Immunophenotype of myeloid granulocytes in Chinese patients with BCR::ABL1-negative myeloproliferative neoplasms

Abstract

Typical BCR::ABL1-negative myeloproliferative neoplasms (MPN) are mainly referred to as polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofbrosis (PMF). Granulocytes in MPN patients are involved in their inflammation and form an important part of the pathophysiology of MPN patients. It has been shown that the immunophenotype of granulocytes in MPN patients is altered. We used flow cytometry to explore the immunophenotype of MPN patients and correlate it with clinical parameters. The results showed that PMF patients and PV patients had higher CD15+CD11b+ granulocytes than ET patients and normal controls. When grouped by gene mutation, changes in the granulocyte immunophenotype of MPN patients were independent of the JAK2V617F and CALR mutations. There was no significant heterogeneity in immunophenotype between ET patients and Pre-PMF, and between Overt-PMF and Pre-PMF patients. Granulocytes from some MPN patients showed an abnormal CD13/CD16 phenotype with a significant increase in mature granulocytes on molecular and cytomorphological grounds, and this abnormal pattern occurred significantly more frequently in PMF patients than in ET patients. CD15-CD11b- was negatively correlated with WBC and Hb and positively correlated with DIPSS score, whereas high CD10+ granulocytes were significantly and negatively associated with prognostic system IPSS and DIPSS scores in PMF patients. In conclusion, this study demonstrates the landscape of bone marrow granulocyte immunophenotypes in MPN patients. MPN patients, especially those with PMF, have a significant granulocyte developmental overmaturation phenotype. CD10+ granulocytes may be involved in the prognosis of PMF patients.

Keywords: Granulocyte; Immunophenotype; Myeloproliferative neoplasm.

Fig. 1

Comparison of individual antigens in MPN patients and healthy controls. Individual patients may lack data for a particular antigen

Fig. 2

Dual antigen combinations of granulocytes from MPN patients and controls

Fig. 3

Comparison of granulocyte immuneophenotypes in patients with ET and Pre-PMF and Overt-PMF. 15 patients with Pre-PMF and 36 patients with Overt-PMF

Fig. 4

A Graphical representation of the CD13/CD16 phenotype in a case of HD and a case of MPN and a histogram of CD56. B Statistical differences comparing CD10 or CD16 in MPN patients with normal CD13/CD16 phenotype and abnormal CD13/CD16 phenotype, respectively

Fig. 5

A Comparison of the proportions of cytomorphologically segmented granulocytes, band granulocytes and metagranulocytes to the whole bone marrow in patients with PMF with abnormal CD13/CD16 phenotype and patients with PMF with normal CD13/CD16 phenotype, respectively. B Analysis of the correlation between the proportion of segmented granulocytes to whole bone marrow and CD16 or CD10 on bone marrow cell morphology in patients with PMF. C Analysis of the correlation between the proportion of segmented granulocytes to whole bone marrow and CD16 or CD10 on bone marrow cell morphology in patients with ET

Fig. 6

Comparisons were made only between patients with concomitant JAK2 mutations or between different mutations in the same disease. JAK2^mut refers to the JAK2V617F mutation. JAK2^mut in PMF, JAK2^mut in PV and JAK2^mut in ET were compared using Dunn’s multiple comparison method. The Mann Whitney test was used to compare the two groups. P < 0.05 was considered statistically significant. NS indicates no statistical significance. CD16+ granulocytes were not statistically significant between groups