| Objective:To analysis of the work character of lymphocytes separated from Plasmodium vivax healing cases, further in order to reveal the specific liver period anti-body product capabilities of host plasma cells suffering from Plasmodium vivax and to provide a research base for screening erythrocyte stage vaccine candidate antigens.Methods:Select the cases who have following conditions suffering from Plasmodium and healing during 2009 in malaria-endemic areas in Ximeng County of Yunnan Province.①Plasmodium vivax malaria cases were diagnosed by microscope in 2009.②The cases were no recrudesce malaria history before chosen.③Plasmodium vivax malaria was negative for microscopy before sampling. Plasmodium vivax convalescent patients were chosen according with conditions as previously stated. To choose healthy volunteers according with following conditions in Yunnan Province Simao District,①No Plasmodium vivax was found in blood-smear before sampling;②The cases were no malaria history in 2009. The persons were chosen as Healthy volunteers according with conditions as previously stated. To collect 20ml heparin anticoagulate venous blood respectively of Plasmodium vivax malaria convalescent patients and Plasmodium vivax malaria cases. To separate lymphocyte by density gradient centrifugation and count, then culture addition of PHA-M. To centrifugate (1000rpm/min) 10 minutes after culture finishing, and then discard all of supernatant. As a spot of residual liquid on tube wall was precipitation on the tube bottom while beating upon to disperse the precipitating cell by disposable pipette. Try to discard the supernatant and suspend the sediment, then take a drop of the sediment to slide it into a blood cell membrane (head, body, tail clear). After fixing with methanol, it should be stained with Giemsa dye.30 minutes later, wash it twice with ultrapure water and out cells under the oil microscope when blood cell membrane is dry.Results:In this study, the lymphocytes were detected from a total of nine cases of Plasmodium vivax malaria healing persons and 8 healthy volunteers.345ml venous blood were collected from 17 selected objects and obtain 74.4×107 lymphocytes,107 ampoules of cells suspension were frozen (1 ml/support). There into,205ml blood,40.2 x 107 lymphocytes and 59 ampoules were collected from nine Plasmodium vivax healing persons. The lymphocytes separated from nine Plasmodium vivax healing persons and eight healthy volunteers were employed conversion experiment for 24h,48h, 58h,68h,70h and 72h respectively. The lymphocytes from Plasmodium vivax healing persons showed transformation after 48h when stimulated. Then the transformed cells proportion was significantly higher at 72h, the conversion rate was 51.78%. After the lymphocytes from healthy human was stimulated for 68h, the proportion of transformed cells significantly increased. The highest conversion rate was 70.37% at 72h. The lymphocyte transformation rate between Plasmodium vivax healing persons with healthy persons was no significant different stimulated for the groups before 58h (P> 0.05),68h after the health of human lymphocyte transformation rate significantly higher than that vivax malaria patients (P<0.05).,The rates of transitional lymphocyte were no significant difference between the two groups.Conclusion:Healthy rate of lymphocyte transformation after PHA stimulation than 68h vivax malaria patients, suggesting that vivax malaria patients the sensitivity of lymphocytes to the PHA weaker than healthy people. |
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