interleukin-8 and Primary-Myelofibrosis

Induced pluripotent stem cells (iPS) derived from disease cells are expected to provide a new experimental material, especially for diseases from which samples are difficult to obtain. In this study, we generated iPS from samples from patients with primary and secondary myelofibrosis. The primary myelofibrosis cells had chromosome 13q deletions, and the secondary myelofibrosis (SMF) cells had JAK2V617F mutations. The myelofibrosis patient cell-derived iPS (MF-iPS) were confirmed as possessing these parental disease-specific genomic markers. The capacity to form three germ layers was confirmed by teratoma assay. By co-culture with specific feeder cells and cytokines, MF-iPS can re-differentiate into blood progenitor cells and finally into megakaryocytes. We found that mRNA levels of interleukin-8, one of the candidate cytokines related to the pathogenesis of myelofibrosis, was elevated predominantly in megakaryocytes derived from MF-iPS. Because megakaryocytes from myelofibrosis clones are considered to produce critical mediators to proliferate fibroblasts in the bone marrow and iPS can provide differentiated cells abundantly, the disease-specific iPS we established should be a good research tool for this intractable disease.

Topics: Adult; Amino Acid Substitution; Cells, Cultured; Chromosome Deletion; Chromosome Disorders; Chromosomes, Human, Pair 13; Coculture Techniques; Female; Fibroblasts; Humans; Induced Pluripotent Stem Cells; Interleukin-8; Janus Kinase 2; Male; Megakaryocytes; Middle Aged; Mutation, Missense; Primary Myelofibrosis; Teratoma

Cytokine-phenotype associations have recently been described in primary myelofibrosis and increased levels of IL-8, sIL-2R, IL-12, and IL-15 were found to be independently predictive of inferior survival. Pomalidomide therapy is effective for alleviating anemia in myelofibrosis; we examined the relationship between plasma cytokine/chemokine levels and response to treatment with pomalidomide. The study population included 32 Mayo Clinic patients (median age 66 years) who participated in two consecutive clinical trials of pomalidomide therapy for myelofibrosis-associated anemia. Ten (31%) patients achieved anemia response per International Working Group criteria. Anemia response was seen only in the presence of JAK2V617F (P = 0.04) and, in addition, predicted by lower circulating levels of MCP-1 (P = 0.003), IL-2R (P = 0.008), IL-15 (0.01), and IL-8 (P = 0.02). Marked splenomegaly and increased serum LDH level were associated with poor response (P = 0.02 and 0.03, respectively) and with each other (P = 0.02), but not with JAK2V617F. The aforementioned cytokines were not significantly associated with JAK2V617F but increased levels of sIL-2R (P = 0.01), IL-15 (P = 0.06), and MCP-1 (P = 0.07) clustered with marked splenomegaly. Current data suggest that, in the context of pomalidomide treatment, response is more likely in the presence of JAK2V617F and further predicted by the absence of marked splenomegaly or increased levels of proinflammatory cytokines.

Topics: Adult; Aged; Aged, 80 and over; Anemia; Chemokine CCL2; Cytokines; Female; Humans; Interleukin-15; Interleukin-8; Janus Kinase 1; Janus Kinase 2; Male; Middle Aged; Mutation; Predictive Value of Tests; Primary Myelofibrosis; Receptors, Interleukin-2; Thalidomide; Treatment Outcome

Topics: Animals; Emperipolesis; GATA1 Transcription Factor; Humans; Interleukin-8; Megakaryocytes; Mice; Neutrophils; Primary Myelofibrosis; Transforming Growth Factor beta

Chronic myeloproliferative neoplasms are characterized by clonal hematopoiesis and persistent inflammatory reaction. In this study, the clinical significance and prognostic impact of several inflammatory markers were evaluated in patients with BCR/ABL-negative myeloproliferative malignancies.. Serum levels of interleukin-8 (IL-8) and lymphoid-associated activation markers - soluble interleukin-2 receptor (sIL-2R) and immunoglobulin-free light chains (FLC) - were evaluated in patients with primary myelofibrosis (MF), post-polycythemia vera MF, and post-essential thrombocythemia MF, and compared with the levels in healthy donors.. In 57 MF patients, sIL-2R excess correlated with transfusion-dependent anemia (p = 0.03) and splenomegaly (p = 0.02). There were no statistically significant correlations between sIL-2R and IL-8 levels, but the plasma concentration of κ-FLC positively correlated with the IL-8 level (p = 0.027). In univariate analysis, increased levels of IL-8 (p = 0.016) and sIL-2R (p = 0.010) significantly reduced 1-year overall survival. Only elevated sIL-2R rate retained significance (p = 0.02) in multivariate analysis when Dynamic International Prognostic Scoring System plus (DIPSSplus) risk stratification was added.. We observed an association between FLC and proinflammatory cytokine hyperexpression. Serum cytokine levels and FLC might be a promising approach to predicting and monitoring treatment response in MF patients.

Topics: Aged; Anemia; Female; Humans; Immunoglobulin kappa-Chains; Immunoglobulin Light Chains; Inflammation Mediators; Interleukin-8; Male; Middle Aged; Polycythemia Vera; Primary Myelofibrosis; Prognosis; Receptors, Interleukin-2; Reference Values; Statistics as Topic; Survival Analysis; Thrombocythemia, Essential

Autoimmune phenomena and cytokines were investigated in 100 patients with myelofibrosis (MF) and related to marrow fibrosis and clinical risk. Anti-erythrocyte antibodies by mitogen-stimulated direct antiglobulin test (MS-DAT) were positive in 45%, anti-platelets in 15% and organ/non organ-specific in 57% of cases, without clinically overt disease, and mostly in low-risk/intermediate-risk-1 and MF-0/MF-1. TGF-β and IL-8 were increased in MS-DAT positive cases, and IFN-γ in patients with serological autoantibodies. TGF-β and IL-17 were elevated in early clinical and morphological stages, while IL-8 increased in advanced stages. These data suggest that autoimmune phenomena and cytokine disregulation are particularly relevant in early MF.

Topics: Adult; Aged; Aged, 80 and over; Antibodies, Anti-Idiotypic; Autoimmunity; Coombs Test; Cytokines; Erythrocytes; Female; Follow-Up Studies; Humans; Interleukin-17; Interleukin-8; Male; Middle Aged; Prevalence; Primary Myelofibrosis; Prognosis; Transforming Growth Factor beta

Abnormal cytokine expression accompanies myelofibrosis and might be a therapeutic target for Janus-associated kinase (JAK) inhibitor drugs. This study describes the spectrum of plasma cytokine abnormalities in primary myelofibrosis (PMF) and examines their phenotypic correlates and prognostic significance.. Patients included in this study were required to have archived plasma, bone marrow biopsy, and cytogenetic information available at the time of first referral to the Mayo Clinic. Multiplex biometric sandwich immunoassay was used to measure plasma levels of 30 cytokines.. In total, 127 PMF patients were studied; comparison with normal controls (n = 35) revealed significantly increased interleukin-1β (IL-1β), IL-1RA, IL-2R, IL-6, IL-8, IL-10, IL-12, IL-13, IL-15, tumor necrosis factor α (TNF-α), granulocyte colony-stimulating factor (G-CSF), interferon alfa (IFN-α), macrophage inflammatory protein 1α (MIP-1α), MIP-1β, hepatocyte growth factor (HGF), IFN-γ-inducible protein 10 (IP-10), monokine induced by IFN-γ (MIG), monocyte chemotactic protein 1 (MCP-1), and vascular endothelial growth factor (VEGF) levels and decreased IFN-γ levels. In treatment-naive patients (n = 90), increased levels of IL-8 (P < .001), IL-2R (P < .001), IL-12 (P < .001), IL-15 (P = .001), and IP-10 (P = .003) were independently predictive of inferior survival. A similar multivariable analysis that included all 127 study patients confirmed the prognostic value of these five cytokines, and IL-8, IL-2R, IL-12, and IL-15 remained significant when risk stratification, according to the recently revised Dynamic International Prognostic Scoring System (DIPSS plus), was added to the multivariable model. Leukemia-free survival was predicted by IL-8, which was also the only cytokine associated with ≥ 1% circulating blasts. Other cytokine-phenotype associations included increased IL-8 and constitutional symptoms; IL-2R, IL-12, and transfusion need; IL-2R, IL-8, and leukocytosis; IP-10 and thrombocytopenia; HGF, MIG, IL-1RA, and marked splenomegaly; and IL-1RA, IL-2R, IP-10, MIP-1β, and JAK2V617F. A two-cytokine (IL-8/IL-2R) -based risk categorization delineated prognostically different groups within specific DIPSS plus risk categories.. This study signifies the presence of specific cytokine-phenotype associations in PMF and a prognostically relevant plasma cytokine signature that might prove useful as a laboratory tool for predicting and monitoring treatment response.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Biopsy; Bone Marrow Examination; Case-Control Studies; Chi-Square Distribution; Cytogenetic Analysis; Enzyme-Linked Immunosorbent Assay; Female; Genotype; Humans; Interleukin-12; Interleukin-15; Interleukin-8; Janus Kinase 2; Kaplan-Meier Estimate; Male; Middle Aged; Minnesota; Mutation; Phenotype; Predictive Value of Tests; Primary Myelofibrosis; Prognosis; Proportional Hazards Models; Protein Array Analysis; Receptors, Interleukin-2; Risk Assessment; Risk Factors; Up-Regulation

Myeloproliferation, myelofibrosis, and neoangiogenesis are the 3 major intrinsic pathophysiologic features of myeloid metaplasia with myelofibrosis (MMM). The myeloproliferation is characterized by an increased number of circulating CD34+ progenitors with the prominent amplification of dystrophic megakaryocytic (MK) cells and myeloid metaplasia in the spleen and liver. The various biologic activities of interleukin 8 (IL-8) in hematopoietic progenitor proliferation and mobilization as well as in neoangiogenesis prompted us to analyze its potential role in MMM. We showed that the level of IL-8 chemokine is significantly increased in the serum of patients and that various hematopoietic cells, including platelets, participate in its production. In vitro inhibition of autocrine IL-8 expressed by CD34+ cells with either a neutralizing or an antisense anti-IL-8 treatment increases the proliferation of MMM CD34(+)-derived cells and stimulates their MK differentiation. Moreover, addition of neutralizing anti-IL-8 receptor (CXC chemokine receptor 1 [CXCR1] or 2 [CXCR2]) antibodies to MMM CD34+ cells cultured under MK liquid culture conditions increases the proliferation and differentiation of MMM CD41+ MK cells and restores their polyploidization. Our results suggest that IL-8 and its receptors participate in the altered MK growth that features MMM and open new therapeutic prospects for this still incurable disease.

Topics: Aged; Antibodies; Antigens, CD34; Blood Platelets; Cell Differentiation; Cell Division; Gene Expression; Humans; Interleukin-8; Megakaryocytes; Middle Aged; Neutralization Tests; Ploidies; Primary Myelofibrosis; Receptors, Interleukin-8A; Receptors, Interleukin-8B