scholarly journals P423 NF-kB as a prognostic marker of response to biologic therapy in children with IBD

2020 ◽  
Vol 14 (Supplement_1) ◽  
pp. S386-S387
Author(s):  
T Radigina ◽  
A Illarionov ◽  
D Kuptsova ◽  
A Potapov ◽  
S Petrichuk ◽  
...  
Keyword(s):  
Nk Cells ◽  
Statistical Evaluation ◽  
Biologic Therapy ◽  
Positive Outcome ◽  
Healthy Children ◽  
Innate And Adaptive Immunity ◽  
Tnf Blockers ◽  
Patient Status ◽  
Endoscopic Remission ◽  
Lymphocyte Populations

Abstract Background The transcription factor NF-kB is a regulator of innate and adaptive immunity through the activation of various pro and anti-inflammatory mediators. The aim of this study was to assess the prognostic value of changes in the level of NF-kB translocation in lymphocyte populations in response to infusion of TNF blockers in children with IBD. Methods There were examined 44 children with IBD (27-CD, 17-UC) and 20 conditionally healthy children aged 3–18 years. Patient status (exacerbation, remission) was assessed by the PUCAI (UC) and PCDIA (CD) indices. There were determined the percentage of cells with NF-kB translocation in populations CD3+CD4+ (Th), CD3+CD8+(Tc), CD3-CD19+ (B cells), CD3-CD16 / 56+(NK cells), CD3+CD4+CD161+ (Th17), CD3+CD4+CD25highCD127low (Tregs) with NF-kB translocation kit using flow cytometry ImageStreamX MKII (Amnis) before infusion of TNF blockers (infliximab, adalimumab), one day after infusion and after 6 months therapy. Statistical evaluation was performed using a nonparametric Mann–Whitney test. The results are presented as the median per cent of cells with NF-kB translocation (Me [Q 0.25-Q 0.75]). Results It was observed an increased level of NF-kB translocation in B-lymphocytes (Me 64 [53–81] – Me 38 [33–45]; p = 6 × 10−5), NK cells (Me 40 [33 -55] – Me 21 [18–26]; p = 4x10-4), Tc (Me 20 [15–24] – Me 16 [13–17]; p = 0.03), Th17 (Me 23 [21–28] – Me 18 [16–19]; p = 0.005) in children with exacerbation compared with remission. The level of NF-kB translocation in populations of lymphocytes during the period of clinical endoscopic remission did not differ from conditionally healthy children. The correlation between the level of NF-kB translocation and the number of cells in the studied lymphocyte populations was not found. In children with IBD after infusion of TNF blockers, a decrease the level of NF-kB translocation in NK cells was observed (Me 27.3 [23.5–39.1] – Me 17.7 [16.5–26.5]; p = 0,03) and an increase in Tregs (Me 18.9 [16.91–20.8] – Me 26.9 [19.4–31.9]; p = 0.0015). Conclusion The level of NF-kB translocation reflects the functional activity of major and small populations of lymphocytes. An increase in activity of Tregs in response to the administration of TNF blockers allows predicting a positive outcome from therapy over the next 6 months. If there is no Tregs reaction in response to the administration of TNF blockers, treatment tactics should be reassessed.

scholarly journals Does massive  bowel resection in newborns  affect further  immunity in children ?

2020 ◽  
Author(s):  
Katarzyna Sznurkowska ◽  
Anna Borkowska ◽  
Agnieszka Jankowska ◽  
Magdalena Malanowska ◽  
Maciej Zagierski ◽  
...  
Keyword(s):  
T Cells ◽  
Nk Cells ◽  
Bowel Resection ◽  
Clinical Signs ◽  
Peripheral Lymphocyte ◽  
Control Group ◽  
Healthy Children ◽  
Activated T Cells ◽  
Serum Immunoglobulins ◽  
Lymphocyte Populations

Abstract Background Short bowel syndrome (SBS) is defined as the a malabsorptive condition most often caused by massive resection of the small intestine. In children most cases of SBS originate in the newborn period and result from congenital anomalies or necrotizing enterocolitis. Loss of gut mucosa during resection does not only mean loss of absorption surface, but also deprives organism of many immunocompetent cells concentrated in gut associated lymphoid tissue, which is regarded the largest immune organ in humans. Aim of the study: We have aimed to access the influence of bowel resection on adaptive immunity in children, basing on peripheral lymphocyte populations and serum immunoglobulins. Patients and methods: 18 children, who underwent bowel resection in the first month of life and required further home parenteral nutrition were enrolled into the study. 12 healthy children, constituted control group. Based on flow cytometry the following subpopulations of lymphocytes were evaluated: T, B, NK, CD4+, C8 + and activated T cells. Serum immunoglobulins were determined with the use of immunoturbidimetric method. Results The percentage of B lymphocytes was reduced, while the rates of lymphocytes T and CD8 + lymphocytes were higher compared to healthy children. We documented significantly lower absolute count and proportion of NK cells in SBS group than in the control group. Absolute counts of lymphocytes, lymphocytes B, T, CD4 + and percentages of lymphocytes CD4+, and activated T cells inversely correlated with the time after resection. No statistically significant differences were found between the levels of IgA, IgM and IgG in the studied and the control group Conclusions Children with SBS do not present with clinical signs of immunodeficiency as well as deficits in peripheral lymphocyte populations and serum immunoglobulins. Lower number of NK cells in SBS patients compared to healthy children needs to be verified in larger cohort. The tendency of the lymphocyte subpopulations to decrease over time after resection points out the necessity for longer follow- up.

scholarly journals Immune response against HtrA proteases in children with cutaneous mastocytosis

2018 ◽  
Vol 65 (3) ◽  
Author(s):  
Joanna Zofia Renke ◽  
Sabina Kędzierska-Mieszkowska ◽  
Magdalena Lange ◽  
Bogusław Nedoszytko ◽  
Anna Liberek ◽  
...  
Keyword(s):  
Mast Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Healthy Children ◽  
Innate And Adaptive Immunity ◽  
Cellular Processes ◽  
Clonal Proliferation ◽  
Frequent Form ◽  
Cutaneous Mastocytosis ◽  
Igg Level

Mast cells play important role in both innate and adaptive immunity but clonal proliferation of abnormal mast cells in various organs leads to mastocytosis. The skin variant of the disease, cutaneous mastocytosis (CM) is the most frequent form of mastocytosis in children. The HtrA proteases are modulators of important cellular processes, including cell signaling and apoptosis, and are connected with development of many pathologies. The above and the observation that mast cells constitutively release the HtrA1 protein, prompted us to investigate a possible involvement of the HtrA proteins in pediatric CM.We assayed the levels of the serum autoantibodies (IgG) against the recombinant HtrA proteins (HtrA1-4) in children with CM (n= 36) and in healthy controls (n= 62). The anti-HtrA IgGs were detected using enzyme linked immunosorbent assay (ELISA) and Western-blotting. In the CM sera the levels of the anti-HtrA1 and anti-HtrA3 autoantibodies were significantly increased compared to the control group while the HtrA proteins’ levels were comparable. No significant differences in the anti-HtrA2 IgG level were found, and the anti-HtrA4 IgGs had a tendency to decrease. In healthy children, the IgG levels against the HtrA1, -3 and -4 increased significantly with the age of children; no significant changes were observed for the anti-HtrA2 IgG. Our results suggest involvement of the HtrA1 and HtrA3 proteins in pediatric CM; the involvement of the HtrA4 protein is possible but needs to be investigated further. In healthy children, the autoantibody levels against HtrA1, -3 and -4 but not against HtrA2 increase with age.

scholarly journals Decreased glycolysis induced dysfunction of NK cells in Henoch-Schonlein purpura patients

2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Wenjia Chai ◽  
Xiaolin Wang ◽  
Wei Wang ◽  
Hui Wang ◽  
Wenjun Mou ◽  
...  
Keyword(s):  
Nk Cells ◽  
Systemic Vasculitis ◽  
Metabolic Reprogramming ◽  
Healthy Children ◽  
Henoch Schonlein Purpura ◽  
Reduced Frequency ◽  
Activating Receptors ◽  
Innate Lymphocytes ◽  
Schonlein Purpura ◽  
Henoch Schönlein Purpura

Abstract Background Henoch-Schonlein purpura (HSP) is the most common systemic vasculitis of the childhood. However, its mechanisms and pathogenesis still need more exploration. Natural killer (NK) cells are innate lymphocytes, and there is a growing appreciation that cellular metabolism is important in determining the immune responsiveness of lymphocytes. Thus, we aimed to analyze the NK cells phenotype and explore the association between glucose metabolism and NK cells function in HSP patients. Results A total number of 64 HSP patients and 34 healthy children were included. The HSP patients were divided into two groups according to whether accompanied with nephritis or not. NK cells in HSP patients without nephritis showed a reduced frequency in peripheral blood, a down-regulated expression of activating receptors both NKp30 and NKp46, and an attenuated cytotoxic function against tumor cells. In addition, the function impairment of NK cells was shown to exacerbate in HSPN. Our data further revealed an aberrant metabolic reprogramming of NK cells in HSP patients. Upon stimulation with cytokines (IL-15, IL-12 and IL-2), NK cells from healthy controls switched to an elevated glycolysis rate to support their effector function. By contrast, the glycolysis rate of activated NK cells in HSP group was not significantly up-regulated from the resting level possibly owing to the inhibition of mTORC1. Conclusions Our study found that HSP patients were accompanied with dysfunction of NK cells. We concluded that the dysfunction of NK cells in HSP patients was induced with a decreased glycolysis rate and suggested that metabolic reprogramming of NK cells might be a player in the pathogenesis of HSP.

NK Receptor (NKR)-Mediated Signaling Pathways in Cord Blood (CB) CD56dim Versus Peripheral Blood (PB) CD56dim NK Cells: Implication for Immaturity in Cord Blood NK CD56dim Innate Immunity

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4897-4897
Author(s):  
Nancy Day ◽  
Evan Shereck ◽  
Janet Ayello ◽  
Catherine McGuinn ◽  
Prakash Satwani ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cord Blood ◽  
Cytotoxic Activity ◽  
Nk Cells ◽  
Signaling Pathways ◽  
Adaptive Immunity ◽  
Innate And Adaptive Immunity ◽  
Differential Protein ◽  
Significant Difference

Abstract Background: NK cells are characterized by absent CD3 but expression of CD56dim (90%, cytotoxic) and CD56bright (10%, mediator). NK cells may contribute to the immaturity in cord blood innate and adaptive immunity, and play an important role in the GVL effect post CBT. However, little is known regarding the NKR signaling pathways in CB vs PB CD56dim NK cells and its relationship to the cytotoxic activity. We previously demonstrated the ability to ex-vivo expand CB into NK subsets with profound NK in-vitro and in-vivo cytotoxic activity (Ayello/Cairo BBMT 2006). We further observed that there were 33 and 37 proteins over and under expressed by proteomic expression profiling studies of CB vs PB CD56dim (Shereck/Cairo, ASH 2007; ASPHO 2007; AACR 2007). The differential protein expressions included NKG2A, IP3R type 3, NCR3, MAPKAPK5, Notch 2, PLEK, and NF-X1 using both immunophenotype and proteomic profiling studies. Objectives: To understand the importance of NKR signaling pathways in mediating the differential protein expression and thus in regulating the NK cytotoxic activities in CB vs PB CD56dim, we compared the genomic expression pattern in CB vs PB CD56dim. Methods: For CD56dim isolation, first, NK cells were isolated indirectly by magnetic separation from non-NK cells. Second, the pre-enriched NK cells (CD56+/CD3−) from CB and PB were directly labeled with CD16 (FCGR3) MicroBeads, and the CD56+ CD16+ NK cells (CD56dim) were eluted after removing the column from the magnetic field (Miltenyi). Purity of CD56dim NK cells were then examined by flow cytometry (BD FACScan). For genomic studies, total RNA was isolated and reverse transcribed to cDNA using T7-Oligo (dT) primer. cRNA was Biotin-labeled by in vitro transcription. Fragmented biotin-labeled cRNA was hybridized to GeneChip U133A_2 in GCOS-operated Fluidics Station 450, and then scanned by GeneChip Scanner 3000 (Affymetrix). Data were analyzed using Agilent GeneSpring. Signal intensities were compared using one way ANOVA and Welch Test for statistical analysis. Results: There were 193 and 222 genes over and under expressed at the genomic level between CB vs PB CD56dim NK cells, respectively. CB vs PB CD56dim significantly overexpressed NKG2A (2.14F), CD16b (2.46F), KIR2D (2.13F), NKp44 (NCR2; 2.62F), PBX1 (4.29F), ENPEP (3.93F). There was no significant difference in NKR gene expression of CD16a, CD161, NKG2C, and NKp46 in CB vs PB CD56dim. CB vs PB CD56dim underexpressed the following NK genes: IP3R (1.32F), MAPKAPK5 (1.77F), NCR3 (1.24F), ACACB (3.23F), BBS1 (2.00F). Conclusion: CB vs PB CD56dim overexpressed NKG2A, CD16b, KIR2D, and NKp44 genes compared to only NKG2A was overexpressed at the protein level. These results suggest that NKR protein product levels in CB CD56dim may be directly regulated at the translation level, but not the transcription level. The discrepancy of IP3R, ENPEP, PBX1, and MAPKAPK5 gene expression suggest the involvements of IP3 and calcium ions in NKR signaling pathways. Since the Notch2, PLEK, and NF-X1 gene expression patterns were not increased, the augmented protein levels may result from the regulation of protein translation. The potential regulators of this process may include PBX1, ENPEP, ACACB, and BBS1 though the roles of these regulators need to be defined. We conclude that genomic differences between CB vs PB CD56dim may play an important role in regulating NKR signaling pathway, and thus contribute to disparate cytotoxic activity between CB vs PB and suggest a possible explanation for immaturity of cord blood innate and adaptive immunity.

scholarly journals Listeria meningitis complicated by hydrocephalus in an immunocompetent child: case report and review of the literature

2020 ◽  
Vol 46 (1) ◽  
Author(s):  
Giacomo Brisca ◽  
Alberto La Valle ◽  
Claudia Campanello ◽  
Mattia Pacetti ◽  
Mariasavina Severino ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Early Recognition ◽  
Treatment Options ◽  
Cranial Nerves ◽  
Positive Outcome ◽  
Healthy Children ◽  
Gram Positive Bacteria ◽  
Brain Abscesses ◽  
Uncommon Case ◽  
Listeria Meningitis

Abstract Background Listeria monocytogenes is a Gram-positive bacteria transmitted to human by animal stools, contaminated water and food. In children, Listeria monocytogenes typically affects newborns and immunocompromised patients often leading to invasive syndromes including sepsis, brain abscesses, meningitis, meningoencephalitis and rhombencephalitis. In healthy and immunocompetent children, Listeria meningitis is rare, but can progress rapidly and may be associated with severe complications (hydrocephalus, ventriculitis, cranial nerves palsy and cerebrospinal abscesses) and high mortality rate. Case presentation We describe a very uncommon case of meningoencephalitis due to Listeria monocytogenes in a 11-month-old immunocompetent girl. Cerebrospinal fluid (CSF) culture was positive on the second day. Antibiotic therapy was promptly started but the disease was complicated by neurological deterioration and decompensated hydrocephalus. The child required a very demanding pediatric and neurosurgical management and was discharged after 40 days without major sequelae. Conclusion Listeria is difficult to isolate and it is not susceptible to first-line treatment for bacterial meningitis with extended-spectrum cephalosporins. Early recognition is therefore crucial for a positive outcome. Pediatricians have to perform close clinical monitoring of these children and be aware of possible complications. A review of all cases of Listeria meningitis complicated by hydrocephalus in healthy children has been performed, to provide an overview on clinical features, treatment options and outcome.

scholarly journals Interactions of Human Myeloid Cells with Natural Killer Cell Subsets In Vitro and In Vivo

2011 ◽  
Vol 2011 ◽  
pp. 1-7 ◽  
Author(s):  
Obinna Chijioke ◽  
Christian Münz
Keyword(s):  
Nk Cells ◽  
Natural Killer ◽  
Nk Cell ◽  
Killer Cell ◽  
Experimental Models ◽  
Human Pathogens ◽  
Innate And Adaptive Immunity ◽  
Antigen Presenting

In both human and mouse it has been recently realized that natural killer (NK) cells do not emerge from the bone marrow with full functional competence but rather acquire functions in interaction with antigen-presenting cells (APCs), primarily dendritic cells (DCs). Here we review the mechanisms and the consequences of this NK-cell preactivation, as well as discuss new experimental models that now allow investigating these interactions for human NK cells and their response to human pathogens in vivo. These investigations will allow harnessing NK cells during vaccination for improved innate and adaptive immunity.

Both Intermittent and Continuous Dosing Regimens Of Pomalidomide Mediate Broad Activation Of Innate and Adaptive Immunity In Relapsed/Refractory Myeloma: Results Of A Phase II Randomized Trial

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 1949-1949
Author(s):  
Kavita Dhodapkar ◽  
Lin Zhang ◽  
Rakesh Verma ◽  
Kartik Sehgal ◽  
Mehmet H. Kocoglu ◽  
...  
Keyword(s):  
T Cells ◽  
Nk Cells ◽  
Adaptive Immunity ◽  
Immune Activation ◽  
Drug Induced ◽  
Innate And Adaptive Immunity ◽  
Initiation Of Therapy ◽  
Induced Changes ◽  
The Impact

Abstract Pomalidomide (POM) is a novel IMiD® immunomodulatory agent with clinical activity in several settings including in relapsed / resistant myeloma (RRMM). Several preclinical studies have documented the immunologic effects of IMiD® immunomodulatory drugs. IMiD® now form the backbone of several emerging combination therapies in hematologic malignancies. In prior studies, the clinical activity of POM in relapsed myeloma has been demonstrated using both continuous and intermittent dosing regimens with and without steroids. However the impact of the specific POM dosing regimen and the effect of concurrent steroids (as commonly utilized in most combination regimens) on POM-induced immune activation in vivo is unknown. In order to evaluate these issues more directly, we analyzed samples from patients enrolled in a randomized phase II clinical trial comparing two POM dosing schedules. Comparison of drug-induced immune activation between the two dosing schedules was one of the pre-specified endpoints in this study. Patients (n=39) with RRMM documented to be refractory to lenalidomide were randomized to therapy with POM 2 mg/day for 28/28 days (Arm A, n=19) or POM 4 mg/day for 21/28 days (Arm B, n=20) of a 28 day cycle. All patients (pts) received POM alone for cycle 1, followed by the addition of dexamethasone (DEX) at 40 mg weekly in subsequent cycles in both arms. In recent studies, we have shown that immune effects of lenalidomide are manifest early, within a week of initiation of therapy (Richter et al, Blood 121:423, 2013). Therefore each patient was analyzed 1 week after initiation of POM alone at 2 or 4 mg (cycle 1) or POM + DEX (cycle 2). POM therapy led to an increase in T cells at day 7 after initiation of therapy (mean percent increase compared to baseline 47%, p <0.01). This POM-induced T cell expansion consisted predominantly of CD8+ T cells (mean increase compared to baseline 40%, p<0.01). POM therapy also led to an increase in circulating natural killer (NK) cells (mean increase compared to baseline 92%, p<0.01) which peaked at 7 days post-initiation of therapy. POM-induced NK expansion was associated with an increase in the expression of NKG2D as well as CD16 and CD56, consistent with NK activation in vivo. Changes in T and NK cells following cycle 2 (POM+DEX) were compared with the data on cycle 1 (POM alone) in the same patient to evaluate the impact of DEX. Addition of DEX led to dampening of drug-induced T cell activation in vivo, but surprisingly did not dampen drug induced NK activation. Comparison of data from the two cohorts demonstrated that both 2 mg and 4 mg dose of POM led to comparable degree of immune activation in vivo, particularly for NK cells. In selected patients, post-treatment bone marrow samples were available after completion of at least 2 cycles of therapy. Comparison of the transcriptome of the CD138-depleted fraction from these marrows (representing bulk tumor microenvironment) identified nearly 300 genes upregulated in the bone marrow microenvironment post therapy compared to paired baseline samples. Interestingly, these genes were highly enriched in genes associated with T and NK activation. As POM led to activation of both innate and adaptive immunity, we also analyzed drug-induced changes in the myeloid compartment. Comparison of gene expression profiles (GEP) of purified CD14+ monocytes before and after POM indicated that POM therapy leads to a distinct GEP signature in myeloid cells in vivo. Finally, correlation of drug-induced changes in immune cells with clinical response demonstrated that an increase in post-therapy T cells correlated with clinical response, but post-treatment increase in NK cells did not. Together these data demonstrate that both 2 mg and 4 mg dose of POM can mediate broad activation of both innate and adaptive immunity in vivo, even in the setting of immune paresis associated with advanced, heavily pre-treated disease. Both continuous and intermittent dosing schedules of POM have comparable pharmacodynamic effects on immune activation. POM-induced immune activation is detected even with concurrent steroids and immune-related genes constitute a dominant component of drug induced changes in tumor microenvironment. POM-mediated immune activation may thus be exploited in the context of combination with immune therapies as well as monoclonal antibodies. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Assessment of modified Forssell’s myectomy success rate in the treatment of crib biting in horses

2015 ◽  
Vol 84 (1) ◽  
pp. 63-69 ◽  
Author(s):  
Šárka Krisová ◽  
Zdeněk Žert ◽  
Kristína Žuffová
Keyword(s):  
Retrospective Study ◽  
Surgical Treatment ◽  
Success Rate ◽  
Early Treatment ◽  
Statistical Evaluation ◽  
Positive Outcome ◽  
Term Outcome ◽  
Long Term Outcome ◽  
Significance Level

The objective of this retrospective study was to assess the success rate of Forssell’s modified myectomy – surgical treatment of crib biting on 33 horses of various breeds identified as stereotypical cribbers which were referred to the Equine Clinic in Brno between January 2001 and December 2010. The long term outcome was available for all horses. The overall success of the surgery without considering influencing factors was 61%. Thirteen horses (39%) returned to the stereotypical behaviour sooner or later after the surgery. In this retrospective study we confirmed the dependence of success of the surgery on the necessity of early treatment of the problem (up to 6 months after the appearance of initial signs), with the significance level of P = 0.037. Any connection between age, breed or sex, and positive outcome of the surgery were not confirmed during statistical evaluation. When the horses were diagnosed early enough, an 80% success rate in surgical treatment performed within 6 months from the initial signs of crib biting was recorded, which seems to be very promising. It can be concluded that surgical treatment of crib biting by modified Forsell’s myectomy is the recommended option in the therapy of this oral stereotype.

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