| Objective and BackgroundPatients with digestive tract malignancy often accompanied by anemia causing by bleeding and many other reasons. Many of them have to be treated by blood transfusion therapy. In 1937, Opelz G discovered that blood transfusion can increase the survival rate of renal transplantation recipient , which suggested that blood transfusion might inhibit the immune response of the patients,from then on many reports about allogeneic blood transfusion cause tumor metastasis and reoccurrence, decrease 5 year survival rate , increase the complication of infection and et al. Transfusion mediated immunosuppression has been comprehended. How to treat the patients who need blood transfusion has become a problem for clinic doctors. Now it has been known that the main component damaging patients' immune function is in plasma, white blood cell and its products. It is suggested that RBC component transfusion may be avoid immunosuppression for tumor patients who need transfusion indeed. But it has been no systemic and specific study about the effect of RBC component transfusion on thetumor patients. The RBC has been realized as an unprofessional innate immune cell. It can recognize ,adhere and kill antigen, clear the circulating immunocomplexes, and regulate adaptive immunity. Recently, the changes of it's innate immune function have been focused by playing a great important role in diagnosis and prognosis of SARS. To evaluate the effect of RBC component transfusion on the immune function of tumor patients, we observed the EIIAF(erythrocyte innate immune adhesion function) of patients for a series studies as follows 1.The comparison of EIIAF and its regulatory factors in digestive tract malignancy patients and in healthy control,and the effect of patients' RBC on the EIIAF and its regulatory factors in vitro. 2. The effect of allogeneic RBC on the EIIAF of patients with digestive tract malignancy in vitro. 3. The changes of EIIAF and its regulatory factors of the patients before and after whole blood transfusion or RBC(leukoreducted) component transfusion. Methods 1. The EIIAF and its regulatory factors measured by RBC-Rosette test(1)The adhesion capacity of RBC to yeast cellRCR 1.25×10~7/ml RBC suspensions(washed 3 times with normal solution 2000r/min. centrifuged 5min) 0.05ml and 1.25×10~8/ml complement sensitized yeast cells (washed 1 times with normal solution 2000r/min. centrifuged 10min)incubated in a water bath at 37℃ for 30 min. then transfered into a slide, observed under microscope. 200 RBCwere counted randomly and the percentage of RBC- Rosette, which adhere at least 2 yeast, is determined. That is the RCR(the Rate of RBC- CR1 Rosette)RICR 1.25x107/ml RBC suspensions(washed 3 times with normal solution 2000r/min, centrifuged 5min) 0.05ml and 1.25><108/ml complement unsensitized yeast cells (washed 1 times with normal solution 2000r/min, centrifuged 10min)incubated in a water bath at 37 °C for 30 min, then transfered into a slide, observed under microscope. 200 RBC was counted and the percentage of RBC- Rosette, which adhere at least 2 yeast, is determined. That is the RICR(the Rate of RBC- Immune Complex Rosette)(2) The adhesion capacity of RBC to tumor cellTRR lxlO6/ml Ehrlich ascites carcinoma cell suspensions(washed 1 time with normal solution lOOOr/min, centrifuged lOmin) 0.1ml and normal serum 0.1ml incubated in a water bath at 37°C for 30 min, wash 1 time with normal solution lOOOr/min, centrifuged lOmin; add l*108/ml RBC suspensions(washed 3 times with normal solution 2000r/min, centrifuged 5min) 0.05ml , incubated in a water bath at 37 °C for 30 min again, 2000r/min, centrifuged lOmin, then transfered into a slide, observed under microscope. 100 tumor cells were counted and the percentage of Tumor-RBC Rosette, which adhere at least 3 RBC, is determined. That is the TRR(Tumor -RBC Rosette Rate). (3) The serum RBC innate immune regulatory factorsSRFIR (the Serum Rosette Forming Inhibitory Rate of RBC - CR1 rosette ) Put the awaiting -test serum 0.075ml in two tubesrespectively, put tubel in a water bath at 58°C for 30 min and tube2 at room temperature, add 1.25xl07/ml healthy "O" type RBC suspensions(washed 3 times with normal solution 2000r/min, centrifuged 5min) 0.05ml and 1.25xlO8/ml complement sensitized yeast cells (washed 1 time with normal solution 2000r/min, centrifuged 10min)incubated in a water bath at 37°C for 30 min, then transfered into a slide,stained , observed under microscope. 200 RBC were counted and the percentage of RBC- Rosette, which adhere at least 3 yeast, is determined. SRFIR=(the Rosette rate of tubel- the Rosette rate of tube2)/ the Rosette rate of tubel x 100%SRFER (the Serum Rosette Forming Enhancing Rate of RBC - CR1 rosette) Put the serum 0.075ml in tubel and Normal Solution0.075ml in tube2, add 1.25xl07/ml healthy O type RBC suspensions(washed 3 times with normal solution 2000r/min, centrifuged 5min) 0.05ml and 1.25xlO8/ml complement sensitized yeast cells (washed 1 times with normal solution 2000r/min, centrifuged 10min)respectively incubated in a water bath at 37°C for 30 min, then transfered into a slide, observed under microscope. 200 RBC was counted and the percentage of RBC- Rosette, which adhere at least 2 yeast, is determined. SRFER=(the Rosette rate of tubel- the Rosette rate of tube2)/ the Rosette rate of tubel x 100%2. The test of autologous RBC enhance EIIAF in vitroPut the patients' RBC lysates, which prepared by osmotic lysis. 0.075ml in tubes before the test of RCR, RICR, TRR, SRFIR, SRFER. And the resulted Rosette rates were compaired with the patients' original Rosette rates, respectively.3. The test of allogeneic RBC enhance EIIAF in vitroPut the healthy people "O" type RBC lysates, which prepared by osmotic lysis, in tubes before the test of RCR, RICR. and TRR. And the resulted Rosette rates were compaired with the patients' original Rosette rates.Then put the "O" type RBC lysates with serum , the "O" type RBC lysates with serum inactivated by 58°C 30 min in tubes respectively before the test of RCR, RICR, and TRR. And the resulted Rosette rates were compaired with the patients' original Rosette rates with RBC lysates only .4.The effect of whole blood transfusion or RBC(leukoreduced) component transfusion on the EIIAF and its regulatory factors of the patients with digestive tract malignancy in vivo16 patients with digestive tract malignancy received RBC(leukoreduced) component transfusion therapy and other 14 patients received whole blood transfusion therapy. Before and after treatment in 3~5 days, the blood samples of the patients were taken to detect the RCR, RICR, TRR, SRFIR, and SRFER. The relationship between the changes of RCR> TRR and the changes of SRFER, SRFIRwas analyzed. Results1. The RCR% TRR of the patients with digestive tract malignancy are much lower than that of normal control, The SRFIR is higher than that of normal control. No differences among the groups of esophageal carcinoma, gastric carcinoma and colorectal carcinoma. Autologous red blood cell can increasing the Rosette Forming Enhancing Rate of RBC -CR1 rosette greatly.2. The allogeneic red blood cell increases the RCR and TRR of the patients with digestive tract malignancy greatly in vitro. Adding the allogeneic serum can inhibit this increasing action of the patients' RCR and TRR. After adding the allogeneic serum inactivated by 58°C 30 min , the RCR and TRR of the patients increase much more .3.The RCRn TRR of the patients with digestive tract malignancy decreased, the SRFIR of the patients increased after whole blood transfusion. The changes of the patients' RCR> TRR are closely related with the changes of the patients' SRFIR.4. The RCR and TRR of the patients with digestive tract malignancy increased after RBC(leukoreducted) component transfusion .The changes of the patients' RCR, TRR are not relate with the changes of the patients' SRFIR. Conclusion 1. The EIIAF of patients with digestive tract malignancy decrease and... |
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