Myeloid cell tropism enables MHC-E-restricted CD8+ T cell priming and vaccine efficacy by the RhCMV/SIV vaccine

Science immunology

2022 Jun 24

Hansen, SG;Hancock, MH;Malouli, D;Marshall, EE;Hughes, CM;Randall, KT;Morrow, D;Ford, JC;Gilbride, RM;Selseth, AN;Trethewy, RE;Bishop, LM;Oswald, K;Shoemaker, R;Berkemeier, B;Bosche, WJ;Hull, M;Silipino, L;Nekorchuk, M;Busman-Sahay, K;Estes, JD;Axthelm, MK;Smedley, J;Shao, D;Edlefsen, PT;Lifson, JD;Früh, K;Nelson, JA;Picker, LJ;
PMID: 35714200 | DOI: 10.1126/sciimmunol.abn9301

The strain 68-1 rhesus cytomegalovirus (RhCMV)-based vaccine for simian immunodeficiency virus (SIV) can stringently protect rhesus macaques (RMs) from SIV challenge by arresting viral replication early in primary infection. This vaccine elicits unconventional SIV-specific CD8+ T cells that recognize epitopes presented by major histocompatibility complex (MHC)-II and MHC-E instead of MHC-Ia. Although RhCMV/SIV vaccines based on strains that only elicit MHC-II- and/or MHC-Ia-restricted CD8+ T cells do not protect against SIV, it remains unclear whether MHC-E-restricted T cells are directly responsible for protection and whether these responses can be separated from the MHC-II-restricted component. Using host microRNA (miR)-mediated vector tropism restriction, we show that the priming of MHC-II and MHC-E epitope-targeted responses depended on vector infection of different nonoverlapping cell types in RMs. Selective inhibition of RhCMV infection in myeloid cells with miR-142-mediated tropism restriction eliminated MHC-E epitope-targeted CD8+ T cell priming, yielding an exclusively MHC-II epitope-targeted response. Inhibition with the endothelial cell-selective miR-126 eliminated MHC-II epitope-targeted CD8+ T cell priming, yielding an exclusively MHC-E epitope-targeted response. Dual miR-142 + miR-126-mediated tropism restriction reverted CD8+ T cell responses back to conventional MHC-Ia epitope targeting. Although the magnitude and differentiation state of these CD8+ T cell responses were generally similar, only the vectors programmed to elicit MHC-E-restricted CD8+ T cell responses provided protection against SIV challenge, directly demonstrating the essential role of these responses in RhCMV/SIV vaccine efficacy.

The multifaceted melanocortin receptors

Endocrinology

2022 Jun 14

Laiho, L;Murray, JF;
PMID: 35700124 | DOI: 10.1210/endocr/bqac083

The five known melanocortin receptors (MCs) have established physiological roles. With the exception of MC2, these receptors can behave unpredictably and since they are more widely expressed than their established roles would suggest, it is likely that they have other poorly characterized functions. The aim of this review is to discuss some of the less well-explored aspects of the four enigmatic members of this receptor family (MC1,3-5) and describe how these are multifaceted G-protein coupled receptors (GPCRs). These receptors appear to be promiscuous in that they bind several endogenous agonists (products of the proopiomelanocortin gene) and antagonists but with inconsistent relative affinities and effects. We propose that this is a result of post-translational modifications that determine receptor localization within nanodomains. Within each nanodomain there will be a variety of proteins, including ion channels, modifying proteins and other GPCRs,that can interact with the MCs to alter the availability of receptor at the cell surface as well as the intracellular signalling resulting from receptor activation. Different combinations of interacting proteins and MCs may therefore give rise to the complex and inconsistent functional profiles reported for the MCs. For further progress in understanding this family, improved characterization of tissue-specific functions is required. Current evidence for interactions of these receptors with a range of partners resulting in modulation of cell signalling suggests that each should be studied within the full context of their interacting partners. The role of physiological status in determining this context also remains to be characterized.

Transcriptome and unique cytokine microenvironment of Castleman disease

Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc

2021 Oct 22

Wing, A;Xu, J;Meng, W;Rosenfeld, AM;Li, EY;Wertheim, G;Paessler, M;Bagg, A;Frank, D;Tan, K;Teachey, DT;Lim, MS;Prak, EL;Fajgenbaum, DC;Pillai, V;
PMID: 34686774 | DOI: 10.1038/s41379-021-00950-3

Castleman disease (CD) represents a group of rare, heterogeneous and poorly understood disorders that share characteristic histopathological features. Unicentric CD (UCD) typically involves a single enlarged lymph node whereas multicentric CD (MCD) involves multiple lymph node stations. To understand the cellular basis of CD, we undertook a multi-platform analysis using targeted RNA sequencing, RNA in-situ hybridization (ISH), and adaptive immune receptor rearrangements (AIRR) profiling of archived tissue from 26 UCD, 14 MCD, and 31 non-CD reactive controls. UCD showed differential expression and upregulation of follicular dendritic cell markers (CXCL13, clusterin), angiogenesis factors (LPL, DLL4), extracellular matrix remodeling factors (TGFβ, SKIL, LOXL1, IL-1β, ADAM33, CLEC4A), complement components (C3, CR2) and germinal center activation markers (ZDHHC2 and BLK) compared to controls. MCD showed upregulation of IL-6 (IL-6ST, OSMR and LIFR), IL-2, plasma cell differentiation (XBP1), FDC marker (CXCL13, clusterin), fibroblastic reticular cell cytokine (CCL21), angiogenesis factor (VEGF), and mTORC1 pathway genes compared to UCD and controls. ISH studies demonstrated that VEGF was increased in the follicular dendritic cell-predominant atretic follicles and the interfollicular macrophages of MCD compared to UCD and controls. IL-6 expression was higher along interfollicular vasculature-associated cells of MCD. Immune repertoire analysis revealed oligoclonal expansions of T-cell populations in MCD cases (2/6) and UCD cases (1/9) that are consistent with antigen-driven T cell activation. The findings highlight the unique genes, pathways and cell types involved in UCD and MCD. We identify potential novel targets in CD that may be harnessed for therapeutics.

The wheat stem rust (Puccinia graminis f. sp. tritici) population from Washington contains the most virulent isolates reported on barley

Plant disease

2021 Sep 21

Upadhaya, A;Gc Upadhaya, S;Brueggeman, RS;
PMID: 34546770 | DOI: 10.1094/PDIS-06-21-1195-RE

A diverse sexual population of wheat stem rust, Puccinia graminis f. sp. tritici (Pgt), exist in the Pacific Northwest (PNW) region of the United States due to the natural presence of Mahonia spp. that serve as alternate hosts to complete its sexual life cycle. The region appears to be a center of stem rust diversity in North America where novel virulence gene combinations can emerge that could overcome deployed barley and wheat stem rust resistances. A total of 100 single pustule isolates derived from stem rust samples collected from barley in Eastern Washington during the 2019 growing season were assayed for virulence on the two known effective barley stem rust resistance genes/loci, Rpg1 and the rpg4/5-mediated resistance locus (RMRL) at the seedling stage. Interestingly, 99% of the Pgt isolates assayed were virulent on barley variety Morex carrying the Rpg1 gene, and 62% of the isolates were virulent on the variety Golden Promise transformant (H228.2c) that carries a single copy insertion of the Rpg1 gene from Morex and is more resistant than Morex to many Rpg1 avirulent isolates. Also, 16% of the isolates were virulent on the near isogenic line HQ-1, that carries the RMRL introgression from the barley line Q21861 in the susceptible Harrington background. Alarmingly, 10% of the isolates were virulent on barley line Q21861 that contains both Rpg1 and RMRL. Thus, we report on the first Pgt isolates worldwide with virulence on both Rpg1 and RMRL when stacked together representing the most virulent Pgt isolates reported on barley.

Intrinsic and growth-mediated cell and matrix specialization during murine meniscus tissue assembly

FASEB journal : official publication of the Federation of American Societies for Experimental Biology

2021 Aug 01

Tsinman, TK;Jiang, X;Han, L;Koyama, E;Mauck, RL;Dyment, NA;
PMID: 34314047 | DOI: 10.1096/fj.202100499R

The incredible mechanical strength and durability of mature fibrous tissues and their extremely limited turnover and regenerative capacity underscores the importance of proper matrix assembly during early postnatal growth. In tissues with composite extracellular matrix (ECM) structures, such as the adult knee meniscus, fibrous (Collagen-I rich), and cartilaginous (Collagen-II, proteoglycan-rich) matrix components are regionally segregated to the outer and inner portions of the tissue, respectively. While this spatial variation in composition is appreciated to be functionally important for resisting complex mechanical loads associated with gait, the establishment of these specialized zones is poorly understood. To address this issue, the following study tracked the growth of the murine meniscus from its embryonic formation through its first month of growth, encompassing the critical time-window during which animals begin to ambulate and weight bear. Using histological analysis, region specific high-throughput qPCR, and Col-1, and Col-2 fluorescent reporter mice, we found that matrix and cellular features defining specific tissue zones were already present at birth, before continuous weight-bearing had occurred. These differences in meniscus zones were further refined with postnatal growth and maturation, resulting in specialization of mature tissue regions. Taken together, this work establishes a detailed timeline of the concurrent spatiotemporal changes that occur at both the cellular and matrix level throughout meniscus maturation. The findings of this study provide a framework for investigating the reciprocal feedback between cells and their evolving microenvironments during assembly of a mechanically robust fibrocartilage tissue, thus providing insight into mechanisms of tissue degeneration and effective regenerative strategies.

ASICs are Required for Immediate Exercise-Induced Muscle Pain (IEIP), but not Delayed Onset Muscle Soreness (DOMS)

The Journal of Pain

2021 May 01

Khataei, T;Sluka, K;Harding, A;Benson, C;
| DOI: 10.1016/j.jpain.2021.03.056

IEIP and DOMS are two types of muscle pain induced by exercise. IEIP occurs during whereas DOMS appears days after exercise. Acid sensing ion channels (ASICs) expressed in muscle afferents, play a role in different pain conditions. ASICs are regulated via chemicals released during intense muscle contraction and microinjuries. Recently, we showed that ASICs are required for IEIP. Here, we tested if ASICs are also required for DOMS. Wild type (WT) and ASIC3-/- mice were divided into control and exercise groups. Exercise group underwent an exhaustive exercise; control group were placed on a non-moving treadmill. Locomotor movement was measured via Open field test to examine fatigue and/or soreness immediately and 24h after exercise. Next, exercise-induced muscle pain was assessed by muscle withdrawal threshold (MWT) at baseline, immediately and 24h after exercise. Our results showed; ASIC3-/- had similar baseline locomotor activity, muscle pain responses and exercise performance as WT. However, ASIC3-/- showed different responses after exercise compared to WT. WT had a lower MWT immediately and 24h after exercise compared to baseline. On the other hand, ASIC3-/- showed a lower MWT only 24h after exercise, but not immediately after. In addition, ASIC3-/- had significant lower locomotor activity than WT immediately after exercise. Also, ASIC3-/- had significant lower movement 24h after exercise compared to control ASIC3-/-, however this difference was not significant for WT mice. In summary, while WT and ASIC3-/- had a same exercise performance, they had different pain perception and fatigue immediately, but not 24h after exercise. Unlike WT, ASIC3-/- did not develop IEIP, even though ASIC3-/- had a higher fatigue level than WT as measured by diminished locomotion after exercise. On the other hand, ASIC3-/- developed DOMS a day after exercise like WT. These results show ASICs are required for immediate exercise-induced pain, but not exercise-induced fatigue and DOMS. Supported by the Department of Veterans Affairs Merit Award (5I01BX000776).

Parvalbumin interneuron inhibition onto anterior insula neurons projecting to the basolateral amygdala drives aversive taste memory retrieval

Current biology : CB

2021 Apr 23

Yiannakas, A;Kolatt Chandran, S;Kayyal, H;Gould, N;Khamaisy, M;Rosenblum, K;
PMID: 33930301 | DOI: 10.1016/j.cub.2021.04.010

Memory retrieval refers to the fundamental ability of organisms to make use of acquired, sometimes inconsistent, information about the world. Although memory acquisition has been studied extensively, the neurobiological mechanisms underlying memory retrieval remain largely unknown. Conditioned taste aversion (CTA) is a robust associative paradigm, through which animals can be trained to express aversion toward innately appetitive tastants. The anterior insula (aIC) is indispensable in the ability of mammals to retrieve associative information regarding tastants that have been previously linked with gastric malaise. Here, we show that CTA memory retrieval promotes cell-type-specific activation in the aIC. Using chemogenetic tools in the aIC, we found that CTA memory acquisition requires activation of excitatory neurons and inhibition of inhibitory neurons, whereas retrieval necessitates activation of both excitatory and inhibitory aIC circuits. CTA memory retrieval at the aIC activates parvalbumin (PV) interneurons and increases synaptic inhibition onto activated pyramidal neurons projecting to the basolateral amygdala (aIC-BLA). Unlike innately appetitive taste memory retrieval, CTA retrieval increases synaptic inhibition onto aIC-BLA-projecting neurons that is dependent on activity in aIC PV interneurons. PV aIC interneurons coordinate CTA memory retrieval and are necessary for its dominance when conflicting internal representations are encountered over time. The reinstatement of CTA memories following extinction is also dependent on activation of aIC PV interneurons, which increase the frequency of inhibition onto aIC-BLA-projecting neurons. This newly described interaction of PV and a subset of excitatory neurons can explain the coherency of aversive memory retrieval, an evolutionary pre-requisite for animal survival.

ABCA1 activity in the RPE is unnecessary for RPE reverse cholesterol transport (RCT) and AMD pathophysiology

Investigative Ophthalmology & Visual Science

2023 Jan 01

Coble, M;Aranda, J;Demirs, JT;Esterberg, R;Hanks, S;Jose, S;Leehy, B;Liao, S;Niu, YZ;Qiu, Y;Yang, J;

METHODS : Gene expression of ABCA1 and ApoA1 on human donor tissue and iPSC-RPE were examined by qPCR (n=3). Bulk RNAseq examined transcript changes in key RCT genes on donor retinas across different stages of disease progression. RNAscope probes (ACDBio) were designed against abca1 transcripts with appropriate mismatch controls. Neutral lipid stain with oil-red O on 10um cryo-sections of abca1 KO and wild type (WT) eyes (N= 5). Two siRNAs knocked down abca1 in iPSC-RPE cells to assess abca1 contribution to cholesterol efflux (n=3). Samples were analyzed with the cholesterol efflux kit (ab196985) and compared to non-targeting control siRNAs. Histological analysis of ABCA1 protein using anti-ABCA1 (Invitrogen-MA516026) on human donor retinas (AMD1 vs AMD3).

CAR-T cell therapy exerts effective antitumor efficacy under immunosuppressive conditions in adrenocortical carcinoma

Endocrine Abstracts

2023 May 02

Philipp, S;Landwehr, L;Justus, W;Altieri, B;Rodrigo, R;Tanja, M;Daniel, O;Kroiss, M;Sbiera, S;Michael, H;Fassnacht, M;
| DOI: 10.1530/endoabs.90.oc6.4

Background: Adrenocortical carcinoma (ACC) is a very rare and aggressive, endocrine malignancy with still limited treatment options. Approximately 60% of patients with ACC show endogenous glucocorticoid excess which could be one potential cause, why first clinical trials with immunotherapies, like immune checkpoint inhibitors, showed only modest results. Due to the lack of an ACC-specific antigen structure, other immunotherapeutic approaches, like specialized cancer treatments using chimeric antigen receptor (CAR) therapy in ACC, have not been tested so far. In this study, we evaluated the expression of a new enticing tumor antigen (TA*) structure and investigated the effect of TA-specific CAR-T cells _in vitro_.

The Role of Non-coding RNAs in Cerebellar Development

Contemporary Clinical Neuroscience

2023 Feb 25

Rahimi-Balaei, M;Ramirez, M;Gupta, I;Goldowitz, D;
| DOI: 10.1007/978-3-031-23104-9_6

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T028: Single-cell RNA sequencing reveals the interplay between circulating CD4 T cells, B cells and cancer-associated monocytes in classic Hodgkin lymphoma treated with PD-1 blockade

HemaSphere

2022 Oct 03

Paczkowska, J;Tang, M;Wright, K;Song, L;Shanmugam, V;Luu, K;Welsh, E;Cader, F;Mandato, E;Ouyang, J;Bai, G;Lawton, L;Rodig, S;Liu, X;Shipp, M;
| DOI: 10.1097/01.hs9.0000890680.82329.6b

The most abundant circulating CD3- population in patients with cHL was a newly identified monocyte subset with increased expression of multiple immunosuppressive and tumorigenic cytokines and chemokines, PD-L1 and SIRPa. This newly identified monocytic population was virtually absent from the blood of healthy donors. RNAscope analysis of the intact tumor microenvironment localized these tumor-infiltrating monocytes/macrophages to the immediate proximity of HRS cells. Monocytes from patients whose disease progressed following PD-1 blockade expressed significantly higher levels of immunosuppressive cytokine/chemokine signature which led to the development of a predictive transcriptional assay. We identified a comparable circulating monocyte population and transcriptional signature associated with unresponsiveness to PD-1 blockade in an additional solid tumor underscoring the broad-based significance of these findings.

Gene Expression Signatures of the Tumor Microenvironment: Relation to Tumor Phenotypes and Progress in Breast Cancer

Biomarkers of the Tumor Microenvironment

2022 Jul 13

Wik, E;Ingebriktsen, L;Akslen, L;
| DOI: 10.1007/978-3-030-98950-7_23

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	Description
sense Example: Hs-LAG3-sense	Standard probes for RNA detection are in antisense. Sense probe is reverse complent to the corresponding antisense probe.
Intron# Example: Mm-Htt-intron2	Probe targets the indicated intron in the target gene, commonly used for pre-mRNA detection
Pool/Pan Example: Hs-CD3-pool (Hs-CD3D, Hs-CD3E, Hs-CD3G)	A mixture of multiple probe sets targeting multiple genes or transcripts
No-XSp Example: Hs-PDGFB-No-XMm	Does not cross detect with the species (Sp)
XSp Example: Rn-Pde9a-XMm	designed to cross detect with the species (Sp)
O# Example: Mm-Islr-O1	Alternative design targeting different regions of the same transcript or isoforms
CDS Example: Hs-SLC31A-CDS	Probe targets the protein-coding sequence only
EnEm	Probe targets exons n and m
En-Em	Probe targets region from exon n to exon m
Retired Nomenclature
tvn Example: Hs-LEPR-tv1	Designed to target transcript variant n
ORF Example: Hs-ACVRL1-ORF	Probe targets open reading frame
UTR Example: Hs-HTT-UTR-C3	Probe targets the untranslated region (non-protein-coding region) only
5UTR Example: Hs-GNRHR-5UTR	Probe targets the 5' untranslated region only
3UTR Example: Rn-Npy1r-3UTR	Probe targets the 3' untranslated region only
Pan Example: Pool	A mixture of multiple probe sets targeting multiple genes or transcripts

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