Enhancement of antibody-dependent cellular cytotoxicity is associated with treatment response to extracorporeal photopheresis in Sézary syndrome

Sézary syndrome (SS) is a rare, leukemic type of cutaneous T-cell lymphoma (CTCL), for which extracorporeal photopheresis (ECP) is a first-line therapy. Reliable biomarkers to objectively monitor the response to ECP in patients with SS are missing. We examined the quantitative and qualitative impact of ECP on natural killer (NK) cell activity in SS patients, and especially their functional ability for antibody-dependent cell-mediated cytotoxicity (ADCC). Further, we addressed the question whether the magnitude of the effect on ADCC can be associated with the anti-cancer efficacy of ECP in SS patients. We assessed numbers of NK cells, ADCC activity, and treatment response based on blood tumor staging in a cohort of 13 SS patients (8 women, 5 men) treated with ECP as a first-line therapy. Blood samples were collected before treatment start and after an average of 9 months of uninterrupted ECP treatment. NK cell numbers were reduced in SS patients compared to healthy individuals and showed a tendency of recovery after long-term ECP treatment, independent of the clinical response to treatment. Patients with marginal increase (≤1.5 AU-fold) or lack of increase in ADCC activity failed to respond clinically to treatment, while patients with an increased ADCC activity showed a reduction in blood tumor burden. NK-mediated ADCC is selectively enhanced and might be a mechanism underlying the effect of ECP while in addition it can possibly serve as a reliable biomarker to objectively monitor response to ECP in patients with SS

mRNA-Based Anti-TCR CDR3 Tumour Vaccine for T-Cell Lymphoma

Efficient vaccination can be achieved by injections of in vitro transcribed mRNA (ivt mRNA) coding for antigens. This vaccine format is particularly versatile and allows the production of individualised vaccines conferring, T-cell immunity against specific cancer mutations. The CDR3 hypervariable regions of immune receptors (T-cell receptor, TCR or B-cell receptor, BCR) in the context of T- or B-cell leukaemia or lymphoma are targetable and specific sequences, similar to cancer mutations. We evaluated the functionality of an mRNA-based vaccine designed to trigger immunity against TCR CDR3 regions in an EL4 T-lymphoma cell line-derived murine in vivo model. Vaccination against the hypervariable TCR regions proved to be a feasible approach and allowed for protection against T-lymphoma, even though immune escape in terms of TCR downregulation paralleled the therapeutic effect. However, analysis of human cutaneous T-cell lymphoma samples indicated that, as is the case in B-lymphomas, the clonotypic receptor may be a driver mutation and is not downregulated upon treatment. Thus, vaccination against TCR CDR3 regions using customised ivt mRNA is a promising immunotherapy method to be explored for the treatment of patients with T-cell lymphomas

Gene Expression Programs in Response to Hypoxia: Cell Type Specificity and Prognostic Significance in Human Cancers

BACKGROUND: Inadequate oxygen (hypoxia) triggers a multifaceted cellular response that has important roles in normal physiology and in many human diseases. A transcription factor, hypoxia-inducible factor (HIF), plays a central role in the hypoxia response; its activity is regulated by the oxygen-dependent degradation of the HIF-1α protein. Despite the ubiquity and importance of hypoxia responses, little is known about the variation in the global transcriptional response to hypoxia among different cell types or how this variation might relate to tissue- and cell-specific diseases. METHODS AND FINDINGS: We analyzed the temporal changes in global transcript levels in response to hypoxia in primary renal proximal tubule epithelial cells, breast epithelial cells, smooth muscle cells, and endothelial cells with DNA microarrays. The extent of the transcriptional response to hypoxia was greatest in the renal tubule cells. This heightened response was associated with a uniquely high level of HIF-1α RNA in renal cells, and it could be diminished by reducing HIF-1α expression via RNA interference. A gene-expression signature of the hypoxia response, derived from our studies of cultured mammary and renal tubular epithelial cells, showed coordinated variation in several human cancers, and was a strong predictor of clinical outcomes in breast and ovarian cancers. In an analysis of a large, published gene-expression dataset from breast cancers, we found that the prognostic information in the hypoxia signature was virtually independent of that provided by the previously reported wound signature and more predictive of outcomes than any of the clinical parameters in current use. CONCLUSIONS: The transcriptional response to hypoxia varies among human cells. Some of this variation is traceable to variation in expression of the HIF1A gene. A gene-expression signature of the cellular response to hypoxia is associated with a significantly poorer prognosis in breast and ovarian cancer

Lactic Acidosis Triggers Starvation Response with Paradoxical Induction of TXNIP through MondoA

Although lactic acidosis is a prominent feature of solid tumors, we still have limited understanding of the mechanisms by which lactic acidosis influences metabolic phenotypes of cancer cells. We compared global transcriptional responses of breast cancer cells in response to three distinct tumor microenvironmental stresses: lactic acidosis, glucose deprivation, and hypoxia. We found that lactic acidosis and glucose deprivation trigger highly similar transcriptional responses, each inducing features of starvation response. In contrast to their comparable effects on gene expression, lactic acidosis and glucose deprivation have opposing effects on glucose uptake. This divergence of metabolic responses in the context of highly similar transcriptional responses allows the identification of a small subset of genes that are regulated in opposite directions by these two conditions. Among these selected genes, TXNIP and its paralogue ARRDC4 are both induced under lactic acidosis and repressed with glucose deprivation. This induction of TXNIP under lactic acidosis is caused by the activation of the glucose-sensing helix-loop-helix transcriptional complex MondoA:Mlx, which is usually triggered upon glucose exposure. Therefore, the upregulation of TXNIP significantly contributes to inhibition of tumor glycolytic phenotypes under lactic acidosis. Expression levels of TXNIP and ARRDC4 in human cancers are also highly correlated with predicted lactic acidosis pathway activities and associated with favorable clinical outcomes. Lactic acidosis triggers features of starvation response while activating the glucose-sensing MondoA-TXNIP pathways and contributing to the “anti-Warburg” metabolic effects and anti-tumor properties of cancer cells. These results stem from integrative analysis of transcriptome and metabolic response data under various tumor microenvironmental stresses and open new paths to explore how these stresses influence phenotypic and metabolic adaptations in human cancers

CD8α, a surface marker of cytotoxic T lymphocyte from orange-spotted grouper (Epinephelus coioides),cDNA cloning and characterization

CD8是表現在毒殺性T淋巴球表面的醣蛋白分子，可連接第一型主要組織相容複合體(MHC class I)，幫助毒殺T淋巴球摧毀被病毒感染或腫瘤細胞。 從點帶石斑魚的胸腺選殖出CD8α cDNA，包括43 bp的5’端非載碼區、684 bp的載碼區，和不同長度333、534和853 bp的3’端非載碼區，分別命名為CD8α S、CD8α M和CD8α L。載碼區轉譯出含227個胺基酸的蛋白質，含一段訊息胜肽、一段免疫球蛋白家族區塊、一段免疫球蛋白關節區塊、一段穿膜區塊和一段細胞質尾區塊；其中細胞質尾區塊有一段和酪胺酸激酶p56lck的結合位。蛋白質分子量推測為24 kDa、等電點9.62。系統分類與分子演化分析顯示石斑魚和其他海水魚的CD8α一致性在50~60%。Q-PCR 檢測CD8α分佈在胸腺、頭腎、鰓、脾臟和腸，不論 S、M或L型都以胸腺表現最多，且以M型的表現最多。石斑魚注射神經壞死病毒後四天，Q- PCR檢測CD8α在脾臟的表現顯著高於未注射組，屬於可誘發基因。利用昆蟲細胞桿狀病毒表現系統合成CD8α重組蛋白，經SDS-PAGE和西方墨點轉漬法檢驗，分子量約為22.5 kDa。CD8α重組蛋白免疫紐西蘭大白兔九週後取得抗血清，使用免疫細胞化學染色，抗血清可辨識部分週邊白血球，細胞質呈環型戒指狀、高核質比、直徑約7 μm，推測為淋巴球次群。石斑魚注射神經壞死病毒一週後，抽血製備週邊白血球，再利用磁珠正篩選計數CD8α+淋巴球從免疫前3.6~5.5% 增加至免疫後4.2~6.68%。石斑魚注射神經壞死病毒一週後，CD8α+淋巴球會毒殺被神經壞死病毒感染、相同MHC基因型鰭細胞，毒殺活性顯著高於被嘉納魚虹彩病毒感染、相同MHC基因型的鰭細胞或是神經壞死病毒感染、不同MHC基因型的鰭細胞。綜合以上結果顯示被CD8α抗體辨識的淋巴球具專一毒殺活性，受主要組織相容複合體限制，推測屬細胞毒殺性T淋巴球。點帶石斑魚CD8α抗體不會辨識黑鯛、日本鰻和吳郭魚的週邊白血球。此CD8α抗體可作為研究點帶石斑魚細胞性免疫反應的工具。CD8 is a heterodimer membrane glycoprotein found primarily on the surface of cytotoxic T lymphocytes. It helps the CTLs to kill the infected cells by binding to MHC class I protein that present antigens on the infected-cell surface. In the present study, we cloned the full length cDNAs from orange-spotted grouper (Epinephelus coioides), which included 43 bp 5’UTR, 684 bp ORF and varied length of 333, 534 and 853 bp 3’UTR, and were named as CD8α S, M and L, respectively. The deduced protein of CD8α is 227 amino acid residues long and included one signal peptide, Ig superfamily domain, hinge region, transmembrane domain and cytoplasmic tail and a conserved binding motif associate with tyrosine kinase p56lck . The MW is estimated as 24 kDa with a pI of 9.62. Phylogenetically, the predicted grouper CD8α protein is similar to CD8α from other marine fish species, and the identity is 50-60％. Q-PCR revealed that CD8α is constitutively express in thymus, head kidney, gill, spleen and gut. Optimal expression was found in thymus for both CD8α S, M and L forms, and the CD8α M transcript is the most abundant. The Q-PCR results revealed that the CD8α in the spleen of NNV-injected fish was significantly up-regulated in 4 days post-injection compared to the untreated fish. Moreover, the recombinant CD8α protein was biosynthesized in insect cell-baculovirus expression system examined the MW was 22.5 kDa via the SDS-PAGE and Western blotting . The antiserum prepared from New Zealand white rabbit against biosynthesized recombinant CD8α protein could recognize the subset lymphocytes specifically from PBL with characteristics of 7μm in diameter, high nucleus/cytoplasm ratio and the ring-shaped cytoplasm. After NNV injection for one week, the CD8α+ lymphocytes were proliferated from 3.6~5.5% to 4.2~6.68% which was counted after dynabeads positive selecton. The cytotoxic activity of CD8α+ lymphocytes at one-week post-NNV injection enhanced significantly against NNV-infected syngeneic fin cells in comparison with NNV-infected allogeneic or RSIV-infected syngeneic fin cells. These results revealed that the specific cytotoxicity and MHC restriction of the CD8α antibodies recognized-lymphocytes are the CTLs. The CD8α antibodies can’t recognize the PBLs of black sea bream, japanese eel and tilapia, however it can only specifically recognize the PBLs of the spotted grouper. Conclusively, the CD8α antibodies can provide a tool to advance the research in cellular immune response of spotted grouper

Effect of 6-week shoulder and neck exercises on improving neck disability of middle-aged and older adults with chronic neck pain

This study explored the effectiveness of 6-week programme involving shoulder and neck exercises on improving the neck disability of middle-aged and older adults with chronic neck pain. Participants in the Intervention Group (IG) were asked to participate in a 6-week training programme of shoulder and neck exercises, thrice a week, in addition to receiving passive modalities. The Control Group (CG) received passive modalities only. A primary outcome measure was change in Neck Disability Index (NDI) and secondary outcome measures involved changes in pain scores on a visual analogue scale and Cervical Range of Motion (CROM). Assessments were conducted at baseline and after 6-week intervention. In total, 72 participants were recruited for randomised controlled study. After the intervention, the pain scores, NDI and CROM of the IG displayed more significant improvement in the post-test than did the CG. The IG achieved significant improvement in NDI (-7.15 scores; p<0.001), pain scores (-27.97mm; p<0.001) and CROM (p<0.001) after the intervention. The intervention can reduce NDI scores, reduce the self-reported perception of pain scores, and improve CROM of middle-aged and older adults with chronic neck pain