Elevated IgA and IL-10 levels in very-early-onset inflammatory bowel disease secondary to IL-10 receptor deficiency

ABSTRACT Objective: To report two patients with very-early-onset inflammatory bowel disease (VEOIBD) secondary to interleukin-10 receptor (IL-10R) mutations, explore immunophenotyping data and plasma cytokine profile on these cases compared to healthy controls, and describe the phenotype of IL-10/IL-10R mutations based on a literature review. Case description: We report on two female infants referred to our tertiary center at the age of ten months, with severe colonic and perianal disease, as well as significant malnutrition, who had shown limited response to usual inflammatory bowel disease (IBD) therapy agents. In the first case, whole-exome sequencing (WES) revealed a homozygous (c.537G>A/p.T179T) mutation in exon 4 of the IL-10RA gene, while in the second patient, compound heterozygosity was identified, also in the IL-10RA gene (chr11:117.859.199 variant A>G/p.Tyr57Cys and chr11: 117.860.335 variant G>T/p.Val123Leu). Both patients underwent hematopoietic cell transplantation (HCT). Immunological work-up of these patients revealed increased IL-10 plasma levels and increased IgA. Comments: Our case reports disclose novel findings on plasma cytokine profile in IL-10R deficiency, and we describe the severe phenotype of IL-10/IL-10R deficiency that should be recognized by physicians.

Cytokine Changes in Cerebrospinal Fluid and Plasma Post-Emergency Orthopaedic Surgery

Background: The process of neuroinflammation after surgery and how it may contribute to post-operative neurocognitive disorders (PND) is not well understood. Studying the association between central and peripheral cytokines and neuroinflammation is a prelude to the development of treatments for PND. Here, we investigate the hypotheses that there is a greater cytokine response in cerebrospinal fluid (CSF) than plasma after orthopaedic surgery, and that plasma cytokine levels are directly related to CSF cytokine levels, enabling plasma cytokine levels to be used as markers of neuroinflammation. Methods: Patients admitted with a fractured neck of femur were invited to participate in this study. Participants had a spinal catheter inserted just prior to induction of anaesthesia. Samples of blood and CSF were taken before, immediately after, and on the first day following emergency surgery. The catheter was then removed. Samples were analysed for the presence of ten cytokines by immunoassay. Results: A spinal catheter was successfully inserted in 11 participants during the 18-month study period. Five plasma cytokines (IL-4, IL-6, IL-10, IL-12p70 and IL-13) rose significantly following surgery, whereas all ten CSF cytokines rose significantly (IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13, IFN-γ and TNF-α) (adjusted-p <0.05). Central (CSF) cytokine levels were consistently higher than their peripheral (plasma) counterparts after surgery, with some patients having a particularly marked neuroinflammatory response. The greatest increases occurred in IL-8 in CSF and IL-6 in plasma. There were significant, strong positive correlations between several of the measured cytokines in the CSF after surgery, but far fewer in plasma. There was no significant correlation between cytokine levels in the plasma and CSF at each of the three time points.Conclusions: To our knowledge, this is the first study to analyse paired samples of plasma and CSF for cytokine levels before and after emergency orthopaedic surgery. This study demonstrates that following surgery for a fractured neck of femur, there is a far greater rise in cytokines in the CSF compared to plasma. The lack of correlation between peripheral and central cytokines suggests measurement of peripheral cytokines are not necessarily related to which patients may have a large neuroinflammatory response.

371 Analysis of changes in plasma cytokine levels in response to IL-12 therapy in three clinical trials

BackgroundThe ability of IL-12 to stimulate NK and T cell anti-tumor activity has made it an attractive candidate for overcoming immunosuppressive tumor microenvironments. Our group has demonstrated in pre-clinical models that IL-12 will enhance IgG receptor-mediated NK cell responses to antibody-coated tumor cells and conducted three studies where IL-12 was used in combination with an anti-tumor monoclonal antibody. These were OSU-9968, Phase 1 study of IL-12 + trastuzumab; OSU-1067, Phase 1 study of IL-12 + trastuzumab + paclitaxel in HER2-positive cancers and OSU-11010, Phase I/II study of IL-12 + cextuximab in head and neck cancer.1–3 Cytokine levels were measured in patients with varying responses in an effort to better characterize IL-12-induced immunity.MethodsPlasma cytokine levels in 21 patients across 3 studies were measured at baseline and at 4 time points after IL-12 administration. 2 patients had complete responses, 1 had a partial response, 9 patients had stable disease > 60 days and 9 had progressive disease. A combination of 7 U-PLEX, V-PLEX, and R-PLEX Human Biomarker Assays (Meso Scale Discovery) were performed to monitor levels of 23 cytokines: GM-CSF, IFN-gamma, IL-10, IL-8, IP-10, MCP-1, MDC, MIP-1alpha, MIP-1ß, TNF-alpha, IL-15, IL-18, MCP-2, MIG, IL-13, IL-17, IL-1ß, IL-4, IL-5, IL-6, IL-1alpha, TGFß, VEGF. Student’s t-test on GraphPad Prism 9.0.0 was used for statistical analyses.ResultsNine cytokines were significantly upregulated following IL-12 therapy. IFN-gamma levels increased from a mean of 27.42 pg/mL at baseline to 1764 pg/mL after IL-12 treatment (p=0.0246). GM-CSF, TNF-alpha and IL-10 also increased following IL-12 therapy (p=0.0199, 0.0004, 0.0003). Several chemotactic factors including MCP-1, MDC, MIP-1alpha, and MIP-1ß increased from means of 483.1 pg/mL to 695.7 pg/mL, 3112 pg/mL to 4305 pg/mL, 62.44 pg/mL to 130.3 pg/mL and 263.1 to 487.4 pg/mL, respectively (p-values all < 0.013). Levels of IL-18 increased from a baseline mean of 2059 pg/mL to 3952 pg/mL (p=0.0003). Several cytokines were also differentially induced across response groups with MCP-1 and GM-CSF increased in responding patients (p=0.02, p=0.04) and IL-10, MIP-1ß and IL-6 increased in progressive disease patients (p=0.02, p=0.01, p=0.03).ConclusionsThe ability to detect significant changes in cytokines as a result of IL-12 therapy across three separate clinical trials supports the broad effects of IL-12 on NK cells and other immune compartments. The additional differential effect in responders vs. progressive disease patients indicates that these cytokines likely affect patient outcome and will be further evaluated as possible markers of response.ReferencesParihar R, et al. A phase I study of interleukin 12 with trastuzumab in patients with human epidermal growth factor receptor-2-overexpressing malignancies. Clin Cancer Res 2004;10:5027 LP–5037.Bekaii-Saab TS, et al. A phase I trial of paclitaxel and trastuzumab in combination with interleukin-12 in patients with HER2/neu-expressing malignancies. Mol Cancer Ther 2009;8:2983–2991.McMichael EL, et al. A phase I/II trial of cetuximab in combination with interleukin-12 administered to patients with unresectable primary or recurrent head and neck squamous cell carcinoma. Clin Cancer Res 2019;25:4955 LP–4965.Ethics ApprovalThese studies were approved by the Human Institutional Review Board at The Ohio State University Medical Center; approval numbers 99H0185, 1999C0326 and 2011c0019, respectively.

Alterations in Saliva and Plasma Cytokine Concentrations During Long-Duration Spaceflight

Long-duration spaceflight is known to cause immune dysregulation in astronauts. Biomarkers of immune system function are needed to determine both the need for and effectiveness of potential immune countermeasures for astronauts. Whereas plasma cytokine concentrations are a well-established biomarker of immune status, salivary cytokine concentrations are emerging as a sensitive indicator of stress and inflammation. For this study, to aid in characterizing immune dysregulation during spaceflight, plasma and saliva cytokines were monitored in astronauts before, during and after long-duration spaceflight onboard the International Space Station. Blood was collected from 13 astronauts at 3 timepoints before, 5 timepoints during and 3 timepoints after spaceflight. Saliva was collected from 6 astronauts at 2 timepoints before spaceflight, 2 timepoints during and 3 timepoints following spaceflight. Samples were analyzed using multiplex array technology. Significant increases in the plasma concentration of IL-3, IL-15, IL-12p40, IFN-α2, and IL-7 were observed during spaceflight compared to before flight baseline. Significant decreases in saliva GM-CSF, IL-12p70, IL-10 and IL-13 were also observed during spaceflight as compared to compared to before flight baseline concentrations. Additionally, plasma TGFβ1 and TGFβ2 concentrations tended to be consistently higher during spaceflight, although these did not reach statistical significance. Overall, the findings confirm an in-vivo hormonal dysregulation of immunity, appearing pro-inflammatory and Th1 in nature, persists during long-duration orbital spaceflight. These biomarkers may therefore have utility for monitoring the effectiveness of biomedical countermeasures for astronauts, with potential application in terrestrial research and medicine.