STAT1 contributes to HLA class I upregulation and CTL reactivity after anti-EGFR mAb cetuximab therapy in head and neck cancer patients

Squamous cell carcinoma of head and neck (HNSCC) cells express low HLA class I and antigen processing machinery (APM) components, such as transporter TAP-1/2, which is associated with the reduced sensitivity to cytotoxic T lymphocyte (CTL) mediated lysis. Epidermal growth factor receptor (EGFR) is overexpressed in HNSCC and is associated with the poor prognosis. FDA approved anti-EGFR blockade mAb cetuximab inhibits HNSCC proliferation, and induces EGFR-specific CTL. However, the molecular mechanism(s) underlying the EGFR-specific CTL recognition of HNSCC in the therapeutic efficacy of anti-EGFR mAb is still emerging. We show that cetuximab or EGFR knockdown enhanced expression of HLA class I antigens, which is associated with the EGFR expression level on HNSCC. These findings were validated in a prospective trial of neoadjuvant cetuximab therapy. Interestingly, upregulation of HLA-B/C alleles were more pronounced than HLA-A alleles after cetuximab or EGFR knockdown treatment. EGFR signaling blockade or EGFR depletion also enhanced IFN gamma receptor (IFNAR) on HNSCC and augmented induction of HLA class I and TAP-1/2 caused by IFN gamma treatment. Cetuximab or EGFR knockdown enhanced the level of HLA class I, STAT-1, TAP-1/2 in a STAT-1+/+ cell line but not in STAT-1-/- cell line, documenting the STAT-1 dependence of this effect. We also found that Src homology domain-containing phosphatase 2 (SHP-2), which is downstream of EGFR and also overexpressed in SCCHN, can suppress the immunostimulatory effect of cetuximab treatment on HLA class I/STAT-1 upregulation, and dual targeting of EGFR and SHP-2 co-operates in the most efficient reversal of immune escape phenotype. In addition, cetuximab-based EGFR inhibition and SHP-2 depletion enhanced the recognition of HNSCC cells by EGFR 853-861 specific CTL, and enhanced surface presentation of non-EGFR TA, such as MAGE-3 271-279 , indicating that a broad tumor antigen repertoire is processed and presented by HLA/APM upregulation. These findings elucidate a novel immune escape mechanism associated with EGFR signaling through STAT1 suppression and the reversal with cetuximab, which may provide additional targets for on-going mAb-based immunotherapy

TIM-3+ T cells are not exhausted but activated cells in the tumor microenvironment

Although T-cell immunoglobulin mucin 3 (TIM-3) does not contain inhibitory or death signaling motifs in its cytoplasmic domain, it has been proposed to be associated with T cell suppression and/or exhaustion. However, several lines of evidence suggest that TIM-3 can stimulate T cells as a costimulatory molecule by coupling Src family tyrosine kinase Fyn and the p85 phosphatidylinositol 3-kinase (PI3K) adaptor to TCR signaling. We examined the expression pattern and function of TIM-3 and other immune checkpoint receptors, CTLA-4 and PD-1 on tumor infiltrating lymphocytes (TIL), compared to those of peripheral blood T lymphocytes (PBL) in patients with head and neck cancer (HNC). Here, we report that TIM-3+CD8+ TIL express higher granzyme B/perforin, more actively proliferate under anti-CD3/-CD28 stimulatory conditions, and are more resistant to activation induced cell death than TIM-3-CD8+ TIL, indicating TIM-3 can positively regulate T cell responses. Analysis of downstream signaling molecules including phosphorylated JAK/STAT-1, PD-1/SHP-2, and costimulatory CD137 in CD8+ TIL subsets supports our observation that TIM-3+CD8+ TIL are activated cells in HNC patients. However, PD-1 and CTLA-4 can negatively regulate immune responses of TIM-3+CD8+ and TIM-3+CD4+ TIL respectively. More importantly, neoadjuvant immunotherapy of HNC patients with the EGFR-specific mAb cetuximab increased both TIM-3 and PD-1 expression on CD8+ TIL, which was correlated with higher granzyme B/perforin expression in TIM-3+CD8+ TIL. Taken together, these findings suggest that TIM-3 functions as a positive regulator of activated T cells in the tumor microenvironment while CTLA-4 and PD-1 modulate the function of activated TIM-3+ TIL. We therefore suggest that TIM-3 can be used as a biomarker to indicate activation status of T cells in the tumor microenvironment depending on PD-1 co-expression, particularly in response to cancer therapy including cetuximab-based immunotherapy

Protective Effects of Gabapentin on Allodynia and α2δ1-Subunit of Voltage-dependent Calcium Channel in Spinal Nerve-Ligated Rats

This study was designed to determine whether early gabapentin treatment has a protective analgesic effect on neuropathic pain and compared its effect to the late treatment in a rat neuropathic model, and as the potential mechanism of protective action, the α2δ1-subunit of the voltage-dependent calcium channel (α2δ1-subunit) was evaluated in both sides of the L5 dorsal root ganglia (DRG). Neuropathic pain was induced in male Sprague-Dawley rats by a surgical ligation of left L5 nerve. For the early treatment group, rats were injected with gabapentin (100 mg/kg) intraperitoneally 15 min prior to surgery and then every 24 hr during postoperative day (POD) 1-4. For the late treatment group, the same dose of gabapentin was injected every 24 hr during POD 8-12. For the control group, L5 nerve was ligated but no gabapentin was administered. In the early treatment group, the development of allodynia was delayed up to POD 10, whereas allodynia was developed on POD 2 in the control and the late treatment group (p<0.05). The α2δ1-subunit was up-regulated in all groups, however, there was no difference in the level of the α2δ1-subunit among the three groups. These results suggest that early treatment with gabapentin offers some protection against neuropathic pain but it is unlikely that this action is mediated through modulation of the α2δ1-subunit in DRG

Prevalence, associated factors and outcomes of pressure injuries in adult intensive care unit patients: the DecubICUs study

Funder: European Society of Intensive Care Medicine; doi: http://dx.doi.org/10.13039/501100013347Funder: Flemish Society for Critical Care NursesAbstract: Purpose: Intensive care unit (ICU) patients are particularly susceptible to developing pressure injuries. Epidemiologic data is however unavailable. We aimed to provide an international picture of the extent of pressure injuries and factors associated with ICU-acquired pressure injuries in adult ICU patients. Methods: International 1-day point-prevalence study; follow-up for outcome assessment until hospital discharge (maximum 12 weeks). Factors associated with ICU-acquired pressure injury and hospital mortality were assessed by generalised linear mixed-effects regression analysis. Results: Data from 13,254 patients in 1117 ICUs (90 countries) revealed 6747 pressure injuries; 3997 (59.2%) were ICU-acquired. Overall prevalence was 26.6% (95% confidence interval [CI] 25.9–27.3). ICU-acquired prevalence was 16.2% (95% CI 15.6–16.8). Sacrum (37%) and heels (19.5%) were most affected. Factors independently associated with ICU-acquired pressure injuries were older age, male sex, being underweight, emergency surgery, higher Simplified Acute Physiology Score II, Braden score 3 days, comorbidities (chronic obstructive pulmonary disease, immunodeficiency), organ support (renal replacement, mechanical ventilation on ICU admission), and being in a low or lower-middle income-economy. Gradually increasing associations with mortality were identified for increasing severity of pressure injury: stage I (odds ratio [OR] 1.5; 95% CI 1.2–1.8), stage II (OR 1.6; 95% CI 1.4–1.9), and stage III or worse (OR 2.8; 95% CI 2.3–3.3). Conclusion: Pressure injuries are common in adult ICU patients. ICU-acquired pressure injuries are associated with mainly intrinsic factors and mortality. Optimal care standards, increased awareness, appropriate resource allocation, and further research into optimal prevention are pivotal to tackle this important patient safety threat

Correction to: Prevalence, associated factors and outcomes of pressure injuries in adult intensive care unit patients: the DecubICUs study

The original version of this article unfortunately contained a mistake

Prevalence, associated factors and outcomes of pressure injuries in adult intensive care unit patients: the DecubICUs study

Funder: European Society of Intensive Care Medicine; doi: http://dx.doi.org/10.13039/501100013347Funder: Flemish Society for Critical Care NursesAbstract: Purpose: Intensive care unit (ICU) patients are particularly susceptible to developing pressure injuries. Epidemiologic data is however unavailable. We aimed to provide an international picture of the extent of pressure injuries and factors associated with ICU-acquired pressure injuries in adult ICU patients. Methods: International 1-day point-prevalence study; follow-up for outcome assessment until hospital discharge (maximum 12 weeks). Factors associated with ICU-acquired pressure injury and hospital mortality were assessed by generalised linear mixed-effects regression analysis. Results: Data from 13,254 patients in 1117 ICUs (90 countries) revealed 6747 pressure injuries; 3997 (59.2%) were ICU-acquired. Overall prevalence was 26.6% (95% confidence interval [CI] 25.9–27.3). ICU-acquired prevalence was 16.2% (95% CI 15.6–16.8). Sacrum (37%) and heels (19.5%) were most affected. Factors independently associated with ICU-acquired pressure injuries were older age, male sex, being underweight, emergency surgery, higher Simplified Acute Physiology Score II, Braden score 3 days, comorbidities (chronic obstructive pulmonary disease, immunodeficiency), organ support (renal replacement, mechanical ventilation on ICU admission), and being in a low or lower-middle income-economy. Gradually increasing associations with mortality were identified for increasing severity of pressure injury: stage I (odds ratio [OR] 1.5; 95% CI 1.2–1.8), stage II (OR 1.6; 95% CI 1.4–1.9), and stage III or worse (OR 2.8; 95% CI 2.3–3.3). Conclusion: Pressure injuries are common in adult ICU patients. ICU-acquired pressure injuries are associated with mainly intrinsic factors and mortality. Optimal care standards, increased awareness, appropriate resource allocation, and further research into optimal prevention are pivotal to tackle this important patient safety threat

Feasibility and performance of a thin-film composite seawater reverse osmosis membrane fabricated on a highly porous microstructured support

Although a highly porous support membrane has attracted increasing attention as an alternative to enhance the water permeability of a thin-film composite (TFC) membrane without compromising salt rejection, its feasibility has not ever been tested in seawater desalination. This study explored the availability and potential of a highly porous microstructured (HPμS) support membrane as a support for a seawater reverse osmosis (SWRO) membrane. Our lab-made membranes, TFC-HPμS, exhibited a higher water permeability of 1.62 L m−2 h−1 bar−1 as compared with most of the state-of-the-art SWRO membranes recently reported in the literature, while achieving comparable NaCl rejection (99%) in SWRO test condition (55 bar, 35,000 mg L−1 of NaCl). This excellent performance is thought to stem from the HPμS support endowing a TFC membrane with comparable mechanical properties to that of existing support used for conventional SWRO membrane and shortened effective diffusion pathway of water molecules over the active layer. The robustness and enhanced mechanical strength of the TFC-HPμS membrane are attributed to its narrow and regularly arranged finger-like structure ensuring the even distribution of local stresses, thereby eliminating the presence of stress convergence points. The shortened effective diffusion pathway was estimated to be achieved mainly by less localized surface pores due to the HPμS support's highly porous surface with a larger number of even distributed surface pores. This study potentially opens up another workable pathway in the fabrication of SWRO membranes with enhanced performance without significant sacrifice of the selectivity.National Research Foundation (NRF)This research grant was supported by the Singapore National Research Foundation under its Environment and Water Research Pro- gram and administered by PUB, Singapore’s National Water Agency (grant number: 1501-IRIS-04)