MedPath

Postprandial Monocyte Study

Not Applicable
Completed
Conditions
Inflammation
Immune System
Nutrition
Interventions
Other: High saturated fat challenge meal
Other: High mono unsaturated fat challenge meal
Registration Number
NCT05792137
Lead Sponsor
USDA, Western Human Nutrition Research Center
Brief Summary

The purpose of this research is to determine the role of a type of immune cell in blood, called a non-classical monocytes (NCMs), following consumption of a high-fat meal. Previous studies have found that monocytes are important for blood vessel health. In this study, two different high-fat meals will be used to study the effect of different types of dietary fat on postprandial NCMs. The investigators will characterize NCMs in both fasting conditions and following consumption of two different high-fat meals, and will evaluate whether the type of fat in a meal affects NCMs in blood.

Detailed Description

Monocytes are a heterogeneous population of circulating blood cells that contribute to tissue integrity as well as to innate and adaptive immune defense. There are three well-characterized subsets based on their relative expression of surface antigens, cluster of differentiation 14 (CD14) and cluster of differentiation 16 (CD16). Monocytes originate from myeloid precursors in the bone marrow and enter the circulation as classical monocytes (CLMs). CLMs represent a transient cell population with a diverse differentiation potential. CLMs comprise 80-90% of the circulating blood monocyte pool and remain in circulation for approximately one day before either migrating into tissue to repopulate the tissue resident macrophage population or maturing into non-classical monocytes (NCMs). NCMs comprise only 5-10% of the circulating blood monocyte pool but have a much longer circulating lifespan of approximately 7 days. NCMs exhibit conflicting functions as anti-inflammatory caretakers of vascular tissue and as contributors to the pathogenesis of disease.

Metabolic responses to food consumption influence the risk of cardiometabolic disease. Postprandial glycemia and lipemia modulate vascular health by altering endothelial function and inducing oxidative stress, inflammation, and apoptosis. Consumption of a single high-fat meal increases circulating interleukin 6 (IL-6), enhances expression of monocyte adhesion molecules, reduces flow-mediated dilation, and increases markers of oxidative stress in human subjects. Although NCMs are described as vascular housekeepers with distinct motility and crawling patterns allowing them to actively surveil endothelium and scavenge luminal debris, their role in the postprandial state is currently unknown.

To better understand the function of postprandial NCMs following consumption of a single high-fat mixed macronutrient challenge meal, the investigators propose a study following a crossover design in which participants will consume one of two isocaloric high-fat challenge meals spaced two-weeks apart, a high-saturated fat mixed macronutrient challenge meal or a high-monounsaturated fat mixed macronutrient challenge meal. Blood at fasting and at six hours postprandial will be collected and the proportion of NCMs and their integrin expression will be analyzed by flow cytometry while changes in global gene expression will be measured by RNA-sequencing.

Recruitment & Eligibility

Status
COMPLETED
Sex
All
Target Recruitment
32
Inclusion Criteria
  • BMI 18.5 - 29.9 kg/m²
  • have a bank account and social security number or taxpayer identification for financial compensation
Exclusion Criteria
  • Pregnant or lactating women
  • Known allergy or hindering intolerance to study meal ingredients
  • Systolic blood pressure greater than 140 mmHg or diastolic blood pressure greater than 90 mmHg measured
  • Fasting glucose above 105 mg/dL
  • Triglycerides above 150 mg/dL
  • HDL cholesterol less than 40 mg/dL (men) and 50 mg/dL (women)
  • Self-reported history of difficulties with blood drawing procedures including prior fainting or dizziness, or veins assessed as not suitable for four separate venipunctures by licensed phlebotomist
  • Diagnosed active chronic diseases for which the individual is currently taking daily medication, including but not limited to Diabetes mellitus, Cardiovascular disease, Cancer, Gastrointestinal disorders, Kidney disease, Liver disease, Bleeding disorders, Asthma, Autoimmune disorders, Hypertension, Osteoporosis
  • Recent minor surgery (within 4 wk) or major surgery (within 16 wk)
  • History of gastrointestinal surgery, including gastric bypass surgery or resection
  • Recent antibiotic therapy (within 4 wk)
  • Known gallbladder disease or history of cholecystectomy
  • Recent hospitalization (within 4 wk)
  • Use of prescription medications at the time of the study that directly affect endpoints of interest (e.g. hyperlipidemia, glycemic control, steroids, statins, anti-inflammatory agents, and over-the-counter weight loss aids)
  • Current participation in another research study
  • Less than 18 and over 39 years old
  • BMI less than 18.5 and above 29.9 kg/m²
  • Has HIV/AIDS or another disease that affects the immune system
  • Unable to fast for 12 hours
  • Gives regular blood donations and is unwilling to stop during the study
  • Has monocytosis (>0.8 x 10³/microliter) or other abnormalities in hematologic parameters based on a screening complete blood count (CBC) with differential

Study & Design

Study Type
INTERVENTIONAL
Study Design
CROSSOVER
Arm && Interventions
GroupInterventionDescription
High mono-unsaturated fat challenge meal followed by high saturated fat challenge mealHigh mono unsaturated fat challenge mealHigh mono-unsaturated fat mixed macronutrient challenge meal with olive oil followed by high saturated fat mixed macronutrient challenge meal with palm oil two weeks later
High saturated fat challenge meal followed by high mono-unsaturated fat challenge mealHigh saturated fat challenge mealHigh saturated fat mixed macronutrient challenge meal with palm oil followed by high mono-unsaturated fat mixed macronutrient challenge meal with olive oil two weeks later
High mono-unsaturated fat challenge meal followed by high saturated fat challenge mealHigh saturated fat challenge mealHigh mono-unsaturated fat mixed macronutrient challenge meal with olive oil followed by high saturated fat mixed macronutrient challenge meal with palm oil two weeks later
High saturated fat challenge meal followed by high mono-unsaturated fat challenge mealHigh mono unsaturated fat challenge mealHigh saturated fat mixed macronutrient challenge meal with palm oil followed by high mono-unsaturated fat mixed macronutrient challenge meal with olive oil two weeks later
Primary Outcome Measures
NameTimeMethod
Change in Monocyte subsetsMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Monocyte subsets will be analyzed using flow cytometry. Subset analysis will be performed by labeling immune cells with anti-cluster of differentiation antigen 45 (anti-CD45), cluster of differentiation antigen 91 (anti-CD91), anti-CD14, and anti-CD16 fluorescently labeled antibodies.

Secondary Outcome Measures
NameTimeMethod
Change in eosinophil countMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Eosinophil (EO) count will be measured by a DxH 520 Hematology analyzer.

Change in basophil countMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Basophil (BA) count will be measured by a DxH 520 Hematology analyzer.

Change in red blood cell countMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Red blood cell (RBC) count will be measured by a DxH 520 Hematology analyzer.

Change in hematocritMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Hematocrit (HCT) will be measured by a DxH 520 Hematology analyzer.

Change in mean corpuscular hemoglobinMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Mean corpuscular hemoglobin (MCH) will be measured by a DxH 520 Hematology analyzer.

Change in CD16 gene expressionMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Transcriptional changes in non-classical and classical monocytes will be measured by RNA-sequencing following the isolation of non-classical monocytes from peripheral blood by fluorescence-activated cell sorting (FACS) using anti-CD16 fluorescently labeled antibodies.

Change in white blood cell countMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

White blood cell (WBC) count will be measured by a DxH 520 Hematology analyzer.

Change in mean platelet volumeMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Mean platelet volume (MPV) will be measured by a DxH 520 Hematology analyzer.

Change in CD45 gene expressionMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Transcriptional changes in non-classical and classical monocytes will be measured by RNA-sequencing following the isolation of non-classical monocytes from peripheral blood by fluorescence-activated cell sorting (FACS) using anti-CD45 fluorescently labeled antibodies.

Change in monocyte countMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Monocyte (MO) count will be measured by a DxH 520 Hematology analyzer.

Change in hemoglobinMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Hemoglobin (HGB) will be measured by a DxH 520 Hematology analyzer.

Change in mean corpuscular hemoglobin concentrationMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Mean corpuscular hemoglobin concentration (MCHC) will be measured by a DxH 520 Hematology analyzer.

Change in platelet countMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Platelet (PLT) count will be measured by a DxH 520 Hematology analyzer.

Change in levels of interleukin-6Measured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Plasma markers of systemic inflammation including interleukin-6 will be measured by ELISA.

Change in soluble cluster of differentiation antigen 146Measured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Endothelial activation including soluble cluster of differentiation antigen 146 (CD146) will be measured by ELISA.

Change in levels of C-reactive proteinMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Acute phase reactants including C-reactive protein (CRP) will be measured by ELISA.

Change in expression of very late antigen-4Measured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Monocyte adhesion molecule expression of very late antigen-4 (VLA-4) will be assessed using flow cytometry.

Change in lymphocyte countMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Lymphocyte (LY) count will be measured by a DxH 520 Hematology analyzer.

Change in change in neutrophil granulocyte countMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Neutrophil granulocyte (NE) count will be measured by a DxH 520 Hematology analyzer.

Change in mean corpuscular volumeMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Mean corpuscular volume (MCV) will be measured by a DxH 520 Hematology analyzer.

Change in red blood cell distribution widthMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Red blood cell distribution width (RDW) will be measured by a DxH 520 Hematology analyzer.

Change in red blood cell distribution width standard deviationMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Red blood cell distribution width standard deviation (RDW-SD) will be measured by a DxH 520 Hematology analyzer.

Change in CD91 gene expressionMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Transcriptional changes in non-classical and classical monocytes will be measured by RNA-sequencing following the isolation of non-classical monocytes from peripheral blood by fluorescence-activated cell sorting (FACS) using anti-CD91 fluorescently labeled antibodies.

Change in CD14 gene expressionMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Transcriptional changes in non-classical and classical monocytes will be measured by RNA-sequencing following the isolation of non-classical monocytes from peripheral blood by fluorescence-activated cell sorting (FACS) using anti-CD14 fluorescently labeled antibodies.

Change in expression of colony stimulating factor 1 receptorMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Monocyte adhesion molecule expression of colony stimulating factor 1 receptor (CSFR1) will be assessed using flow cytometry.

Change in expression of C-X3-C motif chemokine receptor 1Measured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Monocyte adhesion molecule expression of C-X3-C motif chemokine receptor 1 (CX3CR1) will be assessed using flow cytometry.

Change in expression of Notch2Measured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Monocyte adhesion molecule expression of Notch2 will be assessed using flow cytometry.

Change in expression of scavenger receptor class B, member 3Measured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Monocyte adhesion molecule expression of scavenger receptor class B, member 3 (CD36) will be assessed using flow cytometry.

Change in levels of total cholesterolMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Lipid-related markers including total cholesterol will be measured by auto-analyzer, Cobas Integra 400+ instrument

Change in intensity of filamentous-actinMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Filamentous-actin (F-actin) intensity will be assessed using phalloidin.

Change in levels of interleukin-8Measured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Plasma markers of systemic inflammation including interleukin-8 will be measured by ELISA.

Change in levels of 8-isoprostane F2alphaMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Plasma markers of oxidative stress including 8-isoprostane F2alpha will be measured by ELISA.

Change in levels of triglyceridesMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Lipid-related markers including triglycerides will be measured by auto-analyzer, Cobas Integra 400+ instrument

Change in levels of HDL-cholesterolMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Lipid-related markers including HDL-cholesterol (HDL-C) will be measured by auto-analyzer, Cobas Integra 400+ instrument

Change in levels of LDL-cholesterolMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Lipid-related markers including LDL-cholesterol (LDL-C) will be measured by auto-analyzer, Cobas Integra 400+ instrument

Change in levels of serum amyloid AMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Acute phase reactants including serum amyloid A (SAA) will be measured by ELISA.

Change in levels of chemokine ligand 2Measured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Chemokines including chemokine ligand 2 will be measured by ELISA.

Change in levels of glucoseMeasured from samples taken at 0 hours (fasting) and 6 hours (postprandial) on 2 test days

Plasma glucose will be measured by auto-analyzer, Cobas Integra 400+ instrument

Trial Locations

Locations (1)

USDA Western Human Nutrition Research Center

🇺🇸

Davis, California, United States

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