bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 548: Scavenger Receptor Bi-deficiency-induced Hypercholesterolemia Impairs Lymphocyte Homeostasis
Authors: Zheng Zhong , Ai Junting , Guo Ling , Howatt Deborah D , Daugherty Alan , Bondada Subbarao , Li Xiang-An 
Published: March 2022

Objective-
Deficiency in Scavenger receptor BI (SR-BI), an HDL receptor, leads to a specific HDL-c-elevated hypercholesterolemia and impairs lymphocyte homeostasis in mice. However, how SR-BI-deficiency-induced hypercholesterolemia contributes to the adaptive immunity defects remains unknown.

Hypothesis-
SR-BI-deficiency-induced hypercholesterolemia leads to the impaired lymphocyte homeostasis.

Methods and Results-

We utilized two animal models combined with cell culture to test our hypothesis. First, in a high fat/high cholesterol (HF) diet mouse model (n>10 per group), we introduced a 2-fold increase in plasma cholesterol concentration. For HF diet SR-BI
+/+
mice, the increased cholesterol was distributed in VLDL/LDL fraction, but the lymphocyte homeostasis was not affected. In contrast, for HF diet SR-BI
-/-
mice, in addition to VLDL/LDL-c, HDL-c was also further elevated. These mice exhibited marked disruption in lymphocyte homeostasis, as characterized by a 1.6-fold increase in T cell number (2.7-fold vs SR-BI
+/+
group), a 2.1-fold increase in B cell number (4.2-fold vs SR-BI
+/+
group), exacerbated T cell activation and further accelerated B cell proliferation compared with control diet SR-BI
-/-
mice. Then, in a bone marrow transplantation model (n=5~6 per group), we found that HF diet SR-BI
-/-
recipients whether receiving SR-BI
+/+
or SR-BI
-/-
BMCs displayed higher lymphocyte numbers, greater lymphocyte activation and more lymphocyte proliferation than respective HF diet SR-BI
+/+
controls, indicating that non-hematopoietic SR-BI-deficiency environment was the dominant factor leading to the impaired adaptive immunity. Finally, in a series of
in vitro
lymphocyte activation experiments (n=4 per group, repeated twice), we found that HDL from control diet or HF diet SR-BI
+/+
mice exhibited an inhibitory effect on T cell or B cell proliferation under TCR or BCR stimulation, but HDL from HF diet SR-BI
-/-
mice completely lost that function, indicating that the dysfunctional HDL in HF diet SR-BI
-/-
mice was responsible for impaired lymphocyte homeostasis.

Conclusion-

The SR-BI-deficiency-induced hypercholesterolemia is the leading factor for impaired lymphocyte homeostasis in SR-BI
-/-
mice.
Abstract 558: A Role for Syndecan-1 in the Motility of Alternatively Activated Macrophages
Authors: Haller Carolyn , Angsana Julianty , Chaikof Elliot 
Published: March 2022

Background
Syndecan-1 (Sdc-1) is a member of a family of cell surface proteoglycans that has been reported to participate in the regulation of events relevant to tissue repair and chronic injury responses, including cell-substrate interactions, matrix remodeling and cell migration and proliferation. We have previously demonstrated that macrophage Sdc-1 expression is up-regulated and protective in 2 clinically relevant models of chronic inflammation: elastase-induced AAA and Western diet-induced atherosclerosis. In this study, we aim to define the subtype and functional consequence of Sdc-1 expressing macrophages.

Methods
Human and mouse Sdc-1 macrophages were enriched and characterized against defined macrophage subtypes, including classically activated M1(IFN-γ/LPS), M2 (IL-4), and M2 (IL-10). Endpoints included (i) surface expression of MHCII, CD86, PD-L2, and Sdc-1; (ii) iNOS and arginase activity; and (iii) ELISA or qPCR cytokine and chemokine analysis. Motility was examined using boyden chamber chemotaxis and adhesion to ECM was measured using CSFE labeled macrophages.

Results

In vitro characterization of Sdc-1 expressing macrophages, which can be generated through the activation of cAMP/PKA signaling pathway or through exposure to adenosine, indicates that Sdc-1 expression defines an anti-inflammatory population with high intrinsic motility. Sdc-1 macrophages displayed reduced expression of pro-inflammatory cytokines and chemokines and displayed limited potential to mediate monocyte or CD4 T cell chemotaxis. Using Sdc-1
-/-
macrophages, we found that the absence of Sdc-1 is associated with defective actin reorganization, resulting in dampened macrophage migration and enhanced adhesion to extracellular matrix. Analysis with human macrophages confirmed Sdc-1 expression in an alternatively activated, motile population.

Conclusions
Collectively, these results reinforce the physiological significance of macrophage motility as an endogenous modulator of the inflammatory response and further suggest Sdc-1 as a marker of alternatively activated macrophages in chronic inflammation.
Abstract 560: Role of Paraoxonase 2 In Innate Immune Response in an Acute Infection Model
Authors: Devarajan Asokan , Bourquard Noam , Gao Feng , Ganapathy Ekambaram , Verma Jitendra , Reddy Srinivasa T 
Published: March 2022

Background

(3-oxododecanoyl)-L-homoserine lactone (3OC12-HSL) is a quorum-sensing molecule produced by certain classes of gram-negative microbial pathogens, such as
Pseudomonas aeruginosa
(PAO1), is involved in the regulation of virulence factors and alters the function of the host cells. Previously, We have shown that paraoxonase 2 (PON2) hydrolyzes 3OC12-HSL in murine tracheal epithelial cells. In this study, we examined the physiological role of PON2 in innate host defense using the PA01-induced acute infection mouse model.

Materials and Results

We demonstrates that both intact cells and membrane-enriched protein lysates obtained from PON2-deficient (PON2-def) macrophages reveal a marked impairment in their ability to hydrolyze 3OC12-HSL. PON2 is neither up-regulated nor down-regulated in response to 3OC12-HSL in control macrophages. In an
ex vivo
model, macrophages derived from PON2-def mice show an increase in ER stress-mediated oxidative stress and defective PI3 kinase/AKT activation, leading to reduced phagocytes function compared to control macrophages following 3OC12-HSL treatment. Moreover, p85, a regulatory subunit of PI3-kinase, shows higher tyrosine nitration in the macrophage of PON2-def mice which is associated with phagocytic function. Restoration of PON2 in the macrophages reduces the ER stress mediated oxidative stress and improves the phagocytosis function in response to 3OC12-HSL treatment. Furthermore, following administration of 1.6x10
7
CFU of PAO1, a decrease in bacterial clearance is found in the lungs, and liver of PON2-def mice by 5.7, and 14.8 fold, respectively.

Conclusion
Increased ER stress mediated oxidative stress decreases phagocytosis function in the macrophages of PON2-def mice in response to 3OC12-HSL may explain the decreased bacterial clearance observed in PON2-def mice following PAO1 infection.
Abstract 561: Serpin Suppression of Macrophage and T Helper Cell Response in Temporal Artery Biopsy Xenograft Implants from Patients with Giant Cell Inflammatory Arteritis
Authors: Chen Hao , Zheng Donghang , Ryden Sally , Hazem Sami , Bartee Mee Y , Davids Jennifer , Sobel Eric , Dai Erbin , Liu Liying , Macaulay Colin , Weyands Cornelia , Thoburn Robert , Lucas Alexandra 
Published: March 2022

Background

Glucocorticoids are a principal treatment for giant cell arteritis (GCA), with some studies reporting predominate effect on T helper17 (Th17) cell activity in GCA, with lesser effects on other inflammatory cells. Serp-1 is a 55kDa myxomaviral
serpin
that modifies macrophage, Th1, and Th17 responses in aortic transplants in animal models.

Objectives

We have analyzed the effects of a virus-derived anti-inflammatory
ser
ine
p
rotease
in
hibitor (
serpin
) on human temporal artery (TA) biopsies isolated from patients with suspected GCA.

Methods
Using a newly developed “Aortic Window Xenopatch” model, human TA biopsy sections were divided implanted as full thickness grafts into the abdominal aorta of SCID mice in parallel and tested for response to Serp-1, with and without human PBMC infusion (N = 32).

Results

TA sections positive for arteritis (GCA
pos
) displayed significantly increased inflammatory plaque when compared to negative sections (GCA
neg
). Serp-1 reduced plaque in GCA
pos
sections (<0.01) with concommitant reduction in Th1, and Th17 splenocytes on flow cytometry (P < 0.01), with decreased IFNγ and IL-17 gene expression by RT-PCR array analysis. Serp-1 also reduced arterial CD11b
+
cells after PBMC infusion with reduced TNF-α expression in spleen, but not CD3
+
, CCR6
+
, nor CD86
+
. Splenocytes from mice with GCA
pos
grafts had increased interleukin-1beta (Il-1β), IL-17, and CD25 expression, while IL-1β was significantly reduced by Serp-1.Serp-1 also markedly reducedt gene expression for the FII, FXa-PAR2, tPA, uPA in splenocytes (P <0.05).

Conclusions

Treatment with a virus-derived serpin, significantly reduces inflammation and plaque thickness in human GCA
pos
TA xenograft implants with associated reductions in macrophage and Th1 and Th17 responses in mice.
Abstract 563: Axl Regulates Major Histocompatibility Complex Class II (MHC-II) Expression through STAT1 Pathway in Smooth Muscle Cells
Authors: Batchu Sri Nagarjun , Gerloff Janice , Korshunov Vyacheslav 
Published: March 2022

Introduction

Major histocompatibility complex class II (MHC-II) is upregulated in vascular smooth muscle cells (VSMCs) in response to injury. Previously we found that Axl, a receptor tyrosine kinase, is important for vascular remodeling via regulation of MHC-II
in vivo
. The aim of this study was to identify Axl-mediated intracellular signaling that is responsible for upregulation of MHC-II in VSMCs.

Methods and Results

Mouse aortic smooth muscle cells (MASMCs) were isolated from Axl wild type (Axl+/+) or knockout (Axl-/-) littermates from our colony. Cells were treated with interferon gamma (IFNγ; 1,000U/ml) or PBS for various times (10min-24hrs). There was no difference in apoptosis as measured by cleaved caspase-3 in Axl-/- vs. Axl+/+ after treatment with IFNγ. Likewise, phosphorylation levels of Akt were similar between the groups. Expression of MHC-II mRNA and protein was significantly lower in Axl-/- vs. Axl+/+ MASMCs. Levels of major histocompatibility class II transactivator (CIITA) mRNA was also significantly reduced in Axl-/- MASMCs. Reduced expression of number of cytokines and chemokines was observed in Axl-/- vs. Axl+/+ MASMCs after treatment with IFNγ. Similar to our
in vivo
results, a complement component 3 (C3) expression was drastically reduced in Axl-/- vs. Axl+/+ MASMCs. We found that STAT1, a major regulator for CIITA-MHC-II and C3 pathways, was not phosphorylated in Axl-/- vs. Axl+/+ MASMCs upon IFNγ treatment.

Conclusions
This is the first study that demonstrates that Axl affects STAT1-signaling in VSMCs. Our findings suggest that Axl contributes to vascular remodeling via enhancement of pro-inflammatory signals in smooth muscle cells.
Abstract 569: M-RNA Expressions in Carotid Atherosclerotic Lesions Related to Plasma Levels of Lipoprotein (a)
Authors: Bergmark Claes 
Published: March 2022

Objective
High plasma levels of Lipoprotein (a), (Lp(a)) is an atherosclerotic risk factor. Individual levels of Lp(a) are genetically determined and constant over lifetime. Thus, individuals with high Lp(a) levels are exposed to this risk factor over a long time. In vitro, Lp(a) has many biochemical and signalling effects which could act proatherogenic, but until now, nothing has been possible to confirm in vivo. Plasma levels of Lp(a) correlate with lesion levels of Lp(a).

Methods
A consecutive series of 153 patients operated for Carotid stenosis by Trombendarterectomy (TEA) had plasma samples taken preoperatively. TEA-specimens were analysed for gene expression by Affymetrix. Plasma samples were analysed for levels of Lp(a) with routine antibody-methods. 20000 different m-RNA expression levels were correlated to 10log levels of Lp(a). Patients with plasma levels of Lp(a) above 60mg/dl were compared with those below 20mg/dl

Results
The most up regulated gene in patients with high Lp(a) compared to normal/low Lp(a) is MIAT (myocardial infarction associated transcript), also known as RNCR2 (retinal non-coding RNA) or Gomafu, a long non-coding RNA. MIAT is associated with risk of myocardial infarction. The most down regulated gene is Glypican 4, a member of heparane sulfate proteoglycans. Lp(a) binds to proteoglycans. Gene enrichment analysis revealed Lp(a) association with cholesterol efflux, oxygen binding and proteasome complex, each with a role in atherosclerosis.
m-RNA from the LPA-gene could not be detected, confirming previous assumption that apo (a) is not synthesised in vascular tissue.

Conclusions
High levels of Lp(a) is associated with gene expression in carotid lesions, some suggestive to a proatherosclerotic pathway. The new finding of association with MIAT and the proteasome complex deserves further studies.
Abstract 74: Crucial Role of ROCK2 in Vascular Smooth Muscle Cells in Hypoxia-Induced Pulmonary Hypertension in Mice
Authors: Shimizu Toru , Fukumoto Yoshihiro , Tanaka Shin-ichi , Satoh Kimio , Ikeda Shohei , Shimokawa Hiroaki 
Published: March 2022

Background
We have previously demonstrated that Rho-kinase (ROCK) pathway in vascular smooth muscle cells (VSMC) plays an important role in the pathogenesis of cardiovascular disease including pulmonary arterial hypertension (PAH). Rho-kinase has 2 isoforms; ROCK1 for inflammatory cells and ROCK2 for VSMC. We thus examined the specific role of ROCK2 in VSMC in the pathogenesis of PAH.

Methods and Results

In patients with PAH as compared with controls, the immunoreactivity of ROCK2 and phosphorylated ERK1/2 co-expressed with αSMA in pulmonary arterial media were markedly increased. Thus, we developed VSMC-specific ROCK2-deficient (ROCK2
+/-
) and VSMC-specific ROCK2-overexpressing (ROCK2-Tg) mice. The extent of hypoxia-induced pulmonary hypertension (10% O
2
for 4 weeks), as evaluated by pulmonary vascular remodeling, right ventricular (RV) systolic pressure and RV hypertrophy, was significantly reduced in ROCK2
+/-
mice and was enhanced in ROCK2-Tg mice compared with the littermates (n=10 each, all P<0.05). The expression of ROCK activity and phosphorylated ERK1/2 in lung tissues analyzed by Western blotting and the numbers of Ki67
+
proliferating VSMC and CD45
+
inflammatory cells in pulmonary arterioles revealed by immunostaining were significantly reduced in the ROCK2
+/-
mice and were increased in the ROCK2-Tg mice compared with the littermates (n=4 each, all P<0.05). In cultured murine aortic VSMC, both scratch assay and Boyden chamber assay showed that migration activity was significantly reduced in the ROCK2
+/-
VSMC and was increased in the ROCK2-Tg VSMC compared with littermates (n=5 each, both P<0.01). In addition, in primary pulmonary arterial VSMC (PASMC) cultured from patients with PAH, siRNA knockdown (-81%) of ROCK2 expression (ROCK2-siRNA) significantly suppressed the expression of ROCK activity and PDGF-induced phosphorylated ERK1/2 (n=4 each, both P<0.05). Compared with control-siRNA, PASMC transfected with ROCK2-siRNA exhibited significant reduction in both migration activity by scratch assay and proliferation activity by growth curve (n=3 each, both P<0.01).

Conclusions
ROCK2 in VSMC plays a crucial role in the pathogenesis of PH through enhanced VSMC migration and proliferation in mice in vivo.
Abstract 78: Inhibition of Mir-33 Overcomes the Deleterious Effects of Diabetes on Atherosclerosis Regression.
Authors: Distel Emilie , Chung Kellie , Murphy Andrew J , Barrett Tessa , Parathath Sajesh , Essau Christine C , Moore Kathryn J , Fisher Edward A 
Published: March 2022

Objective
Diabetes increases the risk of cardiovascular morbidity and mortality which remains elevated with conventional therapies, such as statins. We have shown that diabetes also impairs plaque regression following LDL reduction in mice, as evidenced by a higher macrophage (CD68+ cells) plaque content and a higher inflammatory state of these cells compared to non-diabetic mice. Type-2 diabetics have dyslipidemia characterized by elevated triglyceride levels and low HDL, the latter is thought to contribute to their risk, based on epidemiologic studies. In non-diabetic atherosclerotic mice, inhibition of miR-33, an intronic micro-RNA located within the SREBF2 gene, increases the plasma level of apoAI and HDL-cholesterol (HDL-C) and promotes plaque regression. We hypothesize that elevation of HDL through the inhibition of miR-33 will overcome the deleterious effect of diabetes on macrophage content and inflammatory phenotype in plaques in a regression evironment.

Methods and Results
Diabetic (STZ-injected) and non-diabetic Reversa (LdLr-/-Apob100/100Mttpfl/flMx1-Cre+/+) mice, a model in which diet-induced hypercholesterolemia can be quickly and efficiently reversed, were fed a western diet for 16 weeks and then treated with anti-miR-33 oligonucleotide or control anti-miR for 4 weeks. Treatment with anti-miR-33 increased HDL-cholesterol in both diabetic (17%) and non-diabetic mice (30%). Anti-miR-33 treated diabetic mice showed an improvement in plaque regression as evidenced by a 26% reduction in CD68 content compared to diabetic mice receiving control anti-miR, and an enrichment in anti-inflammatory M2 macrophages (assessed by MR 1 and YM1). Monocyte tracking with latex beads suggests that the reduction of macrophage content is due to a decrease in monocyte recruitment to the plaque, rather than an increase in macrophage egress. This reduction in recruitment correlated with a decrease in the monocytosis associated with diabetes in the anti-miR-33 treated diabetic mice.

Conclusion
These findings suggest that miR-33 inhibition is able to overcome the deleterious effects of diabetes on atherosclerosis regression, by decreasing monocytosis, monocyte recruitment and improving the inflammatory status of plaque macrophages.
Abstract 9: Apelin Protects Against Arterial Thrombosis in Mice with Diet-induced Obesity
Authors: Jin Hong , Wilson Katina M , Chauhan Anil K , Tsao Philip S , Lentz Steven R 
Published: March 2022

Background and Hypothesis
The prevalence of obesity and related thrombotic diseases is increasing steadily, but the underlying mechanisms of thrombosis are not well elucidated. Apelin is a peptide adipokine that is upregulated in obesity and has protective effects on cardiovascular function. We tested the hypothesis that apelin protects from thrombosis in a mouse model of diet-induced obesity.

Methods

To induce obesity, male apelin null (
Apln -/y
) mice and wild-type (WT) littermates were fed a high fat (HF) diet with 60% of Kcal from fat (TD.06414) for 12 weeks. Susceptibility to thrombosis of the carotid artery was assessed using a photochemical injury (rose bengal) method. Expression of plasminogen activator inhibitor-1 (PAI-1) was measured by ELISA and real-time PCR.

Results

Body weight and fasting glucose were higher in mice fed the HF diet than in mice fed a control diet (P<0.05) but did not differ between
Apln -/y
and WT mice (P>0.05). The time to stable occlusion of the carotid artery was significantly accelerated in
Apln -/y
mice compared with WT mice (24.8±1.5 vs 43.9±6.8 min, P=0.04) fed the HF diet, but not a control diet. Because apelin is known to regulate the expression of the antifibrinolytic serpin PAI-1, we investigated the role of PAI-1 in the antithrombotic mechanism of apelin. Plasma levels of total PAI-1 were elevated 3-fold in WT mice fed the HF diet (P<0.05 vs. control diet). An even larger 5-fold elevation of plasma PAI-1 was seen in
Apln -/y
mice fed the HF diet (P<0.05). Levels of PAI-1 mRNA in lung also were significantly elevated in
Apln -/y
mice (P<0.01 vs. WT mice). To directly determine the role of PAI-1 in the prothrombotic phenotype of
Apln -/y
mice, a PAI-1-blocking antibody was administered prior to inducing thrombosis of the carotid artery. The PAI-1 antibody nearly completely reversed the shortened time to stable occlusion (P=0.03) in obese
Apln -/y
mice.

Conclusions
Endogenous apelin protects from arterial thrombosis in obesity, in part by dampening obesity-induced upregulation of PAI-1. These findings may have pharmacological implications for the prevention of thrombotic complications of obesity and metabolic syndrome.
Abstract 103: Cross-Talk Between Macrophages, Smooth Muscle Cells and Endothelial Cells in Response to Cigarette Smoke: The Effects on MMP2 and -9
Authors: Ghosh Abhijit , Pechota Angela , Neevel Andrew , Golota Natalie , Upchurch Gilbert R , Eliason Jonathan L 
Published: March 2022

Introduction:
Abdominal Aortic Aneurysm (AAA) development is linked to smoking and is characterized by extracellular matrix remodeling by MMP2 and -9. We hypothesized that MMP2 and -9 secretions and regulation in response to cigarette smoke is modulated by interactions between resident cells within the aorta, namely, aortic smooth muscles, endothelial cells, and infiltrating macrophages and could influence AAA formation and/or progression.

Methods:
Aqueous extract of cigarette smoke (AEC) was applied to male rat aortic smooth muscle (RASMC), endothelial (RAEC) or RAW cells in serum free medium (SFM). After 24h the cells were washed, fresh SFM was added and collected after 24h. This conditioned media (CM) was applied to the cells for 24h, again washed, fresh SFM added, and then incubated for 72h, at which point the media was saved for MMP2 and -9 analysis by zymography while the cellular protein was used to determine the ratio (pS/pJ) of phospho-Stat3 (pStat3) and phospho-Jak2 (pJak2) by western blot.

Results:
Unstimulated CM from RAW cells increased MMP2 by 24% (p=0.0357), MMP9 by 282.1% (p=0.0004) and pS/pJ by 136.5% in RASMC. AEC-CM from RAW and RAEC cells increased MMP9 by 200.5% (p=0.0004) and 17.1% (p=0.0301) respectively in RASMC and corresponding increases in pS/pJ (305.7% and 227.6%, respectively).

AEC-CM derived from RAW cells induced RAEC to produce moderate amounts of MMP2 (16.5%, p=0.0140) and MMP9 (29.5%, p=0.0207) and a 137.0% increase in pS/pJ.
RAW cells receiving unstimulated CM from RASMC and RAEC cells produced significant amounts of MMP9 (127.7%, p=0.0026 and 236.8%, p=0.0020, respectively) and increased levels of pS/pJ ratios (45.2% and 1282.8%, respectively). Moreover, AEC-CM from RASMC and RAEC induced significant production of MMP9 from RAW cells (154.8%, p=0.0001 and 162.4%, p=0.0022, respectively) and increase in pS/pJ ratios (1348.2%, and 1493.9%, respectively) over baseline.

Conclusion:
This is the first in vitro study demonstrating that cigarette smoke results in differential interactions between aortic smooth muscle cells, endothelial cells, and macrophage lineage cells resulting in enhanced levels of MMP2, MMP9, pStat3 and pJak2. These interactions may play an important role in AAA formation under in vivo conditions.