Reverse Migration Of Neutrophils And Its Relationship With CD8~+T Cells In Patients With MDS

ObjectiveThis study was designed to examine the distribution of neutrophils in peripheral blood of patients with myelodysplastic syndrome（MDS）and their effect on the function of CD8⁺T cellsThe experimental group consisted of 30 patients with myelodysplastic syndromes（MDS）hospitalized in the Department of Hematology,General Hospital of Tianjin Medical University。Twelve people in the control group,including 7 healthy control and 5 iron deficiency anemia（IDA）patients.The PE-Cy7-CD33、APC-CD11b、PE-CD54 and FITC-CD181 antibodies were labeled and flow cytometry was used to determine the proportion of cells.CD8⁺T cells isolated by immunomagnetic beads were cultyred with ICAM-1^highCXCR1^low neutrophils that were isolated by flow cytometry,using RT-PCR to detecte the mRNA relative expression of perforin and Granzyme-B.Results1.The expression of ICAM-1 in MDS patients（9984.09±5046.57）s was significantly higher than the control group（3298.66±996.69）（P<0.05）.2.The proportion of neutrophils(CD54^highCD181^low)in patients with MDS increased.The MDS high risk group（6.29±1.60）was significantly higher than the low risk group（3.57±1.45,P=0.01）;it was significantly higher than the normal control group（1.47±1.07,P=0.01）.3.CD8⁺T cell function is inhibited by neutrophils,as the culture system shown in Table 1.（1）The expression level of granzyme B in group 1（4.19±1.48）was higher than that in group 4（2.09±1.43,p<0.05）,and that in group 3（3.45±1.07）was higher than that in group 4（2.09±1.43,p<0.05）too.And group 2（3.14±1.89）was higher than group 6（2.12±0.49,p<0.05）.There was no significant difference between the in other groups（P>0.05）.（2）The perforin expression was higher in group3（0.26±0.10）than in groups6（0.12±0.99,p<0.05）,and the group 2（0.25±0.04）was higher than group 6（p<0.01）.There was no statistical difference between the in other groups（P>0.05）.4.The relationship between CD8⁺T cell function and reverse migration of neutrophils.The relative expression of perforin and granzyme in the experimental group（reverse migrating neutrophils and normal CD8⁺T cells co-cultured group）was higher than that in the control group（normal control neutrophils and normal control CD8⁺T cells were co-cultured）.However,there was no statistical significance（P>0.05）.Conclusion（1）The mean fluorescence intensity of ICAM-1 in neutrophils in patients with MDS increased.The increased proportion of ICAM-1^highigh neutrophils suggests the potential for disseminated inflammation in patients with MDS.（2）Compared with the control group,the proportion of reverse migration neutrophils(ICAM-1^highCXCR1^low)in patients with MDS increased,and the higher the risk,the higher the proportion of patients with MDS.（3）Neutrophil in MDS and normal control neutrophils can inhibit granzyme expression in CD8⁺T cells,and the suppression of CD8⁺T cell granzyme expression in MDS patients by neutrophils in MDS is stronger than in normal neutrophils.Normal neutrophils can inhibit the expression of perforin of CD8⁺T cells in normal controls and MDS patients.It is suggested that MDS neutrophils can inhibit the function of CD8⁺T cells.（4）The reverse migration neutrophils of peripheral blood in patients with MDS may regulate CD8⁺T cell function,but further studies are needed.