The Study Of Quantity And Function Of T Cell Subsets Of The Patients With Paroxysmal Nocturnal Hemoglobinuria

Objective:This study was to investigate the immune state of T cells and the quantity, function of GPI+ T cells or GPI- T cells of the patients with paroxysmal nocturnal hemoglobinuria (PNH).Methods:22 cases of PNH and 18 normal controls were enrolled in this study. Their T lymphocyte subsets, Th lymphocyte subsets were assayed by flow cytometry with the monoclonal antibodies concerned. The proportion of GPI+ T cells or GPF T cells in CD3+ T cells, CD4+ T cells, CD8+ T cells and the expression of CD69 on these T cells was also respectively assayed.Results:1.The variations of T cell subsets①The proportion of CD4+ T cells in CD3+ T cells in PNH [(47.7670±13.91139)%] was lower than that in controls [(54.9592±7.11678)%](p<0.05).CD8+ T cells in CD3+ T cells of PNH cases [(52.2767±13.90395)%] were higher than that of controls [(45.2418±6.75306)%](p<0.05).The ratio of CD4+ T cells to CD8+ T cells was inversion in PNH.②The proportion of CD4+ T cells was decreasing in controls [(54.9592±7.11678)%], classic PNH[(52.9287±13.51782)%],PNH-AA[(41.2212±12.34295)%].The proportion of CD8+ T cells was increasing in controls [(45.2418±6.75306)%],classia PNH[(47.7752±13.53115)%],PNH-AA[(58.7788±12.34295)%]].The ratio of CD4+ T cells to CD8+ T cells was decreasing in concrols(1.26356±0.348878), classic PNH(1.26041±0.663275),PNH-AA(0.77111±0.383226).All above were more significantly in PNH-AA (p<0.05).③The proportion of CD4+ T cells was positively correlated with RBC (r=0.665, p<0.05) and the proliferative degree of bone marrow (r=0.680, p<0.05).The proportion of CD8+ T cells was negatively correlated with RBC (r=-0.665,p<0.05) and the proliferative degree of bone marrow (r=-0.680, p<0.05).The ratio of CD4+ T cells to CD8+ T cells was positively correlated with RBC (r=0.812, p<0.05) and the proliferative degree of bone marrow (r=0.694, p<0.05).2.The variations of Th cell subsets ①The proportion of Thl cells in PNH [(16.9136±6.78899)%] was higher than controls[(4.4600±1.81879)%].It was significantly higher in PNH-AA[(22.8000±5.45244)%]than controls[(4.4600±1.81879)%](p<0.05)and classic PNH[(12.0083±2.20770)%] (p<0.05).And this proportion in classic PNH was significantly higher than controls.The proportion of Th2 cells in PNH [(4.7582±1.98441)%] had no difference with controls[(3.7960±1.13810)%].②The proportion of Th1 T cells was negatively correlated with RBC (r=-0.625, p<0.05).3.The number of GPI+ T cells or GPI-T cells①In classic PNH,the number of GPI- T cells in CD3+ T cells was(10.5240±7.71277)%,that in CD8+ T cells was(14.6797±11.96718)%,that in CD4+T cells was(3.9241±2.46263)%.②In PNH-AA,the number of GPI- T cells in CD3+ T cells was(18.4649±12.41783)%, that in CD8+ T cells was(25.6295±14.39083)%, that in CD4+ T cells was(5.8969±2.20670)%.4. The expression of CD69 on GPI+ T cells or GPI- T cells①The expression of CD69 on both GPI+CD8+ T cells[(17.67881±8.562493)%] and GPI-CD8+ T cells[(15.86575±7.279743)%] in PNH were significantly higher than GPI+CD8+T cells in controls [(4.68038±1.216645)%](p<0.05).The expression of CD69 on both GPI+CD8+T cells[(18.49545±9.014283)%] and GPI-CD8+ T cells [(16.52055±7.49029)%] in classic PNH were significantly higher than controls (p<0.05). The expression of CD69 on both GPI+CD8+T cells [(15.88220±8.119361)%] and GPI-CD8+T cells[(14.42520±7.395645)%] in PNH-AA were significantly higher than controls (p<0.05).The expression on GPI+CD8+T cells was similar to GPI-CD8+ T cells in classic PNH and PNH-AA(p>0.05).②The expression of CD69 on GPI+CD8+ T cells was negatively correlated with N% (r=-0.676, p<0.05) and the percentage of leukocytic lineage in sternal bone marrow (r=-0.851,p<0.05).It was positively correlated with the percentage of erythroid lineage in sternal bone marrow (r=0.817, p<0.05).The expression of CD69 on GPI-CD8+ T cells was negatively correlated with N%(r=-0.778, p<0.05) and the percentage of leukocytic lineage in sternal bone marrow (r=-0.712,p<0.05).It was positively correlated with the percentage of erythroid lineage in sternal bone marrow (r=0.766,p<0.05).③The expression of CD69 on both GPI+CD4+ T cells[(4.65431±1.984378)%]and GPI-CD4+ T cells [(4.93181±1.73001)%] in PNH were significantly higher than GPI+CD4+ T cells in controls[(1.77339±0.645259)%] (p<0.05).The expression of CD69 on either GPI+CD4+ T cells[(5.01455±2.095361)%] or GPI-CD4+ T cells [(5.41600±1.702866)%] in classic PNH was significantly higher than controls (p<0.05). The expression of CD69 on either GPI+CD4+ T cells [(3.86180±1.627353)%] or GPI-CD4+ T cells[(3.86660±1.38237)%]in PNH-AA was significantly higher than controls (p<0.05).The expression on GPI+CD4+ T cells was similar to GPI-CD4+ T cells in classic PNH and PNH-AA (p>0.05)④The expression of CD69 on GPI+CD4+ T cells was negatively correlated with N% (r=-0.750, p<0.05).It was positively correlated with the percentage of erythroid lineage in sternal bone marrow (r=0.690, p<0.05).The expression of CD69 on GPI-CD4+ T cells was negatively correlated with the percentage of leukocytic lineage in sternal bone marrow (r=-0.533,p<0.05).Conclusion:It was concluded that cytoimmunity showed higher function in PNH, which might be responsible for bone marrow failure but not related to the existence of PNH clone in T cell population.