Analysis of cell-specific peripheral blood biomarkers in severe allergic asthma identifies innate immune dysfunction

Asthma is a disease of complex origin and multiple pathologies. There are currently very few biomarkers of proven utility in its diagnosis, management or response to treatment. Recent studies have identified multiple asthma phenotypes following biofluid analysis; however, such findings may be driven by the well-characterised alterations in immune cell populations in asthma. We present a study designed to identify cell type-specific gene signatures of severe allergic asthma in peripheral blood samples. Using transcriptomic profiling of four magnetically purified peripheral blood cell types, we identify significant gene expression changes in monocytes and NK cells but not T lymphocytes in severe asthmatics. Pathway analysis indicates dysfunction of immune cell regulation and bacterial suppression in the NK cells. These gene expression changes may be useful on their own as prognostic peripheral blood cell markers of severe asthma, but also may indicate novel cell pathways for therapeutic intervention.

Correlation Between Changes of Pelvic Bone Marrow Fat Content and Hematological Toxicity in Concurrent Chemoradiotherapy for Cervical Cancer

Objectives: To quantify the pelvic bone marrow (PBM) fat content changes receiving different radiation doses of concurrent chemoradiotherapy for cervical cancer and to determine association with peripheral blood cell counts. Methods: Fifty-four patients were prospectively collected. Patients underwent MRI iterative decomposition of water and fat with echo asymmetrical and least squares estimation (IDEAL IQ) scanning at RT-Pre, RT mid-point, RT end, and six months. The changes in proton density fat fraction (PDFF%) at 5–10 Gy, 10–15 Gy, 15–20 Gy, 20–30 Gy, 30–40 Gy, 40–50 Gy, and >50 Gy doses were analyzed. Spearman’s rank correlations were performed between peripheral blood cell counts versus the differences in PDFF% at different dose gradients before and after treatment. Results: The lymphocytes (ALC) nadirs appeared at the midpoint of radiotherapy, which was only 27.6% of RT-Pre; the white blood cells (WBC), neutrophils (ANC), and platelets (PLT) nadirs appeared at the end of radiotherapy which was 52.4%, 65.1%, and 69.3% of RT-Pre, respectively. At RT mid-point and RT-end, PDFF% increased by 46.8% and 58.5%, respectively. Six months after radiotherapy, PDFF% decreased by 4.71% under 5–30 Gy compared to RT-end; while it still increased by 55.95% compared to RT-Pre. There was a significant positive correlation between PDFF% and ANC nadirs at 5–10 Gy (r = 0.62, P = 0.006), and correlation was observed between PDFF% and ALC nadirs at 5–10 Gy (r = 0.554, P = 0.017). Conclusion: MRI IDEAL IQ imaging was a non-invasive approach to evaluate and track the changes of PBM fat content with concurrent chemoradiotherapy for cervical cancer. The limitation of low-dose bone marrow irradiation volume in cervical cancer concurrent chemoradiotherapy should be paid more attention.

Explore the Predictive Value of Peripheral Blood Cell Parameters in Refractory Mycoplasma pneumoniae Pneumonia in Children Over 6 Years Old

Background: To determine the predictive value of peripheral blood cell parameters for refractory Mycoplasma pneumoniae pneumonia (RMPP) in children over 6 years old.Methods: A retrospective study was conducted in children with RMPP admitted to the respiratory department of Tianjin Children's Hospital from September 2017 to September 2019, and non-refractory Mycoplasma pneumoniae pneumonia (NRMPP) was selected by the propensity score method and matched according to the ratio of 1:1.5. We analyzed the differences in clinical characteristics, peripheral blood cell parameters, imaging findings, and treatments between the two groups, and further determined the predictive value of peripheral blood cell parameters on RMPP.Results: There were 76 patients in the RMPP group and 114 patients in the NRMPP group. We found that the RMPP group has a longer clinical course and a higher incidence of intrapulmonary and extrapulmonary complications (p < 0.01). Moreover, the proportion of children in the RMPP group who received immunotherapy (such as glucocorticoid, gamma immunoglobulin) and fiberoptic bronchoscopy intervention was higher than that in the NRMPP group (p < 0.01). Meanwhile, the level of neutrophil, neutrophil/lymphocyte ratio (NLR), platelet count/lymphocyte ratio (PLR), mean platelet volume/lymphocyte ratio (MPVLR), C-reactive protein (CRP), lactic dehydrogenase (LDH), and interleukin (IL)-6 in the RMPP group was significantly higher (p < 0.01) than those in the NRMPP group. The incidence of pulmonary consolidation, atelectasis, and pleural effusion was also higher in the RMPP group (p < 0.05). ROC curve and binary logistic regression analysis showed that NLR > 3.92 (OR = 3.243; 95% CI = 1.485–7.081; p = 0.003), MPVLR > 5.29 (OR = 2.700; 95% CI = 1.258–5.795; p = 0.011), and pleural effusion (OR = 3.023; 95% CI = 1.424–6.420; p = 0.004) were significant factors in predicting RMPP. Our study showed that NLR had higher accuracy in predicting RMPP than CRP.Conclusions: The parameters of peripheral blood cells might be a predictor of RMPP. NLR > 3.92, MPVLR > 5.29, and pleural effusion might have important predictive value for RMPP in children over 6 years old.

Impaired Left Ventricular Ejection Fraction Induces "Very Early Pre-Diagnostic" Hemopoietic Modifications in Myeloid Leukemia Patients

Background: There is strong association between clonal hematopoiesis of undetermined prognosis (CHIP) and coronary artery disease (CAD) development. Indeed, ischemic heart disease is a leading cause of congestive heart failure (CHF) in western society. CH induces not only enhanced risk for myeloid malignancies, but also significant cardiovascular morbidity in elderly patients (pt). Pt with impaired ejection fraction (EF) and clinical manifestation of heart failure frequently initiates systemic pro-inflammatory state [SPS] characterized by interleukin-6 (IL6) and tumor necrosis factor alpha (TNF-α) upregulation. Previous studies suggest that such an inflammatory state is capable of sustaining and/or accelerating hemopoietic clonal selection/dominance. How these CHF induced inflammatory changes affect "peripheral blood cell count composition' in pt with suspected CHIP is unknown. However, modifications in peripheral blood count data "months before hemopoietic malignancy diagnosis" may inform association between clonal dynamics and hemopoietic output under CHF stress. The primary objective of our study was to investigate effect of heart failure in "pre- AML diagnosis" peripheral blood count data to detect "early" hemopoietic modifications induced by CHF. Methods: After IRB approval, AML pt diagnosed with and without CHF were selected for analysis. Given possible differential inflammatory effect between pt exhibiting CHF with and with low EF, we analyzed peripheral blood cell count in pt with "echocardiographically confirmed" EF <> 50% and those pt with and without clinically documented CHF [cdCHF]. Descriptive statistics was performed for categorical and continues variable with Chi-square and t-test. SAS software was used for analysis. Results: 27/152 (17.7%) and 125/152 (82.2%) pt developed or not EF <50%. Median age was 71.1 years (y) v 61.2 y, in pt with and without EF<50%, p=<0.0001. Aging was negatively correlated with EF, [R= -0.25, p=0.001]. Median age was 73 y v 61 y, for pt with and without cdCHF, p=0.001. 100% and 82% of pt were male among those with and without EF <50%. In pt who had or did not have EF <55% Favorable (fav), Intermediate (Int) and unfavorable (unfav) ELN-2017 subgroups were 3.23% vs 13.6%; 42% vs 46%; and 55% vs 40%, p=0.07. To address our main hypothesis that CHF induces "early pre-diagnostic AML' hemopoietic modifications, CBC data at 6 and 12 months (m) before AML onset in pt with and without EF<50% v those with and without cdCHF was extracted. At 12 m prior AML diagnosis, EF<> 50% detected differential expression for WBC [p=0.07], Hemoglobin [p=0.002], platelets [p=0.05] and absolute lymphocyte count (ALC) [p=0.006] [Fig 1, panel A, C, I, K]. Similarly, EF <>50% at 6 m prior AML diagnosis detected differential expression for platelets [p=0.01], ALC [p=0.04] [Fig 1, panel D, J, L]. In contrast, cdCHF detected only differential hemoglobin at 12 m [p=0.01]. Conclusions: CHF induces quantitative count defects 6 and 12 m before AML diagnosis. Changes are characterized by WBC, hemoglobin, platelets and ALC decline that are directly correlated with objective left ventricular EF impairment, but not with only clinical CHF. Our study adds body of evidence to support the role of CHF with low EF as "hemopoietic cell extrinsic stressor'. CHF associated with low EF seems to be prerequisite for hemopoietic stress in hosts with already initiated clonal hematopoiesis. Our findings have important experimental implications, especially for studies that seek to understand how cell extrinsic stressors facilitate clonal progression. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

Radiotherapy and High-Dose Interleukin-2: Clinical and Immunological Results of a Proof of Principle Study in Metastatic Melanoma and Renal Cell Carcinoma

High-dose interleukin-2 (HD IL-2) has curative potential in metastatic melanoma (MM) and renal cell carcinoma (RCC). Radiotherapy (RT) kills cancer cells and induces immunomodulatory effects. Prospective trials exploring clinical and immunological properties of combined RT/HD IL-2 are still needed. We designed a phase II, single-arm clinical trial for patients with MM and RCC. The treatment schedule consisted of 3 daily doses of 6-12 Gy of RT to 1-5 non-index metastatic fields, before IL-2 at the first and third treatment cycle. HD IL-2 was administered by continuous infusion for 72 hours and repeated every 3 weeks for up to 4 cycles, thereafter every 4 weeks for a maximum of 2 cycles. The primary endpoint was the immunological efficacy of the combined RT/HD IL-2 treatment (assessed by IFN-γ ELISPOT). Nineteen out of 22 patients were evaluable for immunological and clinical response. Partial response occurred in 3 (15.7%) patients and stable disease was observed in 7 (36.8%). The disease control rate was 52.6% after a median follow up of 39.2 months. According to Common Terminology Criteria for Adverse Events 4.0 (CTCAE 4.0), the majority of toxicities were grade 1-2. Immunological responses were frequent and detected in 16 (84.2%) patients. Increased levels of IL-8 and IL-10 in melanoma, circulating effector memory CD4+ and intratumoral CD8+ T cells in both tumor types were detected after therapy. Overall the treatment was well tolerated and immunologically active. Immunomonitoring and correlative data on tumor and peripheral blood cell subsets suggest that this combination treatment could be a promising strategy for patients progressing after standard treatments.

The Involvement of Macrophage Colony Stimulating Factor on Protein Hydrolysate Injection Mediated Hematopoietic Function Improvement

Protein hydrolysate injection (PH) is a sterile solution of hydrolyzed protein and sorbitol that contains 17 amino acids and has a molecular mass of 185.0–622.0 g/mol. This study investigated the effect of PH on hematopoietic function in K562 cells and mice with cyclophosphamide (CTX)-induced hematopoietic dysfunction. In these myelosuppressed mice, PH increased the number of hematopoietic cells in the bone marrow (BM) and regulated the concentration of several factors related to hematopoietic function. PH restored peripheral blood cell concentrations and increased the numbers of hematopoietic stem cells and progenitor cells (HSPCs), B lymphocytes, macrophages, and granulocytes in the BM of CTX-treated mice. Moreover, PH regulated the concentrations of macrophage colony stimulating factor (M-CSF), interleukin (IL)-2, and other hematopoiesis-related cytokines in the serum, spleen, femoral condyle, and sternum. In K562 cells, the PH-induced upregulation of hematopoiesis-related proteins was inhibited by transfection with M-CSF siRNA. Therefore, PH might benefit the BM hematopoietic system via the regulation of M-CSF expression, suggesting a potential role for PH in the treatment of hematopoietic dysfunction caused by cancer therapy.

Lymphocyte monocyte ratio is an effective and simple predictor for nosocomial influenza outbreaks

ObjectivesNosocomial influenza outbreak detection remains challenging. We evaluated the diagnostic utility of blood cell parameters, along with their capacity to differentiate between hospital acquired influenza and coronavirus disease 2019 (COVID-19).MethodsWe retrospectively analyzed patients diagnosed with nosocomial influenza from January 2017 to December 2019, and patients with COVID-19 in early 2020 at a tertiary teaching hospital in Beijing, China. We compared the differences between blood cell count and ratios (lymphocyte-to-monocyte ratio [LMR], neutrophil-to-lymphocyte ratio [NLR], lymphocyte-to-platelet ratio [LPR]) at symptom onset, before (admission), and after (recovery) nosocomial influenza. We also compared the abovementioned parameters between influenza and COVID-19 patients.ResultsLymphocyte count, LMR, and LPR were significantly lower in the symptom onset than in the admission and recovery groups (p < 0.001), while NLR was higher (p < 0.001). LMR and NLR exhibited similar and consistent tendencies among different subgroups of patients with nosocomial influenza (p < 0.001). The area under the receiver operating curve (AUC) of LMR, NLR, LPR, and lymphocyte count were 0.914, 0.872, 0.806, and 0.866, respectively. The optimal LMR cut-off value was 2.50, with specificity and sensitivity of 92.0% and 81.3%, respectively. Peripheral blood cell ratios can help diagnose nosocomial influenza significantly earlier than conventional methods. For differentiating influenza and COVID-19, the AUCs of LMR was 0.825.ConclusionsLMR effectively predicts nosocomial influenza outbreaks, particularly during the COVID-19 pandemic when simultaneous transmission can be a substantial threat.

Leukemic retinopathy, a rare entity: A case report

Chronic myelogenous leukemia (CML), also known as chronic myeloid leukemia, is a myeloproliferative disorder characterized by increased proliferation of the granulocytic cell line without the loss of their capacity to differentiate. Consequently, the peripheral blood cell profile shows an increased number of granulocytes and their immature precursors, including occasional blast cells. CML accounts for 20% of all leukemias affecting adults. Retinal lesions are the most common ocular manifestation of leukemia. They are found most often in adults and in patients with myeloid leukemia. Despite the significant efforts made by different groups to optimize treatment and outcome, there are still unmet needs and unanswered questions. Ophthalmologic manifestations are among the therapeutic challenge. Here we present a case of CML (chronic phase) with ophthalmologic manifestations as initial presentation, trying to shed light on this important type of presentation.

Comparison of the effects between MPL and JAK2V617F on thrombosis and peripheral blood cell counts in patients with essential thrombocythemia: a meta-analysis

To assess the effects between MPL and JAK2V617F on the thrombosis risk and peripheral blood cell counts in patients with essential thrombocythemia (ET), we identified eligible studies from PubMed, Embase, and the Cochrane Library. Seven studies were ultimately included in this meta-analysis. All studies reported the peripheral blood cell counts of ET patients, and three of them reported the eligible thrombotic events. In comparing the effect of MPL versus JAK2V617F on thrombosis, 1257 ET patients (73 MPL + and 1184 JAK2V617F +) were included. MPL-positive (MPL +) ET patients had a higher risk of thrombosis than JAK2V617F-positive (JAK2V617F +) ET patients [RR = 1.80 (1.08–3.01), P = 0.025]. And 3453 ET patients (138 MPL + and 3315 JAK2V617F +) were included in the comparison of peripheral blood cell counts. Platelet counts of MPL + ET patients were higher than that of JAK2V617F + ET patients [WMD = 81.18 (31.77–130.60), P = 0.001]. MPL + ET patients had lower hemoglobin [WMD = − 11.66 (− 14.32 to − 9.00), P = 0.000] and white blood cell counts [WMD = − 1.01 (− 1.47 to − 0.56), P = 0.000] than JAK2V617F + ET patients. These findings indicate that the MPL mutation is a high-risk factor for thrombosis in ET patients, and it may be rational to include MPL mutation in the revised IPSET as a criterion for thrombosis prediction scores. And given the differences in peripheral blood, it is necessary to further study whether MPL + ET patients differ from JAK2V617F + ET patients in bleeding and survival.