Clinical Study Of Lymphocyte Subsets And Cytokines In Patients With BCR/ABL Negative Myeloproliferative Neoplasms

Objective:This research mainly detectsed the expression levels of lymphocyte subsets,T cell activation subsets,regulatory T cells,and cytokines in peripheral blood of patients with newly Ph negative myeloproliferative neoplasms（MPN）.To investigate the clinical significance of abnormal immune function in patients with MPN.Methods:This research included 123 newly diagnosed MPN patients in the First Affiliated Hospital of Nanchang University from October 2020 to February 2023.Multiparameter flow cytometry was used to tests peripheral blood lymphocyte subsets(including CD3⁺CD4⁺T cells,CD3⁺CD8⁺T cells,CD56⁺CD16⁺NK cells,CD19⁺CD3^-B,Treg(CD4⁺CD25⁺CD127^dim)absolute cell count and ratio,CD4⁺/CD8⁺ratio),cytokines（including IL-5、IL-2、IL-6、IL-1β、IFN-γ、IL-8、IL-10、TFN-α、IL-4、IL-12P70）,ratio of activated T cells（including HLA-DR^-CD3⁺resting T cells,HLA-DR⁺CD3⁺activated T Cells,HLA-DR^-CD8⁺resting Ts cells,HLA^-DR⁺CD8⁺activated Ts cells,CD28⁺CD8⁺CTL cells）,ratio of NK cell subsets(including CD16^hiCD56^low,CD16^lowCD56^low,CD16^+/-CD56^hi).Collect clinical data from patients and analyse the correlation between lymphocyte subsets,Treg,cytokines,T cell activation subsets,NK cell subsets and MPN subtypes,clinical features and routine test results in MPN patients.Results:（1）Immunological characteristics of MPN patients:Lymphocyte subsets and cytokine disorders are present in patients with different subtypes of MPN,as reflected by a decrease in the absolute count of the corresponding lymphocytes and an increase in the levels of the associated cytokines.Patients with primary myelofibrosis（PMF）have reduced CD3+T-cell,CD4+T-cell and Terg-cell counts,increased IL-8 levels compared to essential thrombocythemia（ET）.（2）Clinical and immunological features of splenomegaly in patients with MPN:Patients with ET who develop splenomegaly show decreased CD8+T cell counts and increased levels of cytokine IL-8.Patients with polycythemia vera（PV）who develop splenomegaly show elevated levels of CD8+activated T cells,CD28+CD8+CTL cell ratio,and cytokine IL-6.Patients with PMF who develop splenomegaly show increased levels of the cytokines IL-8 and IL-10.（3）Immunological characteristics of MPN patients between different MF degrees:Overt-PMF patients had lower counts of CD3+T cells,CD4+T cells,CD8+T cells,Treg cells and higher levels of cytokines IL-8 and IL-10 compared to pre-PMF.（4）Immunological features between different driver mutations in MPN patients:Patients with CALR mutation had significantly lower CD16⁺CD56⁺NK cell counts compared to controls.JAK2V617F mutation and triple negative patients had significantly lower CD3⁺T,CD8⁺T,CD16⁺CD56⁺NK cell counts compared to controls.（5）Correlation between age,lymphocyte subsets,and cytokines in patients with different MPN subtypes:In ET patients,IL-8 levels were negatively correlated with CD3+T、CD4+T cell、CD8+T cell、Terg cell count.In PMF patients,IL-10levels were negatively correlated with CD3+T cell,CD4+T cell,CD8+T cell,B cell,and Treg cell counts.Conclusion:（1）The immune microenvironment differs between MPN subtypes,with PMF patients having more impaired immune function and cytokine abnormalities than ET and PV patients.（2）Patients with larger spleens,and more severe myelofibrosis have more impaired immune function and cytokine abnormalities.（3）Immune function was impaired differently in MPN patients with different driver mutations,with JAK2V617F mutations and triple-negative patients more severely impaired than CALR mutated patients.（4）Significant lymphopenia is often accompanied by increased levels of the cytokine IL-8 in patients with ET.Significant lymphopenia is often accompanied by increased levels of the cytokine IL-10 in patients with PMF.