| BackgroundCell death mainly features in two forms:apoptosis or necrosis. Apoptosis is an active programmatic and energy conserving cell death, the characteristic of which is formation of apoptotic bodies. Apoptotic cells are cleared very rapidly in a strict package condition, which will not lead to immune response. While necrosis is a passive, non-programmatic and destructive cell death process that is characterized by abrupt stop of energy metabolism and the release of cellular ingredients into adjacent environment, which will cause immune response. Compared with apoptotic cells, the most important difference is necrotic cells can release cellular ingredients into adjacent environment. Many studies demonstrate that necrosis is not totally non-programmatic. Different signal transduction pathways can be activated during the process and many inflammatory molecules were released, which play an important role in the inflammation through a series of signal cascading response. The main roles of necrotic cells in the immune system are:(1) to promote the release of inflammatory factors; (2) to activate the innate immune system to release the innate immune molecules; (3) to resist infection; (4) to induce autoimmune diseases.The research about immune function of necrosis are as follows:(1) cell death are related to the combination of cytokines and death receptors, damaged ion channels, the release of reactive oxygen species and mitochondrial damage, as well as enhanced expression of protein kinase; (2) cell necrosis can release a variety of inflammatory factors to promote inflammation; (3) necrotic cells can release chemokine that will attract a variety of phagocytic cells to reach local sites to play different roles; (4) phagocytic cells can release multiple cytokines to modulate the immune system after phagocytosis of necrotic cells; (5) a variety of inflammatory molecules can activate the death receptor induced programmed cell necrosis. However, the regulation of necrotic cells on different CD4+ T cell subset balance remains unclear.CD4+ T cells are known to have several subsets, such as Thl, Th2, Treg, Th17, Tfh, Th9 and Th22. Regulatory T cells (Treg) are a group of cells which can inhibit the function of other immune cells and play an important role in autoimmune tolerance. Th17 is a new type of T cell subset characterized by the production of IL-17, which plays an important role in immune regulation, anti-infection and autoimmune diseases. The change of CD4+ T cell subsets means the immune abnormalities and may be related to the development of some diseases.It is generally believed that there exists a balance between Thl and Th2 cells. Th cell precursors differentiate into ThO cells in the case of antigen stimulation and ThO cells differentiate into Thl or Th2 cells under the effect of different cytokines indifferent microenvironments.Thl/Th2 cell imbalance involves in the development of many diseases.Some reports show that there also exits a Treg/Th17 immune balance. Because the only existence of TGF-β in the microenvironment can induceFoxp3 and RORyt expression, but Foxp3 inhibits the RORyt activity, which make the naive T cells fail to differentiate into Th17 cells. If TGF-β in the microenvironment persists, and IL-2 and RA exist, naive T cells will differentiate to Treg. On the contrary, if the microenvironment of TGF-P persistence, and IL-6, IL-21 and IL-23 exist, Foxp3 activity will be inhibited and naive T cells will differentiate to Th17 cells.It is known that apoptotic and necrotic cells are both able to release different signals to promote phagocytosis. Either apoptotic cells or necrotic cells are able to promote the phagocytic cells to secrete large amounts of TGF-β. Necrotic cells can also promote the release of IL-6 and other inflammatory cytokines. According to the balance between different CD4+T cell subsets, we guess that different ways of cell death may have different role in regulating the balance between CD4+T cell subsets. Apoptotic cells can stimulate phagocyte to secret TGF-P and simultaneously inhibit the secretion of IL-6 to promote Treg induction. While necrotic cells can stimulate the phagocytic cells to secrete TGF-P and the secretion of IL-6 to promote the induction of Th17. Apoptotic cells/necrotic cells regulate the Thl/Th2 differentiation through their own release or stimulating antigen-presenting cells to secrete inflammatory factors (such as IDO, HMGB1, etc.).It has been confirmed that apoptotic cells can regulate the ratio of different T cell subsets. The infusion of apoptotic cells can induce the generation of Treg cells and inhibit the generation of p-STAT3 which is necessary for Thl7 cell differentiation and development. Necrotic cells stimulate phagocytic cells to secrete IL-6 and greatly enhanced IL-6 secretion after LPS stimulation, while IL-6 can inhibit the generation of Treg. HMGB1, which is inflammatory protein released by necrotic cells can stimulate lymphocyte to differentiate to Thlat high concentrations. The release of IDO after the phagocytosis of apoptotic cells can also affectThl/Th2 balance. However, the regulation of necrotic cells on T cell subset balance remains unclear in vivo.According to the cell death recognition model for the immune system proposed by Professor Sun Er-wei, necrotic cells are critical for the induction of immune responses. Further study about the role of the necrotic cells on different T cell subsets is of great significance to the proof of the cellular and molecular mechanisms of cell death in immune recognition model, the finding of important immune cells and molecules regulating the immune system and explore the mechanisms and prevention of immune-related diseases from the perspective of the immune balance.PurposeThis study focuses on the following contents:1. The effect of necrotic cells on CD4+T cell subsets in normal mice;2. The effect of necrotic cells on CD4+T cell subsets in mice after the pre-infusion of GdCl3;3. The effect of pre-infusion of necrotic cells on CD4+T cell subsets in mice sepsis model;4. The effect of aseptic inflammation in the body (ischemic reperfusion) on CD4+ T cell subsets.5. The effect of necrotic cells on CD4+T cell subsets in EAE mice;Method1.26 SPF Balb/C mice (6-8week old) were randomly divided into three groups. The first group is PBS control group (n=6), with intraperitoneal injection of 200ul of PBS; the second is live cell group (n=10), with intraperitoneal injection of 2×107 live thymocytes; the third group is necrotic cells group (n=10), with intraperitoneal injection of 2×107necrotic cells without any treatment. All experimental mice were sacrificed 2 weeks later and CD4+Foxp3+Treg cells, CD4+IL-17A+Th17 cells, CD4+IFN-y+Th1 cells and CD4+IL-4+Th2 cell ratio were detected by flow cytometry in peripheral blood, spleen, and thymus.(1) Preparation of necrotic cellsThree 6 weeks old SPF Balb/C mice were killed and thymus were quickly removed for grinding. Then the cell suspension was washed with PBS and counted. The cell was induced to necrosis by 1 hour bath in 56℃ water. Necrosis rate was detected with Annexin V FITC and PI staining by flow cytometry.(2) The detection of CD4+T cell subsetsAnti-coagulated by heparin, blood was collected. Spleen and thymus was separated and grinded with the flat side of 5ml syringe. The cells were washed with PBS, filtered with 200 mesh metal meshes and resuspended in complete medium to prepare single with a concentration of 1×107/ml. The cell suspension were added with PMA (final concentration 100ng/ml), Ionomycin (final concentration 1μg/ml) and BFA (10μg/ml) and then placed in the incubator (37℃,5%CO2) for 6 hours. After incubation, cells were washed with PBS and added with 1μl of FITC CD4 antibody. After 60min incubation at 4℃ protected from light, peripheral blood were added with RBC permeabilization buffer and washed 2 times. All samples were washed with PBS and the supernatant was discarded, lml of Foxp3 Permeabilization/Fixation buffer were added and mixed well. The cells were placed at 4℃ and incubated for 60 minutes. Then add 2ml permeabilization solution (diluted to 1 X) to the wash cells and resuspended them. Then each sample was divided into two tubes. One was added with0.5μl anti-Foxp3 antibody and 1μlanti-IL-17A antibody, the other was added with 1μl anti-IL-4 antibody and 1μl anti-IFN-y antibody. An isotype control tube was established with addition of 0.5 u 1 anti-Foxp3 antibody,1 u 1PE IgGl k and PE-Cy7 IgGl k l.The cell suspension were incubated at 4℃for 60 minutes, washed 2 times with 2ml permeabilization working solution, resuspended with cold PBS and then detected on flow cytometry (the same way as described).2.23 SPF Balb/C mice(6-8weekold) were randomly divided into four groups. The first is PBS control group (n=5) with 9 consecutive days of intraperitoneal injection of 200μl PBS; the second group is GdCl3 control group (n=6), with tail vein injection of 200μl of 1mg/ml GdCl3 at day 1, day 3and intraperitoneal injection of 200μl of PBS in the following 9 days; the third group was necrotic cell control group (n= 6), with intraperitoneal injection of 2 × 107necrotic cells (200μl)for 9 consecutive days; the fourth group was experimental group (n=6), with tail vein injection of lmg/ml GdC13 (200μl)at day 1, day 3 and intraperitoneal injection of 2×107 necrotic cells (200μl)for nine consecutive days. All animals were sacrificed at day23. CD4+Foxp3+Treg cells, CD4+IL-17A+Th17 cells, CD4+IFN-γ+Thl cells and CD4+IL-4+Th2 cell ratio were detected by flow cytometry in peripheral blood, spleen, and thymus (the same way as described).3.19 SPF C57BL/6 mice (10 to 12 weeks old) were randomly divided into three groups:renal ischemia-reperfusion group (RIR group, n=7), sham-operated group (Sham group, n=7), normal control group (n=5). RIR mouse abdomen was incised after anesthesia and the left renal pedicle was clamped to block blood flow for 1 hour. Sham group mouse only had abdomen incised after anesthesia and the left renal pedicle separated without other special treatment. The normal control group was the same batch of mice without special treatment. All mice were sacrificed 14 days after surgery and the kidneys were sent for pathology testing. CD4+Foxp3+Treg cells, CD4+IL-17A+Th17 cells, CD4+IFN-γ+Thl cells and CD4+IL-4+Th2 cell ratio were detected by flow cytometry in peripheral blood, spleen, and thymus (the same way as described).Preparation of renal ischemia-reperfusion mouse modelMice were anaesthetized with 200μL of 1% sodium pentobarbital by intraperitoneal injection. Disinfect conventionally with iodine alcohol, incise along ventral midline, retract the gastrointestinal with a sterile cotton swab, and wrap it with preheating (37℃) saline moist gauze. Expose the left renal pedicle and clamp the renal artery and vein with a micro vascular clip. One hour later, remove the clip and observe if the renal blood flow was restored. After successful reperfusion, close the abdominal cavity layer by layer, disinfect with 75%alcohol and inject 500μl of saline subcutaneously.4.34 SPF C57BL/6 mice (10-12 weeks old) were randomly divided into four groups:normal control group (PBS group, n=5), sham-operated group (sham group, n=5), sepsis group (CLP group, n=12) and necrotic cells treated sepsis group (CLP+Nec group, n=12). A normal control was injected intraperitoneal 200μL of PBS while sham group only underwent the separation of the end of cecum without ligation and puncture after anesthesia. Sepsis model was established by cecal ligation and puncture method (CLP). Necrotic cells treated sepsis group was first injected intraperitoneal 2×107 necrotic cells and underwent cecal ligation and puncture at day5. All mice were sacrificed at day 14 and the liver, lung and kidney were sent for pathology testing. CD4+Foxp3+Treg cells, CD4+IL-17A+Th17 cells, CD4+IFN-y+ Thl cells and CD4+IL-4+Th2 cell ratio were detected by flow cytometry in peripheral blood, spleen, and thymus (the same way as described).Preparation of septic miceThe mouse was anesthetized with 200μL of 1% sodium pentobarbital. After conventional disinfection, abdomen was incised along the ventral midline. Separate the distal end of cecum and large intestine mesentery carefully, and then ligature at the center of distal cecum with sterile 4#silk thread. Puncture the cecum thoroughly at the center of ligatured cecum with sterile 7# needle, squeeze out a little contents of the cecum, push back the cecum to abdominal cavity and finally close the abdominal cavity with layer by layer suture. Sham group underwent the separation of the end of cecum without ligation and puncture. After 30 second exposure, close the abdominal cavity with layer by layer suture. Disinfect the incision with 75% alcohol and inject normal saline subcutaneously immediately after operation (50ml/kg).5.26 C57BL/6 mice (8-10 weeks old, female) were randomly divided into three groups:normal control group (PBS group, n=6), EAE model group (n=10), necrotic cells treated EAE group (EAE+Nec group, n=10). The EAE group was treated by intraperitoneal injection of 200μl sterile PBS for 9 days after preparation of the EAE disease model. The EAE+Nec group was treated by intraperitoneal injection of 2×107 necrotic cells (200(0.1) for 9 days after preparation of the EAE disease model. PBS group was treated by intraperitoneal injection of 200μl sterile PBS for normal control. All mice were sacrificed at day 14. CD4+Foxp3+Treg cells, CD4+IL-17A+ Thl7 cells, CD4+IFN-y+Thl cells and CD4+IL-4+Th2 cell ratio were detected by flow cytometry in peripheral blood, spleen, and thymus (the same way as described). Preparation of the EAE mouse model200μg/ml peptide MOG35-55 was mixed well emulsified with same volume of complete Freund’s adjuvant containing the 5mg/ml Mycobacterium tuberculosis H-37 RA. To immune mice,200μl intermixture was multi-point injected subcutaneous. Then mice were treated with 500ng pertussis toxin (200μl) by intraperitoneal injection on dayO and day 2. The status and clinical score of each mouse was continuously observed.6. SPSS 16.0 was used for one-way analysis of variance. The proportion of T cell subsets in CD4+T cells in the peripheral blood, spleen and thymus was compared between the groups. All data were presented as mean ± standard difference (X± SD).Result1. The mouse thymus cells were induced necrosis by 56℃ water bath for 1 hour and detected with flow cytometry. Cell necrosis rate was up to 99% and can be used for further experiments.2. The effect of necrotic cells on normal mice CD4+T cell subsets(1) Compared with the infusion of live cells group, a single infusion of necrotic cells can significantly increase the proportion of Thl and Th2 cells in CD4+T lymphocytes in the peripheral blood. (FTh1=4.671, PTh1 vstheLiving=0.024; FTh2= 5.576, PThavsLiving=0.014). Compared with the infusion of live cells or infusion of PBS, infusion of necrotic cells can also significantly reduce the proportion of Thl cells inCD4+T cells in mouse thymus (FTh1=6.827, Pvs living cells=0.004, As the pbs=0.005). Simple infusion of necrotic cells had no effect on Th1/Th2 and Treg/Th17 in the body of mice.(2) After GdCl3 inhibition of macrophages and 9 consecutive day of infusion of necrotic cells, the infusion of necrotic cells significantly increased Thl and Th2 in CD4+T lymphocytes in peripheral blood(FTh1= 3.883, PTh1= 0.025; FTh2= 3.468, Ptii2= 0.037)compared with other three groups. In addition, compared with the PBS control group, infusion of necrotic cells can significantly inhibit Treg cells in CD4+T cells in peripheral blood (FTreg=4.251,-PTregvsPBS=0.039). Treg cells in CD4+T cells in peripheral blood of GdCB+Necrotic cell group were also significantly lower than PBS group and GdC13 group (FTreg=4.251, PTreg vS pbs=0.004, PTreg vs Gdcn=0.021). Thl and Th2 cells in CD4+T lymphocytes in the spleen of necrotic cell infusion group were significantly increased compared with the other three groups (FTh1=12.112, PTh1=0.000; FTh2=7.169, PTh2=0.002). Compared with the PBS group, the infusion of necrotic cells or GdCb were both able to reduce the proportion of Th1 and Th2 cells inCD4+T cells of mouse thymus significantly (FTh1=30.385, PTh1=0.000; FTh2=63.084, PTh2=0.000). Compared to the simple infusion GdCl3 group,Thl and Th2 in CD4+T cell in mouse thymus of the GdCB+Necrotic cell group were similarly reduced significantly (PTh1 vsGdCl3=0.016,PTh2 vs GdCl3=0.000). Compared with the PBS group, the proportion of Th17 cells in CD4+T cells in mouse thymus of GdCl3 or GdCl3+Necrotic cell groups was significantly decreased (FTh17=18.211, PTh17=0.000). Compared with the PBS group, continuous infusion of necrotic cells can significantly increaseThl/Th2 ratio in mice peripheral blood (F=2.372, PNec vs pbs=0.020). Treg/Thl7 proportion in peripheral blood was significantly reduced after infusion of necrotic cells or GdCl3+Necrotic cells compared with PBS group (F=41.354, PNecVs PBS=0.000, iWodCB vspbs=0.000).3 The CD4+T cell subsets in renal ischemia-reperfusion miceThe proportion of Thl cells in CD4+T cells in peripheral blood was significantly increased in RIR mice compared with the sham group in 14 days (FTh1=3.498, JW vs sham= 0.026). Compared with the sham group, the ratio of Thl7/CD4+T in peripheral blood also significantly increased in sham group (FTh17= 4.942, Prnvsof sham= 0.006). Compared with the PBS group and RIR group, Thl cells of CD4+T lymphocyte ratio in spleen was significantly reduced in sham group (FTh1= 6.236 PTh1= 0.010). The ratio of Th17 in CD4+T in the RIR mice spleen was higher than it in sham mice, showing significant difference (FTh17-6.236-PwRvsSham= 0.024). The ratio of Thl cells in CD4+T cells within the thymus of the RIR mice is higher than the normal control group or the Sham group, there is a statistically significant difference (FTh1= 8.949 PTh1= 0.017). The proportion of Thl 7 in spleen in sham group is lower than these in the normal control group and RIR group, there is a statistically significant difference (Fjhi7= 8.139, Pihn= 0.009). Compared with normal control group, the proportion of Treg subsets in thymus in Sham group and RIR mice was significantly reduced (Fireg=4.566; Prreg=0.027). Calculated the proportion of Thl/Th2 and Treg/Thl7, we found that Thl/Th2 ratio of RIRs group was increased compared with Sham group, the difference was statistically significant (FBlood,Th1/Th2= 5.472, P BloodTh1/Th2= 0.015; Fspleen Th1/Th2= 18.247, P Spleen Th1/Th2= 0.000; F Thymus Th1/Th2= 9.771, P Thymus Th1/Th2= 0.008). In the peripheral blood, the ratio of Treg/Th17 in RIRs group was significantly lower than the Sham group and the normal control group the mice (F Biood,Treg/rh17= 9.993, PBlood,Treg/Th17= 0.002). The ratio of Treg/Thl7 in RIR mice thymus was significantly lower than these in normal control group and Sham group (FThymus Treg/Th17=56.122, PThymus Treg/Th17= 0.000).4. The effect of necrotic cells on CD4+T cell subsets in sepsis mouse model(1) CLP method can prepare septic mice successfully and the total mortality of mice was 54.2%.Pathology can be used to identify if the model was successful.14 days after CLP surgery, the proportion of Thl cells in CD4+T lymphocytes in peripheral blood of sham group, CLP group and CLP+Necrotic cell group were all significantly reduced compared with normal control group(FTh1=14.321, PTh1=0.000). And the Th1/CD4+ratio in CLP+Necrotic cell pre-infusion group was also significantly decreased compared with CLP group (FTh1=14.321,PCLP+Nec VS CLP =0.033). At the same time, Th2/CD4+ratio in CLP+Ne group was also significantly reduced compared to PBS group (FTh2-2.314, PCLP+Nec=0.024). The proportion of Treg cells in CD4+T lymphocytes in the peripheral blood of Sham group, CLP group and CLP plus necrotic cell group were significantly lower compared with PBS group (FTreg=3.201, PTreg=0.050). After the sepsis model surgery, the proportion of CD4+T cells in peripheral blood of CLP group and CLP+ Nee group was lower than PBS control group and Sham control group significantly (FCD4=10.261,PCD4=0.000). The proportion of Thl cells in CD4+T lymphocytes in the spleen of CLP group was also lower than normal control group (FTh1=9.701, PCLP VS PBS=0.007). Compared with the PBS control group, Th2/CD4+ratio of Sham group, CLP group and CLP+Necrosis group in the spleen were all significantly reduced (FTh2=12.137, PTh2= 0.002). Detected the ratio of Th17/CD4+, we found that the ratio is higher in CLP group than it in the other three groups in the spleen, the difference had significance of statistics (FTh17=5.513, PTh17= 0.008). The percentage of Treg in CD4+T in Sham group, CLP group and the CLP+Necrosis group were significantly lower than PBS control group in spleen(FTreg= 12.155,-PTh17= 0.000), but in these three groups, there was no significant difference (P> 0.05). The Th2/CD4+T ratio in thymus in CLP+Necrosis group was significantly less than Sham group mice (FTh2= 2.368,PCLP+Nec vs Sham= 0.039). The percentage of Treg in CD4+T in the thymus of CLP mice was significantly lower than the Sham group mice and CLP+Necrosis mice (FTreg= 3.683, the PCLP vs.of Sham=0.007, PCLP vs CLP+Nec= 0.021). Further calculation in each experimental group and control group mice, peripheral blood and spleen Th1/Th2 and Treg/Th17 ratio, we found that CLP mice peripheral Th1/Th2 ratio is not only higher than the PBS group and Sham group mice also higher than the CLP+Necrosis group, the difference had significance of statistics (FBLoodTh1/Th2= 10.482, PBlood Th1/Th2=0.000). In the spleen, the Th1/Th2 ratio of Sham group and the CLP+Necrosis group was significantly higher than the CLP group and PBS group (FSpleen Th1/Th2=12.273, PSpleen Th1/Th2=0.000). Compared with the PBS group of Sham group, the Treg/Thl7 ratio in spleens of mice in CLP and CLP+necrosis group was significantly lower. The Treg/Th17 ratio in spleens of the CLP group was significantly lower than them in the Sham group and the CLP+necrosis group (FSpleenTreg/Th17=29.641, Pspleen Treg/Th17= 0.000).5. The effect of necrotic cells on CD4+ T cell subsets in EAE mouse model(1) Subcutaneous injection of MOG35-55 antigen peptide can effectively induce EAE mice model. In 14 days of experiments, the incidence of overall was 90% (18/20) in EAE group and EAE+Nec group.(2) In EAE model, the Th17 proportion in mouse peripheral blood was significantly higher than the PBS group (P= 0.005), while the proportion of Th1 and Treg cells in the spleen was significantly decreased (PTh1= 0.026, PTreg= 0.026). The ratio of Th1/Th2 in EAE group spleens was significantly lower than the PBS group (P=0.001). Infusion of necrotic cells in EAE mice, which can be significantly increased in peripheral blood Th17 cell ratio (PNec vs. PBS=0.000, PNec vs EAE= 0.005), it also increased the proportion of Thl7 cells in the spleen (PNec vs EAE= 0.004). Necrotic cells can also be collaborative EAE model to reduce the proportion of Treg cells in the spleen (PNec vs. PBS=0.000). There was no significant difference in the proportion of Treg between the EAE group of EAE+Nec group (PEAE vs Nec=0.070). In the mouse thymuses, the percentage of Thl, Thl7 and Treg cells in CD4+T in EAE+Nec group was significantly lower than it in the EAE group (PTh1=0.034, PTh17=0.002, PTreg= 0.001). Compared with PBS group, the Treg/Thl7 ratio in the spleen of EAE mice was significantly decreased (PNec vs EAE=0.006), and the Thl/Th2 proportion of the thymus were significantly increased (PNec vs EAE=0.003) after infusion of the necrotic cells.Conclusion1. A simple infusion of autologous necrotic cells can increase Thl and Th2 proportion in peripheral blood and shift Thl/Th2 balance to Thl direction. It was more obvious under continued stimulation of necrotic cells, which may be related to macrophage. Because the effect of necrotic cells on Thl and Th2disappeared after the inhibition of macrophage by GdCl3. Necrotic cells and GdCl3can both inhibit Th1 and Th2 in the thymus and they had a synergistic effect. GdCl3 can also inhibit the proportion of Th17 cells in the mouse thymus. Infusion of necrotic cells can also inhibit peripheral blood Treg cells, resulting in Treg/Th17 balance to shift to Th17 direction. The inhibition of GdCl3 on macrophages didn’t affect the inhibition of necrotic cells on Treg. A simple infusion of autologous necrotic cells had no effect on the proportion of Thl 7 cells.2. The surgery could reduce the proportion of Th1 in the spleen and Treg in the thymus in unilateral renal ischemia reperfusion (RIR) model. Compared with sham group, RIR significantly increased Th1 and Th17 ratio in mouse peripheral blood, spleen and thymus, which make Thl/Th2 balance shift to Th1 direction and Treg/Th17 balance shift to Th17 direction.3. Surgery can reduce Th1, Th2 and Treg cell ratio in peripheral blood in sepsis model. Sepsis can shift Th1/Th2 balance to Thl direction in mouse peripheral blood and to Th2 direction in the spleen compared to sham group. At the meantime, Sepsis can increase Th17 proportion in mouse spleen, resulting Treg/Th17 balance to shift to Th17 direction. In addition, the proportion of Treg in thymus of septic mice was significantly decreased compared with sham group. Pre-infusion of necrotic cells could reverse the effect of sepsis on Thl, Th2, Th17 and Treg.4. For preparation of EAE model mice, we used the antigen MOG35-55 with CFA to stimulate mice, and then gave them a continuous stimulation of necrotic cells. The results showed that the proportion of Th17 cells in peripheral blood increased in EAE mice while the proportion of Treg cells reduced in spleen. EAE can reduce the Th1/Th2 ratio in the spleen and shift Th1/Th2 balance to Th2 direction. After infusion of necrotic cells to EAE mice, we found that necrotic cells collaborating with the EAE model can significantly raise the mouse Th17 ratio and reduce the percentage of Treg cells in the thymus, prompting that Treg/Th17 balance to Th17 direction in peripheral blood and spleen. The effect of necrotic cells on Thl, Th2 and Treg cell ratio in mouse peripheral blood and spleen is similar with it in normal mice model experiment.In summary, the continued presence of necrotic cells will increase the proportion of Th1, Th17 through macrophages. Necrotic cells can play a proinflammatory role by down regulating the proportion of Treg cells. Necrotic cells can also play a synergistic role with other mechanisms in inflammation. |
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