Alteration of type I interferon response is associated with subclinical atherosclerosis in virologically suppressed HIV‐1‐infected male patients

Abstract Given human immunodeficiency virus‐1 (HIV‐1)‐infected patients have alterations in the type I interferon (IFN‐I) pathway and are also at elevated risk of atherosclerosis, we evaluated IFN‐I response and subclinical cardiovascular disease (CVD) association in HIV‐1‐infected patients. Transcript levels of IFN‐α/β and IFN‐stimulated gene 56 (ISG56) were evaluated by RT/real‐time PCR in peripheral blood mononuclear cells collected from asymptomatic HIV‐1‐positive male patients at high risk of developing CVD (n = 34) and healthy subjects (n = 21). Stenosis degree (≥ or <50%), calcium volume score, calcium Agatston score, and myocardial extracellular volume were examined by coronary computerized tomography scan. Carotid intima‐media thickness (cIMT), Framingham risk score, atherosclerotic cardiovascular disease (ASCVD) score, and risk score developed by data collection on adverse effects of anti‐HIV drugs (D:A:D) were also measured. Increased IFN‐α, IFN‐β, and ISG56 levels were observed in all HIV‐1‐infected males compared to healthy controls (p < .001 for all genes analyzed). HIV‐1‐infected patients with a stenosis degree ≥50% showed a higher Framingham risk score (p = .019), which was correlated with IFN‐β and ISG56 levels. HIV‐1‐infected males with enhanced IFN‐I levels and stenosis displayed a higher ASCVD calculated risk (p = .011) and D:A:D score (p = .004). Also, there was a trend toward higher IFN‐α and ISG56 mRNA levels in HIV‐1‐positive patients with an increased cIMT (p > .05). Dysregulation of IFN‐I response might participate in the pathogenesis of HIV‐1‐associated CVD.


| INTRODUCTION
Since the introduction of combined antiretroviral therapy (cART), the life quality and survival of people living with human immunodeficiency virus-1 (HIV-1) have considerably enhanced. 1 Nevertheless, as a result of the harmful effects of HIV-1, exposure to prolonged cART and accelerated aging, cardiovascular disease (CVD) increasingly afflicts people living with HIV-1. 2,3 The variety of CVD in HIV-1 is not only restricted to enhanced atherosclerosis, but includes pulmonary hypertension, ventricular dysfunction related to myocarditis, complex cerebrovascular disease, pericardial pathology, endocarditis, and cardiac tumors. 4,5 The pathogenesis behind the cardiovascular HIV-1-associated complications is multifactorial, and involves distinct CVD risk parameters, but also viral and host factors associated with immunological and metabolic dysfunction 6 observed during HIV-1 infection. As a result of these observations, great interest has been directed toward the role of type I interferon (IFN-I) in atherosclerosis. [7][8][9][10][11][12][13] As HIV-1-infected patients have an enhanced activation of IFN-I signalling [14][15][16][17] and are at elevated risk of developing atherosclerosis, 18,19 we hypothesized that persistent upregula-

| Multi-detector computed tomography (MDTC) protocol and coronary angiography (CA)
All HIV-1-infected patients were tested for the severity of CVD using a low-dose prospectively ECG-triggered computerized tomography (CT) coronary angiography protocol, with a 64-slice multidetector MDCT scanner (Somatom Definition Siemens medical Solution, Forchheimen), as previously described. 21 Calcium score was quantified using the Agatston Calcium Score method 22 from 3 mm nonoverlapping sections by using a semiautomated software (calcium scoring CT; Siemens Medical Solutions). The luminal stenosis degree was classified as mild (Grade I: 30%-49%), moderate (grade II: 50%-69%), severe (Grade III: 70%-99%), or coronary occlusion (Grade IV: 100%). A threshold of 50% luminal narrowing in any coronary segment greater than 1.5 mm in diameter was adopted to define clinically significant coronary stenosis. 23 All HIV-1-infected patients with coronary lesions ≥ grade II at MDCT analysis were considered for coronary angiography. This evaluation was performed using the standard technique with angiograms that were examined by an experienced blinded operator to the MDTC results. The American Heart Association (AHA) segmentation model 24 was applied and the degree of stenosis was quantified (quantitative coronary angiography, QCA) (Allura Xper FD 10; Philips Medical Systems).
Significant stenosis was defined as a reduction in diameter ≥50%.  at the myocardial septum and within the left ventricular chamber; the mean area was 3 cm 2 (range, 1.5-5 cm 2 ). Mean attenuation at the ROI was recorded in HU and Myocardial ECV fraction was calculated using the following equation:

| CT image analysis
where the contrast agent partition coefficient (λ) represents the ratio of the change in the blood and myocardial attenuation (ΔHU) and H is the haematocrit level. The change in attenuation (ΔHU) was de- To avoid inter-operator differences, all the measurements of c-IMT were performed by a single operator.

| cIMT level and IFN-I expression relationship in the development of atherosclerosis
cIMT is considered a surrogate measure of subclinical atherosclerosis that can predict CVD events among the general population. 30 Thus, we  (Table 3).
This group of HIV-1-infected patients had higher calcium volume score, calcium Agatston score, and ECV compared to those with a cIMT <0.09 mm. (Table 3

CONFLICT OF INTERESTS
The authors declare that there are no conflict of interests.

PEER REVIEW
The peer review history for this article is available at https://publons. com/publon/10.1002/jmv.27028

DATA AVAILABILITY STATEMENT
The data are not publicly available due to them containing information that could compromise research participant privacy/ consent.