bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of Rab7 in activating NF-kB in B cells to mediate class-switching and plasma cell survival in antibody and autoantibody responses and to promote B cell lymphomagenesis
Authors: Yan Hui , Wang Rui , Lam Tonika , Kulp Dennis , Rivera Carlos Eduardo , Taylor Julia R. , Wang Zhonghua , Zan Hong , Zhong Guangming , Ivanov Dmitri , Casali Paolo , Xu Zhenming 
Published: January 2023

Abstract
NF-kappaB activation mediates B cell class-switching to generate IgG, IgA and IgE as well as survival of plasma cells, as differentiated from B cells to secrete antibodies at a high rate. Here we have identified a B cell-intrinsic role of Rab7, a small GTPase located in endosomes, in activating NF-kappaB for class-switching and plasma cell survival. Mice that are deficient in Rab7 specifically in B cells or treated with CID 1067700, a Rab7-specific small molecule inhibitor, cannot mount T-independent or T-dependent switched antibody responses, despite normal B cell proliferation/survival and T cell/myeloid cells functions. Rab7 co-localizes, on endosomes, with the CD40 receptor that is internalized upon engagement and directly interacts with TRAF6, an adaptor of CD40 and TLR signaling for NF-kB activation, thereby forming complexes that transduce signals. Disruption of such complexes by CID 1067700 hampers NF-kappaB activation in normal B cells and abrogates NF-kappaB dysregulation in lupus B cells and B lymphoma cell lines, in which Rab7 is upregulated. Treatment with CID 1067700 reduces pathogenic IgG autoantibody responses in murine lupus models, leading to life-span extension, and prevents tumorigenesis in nude mice transplanted with B lymphoma cells. These, together with the rescue of class-switching/plasma cell survival in normal B cells and proliferation/survival of B lymphoma cells by enforced NF-kappaB activation, indicate that Rab7 plays an important role in NF-kappaB activation to mediate B cell differentiation and dysregulation and is a potential therapeutic target in lupus and B cell lymphomagenesis.
Supported by NIH grants AI 079705, AI 105813/105813S1 (to P. C.) and AI 124172 (to Z. X.).
ADAM10 is a novel sheddase of Inducible Costimulatory Ligand (ICOSL).
Authors: Luker Andrea , Lownik Joseph Cornelius , Conrad Daniel 
Published: January 2023

Abstract
ICOSL is an important co-stimulatory molecule involved in the development of TH2 responses through germinal center reactions. Although not well defined, the regulation of ICOSL is known to occur through its interaction with ICOS. Evidence suggests surface expression levels are regulated by a catabolic event, and not through internalization. Here we confirm that regulation of ICOSL is mediated by the metalloproteinase ADAM10. Cleavage of recombinant ICOSL results in an amino-terminal fragment of approximately 3kDa, which is blocked by an ADAM10-specific inhibitor. Recombinant and cell-based cleavage assays suggest that ADAM17 can also cleave ICOSL. In vivo, we observed a ten-fold increase in ICOSL expression in ADAM10B−/− mice compared to wildtype, a phenomenon that was absent in ADAM17B−/− mice. This suggests that ADAM10 is the primary physiological sheddase of ICOSL, while ADAM17 may serve as a secondary sheddase. Additionally, ADAM10B−/− mice develop a significantly diminished T follicular helper (TFH) cell population in response to NP-KLH/alum immunization. Together, these data provide insight into the regulatory mechanism between ICOS/ICOSL and suggest ADAM10 may be a potential therapeutic target for attenuating antibody driven diseases.
Alpha(v) Integrins Engage Autophagy Components to Regulate B cell Immune Response
Authors: Acharya Mridu , Sokolovska Anna , Tam Jenny M , Raso Fiona , Stuart Lynda , Jackson Shaun , Rawlings David J , Lacy-Hulbert Adam 
Published: January 2023

Abstract
Signaling through Toll-like receptors (TLR) has been implicated in activation of immune cell populations such as B lymphocytes. TLR ligands associated with self-antigens can drive increased self-reactive B cell responses however TLR ligands present in pathogenic products or vaccines can also enhance protective antibody responses. Our recent work has identified a novel mechanism by which αv integrins, a family of adhesion molecules and the autophagy proteins limit excessive B cell TLR signaling.
Specifically, we have found that αvβ3 integrin regulates TLR signaling (NFKB and IRF7 activation) by directing maturation of TLR containing endosomes, through activation of components of the autophagy pathway (LC3 and Atg5). B cells lacking either αv integrins or autophagy components show enhanced TLR signaling and increased proliferative responses to TLR ligands. Mice with disruption in this pathway develop increased autoantibodies with age and develop accelerated autoimmunity in murine lupus models. Therefore, we propose that αv-mediated regulation of TLR signaling exists to limit excessive B cell responses to self-antigens.
In addition, we find that mice in which αv is specifically deleted from B cells mount stronger antibody responses when immunized with exogenous antigens containing TLR-ligand adjuvants. Immune responses to these antigens are characterized by increased expansion of germinal center cells, increased somatic hyper-mutation and higher levels of high-affinity class-switched antibodies. Therefore this αv mediated regulation of TLR signaling not only regulates B cell responses to self-antigens but it is also important for regulating germinal center B cell responses and production of high-affinity antibodies.
TRPV4 attenuates M1 polarization and subsequent NASH progression by stalling CYP2E1-mediated redox toxicity via eNOS.
Authors: Seth Ratanesh K , Chandrashekaran Varun , Dattaroy Diptadip , Alhasson Firas , Nagarkatti Mitzi , Nagarkatti Prakash , Diehl Anna Mae , Liedtke Wolfgang , Chatterjee Saurabh 
Published: January 2023

Abstract
M1 macrophage polarization in nonalcoholic steatohepatitis (NASH) liver has been impacted by several exogenous or endogenous factors including inflammatory stimuli, oxidative stress, and cytokines. In absence of an active endogenous defense mechanism, the M1 polarization bias potentiates NASH progression by intensified inflammation and intermittent fibrosis in the rodent model of liver injury. We introduce an endogenous defense mechanism in the liver that is mediated by TRPV4, a transient receptor potential calcium-permeable ion channel that responds to the cytotoxic liver environment and negatively regulates CYP2E1. We hypothesized that CYP2E1-mediated oxidative stress causes M1 polarization in experimental NASH and active TRPV4 inhibits CYP2E1 mediated inflammation with concomitant attenuation of M1 polarization. Since CYP2E1 takes center stage in redox toxicity we use a toxin model of NASH which uses pyrazole, a ligand and a substrate of CYP2E1 for inducing liver injury with NASH-like phenotype. Using both cyp2e1−/− and trpv4−/− mice, we show that in the absence of active TRPV4, CYP2E1 induced oxidative stress causes M1 polarization bias, that includes a significant increase in IL-1β, IL-12, IL-6 and IL-23 while CYP2E1 null or diallyl sulfide treated mice prevent it. Recently, we discovered that the TRPV4 modulates eNOS activation and nitric oxide release from Kupffer cell during an early stage of liver injury. Based on the adaptive NO increase in trpv4+/+ mice, NO donor administration in trpv4−/− mice abrogated CYP2E1-induced oxidative stress, M1 polarization, and NASH progression. Thus, a novel endogenous defense molecule TRPV4 can be a promising immunotherapeutic approach against chronic liver injury.
Reduced circulating MAIT cells in obesity
Authors: Liu Jianyun , Brutkiewicz Randy R 
Published: January 2023

Abstract
Obesity is a major risk factor for certain types of cancer and the mechanisms by which obesity contributes to tumor metastasis are not well understood. Whereas the immune system is essential for fighting against tumors, in the context of obesity, systemic chronic inflammation with altered immune responses are found. Recent studies have revealed that the gut microbiota also contributes to obesity and the pathology associated with that disease. Mucosal-associated innate T (MAIT) cells are a unique subpopulation of T cells that are characterized by the expression of a semi-invariant TCR a chain (Vα19 in mice; Vα7.2 in humans). Although the development and maturation of MAIT cells require the gut microbiota and antigen presenting molecule MR1, the actual function of MAIT cells has not been well studied. We have found reduced circulating MAIT cells in obese patients as compared to age- and gender-matched healthy donors. Using real-time PCR to determine the expression levels of the invariant TCR of MAIT cells and classical T cell transcription factors, we found that, although no difference was observed in any gene expression analyzed between samples from obese patients or healthy donors, mRNA expression of the Vα7.2 TCR positively correlated with the transcription factor TBX21 (found in T helper 1 cells) and BCL6 (found in follicular helper T cells). Our work suggests that obesity alters the number of MAIT cells. The correlation of MAIT cell TCR gene expression with transcription factors of classical effector T cells, suggests there may be transcriptional pathways linking MAIT cells to the function of conventional T cells. Future work will focus on determining how obesity impacts the function of MAIT cells.
TWEAK/Fn14 activation is instrumental in Ro52-mediated photosensitization in cutaneous lupus erythematosus
Authors: Liu Yale , Xu Meifeng , Min Xiaoyun , Wu Kunyi , Zhang Ting , Li Ke , Xiao Shengxiang , Xia Yumin 
Published: January 2023

Abstract

Background
TWEAK, a member of the TNF superfamily, binds to the Fn14 receptor, participating in many pathological alterations. Ro52 autoantigen mediates the photosensitization of lupus erythematosus. Recently, TWEAK/Fn14 activation was found in the lesion of cutaneous lupus erythematosus (CLE).

Objectives
To further show the potential role of TWEAK/Fn14 pathway in Ro52-mediated photosensitization.

Methods
The expression of TWEAK and Fn14 was detected in skin tissues from patients with LE and healthy donors. The expression of Ro52 was determined in murine keratinocytes that were treated with TWEAK or plus UVB irradiation. Some cells were pretreated with siRNA transfection or specific inhibitors of signaling pathways. The chemotaxis of J774.2 macrophages was evaluated on TWEAK stimulation of cocultured keratinocytes. The expression of TNF receptor associated factor 2 (TRAF2) and TNF receptors (TNFR) was determined in keratinocytes upon UVB or plus TWEAK treatment.

Results
TWEAK, Fn14, and downstream cytokines were highly expressed in CLE lesions that overexpressed Ro52. Moreover, TWEAK promoted the enhancement effect of UVB on Ro52 expression in keratinocytes. Meanwhile, TWEAK stimulation of keratinocytes favored the migration of macrophages. Furthermore, nuclear factor-kappa B inhibitor or Fn14 siRNA transfection abrogated the TWEAK effect on Ro52 regulation. Similarly, TRAF2 siRNA attenuated the TWEAK effect on Ro52 expression. Interestingly, UVB irradiation increased the expression of TNFR1 but decreased TNFR2 expression in keratinocytes.

Conclusion
TWEAK/Fn14 activation is instrumental in Ro52-mediated photosensitization and involves the function of the Fn14-TRAF2-TNFR axis.
Noncanonical NF-κB signaling is elevated in inflammatory bowel disease patients and may be associated with therapeutic response
Authors: Allen Irving C , Eden Kristin , Nguyen Vu , Knight Kristin , Sorrentino Dario 
Published: January 2023

Abstract
The canonical NF-κB signaling pathway is well characterized in the context of inflammatory bowel disease (IBD). However, noncanonical NF-κB signaling has not been evaluated. This pathway is key for the production of specific chemokines with diverse immunological effects in the gut. To evaluate this pathway, biopsy specimens from control patients and patients with IBD (CD and UC) were collected and pathways associated with inflammation were assessed via custom RT-PCR arrays. These expression data were compared to lesion status, IBD subtype (CD or UC), and treatment. Our analysis revealed significant up-regulation of several key genes associated with noncanonical NF-κB signaling in lesion tissue from IBD patients compared to controls and non-lesion areas. Interestingly, CD patients displayed increased noncanonical signaling compared to both control and UC patients in over 20 genes directly involved in the noncanonical pathway. Treatment with the anti-TNF biologic infliximab resulted in down-regulation of these and other noncanonical NF-κB signaling molecules in CD patients. Subsequent bioinformatics analysis of CD patients revealed that elevation of chemokines associated with noncanonical NF-κB signaling are indicators of infliximab responsiveness, with patients refractory to infliximab showing significantly elevated levels of gene expression. Together, these data indicate that noncanonical NF-κB signaling is elevated in IBD patients and may have links to therapeutic response.
Correlation of sentinel lymph node immune cells with disease-free survival and metastasis in breast cancer patients using 4-color chromogen-based immunohistochemistry and quantitative imaging microscopy
Authors: Blenman Kim RM 
Published: January 2023

Abstract
In this study, we assessed whether the cell densities of in situ sentinel lymph node (SLN) CD3+ T cells, CD20+ B cells, and CD1a+ dendritic cells correlated with cancer cell invasion, disease-free survival, or distant metastasis in breast cancer patients. Multiplexed chromogen-based IHC and quantitative microscopy imaging were performed on SLNs from 115 breast cancer patients.

Results
In our cohort, breast SLN T cells were reduced in microenvironments invaded by cancer cells (positive) compared to microenvironments that were cancer cell free (negative) (p = 0.03). The T cell reduction was dependent on cancer cell invasion (pearson correlation = −0.21, p = 0.03). There were no differences in dendritic cells or B cells between positive and negative microenvironments. Low numbers of T cells and B cells were associated with shorter disease-free survival in univariate analysis (p &lt; 0.0001, HR = 0.43; p = 0005, HR = 0.46, respectively). Logistic regression revealed that B cells were the only immune cells that were associated with an increased odds of metastasis in both univariate and multivariate analysis (p = 0.02, OR = 2.41; p = 0.01, OR = 5.42, respectively). SLN T cells were the only immune cells that were significantly reduced in patients with local metastases (p = 0.04). SLN B cells were the only immune cells that were significantly reduced in patients with distant metastases (p &lt; 0.0001).

Conclusion
In our breast cancer cohort, reduced numbers of SLN T cells predicted cancer cell invasion, shorter disease-free survival, and local metastases. Reduced numbers of SLN B cells predicted shorter disease-free survival and distant metastases.
hetIL-15 treatment of rhesus macaques is safe and systemically increases effector cytotoxic lymphocytes that infiltrate the lymph nodes.
Authors: Watson Dionysios C , Valentin Antonio , Moysi Eirini , Petrovas Costas , Bergamaschi Cristina , Felber Barbara K. , Pavlakis George N 
Published: January 2023

Abstract

Background
Heterodimeric interleukin-15 (hetIL-15) is presently evaluated in humans for the treatment of cancer. hetIL-15 activates and expands cytotoxic T and NK cells, which suggests that the cytokine could be useful for treating diseases such as malignancies and HIV infection.

Methods
Rhesus macaques received six s.c. injections of hetIL-15 over 2 weeks using increasing doses of cytokine (step-dosing). Animals were sacrificed after treatment and hetIL-15 effects on lymphocytes isolated from necropsy tissues were monitored by multiparametric flow cytometry and quantitative multiplexed confocal microscopy imaging (Histo-cytometry).

Results
This regimen was found to be safe in macaques and resulted in systemic proliferation and expansion of CD8+ and NK cells with higher granzyme B content. These expanded cells were found in both effector sites, such as liver, vagina and rectum, and secondary lymphoid tissues. A significant increase in effector memory CD8+ T cells was found in peripheral lymph nodes (LN) from all hetIL-15-treated macaques. Imaging analysis by Histo-cytometry revealed that effector CD8+ T cells infiltrated the B cell follicles where chronically infected follicular helper CD4+ T cells are located.

Conclusions
Step-dose administration of hetIL-15 is a well-tolerated regimen that results in systemic activation and expansion of cytotoxic leukocytes that infiltrate areas where chronic HIV-infected cells reside. These results suggest that hetIL-15 could be useful in disrupting or eliminating long-term viral reservoirs in HIV-1 infected individuals, contributing to a functional cure of the infection. Work assessing the impact of hetIL-15 on the size of the viral reservoir in sampled LN is currently ongoing.
Gemcitabine-generated ROS promotes neutrophil chemotactic activity through CXCL8 production via PKCδ/ERK pathway in a murine 4T1 breast cancer model
Authors: Jeong Jinseon , Kim Yong-Jae , Yang Kwang Hoon , Yoon Sun Young , Sohn Ki-Young , Kim Heung-Jae , Kim Jae Wha 
Published: January 2023

Abstract
Chemotherapy-induced neutropenia (CIN) is a common side effect that necessitate dose reductions during treatment of cancer patients. Preventing CIN is critical in chemotherapy because a rapid decline of neutrophil counts increase susceptibility to infection of cancer patients. In spite of its importance, the mechanism of CIN still remains unclear. This study is aimed to investigate the mechanism of CIN in an aspect of neutrophil activation by chemotaxis in gemcitabine-treated breast tumor model. We hypothesized that gemcitabine-generated ROS promotes CXCL8 or MIP-2 production by upregulating ERK pathway in PKCδ-activated intratumoral macrophages and thus neutrophil recruitment to the chemokine gradient. In 4T1 breast tumor mouse model, we found that the basal level of PKCδ activity was higher than in normal mice. The number of blood and peritoneal leukocytes was analyzed at 24h after intraperitoneal administration of gemcitabine. The neutrophil counts decreased in the blood but increased in the peritoneal cavity. In addition, intratumoral neutrophils were elevated in the gemcitabine-treated group. The mRNA levels of neutrophil attracting chemokines/receptors were increased in the peritoneal and tumoral cells. ROS inhibitors N-Acetyl-L-cysteine and Apocynin blocked gemcitabine-induced neutrophil migration, mRNA expression of the chemokines and receptors and PKCδ/ERK phosphorylation. CXCR1/2 inhibitor reparixin decreased neutrophil recruitment to the peritoneal cavity and the tumor tissue. Together, our results suggest that gemcitabine treatment promotes neutrophil chemotactic activity by increasing ROS/PKCδ/ERK/CXCL8 pathway. These findings will provide new insights for developing a therapeutic for CIN.
Cooperative anti-tumor effect of soluble NKG2D ligand MIC neutralization and PD-1 pathway blockade
Authors: Basher Fahmin , Wu Jennifer 
Published: January 2023

Abstract
Shedding of the human NKG2D ligand MIC from tumor cell surfaces correlates with progression of many epithelial cancers. Soluble MIC enables tumor immune escape through multiple immune suppressive mechanisms, such as disturbing natural killer (NK) cell homeostatic maintenance, impairing NKG2D expression on NK cells and effector T cells, and facilitating the expansion of arginase I+ MDSCs. Our recent studies have demonstrated that sMIC is an effective cancer therapeutic target. We thus sought to investigate whether targeting sMIC using a nonblocking monoclonal antibody would enhance immunotherapies currently in use, such as PD-1 blockade therapy, using genetically engineered transplantable syngeneic sMIC+ tumor models. We demonstrate that combination therapy of a sMIC-neutralizing antibody and anti-PDL1 improved the survival of animals bearing sMIC+ tumors, in comparison to monotherapy. We further demonstrate that combination therapy augmented the activation and functional potential of NK cells, CD8 T cells, and CD4 T cells. Combined therapy significantly enhanced antigen-specific CD8 T and CD4 cell responses. Combined therapy also re-modulated the tumor microenvironment to be more immune activating and reduced neovascularization. Our findings provide a rationale for co-targeting sMIC to enhance the therapeutic efficacy of PD-1 blockade.
Immune modulatory effects of chemotherapy increase the effectiveness of anti-PD1 immunotherapy in a poorly immunogenic murine model of osteosarcoma
Authors: Ladle Brian H , Phillips Maggie J , Yu Colburn , Gamper Christopher J 
Published: January 2023

Abstract
While great excitement surrounds the success of anti-PD1 cancer immunotherapies, few responses are observed treating sarcomas with these agents. The low mutation burdens and minimal T cell infiltrates present in sarcomas, especially pediatric sarcomas, may explain the lack of response to anti-PD1 therapy. We investigated combining chemotherapy treatment with a sarcoma vaccine (GVAX) and anti-PD1 treatment in a mouse model of osteosarcoma. We targeted osteosarcoma as patients treated with standard of care chemotherapy agents (doxorubicin, cisplatin, and methotrexate (MTX)) maintain robust lymphocyte counts, making combination with immunotherapy plausible. Consistent with the poor immunogenicity of sarcomas, mice challenged with the syngeneic osteosarcoma cell line K7M2 showed poor responses to GVAX alone (irradiated K7M2 tumor cells mixed with irradiated GM-CSF-secreting bystander cell line), anti-PD1 alone, or combining GVAX with anti-PD1 treatment. Treating K7M2 tumors with doxorubicin or cisplatin alone had no impact on tumor growth. However, treatment with MTX alone did show shrinkage of tumors in 30–40% of mice with kinetics consistent with an immune-mediated response. Further supporting an immune-mediated effect, MTX alone had no impact on K7M2 tumor growth in NSG immune deficient mice. Combining MTX with GVAX and anti-PD1 resulted in complete rejection of established K7M2 tumors in 70% of mice. These experiments support further investigation of combining chemotherapy agents (such as MTX) with immunotherapies. Our findings also indicate that poorly responsive tumors to anti-PD1 monotherapy can be rendered responsive, broadening the potential use of checkpoint blockade to include poorly immunogenic tumors.