| AimGraft - versus - host disease is the major complication after allo-genic bone marrow transplatation especially acute graft - versus - host disease ( aGVHD) . It reduces the survival rate remarkably . Atpresent immuno-suppressive agents are broadly adopted for inhibiting aGVHD in clinic, especially the combination of CsA with MXT , but their side - effect and toxic effect are serious , and they can lead to severe infection and induce tumor . Therefore it is an acute problem that we seek a new effective measure to prevent aGVHD.We have proved that HBO could prolong the survival time of allo-genic transplanted skin piece , and could suppress the cellular immune fanction. Meanwhile it has less side - effect. So in this study , we observed the effect of HBO on the number of T lymphocytes and their subgroups , the expression of adhesion molecules in skin , lung small intestine in post allogenic bone marrow transplanted mice after they had been treated with HBO or CsA + MTX or neither . We probed the mechanism of HBO antagonizing aGVHD and provided experimental data to clinic application of HBO.Method1 Duplication of the model of aGVHD.Donor mice BALB/C female , receptor mice BALB/C , C57 BL/6male ,10-12 weeks , about 20g . BALB/C mice were sacrificed and the bone marrow and the spleen removed , then spleens cells and bone marrow cells were suspended in RPMI1640 to a concentration of 2 107 ml and 4 106ml respectively . Mix them in proportion of 1:1 . All the processes were operated in sterilization . The recipient mice were exposed to 1800cGy total body radiation (TBI) delivered by a liner ac-celeralor at a close rate of 200cGy/min with in 24 hours before bone marrow transplantation ( BMT) . All recipient mice were given BMC though lateral tail vein.2 Treatment of animals.After Radiation the recipient mice were divided into four groups randomly . A: Both the donor and receptor mice were BALB/C . B : the donor were BALB/C , the receptor were C57 BL/6, C : CsA + MTX group,the recipient mice were injected into CsA and MTX . From 1 day to 10 day post BMT ,the recipient mice of C group were injected CsA (2. 5mg/kg. day) intraperitionally, and on day 1,3,5,9 post BMT the recipient mice of C group were injected MTX(7mg/m2) simultaneous - ly . D : donor and receptor mice of D group were disposed by 99. 9% oxygen with 0.25MPa for 1. 5 hour once a day on day 3, -2 1 before BMT. From 1 day to 10 day the recipient mice of D group were treated with HBO as described above . After BMT all the recipient mice were bred in sterile room.3 T lymphocyte subgroup and adhesion molecule assaysOn day,after BMT ten days skin ,lung , small intestine were extracted from recipient mice , and immediately placed in 10% buffered formaldehyde , fixed tissues were embedded in paraffin , cut into 6 -um tissues sections , mounted on glass sb'des , then stained with a standard hematoxylin and eosin (HE) procedure;or using immunohis-to - chemistry stained with 8ul FITC conjugated monoclonal anti -mouse CD3, CD4, CD8 and PE conjugated monoclonal anti - mouse CD11a,CD18. Then calculate fluorescence dimension (+ 105 um2, LUZEXF, Japan ) to observe the changes of CD3+ , CD4+, CD8+ , CD11a,CD18+ lymphocytes in skin lung and intestine.Result1 Recipient mice were monitored for clinical signs of aGVHD following BMT . The appearance of control group was charact - eristic of aGVHD , including hunched posture ,poor appetite and patchly alopecia . In contrast , CsA + MTX group and HBO group seldom exhibited the signs of aGVHD . But the CsA + MTX group was not as active as the two HBO group and showed diarrhea because of the side effects of immunosuppressive agents .2 Survival of recipients was followed for 11 days post BMT . The survival rate of the A group and D group (9/10 9/11 ) were much higher than central group ( 8/17 ) and CsA + MTX group ( 9/19 ) . There was no change was observed between control group and CsA + MTX group .3 Both HBO and CsA + MTX decreased markedly (P <0. 05) the number of T lym... |
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