本文選題：輸血相關(guān)免疫調(diào)節(jié) + 異體紅細胞��； 參考：《第二軍醫(yī)大學》2012年碩士論文

【摘要】：（一）目的 運用郭氏“系統(tǒng)血液快速固有免疫反應(yīng)分離實驗體系”體外觀察異體紅細胞對IL-8、IL-10、IL-12分泌的影響。 （二）方法 健康成人同型新鮮抗凝全血，常規(guī)分離制備血漿、紅細胞和白細胞，以自體血漿作為反應(yīng)基質(zhì)建立系統(tǒng)血液分離實驗體系，考察異體紅細胞對IL-8、IL-10、IL-12分泌的影響，設(shè)立自體紅細胞作為對照。ELISA法測定上清液中的細胞因子IL-8、IL-10、IL-12，并探討其改變可能存在的意義。 （三）結(jié)果 1、血漿IL-8濃度比較 ⑴異體紅細胞生理鹽水體系組[28.89179±2.674565pg/ml]與異體紅細胞人大腸埃希桿菌體系組[119.05455±31.056229pg/ml）比較，其均值差值[-90.1628±59.37619pg/ml],（p=0.0050.05），IL-8分泌量兩組之間有著顯著的差別。 ⑵自體紅細胞生理鹽水體系組[29.05148±2.972132pg/ml]與自體紅細胞大腸埃希桿菌體系組[132.63303±35.072837pg/ml]比較，其均值差值[-103.5816±59.37619pg/ml],（p=0.0000.05），IL-8分泌量兩組之間有著顯著的差別。 ⑶異體紅細胞生理鹽水體系組與自體紅細胞生理鹽水體系組相比較，其均值差值[-0.1599±59.37619pg/ml]，IL-8分泌量兩組之間沒有明顯的差別（p=0.8190.05）。 ⑷異體紅細胞人大腸埃希桿菌體系組與自體紅細胞大腸埃希桿菌體系組相比較，其均值差值[-13.5785±59.37619]，IL-8分泌量兩組之間沒有明顯的差別（p=0.9970.05）。 2、血漿IL-10濃度比較 ⑴異體紅細胞生理鹽水體系組[27.90894±5.445463pg/ml]與異體紅細胞人大腸埃希桿菌體系組[22.28497±4.010929pg/ml]比較，其均值差值[5.6240±5.37453pg/ml]，（p=0.2960.05） IL-10分泌量兩組之間沒有顯著的差別。 ⑵自體紅細胞生理鹽水體系組[24.15185±3.748592pg/ml]與自體紅細胞大腸埃希桿菌體系組[27.58494±4.740832pg/ml]比較，其均值差值[-3.4331±5.37453pg/ml（p=0.5240.05），IL-10分泌量兩組之間沒有顯著的差別。 ⑶異體紅細胞生理鹽水體系組與自體紅細胞生理鹽水體系組相比較，其均值差值[3.7571±5.37453pg/ml] IL-10分泌量兩組之間沒有顯著的差別（p=0.3250.05）。 ⑷異體紅細胞人大腸埃希桿菌體系組與自體紅細胞大腸埃希桿菌體系組相比較，其均值差值[-5.2999±5.37453pg/ml]，IL-10分泌量兩組之間沒有明顯的差別（p=0.4850.05）。 3、IL-12分泌量的測定值 ⑴異體紅細胞生理鹽水體系組[25.80663±3.872463pg/ml]與異體紅細胞人大腸埃希桿菌體系組[22.97730±2.298994pg/ml]比較，其均值差值[2.8289±4.48095pg/ml]，（p=0.5280.05），IL-12分泌量兩組之間沒有顯著的差別。 ⑵自體紅細胞生理鹽水體系組[37.74982±5.783372pg/ml]與自體紅細胞大腸埃希桿菌體系組[33.32391±2.982255pg/ml]比較，其均值差值[4.4259±4.48095pg/ml]，（p=0.3240.05），IL-12分泌量兩組之間沒有顯著的差別。 ⑶異體紅細胞生理鹽水體系組與自體紅細胞生理鹽水體系組相比較，其均值差值[-11.9436±4.48095pg/ml]，IL-12分泌量兩組之間有著顯著的差別（p=0.0080.05）。 ⑷異體紅細胞人大腸埃希桿菌體系組與自體紅細胞大腸埃希桿菌體系組相比較，其均值[-10.3466±4.48095pg/ml]，（p=0.0220.05），IL-12分泌量兩組之間有著顯差別。 （四）結(jié)論 異體紅細胞實驗組相對于自體紅細胞對照組IL-8、IL-10的分泌量沒有顯著的不同，表明異體紅細胞對單核-巨噬細胞、Th2等細胞分泌細胞因子IL-8、IL-10的功能沒有明顯的調(diào)節(jié)作用；異體紅細胞實驗組分泌IL-12的量顯著低于自體紅細胞對照組，表明異體紅細胞對DC細胞、巨噬細胞等細胞分泌細胞因子IL-12的能力有著一定程度的負向調(diào)控作用。 （一）目的 運用郭氏“系統(tǒng)血液快速固有免疫反應(yīng)分離實驗體系”體外觀察異體白細胞對IL-8、IL-10、IL-12分泌的影響。 （二）方法 健康成人同型新鮮抗凝全血，常規(guī)分離制備血漿、紅細胞和白細胞，以自體血漿作為反應(yīng)基質(zhì)建立系統(tǒng)血液分離實驗體系，考察異體白細胞對IL-8、IL-10、IL-12分泌的影響，設(shè)立自體白細胞作為對照。ELISA法測定上清液中的細胞因子IL-8、IL-10、IL-12，并探討其改變可能存在的意義。 （三）結(jié)果 1、血漿IL-8濃度比較 ⑴異體白細胞生理鹽水體系[31.12373±2.828597pg/ml]組與異體白細胞人大腸埃希桿菌體系組[199.75970±62.743079pg/ml]比較，其均值差值[-168.6360±59.37619pg/ml]，（p=0.0050.05），IL-8分泌量兩組之間有著顯著的差別。 ⑵自體白細胞生理鹽水體系[48.29476±6.511376pg/ml]組與自體白細胞大腸埃希桿菌體系組[465.92697±111.354713pg/ml]比較，其均值差值[-417.6322±59.37619pg/ml],（p=0.0000.05）,IL-8分泌量兩組之間有著顯著的差別。 ⑶異體白細胞生理鹽水體系組與自體白細胞生理鹽水體系組相比較，其均值差值[-17.1710±59.37619pg/ml]，IL-8分泌量兩組之間沒有明顯的差別（p=0.7730.05）。 ⑷異體白細胞人大腸埃希桿菌體系組與自體白細胞大腸埃希桿菌體系組相比較，其均值差值[-266.1673±59.37619pg/ml]，IL-8分泌量兩組之間沒有明顯的差別（p=0.0000.05）。 2、血漿IL-10濃度比較 ⑴異體白細胞生理鹽水體系組[21.37236±3.307657pg/ml]與異體白細胞人大腸埃希桿菌體系組[16.93582±2.346794pg/ml]比較，其均值差值[4.4365±5.37453pg/ml]，（p=0.4100.05）， IL-10分泌量兩組之間沒有顯著的差別。 ⑵自體白細胞生理鹽水體系組[47.65618±7.432838pg/ml]與自體白細胞大腸埃希桿菌體系組[46.16091±9.180075pg/ml]比較，其均值差值[1.4953±5.37453pg/ml],（p=0.7810.05），IL-10分泌量兩組之間沒有顯著的差別。 ⑶異體白細胞生理鹽水體系組與自體白細胞生理鹽水體系組相比較，其均值差值[-26.2838±5.37453pg/ml]， IL-10分泌量兩組之間有顯著的差別（p=0.0000.05）。 ⑷異體白細胞人大腸埃希桿菌體系組與自體白細胞大腸埃希桿菌體系組相比較，其均值差值[-29.2251±5.37453pg/ml]，IL-10分泌量兩組之間有顯著的差別（p=0.0000.05）。 3、IL-12分泌量的測定值 ⑴異體白細胞生理鹽水體系組[47.30961±8.11704pg/ml]與異體白細胞人大腸埃希桿菌體系組[39.69006±6.517480pg/ml]比較，其均值差值[7.6402±4.48095pg/ml]（p=0.6650.05），IL-12分泌量兩組之間沒有明顯的差別。 ⑵自體白細胞生理鹽水體系組[39.95673±4.607648pg/ml]與自體白細胞大腸埃希桿菌體系組[29.00088±3.838735pg/ml]比較，其均值差值[10.9558±4.48095pg/ml],（p=0.0150.05），IL-12分泌量兩組之間有顯著的差別。 ⑶異體白細胞生理鹽水體系組與自體白細胞生理鹽水體系組相比較，其均值差值[7.3529±4.48095pg/ml]，IL-12分泌量兩組之間沒有明顯的差別（p=0.1020.05）。 ⑷異體白細胞人大腸埃希桿菌體系組與自體白細胞大腸埃希桿菌體系組相比較[10.6892±4.48095pg/ml]，，（p=0.0180.05），IL-12分泌量兩組之間有著顯差別。 （四）結(jié)論 異體白細胞實驗組相對于自體白細胞對照組IL-8、IL-10的分泌顯著降低，說明異體白細胞對單核-巨噬細胞及Th2細胞等細胞分泌細胞因子IL-8、IL-10有著明顯的負向調(diào)節(jié)作用；異體白細胞實驗組與自體白細胞對照組相比較IL-8與IL-10的分泌量顯著下降且呈明顯的正相關(guān)性，至少表明這兩種細胞因子之間相互調(diào)節(jié)，以達成細胞因子網(wǎng)絡(luò)的穩(wěn)態(tài)平衡；異體白細胞實驗組相對于自體白細胞對照組IL-12的量顯著降低，表明異體白細胞對DC細胞、巨噬細胞等細胞分泌細胞因子IL-12的能力有著一定程度的負向調(diào)控作用。
[Abstract]:( i ) Purpose

The effects of allogeneic red blood cells on the secretion of IL - 8 , IL - 10 and IL - 12 were observed in vitro by using the " system of rapid innate immune reaction in blood " of Guo ' s system .

( II ) Methodology

The effects of allogeneic red blood cells on the secretion of IL - 8 , IL - 10 and IL - 12 were determined by ELISA . The levels of IL - 8 , IL - 10 , IL - 12 in supernatant were determined by ELISA .

( III ) Results

1 . Comparison of plasma IL - 8 concentration

( 1 ) There was a significant difference between the two groups : ( 1.89179 鹵 2.674565pg / ml ) of allogeneic red blood cell physiological saline group compared with ( 119.05455 鹵 31.056229pg / ml ) .

( 2 ) There was a significant difference between the two groups of autologous red blood cell physiological saline system group ( 29.05148 鹵 2.972132pg / ml ) and autologous red blood cell large intestine Escherichia coli system group ( 132.63303 鹵 35.072837pg / ml ) .

There was no significant difference between the two groups ( p = 0.819 . 05 ) .

( 4 ) There was no significant difference between the two groups ( p = 0.9970 . 05 ) .

2 . Comparison of plasma IL - 10 concentration

The mean difference was 5.6240 鹵 5.37453pg / ml , and there was no significant difference between the two groups ( p = 0 . 2960 . 05 ) .

( 2 ) There was no significant difference between IL - 10 secretion and IL - 10 secretion in autologous red cell physiological saline system group ( 24.15185 鹵 3.748592pg / ml ) and autologous red blood cell large intestine Escherichia coli system group ( 27.58494 鹵 4.740832pg / ml ) .

The mean difference was 3.7571 鹵 5.37453pg / ml and there was no significant difference between the two groups ( p = 0.3250.05 ) .

( 4 ) There was no significant difference between the two groups ( p = 0.4850.05 ) .

3 . Measured value of IL - 12 secretion

The mean difference was 2.8289 鹵 4.48095pg / ml , ( p = 0.5280.05 ) and there was no significant difference between the two groups of IL - 12 secretion .

( 2 ) There was no significant difference between the two groups of autologous red blood cell physiological saline system group ( 37.74982 鹵 5.783372pg / ml ) and autologous red blood cell large intestine Escherichia coli system group ( 33.323 91 鹵 2.982255pg / ml ) , and the mean difference was 4.4259 鹵 4.48095pg / ml respectively , ( p = 0.3240.05 ) and IL - 12 secretion .

( 3 ) There was a significant difference between the two groups ( p = 0.0080.05 ) .

( 4 ) Compared with the autologous red cell E . coli system group , the average value of E . coli was 10.3466 鹵 4.48095pg / ml , ( p = 0.0220 . 05 ) , and there was a significant difference between the two groups of IL - 12 secretion .

( IV ) Conclusion

There was no significant difference in the secretion of IL - 8 and IL - 10 from the experimental group , indicating that allogeneic red blood cells had no obvious regulation effect on the secretion of cytokines IL - 8 and IL - 10 by mononuclear - macrophage , Th2 , etc .
The amount of IL - 12 secreted by allogeneic red blood cells in experimental group was significantly lower than that of autologous red blood cell control group , indicating that the ability of allogeneic red blood cells to secrete cytokines IL - 12 in DC cells , macrophages and other cells has a certain degree of negative regulation .

( i ) Purpose

The effect of allogeneic white blood cell on IL - 8 , IL - 10 and IL - 12 secretion was observed in vitro by using the " system of rapid innate immune reaction in blood " of Guo ' s system .

( II ) Methodology

The effects of allogeneic white blood cells on the secretion of IL - 8 , IL - 10 and IL - 12 were determined by ELISA . The levels of IL - 8 , IL - 10 , IL - 12 in supernatant were determined by ELISA .

( III ) Results

1 . Comparison of plasma IL - 8 concentration

( 1 ) There was a significant difference between IL - 8 secretion and IL - 8 secretion between the two groups . The mean difference of IL - 8.6360 鹵 59.37619pg / ml , ( p = 0.0050.05 ) and IL - 8 secretion were significantly different between the two groups .

( 2 ) The mean value difference of autologous white blood cell physiological saline system ( 48.29476 鹵 6.511376pg / ml ) was significantly different from that of autologous white cell E . coli system group ( 465.92697 鹵 111.354713pg / ml ) , and the mean difference of IL - 417.6322 鹵 59.37619pg / ml ) , ( p = 0.00000.05 ) and IL - 8 secretion .

There was no significant difference between the two groups ( p = 0.777 . 05 ) .

( 4 ) There was no significant difference between the two groups ( p = 0.0000.05 ) .

2 . Comparison of plasma IL - 10 concentration

The mean difference was 4.4365 鹵 5.37453pg / ml , ( p = 0.41 . 05 ) , and there was no significant difference between the two groups .

The mean difference was 1.4953 鹵 5.37453pg / ml , ( p = 0.781 . 05 ) and there was no significant difference between the two groups .

( 3 ) Compared with the normal saline system group , the mean difference of IL - 10 secretion was significantly different between the two groups ( p = 0.0000 . 05 ) .

( 4 ) There was a significant difference between the two groups ( p = 0.0000 . 05 ) .

3 . Measured value of IL - 12 secretion

The mean difference was 7.6402 鹵 4.48095pg / ml ( p = 0.665 . 05 ) and there was no significant difference between the two groups of IL - 12 secretion .

The mean difference was 10.9558 鹵 4.48095pg / ml , ( p = 0.0150.05 ) , and there was a significant difference between the two groups of IL - 12 secretion .

The mean difference was 7.3529 鹵 4.48095pg / ml , and there was no significant difference between the two groups ( p = 0.1020 . 05 ) .

( 4 ) There was a significant difference between the two groups ( 10.6892 鹵 4.48095pg / ml ) , ( p = 0.0180 . 05 ) and IL - 12 secretion .

( IV ) Conclusion

The secretion of IL - 8 and IL - 10 decreased significantly in the experimental group , and the IL - 8 and IL - 10 secretion of allogeneic white blood cells were negatively regulated by IL - 8 and IL - 10 .
The levels of IL - 8 and IL - 10 decreased and positively correlated with IL - 8 and IL - 10 in the experimental group .
Compared with the control group , the amount of IL - 12 decreased significantly in the experimental group , indicating that the ability of allogeneic white blood cells to secrete cytokines IL - 12 in DC cells , macrophages and other cells has a certain degree of negative regulation .

【學位授予單位】：第二軍醫(yī)大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R363

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