and erythrocyte is the determinative factor to make immune response choose cell mediated immunity or humoral-mediated immunity.