interleukin-8 and Graft-vs-Host-Disease

The role of cytokines during transplantation is currently an area of intense research. In particular, cytokines are increasingly appreciated as critical mediators of inflammatory responses to allogeneic tissue. Allogeneic bone marrow transplantation (BMT) involves cytokines not only in their capacity as hematopoietins but also as inducers of systemic inflammation after conditioning for BMT and during graft-versus- host disease (GVHD). GVHD is, in fact, a paradigm disease of systemic cytokine dysregulation, and the onset of severe acute GVHD can be considered a "cytokine storm." Advances in our understanding of the cytokine networks involved in BMT and GVHD should increase our ability to modify the complex interactions among cytokines and their cellular targets that lead to transplant-related morbidity.

Topics: Bone Marrow Transplantation; Cytokines; Graft vs Host Disease; Humans; Interferons; Interleukin-1; Interleukin-2; Interleukin-6; Interleukin-8; Tumor Necrosis Factor-alpha

There is accumulating evidence indicating that endothelial factors are involved in the pathogenesis of GVHD. We have recently shown that steroid-refractory, but not sensitive, GVHD is characterized by higher pretransplantation serum levels of angiopoetin-2 (ANG2), a hormone mediating endothelial vulnerability. To evaluate whether endothelial vulnerability is a risk factor for GVHD per se or becomes important only when noticeable GVHD is established, we measured ANG2 along with additional serum markers of endothelial stress, including soluble thrombomodulin (sTM), IL-8 (CXCL8), and hepatocyte growth factor (HGF), in patients with no, low-grade, or severe GVHD. Patients with refractory GVHD exhibited elevated serum levels of ANG2, sTM, HGF, and IL-8 posttransplantation compared with patients with sensitive GVHD and patients without GVHD. Pretransplantation ANG2 was the only growth factor correlated with the risk of refractoriness and mortality, and then only within the subset of patients who developed grade III-IV GVHD. In contrast, ANG2 was not predictive of GVHD or nonrelapse mortality (NRM) in patients with no GVHD or low-grade GVHD. These findings provide evidence that endothelial function plays an important role in the pathogenesis of steroid refractoriness in ongoing GVHD; however, endothelial vulnerability does not predict incidence of GVHD.

Topics: Adolescent; Adult; Aged; Endothelium, Vascular; Female; Glucocorticoids; Graft vs Host Disease; Hematologic Diseases; Hematopoietic Stem Cell Transplantation; Hepatocyte Growth Factor; Humans; Immunosuppressive Agents; Incidence; Interleukin-8; Male; Middle Aged; Mycophenolic Acid; Prednisolone; Retrospective Studies; Risk Factors; Thrombomodulin; Transplantation, Homologous; Vesicular Transport Proteins

To analyze and compare the tear immunological profile in ocular GVHD (oGVHD) patients with that in non-oGVHD patients and to correlate them with ocular surface parameters based on the International Chronic Ocular GVHD Consensus Group (ICCGVHD) diagnostic criteria.. Tear samples from 20 individuals who underwent allo-hematopoietic stem cell transplantation and were grouped according the presence or absence of oGVHD were analyzed using Bio-Plex assay.. IL-8 and MIP-1α levels were significantly higher in tears from oGVHD patients compared with those in tears from non-oGVHD patients (p<0.001 and p=0.001, respectively). Tear IL-8 levels correlated significantly with OSDI criteria (ρ=0.5159, p=0.001), ocular hyperemia (ρ=0.469, p=0.002), and corneal staining (ρ=0.339, p=0.032), whereas tear Mip-1α levels correlated with OSDI score (ρ=0.358, p=0.023).. We demonstrated higher tear levels of IL-8 and MIP-1α in oGVHD patients and significant correlations between theses cytokines and ocular surface parameters based on the ICCGVHDCG criteria.

Topics: Chemokine CCL3; Dry Eye Syndromes; Eye; Graft vs Host Disease; Humans; Interleukin-8; Tears

The local application of antiinflammatory and immunosuppressive agents is an effective method for the treatment of ocular graft-versus-host disease (oGVHD); however, we noticed that some patients with oGVHD did not respond to topical therapy as well as many others. This study aimed to determine whether tear cytokines were associated with therapeutic effects in oGVHD.. Forty patients with chronic oGVHD were enrolled and grouped as responders (n = 24) and nonresponders (n = 16) based on the clinical response to 1 month of topical treatment. Tear samples were collected from each participant before and after treatment, and the tear concentrations of 7 cytokines (IL-2, IL-6, IL-8, IL-10, IL-17A, TNF-α, and ICAM-1) were measured using microsphere-based immunoassay analysis. Differences between pretreatment and posttreatment tear samples were analyzed using the Wilcoxon test.. No significant differences in ophthalmic symptoms or cytokine levels were observed between responders and nonresponders at baseline. After 1 month of topical treatment, ocular surface parameters (including Ocular Surface Disease Index, National Institutes of Health eye score, best-corrected visual acuity, corneal fluorescein staining score, and fluorescein tear film break-up time) were significantly ameliorated in responders, but not in nonresponders. Moreover, none of the cytokines exhibited significant alteration in nonresponders, whereas the tear levels of IL-6 (P = 0.031) and IL-8 (P = 0.037) exhibited significant decreases in responding patients.. Our results revealed that tear IL-6 and IL-8 levels were significantly altered in response to topical oGVHD treatment.

Topics: Cytokines; Dry Eye Syndromes; Fluoresceins; Graft vs Host Disease; Humans; Interleukin-6; Interleukin-8; Tears

The purpose of this study was to analyze tear cytokine and complement levels in patients diagnosed with acute ocular graft-versus-host disease (oGVHD) and examine the consistency of these levels with the severity of clinical manifestations.. Ten patients with acute oGVHD (20 eyes) were enrolled for the assessment of tear cytokine levels and ocular surface parameters, and 18 healthy people (36 eyes) were selected as the control group. The tear cytokine and complement levels were measured using microsphere-based immunoassay analysis.. The main clinical manifestations of acute oGVHD include eye redness, a large amount of purulent exudate, eye pain, and even false membranes. The levels of intercellular cell adhesion molecule-1, interleukin 6 (IL-6), interleukin 1 beta (IL-1β), interleukin 8, epidermal growth factor (EGF), interleukin 7 (IL-7), B-cell activating factor, granulocyte-macrophage colony-stimulating factor (GM-CSF), and complement in patients with acute oGVHD showed significant differences compared with those in normal people. Furthermore, the levels of IL-6, IL-1β, EGF, GM-CSF, IL-7, and C3a showed a stronger correlation with ocular surface parameters.. Our study was the first to enroll patients with acute oGVHD to assess tear cytokine levels as a method contributing to the diagnosis of acute oGVHD. In addition, it has been demonstrated that certain tear cytokines, including intercellular cell adhesion molecule-1, IL-6, IL-1β, interleukin 8, B-cell activating factor, GM-CSF, IL-7, EGF, and complement, may be new diagnostic biomarkers for acute oGVHD.

Topics: B-Cell Activating Factor; Biomarkers; Cell Adhesion Molecule-1; Cytokines; Epidermal Growth Factor; Graft vs Host Disease; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-1beta; Interleukin-6; Interleukin-7; Interleukin-8; Tears

The purpose of this study was to analyze inflammation- and pain-related molecules in tears of patients suffering from chronic ocular pain associated with dry eye (DE) and/or a previous corneal refractive surgery (RS). Based on history, symptomatology, and clinical signs, the subjects (n = 180, 51.0 ± 14.7 years, 118 females, 62 males) in this cross-sectional study were assigned to one of five groups: DE and chronic ocular pain after RS (P/DE-RS, n = 52); asymptomatic subjects, i.e., without DE and chronic ocular pain, after RS (A-RS, n = 30); DE and chronic ocular pain without previous RS (P/DE-nonRS, n = 31); DE, no pain, and no previous RS (DE-nonRS, n = 35); and asymptomatic subjects with no previous RS (controls, n = 32). The tear concentrations of 20 cytokines and substance P (SP) were analyzed by immunobead-based assay and enzyme-linked immunosorbent assay, respectively. We found that tear levels of interleukin (IL)-10 and SP were increased in the RS groups. There were significant differences in IL-8/CXCL8 among the five groups. Nerve growth factor (NGF) tear levels were significantly higher in P/DE-RS than in DE-nonRS and controls. IL-9 had the highest percentage of detection in the P/DE-RS and P/DE-nonRS groups, while macrophage inflammatory protein (MIP)-1α, IL-2, and interferon (IFN)-γ were higher in the P/DE-RS, A-RS, and P/DE-nonRS groups. IL-17A was detected only in the A-RS group. Moderate correlations were observed in the A-RS, P/DE-nonRS, DE-nonRS and controls groups. A positive correlation was obtained between growth related oncogene concentration and tear break-up time (rho = 0.550; p = 0.012), while negative correlation was found between monocyte chemoattractant protein-3/CCL7 and conjunctival staining (rho = -0.560; p = 0.001), both in the A-RS group. IL-10 correlated positively with ocular pain intensity (rho = 0.513; p = 0.003) in the P/DE-nonRS group. Regulated on Activation Normal T Cell Expressed and Secreted/CCL5 correlated negatively with conjunctival staining (rho = -0.545; p = 0.001) in the DE-nonRS group. SP correlated negatively with corneal staining (rho = -0.559; p = 0.001) in the controls. In conclusion, chronic ocular pain was associated with higher IL-9 tear levels. IL-10, SP, MIP-1α/CCL3, IL-2, and IFN-γ were associated with previous RS. Higher levels of IL-8/CXCL8, MIP-1α/CCL3, IL-2, and IFN-γ were associated with DE-related inflammation, while NGF levels were related to chronic ocular pain and DE in RS patients. Thes

Topics: Chemokine CCL3; Conjunctiva; Cross-Sectional Studies; Cytokines; Dry Eye Syndromes; Female; Graft vs Host Disease; Humans; Inflammation; Interleukin-10; Interleukin-2; Interleukin-8; Interleukin-9; Male; Nerve Growth Factor; Pain; Tears

Mesenchymal stem cells (MSC) are characterized by tolerogenic potential and therefore, are used in the treatment of autoimmune diseases such as graft-versus-host disease (GVHD) reactions after allogeneic hematopoietic cell transplantation to improve the transplant functions, as well as for the therapy and prevention of cytokine storm in COVID-19 patients and some other conditions. However, MSC can exhibit proinflammatory activity, which causes risks for their clinical use. We studied the cytokine profile of bone marrow MSC culture and demonstrate intensive production of IL-6, IL-8, and chemokine MCP-1, which participate in the pathogenesis of cytokine storm and GVHD. At the same time, no anti-inflammatory IL-4 and IL-10 were detected. To reduce the risks of MSC application in the GVHD therapeutic protocols, further studies of the conditions promoting generation of MSC with tolerogenic potential and approved clinical standards of MSC use are required.

Topics: Bone Marrow Cells; Cells, Cultured; Chemokine CCL2; COVID-19; Cytokine Release Syndrome; Cytokines; Graft vs Host Disease; Hematopoietic Stem Cell Transplantation; Humans; Immunosuppressive Agents; Interleukin-6; Interleukin-8; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; SARS-CoV-2; Transplantation, Homologous

Transplant-associated thrombotic microangiopathy (TA-TMA) is a common and poorly recognized complication of hematopoietic stem cell transplantation (HSCT) associated with excessive complement activation, likely triggered by endothelial injury. An important missing piece is the link between endothelial injury and complement activation. We hypothesized that neutrophil extracellular traps (NETs) mechanistically link endothelial damage with complement activation and subsequent TA-TMA. Neutrophil activation releases granule proteins together with double-stranded DNA (dsDNA) to form extracellular fibers known as NETs. NETs have been shown to activate complement and can be assessed in humans by quantification of dsDNA in serum. We measured levels of dsDNA, as a surrogate for NETs in 103 consecutive pediatric allogeneic transplant recipients at day 0, +14, +30, +60, and +100. A spike in dsDNA production around day +14 during engraftment was associated with subsequent TA-TMA development. Peak dsDNA production around day +14 was associated with interleukin-8-driven neutrophil recovery. Increased dsDNA levels at days +30, +60, and +100 were also associated with increased mortality and gastrointestinal graft-versus-host disease (GVHD). NETs may serve as a mechanistic link between endothelial injury and complement activation. NET formation may be one mechanism contributing to the clinical overlap between GVHD and TA-TMA.

Topics: Adolescent; Adult; Child; Child, Preschool; Complement Activation; Demography; DNA; Endothelial Cells; Extracellular Traps; Graft vs Host Disease; Hematopoietic Stem Cell Transplantation; Humans; Infant; Interleukin-8; Longitudinal Studies; Neutrophils; Receptors, Interleukin-8A; Receptors, Interleukin-8B; Thrombotic Microangiopathies; Treatment Outcome; Young Adult

While cord blood transplantation (CBT) is an effective therapy for hematologic malignancies, acute graft-versus-host disease (aGVHD) is a leading cause of transplant-related mortality (TRM). We investigated if biomarkers could predict aGVHD and TRM after day 28 in CBT recipients. Day 28 samples from 113 CBT patients were analyzed. Suppressor of tumorigenicity 2 (ST2) was the only biomarker associated with grades II-IV and III-IV aGVHD and TRM. Day 180 grade III-IV aGVHD in patients with high ST2 levels was 30% (95% confidence interval [CI], 18-43) vs 13% (95% CI, 5-23) in patients with low levels (P = .024). The adverse effect of elevated ST2 was independent of HLA match. Moreover, high day 28 ST2 levels were associated with increased TRM with day 180 estimates of 23% (95% CI, 13-35) vs 5% (95% CI, 1-13) if levels were low (P = .001). GVHD was the most common cause of death in high ST2 patients. High concentrations of tumor necrosis factor receptor-1, interleukin-8, and regenerating islet-derived protein 3-α were also associated with TRM. Our results are consistent with those of adult donor allografts and warrant further prospective evaluation to facilitate future therapeutic intervention to ameliorate severe aGVHD and further improve survival after CBT.

Topics: Adolescent; Adult; Aged; Biomarkers; Child; Child, Preschool; Cord Blood Stem Cell Transplantation; Female; Graft vs Host Disease; Hematologic Neoplasms; HLA Antigens; Humans; Infant; Interleukin-1 Receptor-Like 1 Protein; Interleukin-8; Male; Middle Aged; Neutrophils; Receptors, Cell Surface; Receptors, Tumor Necrosis Factor, Type I; Retrospective Studies; Time Factors; Transplantation Conditioning; Young Adult

No predictive factors are currently available to establish patient-specific GVHD risk. A panel of six serum cytokines (TNF receptor 1, IL-2 receptor alfa (IL-2Rα), hepatocyte growth factor (HGF), monocyte chemo-attractant protein-2, IL-8, IL-12p70) were monitored at established time points (days -1, +1, +7, +14, +21, +28 and +60) in 170 paediatric hematopoietic SCT (HSCT) recipients. We found that higher concentrations of IL-2Rα on days +14 and +21 together with HGF on days +14 and +21 were significantly associated at a higher probability of both grade II-IV GVHD (on day +14 it was: 60% vs 28%, P=0.007) and grade III-IV (on day +14 it was: 40% vs 15%, P=0.001). The higher IL-8 serum concentration on day +28 was associated with a lower probability of chronic GVHD being 4% vs 29% (P=0.01) for patients with higher vs lower IL-8 serum concentration. These findings were confirmed when the analysis was restricted to the the matched unrelated donor group. In conclusion, even if the serum cytokine levels were related to several variables associated with HSCT, we identified two cytokines as predictors of GVHD II-IV and III-IV, translating into a higher TRM risk (17% vs 3%, P=0.004).

Topics: Adolescent; Adult; Biomarkers, Tumor; Child; Child, Preschool; Cytokines; Female; Graft vs Host Disease; Hematopoietic Stem Cell Transplantation; Hepatocyte Growth Factor; Humans; Infant; Infant, Newborn; Interleukin-12; Interleukin-2 Receptor alpha Subunit; Interleukin-8; Male; Neoplasms; Prognosis; Receptors, Tumor Necrosis Factor, Type I; Transplantation Conditioning; Transplantation, Homologous; Young Adult

Acute graft-versus-host disease (GVHD) is the primary limitation of allogeneic hematopoietic cell transplantation, and once it develops, there are no reliable diagnostic tests to predict treatment outcomes. We hypothesized that 6 previously validated diagnostic biomarkers of GVHD (IL-2 receptor-α; tumor necrosis factor receptor-1; hepatocyte growth factor; IL-8; elafin, a skin-specific marker; and regenerating islet-derived 3-α, a gastrointestinal tract-specific marker) could discriminate between therapy responsive and nonresponsive patients and predict survival in patients receiving GVHD therapy. We measured GVHD biomarker concentrations from samples prospectively obtained at the initiation of treatment, day 14, and day 28, on a multicenter, randomized, 4-arm phase 2 clinical trial for newly diagnosed acute GVHD. We found that at each of 3 time points, GVHD onset, 2 weeks into treatment, and 4 weeks into treatment, a 6-protein biomarker panel predicted for the important clinical outcomes of day 28 posttherapy nonresponse and mortality at day 180 from onset. GVHD biomarker panels can be used for early identification of patients at high or low risk for treatment nonresponsiveness or death, and they may provide opportunities for early intervention and improved survival after hematopoietic cell transplantation. The study was registered in clinicaltrials.gov as NCT00224874.

Topics: Acute Disease; Adult; Aged; Biomarkers; Bone Marrow Transplantation; Elafin; Female; Graft vs Host Disease; Hepatocyte Growth Factor; Humans; Immunosuppressive Agents; Interleukin-8; Logistic Models; Male; Middle Aged; Pancreatitis-Associated Proteins; Predictive Value of Tests; Prognosis; Proteins; Randomized Controlled Trials as Topic; Receptors, Interleukin-2; Tumor Necrosis Factor-alpha; Young Adult

No validated biomarkers exist for acute graft-versus-host disease (GVHD). We screened plasma with antibody microarrays for 120 proteins in a discovery set of 42 patients who underwent transplantation that revealed 8 potential biomarkers for diagnostic of GVHD. We then measured by enzyme-linked immunosorbent assay (ELISA) the levels of these biomarkers in samples from 424 patients who underwent transplantation randomly divided into training (n = 282) and validation (n = 142) sets. Logistic regression analysis of these 8 proteins determined a composite biomarker panel of 4 proteins (interleukin-2-receptor-alpha, tumor-necrosis-factor-receptor-1, interleukin-8, and hepatocyte growth factor) that optimally discriminated patients with and without GVHD. The area under the receiver operating characteristic curve distinguishing these 2 groups in the training set was 0.91 (95% confidence interval, 0.87-0.94) and 0.86 (95% confidence interval, 0.79-0.92) in the validation set. In patients with GVHD, Cox regression analysis revealed that the biomarker panel predicted survival independently of GVHD severity. A panel of 4 biomarkers can confirm the diagnosis of GVHD in patients at onset of clinical symptoms of GVHD and provide prognostic information independent of GVHD severity.

Topics: Acute Disease; Adolescent; Adult; Aged; Biomarkers; Child; Child, Preschool; Disease-Free Survival; Female; Graft vs Host Disease; Hematopoietic Stem Cell Transplantation; Hepatocyte Growth Factor; Humans; Infant; Interleukin-2 Receptor alpha Subunit; Interleukin-8; Male; Middle Aged; Predictive Value of Tests; Receptors, Tumor Necrosis Factor, Type I; Retrospective Studies; Survival Rate; Transplantation, Homologous

The principal cause of mortality and morbidity following hematopoietic cell transplantation (HCT) is graft-versus-host disease (GVHD). Studies in murine models have revealed that inflammatory mediators such as tumor necrosis factor-alpha (TNF-alpha) promote destruction of host tissue following HCT. Elevated plasma levels of soluble TNF receptor 1 have been associated with patients with GVHD and blocking TNF-alpha in experimental models has shown a reduced incidence of GVHD. Based on this finding, patients with new onset GVHD were treated with steroids plus the TNF-alpha inhibitor, etanercept, on a previously reported pilot trial (n=20) and a phase 2 trial (n=41) and their outcomes were compared with those of contemporaneous patients with GVHD (n=99) whose initial therapy was steroids alone. Patients treated with etanercept were more likely to achieve CR than were patients treated with steroids alone. Plasma TNFR1 levels, a biomarker for GVHD activity, were elevated at GVHD onset and decreased significantly only in patients with CR. A four protein fingerprint of IL-2Ralpha, TNFR1, IL-8, and HGF in the plasma has been identified to predict whether a patient will be at high-risk for GVHD based on biomarker analysis. In univariate and multivariate analysis, this four-protein fingerprint has shown a strong association with the grade of acute GVHD, and it can stratify patients into low- and high-risk groups and can be used as a laboratory-based screening method, to diagnose and perhaps treat patients preemptively.

Topics: Animals; Biomarkers; Disease Management; Etanercept; Graft vs Host Disease; Hematopoietic Stem Cell Transplantation; Hepatocyte Growth Factor; Humans; Immunoglobulin G; Interleukin-8; Receptors, Interleukin-2; Receptors, Tumor Necrosis Factor; Receptors, Tumor Necrosis Factor, Type I; Steroids

The purpose of this study was to explore the roles of cytokines IL-2, TNF-alpha, IL-10 and IL-8 in the pathogenesis of acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation. The incidence of aGVHD was observed in 33 patients undergoing allogeneic hematopoietic stem cell transplantation. The aGVHD was clinically diagnosed. Sera from the 33 patients were taken before and after allogeneic hematopoietic stem cell transplantation. The IL-2, TNF-alpha, IL-8, IL-10 levels in serum of 33 patients were measured serially by using radioimmuno-assay (RIA). aGVHD occurred in 13 patients including 8 patients with aGVHD I and 5 patients with aGVHD II-IV. The results showed that the circulating levels of IL-2 and TNF-alpha were markedly elevated during aGVHD and strongly correlated with the severity of aGVHD as compared with patients without aGVHD. However, the level of the IL-10 in patients with aGVHD was significantly lower than that in patients without aGVHD. The change of IL-8 level was not significant statistically. It is concluded that IL-2 and TNF-alpha may play important roles in the pathogenesis of aGVHD, and measurement of serum IL-2 and TNF-alpha levels after allo-HSCT can provide predictive indicator for acute GVHD.

Topics: Adolescent; Adult; Female; Graft vs Host Disease; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Humans; Interleukin-10; Interleukin-2; Interleukin-8; Male; Middle Aged; Tumor Necrosis Factor-alpha

Serum levels of interleukin-6 (IL-6), interleukin-8 (IL-8) and tumor necrosis factor (TNF)-alpha were frequently measured during the first 30 days after allogeneic bone marrow transplantation (BMT) in 84 consecutive adult patients. Major transplant-related complications (MTCs) occurred in 33% of cases and included veno-occlusive liver disease, idiopathic pneumonia syndrome, severe endothelial leakage syndrome and >grade II acute graft-versus-host disease. Compared with patients having minor complications, those with MTCs developed higher levels at times of maximal clinical signs (all cytokines, P<0.001), between days 0-5 post-BMT (IL-6 and IL-8, P<0.05) and days 6-10 (L-6, P<0.001; IL-8 and TNF, P<0.01) post-BMT. We could not discriminate patterns of cytokine release that were specific for any subtype of MTC. Higher levels of IL-8 during days 0-5 were associated (P=0.044) with early (<40 days) death. Multivariate analysis including patient and transplant characteristics as well as post-BMT levels of C-reactive protein showed that high average levels of one or more of the cytokines within the first 10 days post-BMT were independently associated with MTC (Odd's ratio: 2.3 [1.2-4.5], P=0.011). This study shows that systemic release of proinflammatory cytokines contributes to the development of MTC and provides a rationale for pre-emptive anti-inflammatory treatment in selected patients.

Topics: Adult; Bacteremia; Bone Marrow Transplantation; C-Reactive Protein; Capillary Leak Syndrome; Female; Graft vs Host Disease; Hepatic Veno-Occlusive Disease; Humans; Interleukin-6; Interleukin-8; Leukemia; Male; Neural Tube Defects; Pneumonia; Risk Factors; Transplantation Conditioning; Transplantation, Homologous; Tumor Necrosis Factor-alpha

The presence of WBCs in blood components is the major factor influencing the immunologic consequences of transfusion. Attempts to ameliorate these responses have used WBC reduction or inactivation by ionizing radiation. PEN110 (Inactine, V. I. Technologies) is a chemical that inhibits the replication of infectious pathogens by modifying their nucleic acids. These experiments compared effects of PEN110 treatment or gamma irradiation on WBC function.. Aliquots of non-WBC-reduced RBC units were treated with PEN110 or gamma irradiation with appropriate controls, and PBMNCs from these units were tested with in vitro assays. The assays included immunophenotyping, activating T cells with phorbol ester, proliferation of cells in response to mitogens or allogeneic cells, and determining the ability of cells to stimulate proliferative responses and to produce IL-8. DNA fragmentation following PEN110 treatment was examined by PCR and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling assays.. Treatment of non-WBC-reduced RBC units with PEN110 functionally inactivated WBC in all in vitro assays used. In contrast, while gamma irradiation inhibited proliferation of the WBCs, it did not or only partially inhibited the ability of WBC to function in the other assays. PEN110, but not gamma irradiation, rapidly induced fragmentation of cellular DNA.. Because PEN110 was as effective as gamma irradiation at inhibiting WBC proliferation, PEN110 treatment could potentially be used to prevent the development of GVHD following transfusion.

Topics: Antigens, CD; Antigens, Differentiation, T-Lymphocyte; Cell Division; DNA Fragmentation; Erythrocyte Transfusion; Gamma Rays; Graft vs Host Disease; Humans; Interleukin-8; Lectins, C-Type; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Neutrophils; Polyamines

Pro-inflammatory (IL-6, TNFalpha and IL-8) and anti-inflammatory (IL-10) cytokines were determined in weekly samples from 52 patients undergoing allogeneic hematopoietic stem cell transplantation (HSCT). IL-6 increased immediately after transplant peaking at week +3, but IL-8 concentrations were elevated only during week +1. After a slight decrease in week +1, TNF-alpha significantly increased from week +2 and peaked at week +3, whereas, IL-10 values started to rise in week +2 and peaked during week +4. IL-6 and TNF-alpha were positively correlated from week +2 to week +4, and IL-6 levels at week +1 were related with fever and severe stomatitis. Serum levels of IL-6 at week +1 and IL-10 at week +4 were significantly higher in patients with early transplant-related complications, such as fever, severe stomatitis or acute GVHD > or = overall grade II than in those without the complications. We conclude that a high serum IL-6 level at week +1 may be an early predictor of transplant-related complications and that it seems to trigger pro- and anti-inflammatory cytokine release. Kinetic patterns of IL-6 and IL-10 were more exaggerated in those with complications after HSCT.

Topics: Acute Disease; Adult; Biomarkers; Cytokines; Female; Fever; Graft vs Host Disease; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Humans; Interleukin-10; Interleukin-6; Interleukin-8; Kinetics; Male; Middle Aged; Mouth Mucosa; Prognosis; Stomatitis; Transplantation, Homologous; Tumor Necrosis Factor-alpha

Acute graft-versus-host disease (aGVHD) is one of the major complications after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Prediction of human aGVHD may be of help in its diagnosis and therapy.. 22 patients undergoing HSCT were included in this study. After transplantation, serum concentrations of interleukin-6, interleukin-8, tumor necrosis factor-alpha, interferon-gamma and sIL-2R were measured periodically by using enzyme-linked immunosorbent assay. Immune reconstitution of CD(+)(3), CD(+)(4), CD(+)(8), CD(+)(25) and CD(+)(69) cells were analyzed with flow cytometry.. All the patients achieved engraftment. 6 patients developed grade I GVHD and 4 patients developed grade III-IV GVHD. Patients with aGVHD demonstrated significantly higher sIL-2R levels than those without. The peak levels of grade III-IV GVHD were (420.3 +/- 59.8) U/L,and the peak levels of grade I GVHD were (221.5 +/- 38.8) U/L. The increase of sIL-2R level in serum preceded the clinical signs of aGVHD. The maximum levels of sIL-2R correlated significantly with the severity of aGVHD. The increase of CD(+)(25) cells was in accordance with that of sIL-2R.. Analysis of serum cytokines and immune reconstitution after HSCT may provide predictive value for aGVHD.

Topics: Adolescent; Adult; Child; Graft vs Host Disease; Hematopoietic Stem Cell Transplantation; Humans; Interferons; Interleukin-6; Interleukin-8; Leukemia; Middle Aged; Receptors, Interleukin-2; T-Lymphocytes; Tumor Necrosis Factor-alpha

A photochemical treatment (PCT) process using a novel psoralen and long wavelength ultraviolet light (UVA, 320-400 nm) has been developed to inactivate bacteria and viruses in platelet concentrates. This study evaluated the efficacy of PCT for inactivation of leukocytes that contaminate platelet preparations. Three psoralens, 8-methoxypsoralen (8-MOP), 4'-aminomethyl 4,5', 8-trimethylpsoralen (AMT), and the novel psoralen S-59, were compared using the following four independent but complementary biological and molecular assays. (1) T-cell viability: Treatment with 150 mumol/L S-59 and 1.0 to 3.0 Joules/cm2 UVA inactivated >5.4 +/- 0.3 log10 of T cells in full-sized single-donor plateletpheresis units. Using 1.0 Joule/cm2 UVA, the lowest dose of S-59, AMT and 8-MOP required to reduce the number of T cells to the limit of detection was 0.05 micromol/L, 1.0 micromol/L, and 10.0 micromol/L, respectively. (2) Cytokine synthesis: Treatment with 1.9 Joules/cm2 UVA and 150 micromol/L S-59 or AMT completely inhibited synthesis of the cytokine IL-8 by contaminating leukocytes during 5 days of platelet storage. After treatment with 75 micromol/L 8-MOP and 1.9 Joules/cm2 UVA, only low levels of IL-8 were detected. (3) Psoralen-DNA adduct formation: The combination of 1.9 Joules/cm2 UVA and 150 micromol/L S-59, AMT, or 8-MOP induced 12.0 +/- 3.0, 6.0 +/- 0. 9, and 0.7 psoralen adducts per 1,000 bp DNA, respectively. (4) Replication competence: Polymerase chain reaction (PCR) amplification of small genomic DNA sequences (242-439 bp) after PCT was inhibited. The degree of PCR amplification inhibition correlated with the level of adduct formation (S-59 > AMT > 8-MOP). In contrast, 2,500 cGy gamma radiation, a dose that inactivates >5 log10 of T cells in blood products, had minimal effect on cytokine synthesis and did not induce sufficient DNA strand breaks to inhibit PCR amplification of the same small DNA sequences. These results demonstrate that leukocytes are sensitive to PCT with psoralens and among the psoralens tested S-59 is the most effective. Therefore, PCT has the potential to reduce the incidence of leukocyte-mediated adverse immune reactions associated with platelet transfusion.

Topics: Adult; DNA Adducts; DNA Damage; Gamma Rays; Graft vs Host Disease; Humans; Interleukin-8; Leukocyte Count; Leukocytes; Methoxsalen; Photochemistry; Photosensitizing Agents; Platelet Transfusion; Plateletpheresis; Polymerase Chain Reaction; T-Lymphocytes; Trioxsalen; Ultraviolet Rays

The relationship between cytokine concentrations and transplant-related complications has been studied in bone marrow transplant patients. The changes in TNF-alpha, IL-1 and IL-6 concentrations after transplantation are well documented in the literature but this is not the case for IL-8. The purpose of the present study was to investigate prospectively the plasma concentration of these cytokines and their relationship to transplant-related complications.. Pro-inflammatory cytokine (TNF-alpha, IL-1, IL-6 and IL-8) levels in plasma were determined in a group of 53 patients undergoing hematopoietic progenitor transplantation. Plasma samples were collected weekly from day -7 to day +35 and stored at -70 degrees C until assayed by ELISA. The major transplant-related toxicities registered were: veno-occlusive disease (VOD), acute graft-versus-host disease (GVHD), infectious episodes, renal failure and mucositis.. In spite of the great variability of plasma cytokine profiles between the different patients, we came to various conclusions. Patients' TNF-alpha and IL-1 concentrations correlated well over time. IL-6 and IL-8 profiles were similar and correlated well with febrile episodes. In some cases, an increase in IL-6 preceded hematologic recovery. In our study, increased levels of TNF-alpha, IL-6 and especially IL-8 correlated with hepatic or renal dysfunction as evaluated by increased bilirubin and creatinine in plasma, while pulmonary complications correlated only with increased IL-6 levels. Allogeneic transplant patients had a tendency to have higher TNF-alpha concentrations than autologous transplant patients, probably because an allogeneic transplant is associated with more transplant-related toxicity. Basal disease usually had no effect on cytokine profiles.. IL-6 and IL-8 were the only cytokines studied whose increase correlated with febrile episodes. High IL-8 values may be a useful predictor of renal dysfunction and pulmonary disease and seems to trigger off high IL-6 levels. Plasma TNF-alpha and IL-1 concentrations during the posttransplant period have not been shown to be predictive of the development of transplant-related complications, and none of the profiles was recognized to be specific for a particular complication in this study.

Topics: Adolescent; Adult; Biomarkers; Bone Marrow Transplantation; Communicable Diseases; Female; Graft vs Host Disease; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Hepatic Veno-Occlusive Disease; Humans; Interleukin-6; Interleukin-8; Male; Middle Aged; Prospective Studies; Renal Insufficiency; Transplantation, Autologous; Transplantation, Homologous

Several recent studies have reported both the generation of cytokines, including interleukin (IL)-1 beta, IL-6, tumor necrosis factor alpha (TNF-alpha), and IL-8, in the supernatants of stored platelet concentrates (PCs) and the implications of this generation in febrile nonhemolytic transfusion reactions. Prestorage filtration is regarded as highly effective in the prevention of cytokine generation.. Studies evaluated 1) the levels of these cytokines in apheresis PCs during storage, 2) the effects of white cell inactivation by ultraviolet B or gamma-radiation on the generation of cytokines, and 3) the effects of poststorage filtration on cytokine levels. The apheresis PCs were treated by either ultraviolet B radiation (20,000 J/m2), gamma-radiation (30 Gy), or filtration. Samples were collected sequentially on various days after storage. Cytokines were determined by enzyme-linked immunosorbent assay.. The average white cell count in 15 PCs tested was 2.58 +/- 0.7 x 10(6) per mL (range, 0.7-10 x 10(6)/mL). A detectable level of IL-8 was found at 3 days of storage, and the levels of this cytokine increased progressively with increasing storage time, ranging from 1.6 to 35,280 pg per mL on Day 5 and from 2.7 to 83,601 pg per mL on Day 8. Reverse transcriptase-polymerase chain reaction analysis showed that the level of IL-8 paralleled the expression of IL-8 transcripts. The levels of IL-1 beta, IL-6, TNF-alpha, and monocyte chemotactic protein-1 were very low, even on Day 8. Ultraviolet B-radiated PCs failed to generate IL-8, even at 8 days of storage, whereas levels of IL-8 in gamma-radiated PCs were similar to those in nonirradiated PCs. Poststorage filtration of PCs with a negatively charged polyester filter, but not with a positively charged one, markedly reduced the levels of IL-8.. Of the cytokines tested, IL-8 had the most evident generation in apheresis PCs during storage. Prestorage inactivation of white cells by ultraviolet B radiation, but not by gamma-radiation, was effective in preventing the generation of cytokines during the storage of PCs.

Topics: Anaphylatoxins; Anemia, Hemolytic; Blood Platelets; Blood Preservation; Filtration; Gamma Rays; Graft vs Host Disease; Humans; Interleukin-8; Plateletpheresis; RNA, Messenger; Time Factors; Transfusion Reaction; Ultraviolet Rays

Serum levels of interleukin-8 (IL-8) were investigated in the perioperative phase of liver transplantation (LTx) in order to help determine whether this cytokine might serve as a parameter for preservation injury. In a study of 45 patients undergoing LTx, systemic IL-8 was estimated at the end of the anhepatic phase, at 30, 60, and 120 min after reperfusion of the graft, and 24 h and 7 days after LTx. A maximum mean concentration of 665 +/- 135 pg/ml was seen 60 min after LTx. The minimum was found on the 1st postoperative day (POD 1): 328 +/- 33 pg/ml. Significant changes were found between 60 min and PODs 1 and 7, as well as between 120 min and POD 1. Differences in cold ischemia time were not found to be significant. We conclude that monitoring of systemic IL-8 levels is not useful in the development of new liver preservation concepts.

Topics: Adult; Cyclosporine; Female; Graft vs Host Disease; Humans; Immunosuppressive Agents; Interleukin-8; Liver; Liver Transplantation; Male; Middle Aged; Postoperative Period; Tacrolimus

We investigated the cytokine profile and peak levels of interleukin (IL) -6, IL-8, IL-10 and tumour necrosis factor (TNF) -alpha levels in 42 patients after allogeneic bone marrow transplantation (BMT). Eleven of them developed veno-occlusive disease (VOD) of the liver. Fourteen patients had moderate-to-severe acute graft-versus-host disease (aGvHD), 10 isolated bacteraemia and 7 had no major complication. Those who developed severe VOD (n=6) showed a short, very high IL-8 peak (median: 6632 pg/ml, range: 5546-10,000 vs. 280 pg/ml, 0-2042 in controls, p<0.01) 1-4 d after diagnosis of the liver disease. Five of these patients had high peak levels of IL-6. Five patients with mild VOD showed a lower increase in the cytokines tested. Bilirubin levels, at day of IL-8 peak, did not differ statistically between mild and severe VOD. The highest levels of IL-10 were found in those with aGvHD. IL-8 levels were also increased, but not to the same extent as in patients with severe VOD (p=0.01 vs. VOD). In patients with bacteraemia, very high levels of IL-6 were seen. In patients without major complications, the levels of cytokines were low. In conclusion, high levels of IL-8 occurred in severe VOD of the liver, which may be of value to determine prognosis.

Topics: Adolescent; Adult; Bacteremia; Biomarkers; Bone Marrow Transplantation; Child; Female; Graft vs Host Disease; Hepatic Veno-Occlusive Disease; Humans; Interleukin-6; Interleukin-8; Male; Prognosis; Transplantation, Homologous

Neutrophil chemotactic and functional defects occur in beta-thalassemia and in patients after bone marrow transplantation (BMT). Interleukin-8 (IL-8) is a novel chemotactic and activating peptide for neutrophils and can be detected in the circulation. IL-8 serum concentrations were evaluated in 30 beta-thalassemic patients before and after BMT. Serial samples from 16 patients were also studied. Fourteen sera from healthy children, 43 patients with chronic viral hepatitis, 16 patients on chronic transfusion treatment for various hematologic disorders, and 28 healthy adults were studied as controls. IL-8 was evaluated by an enzyme-linked immunosorbent assay. Patients with beta-thalassemia had higher IL-8 concentrations than did normal controls, patients with liver disease, and patients on chronic transfusion. beta-Thalassemic patients with severe liver siderosis and fibrosis had the highest IL-8 concentrations. After BMT in patients with successful engraftment, IL-8 concentrations decreased significantly. In contrast, in patients with acute graft-versus-host disease (GVHD), IL-8 concentrations were not statistically different from the concentrations found before BMT and were higher than in patients with no complications and patients with graft rejection. IL-8 may play a part in the immune dysregulation that occurs in beta-thalassemia and may be involved in the immune mechanisms leading to GVHD.

Topics: Adolescent; Adult; beta-Thalassemia; Biomarkers; Biopsy; Blood Transfusion; Bone Marrow Transplantation; Child; Child, Preschool; Enzyme-Linked Immunosorbent Assay; Female; Graft vs Host Disease; Hepatitis, Viral, Human; Humans; Interleukin-8; Liver; Male; Reference Values; Time Factors