Serum cytokine profiles in healthy nonhuman primates are blunted by sedation and demonstrate sexual dimorphism as detected by a validated multiplex immunoassay

AbstractCytokine profiling is a valuable tool for monitoring immune responses associated with disease and treatment. This study assessed the impact of sex and sedation on serum cytokines in healthy nonhuman primates (NHPs). Twenty-three cytokines were measured from serum using a bead-based multiplex assay. Assay validation for precision, sensitivity, recovery, linearity, and stability was performed. Samples from male and female cynomolgus and rhesus macaques either cooperating or sedated were compared. All cytokines except TNFα demonstrated acceptable sensitivity and precision, with variable recovery and linearity. IFNγ, IL-2, IL-5, IL-6, IL-8, IL-12/23 (p40), IL-13, IL-15, MCP-1, TGFα, VEGF met acceptance criteria; G-CSF, IL-4, IL-10, MIP1α, sCD40L were marginal. Higher cytokine levels were observed in females and cytokine levels were blunted in sedated NHPs when compared to awake cooperating NHPs. Significant differences observed in cytokines related to sex, species, or imposed by handling highlight the importance of model design on translational relevance for clinical settings.

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Aerobic exercise attenuates frailty in aging male and female C57Bl/6 mice and effects systemic cytokines differentially by sex

The Journals of Gerontology Series A ◽

10.1093/gerona/glab297 ◽

2021 ◽

Author(s):

Elise S Bisset ◽

Stefan Heinze-Milne ◽

Scott A Grandy ◽

Susan E Howlett

Keyword(s):

Life Expectancy ◽

Aerobic Exercise ◽

Systemic Inflammation ◽

Frailty Index ◽

Biological Age ◽

Beneficial Effects ◽

Serum Cytokines ◽

Cytokine Levels ◽

The Impact ◽

Pro Inflammatory Cytokines

Abstract Aerobic exercise is a promising intervention to attenuate frailty, but preclinical studies have used only male animals. We investigated the impact of voluntary aerobic exercise on frailty, biological age (FRIGHT clock), predicted life expectancy (AFRAID clock) and mortality in both sexes and determined whether exercise was associated with changes in inflammation. Older (21-23 months) male (n=12) and female (n=22) C57Bl/6 mice matched for baseline frailty scores were randomized into exercise (running wheel) and sedentary (no wheel) groups. Frailty index scores were measured biweekly (13 weeks), and 23 serum cytokines were measured at midpoint and endpoint. Exercise levels varied between mice but not between the sexes. Exercise had no effect on mortality, but it attenuated the development of frailty in both sexes (female=0.32±0.04 vs 0.21±0.01; p=0.005; male=0.30±0.02 vs. 0.22±0.02; p=0.042) and reduced frailty in older females after 10 weeks. FRIGHT scores were unaffected by exercise but increased with time in sedentary males indicating increased biological age. Exercise prevented the age-associated decline in AFRAID scores in older females such that exercised females had a longer life expectancy. We investigated whether aerobic exercise was associated with changes in systemic inflammation. Cytokine levels were not affected by exercise in males, but levels of pro-inflammatory cytokines were positively correlated with the frequency of exercise in females. Despite increases in systemic inflammation, exercise reduced frailty and increased lifespan in older females. Thus, voluntary aerobic exercise, even late in life, has beneficial effects on health in both sexes but may be especially helpful in older females.

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Controlled-release mitochondrial protonophore (CRMP) reverses dyslipidemia and hepatic steatosis in dysmetabolic nonhuman primates

Science Translational Medicine ◽

10.1126/scitranslmed.aay0284 ◽

2019 ◽

Vol 11 (512) ◽

pp. eaay0284 ◽

Cited By ~ 9

Author(s):

Leigh Goedeke ◽

Liang Peng ◽

Valle Montalvo-Romeral ◽

Gina M. Butrico ◽

Sylvie Dufour ◽

...

Keyword(s):

Controlled Release ◽

Fatty Liver ◽

Nonhuman Primates ◽

Rhesus Macaques ◽

Density Lipoprotein ◽

Glucose Production ◽

Endogenous Glucose Production ◽

Whole Body ◽

Nonalcoholic Fatty Liver ◽

The Impact

Nonalcoholic fatty liver disease (NAFLD) is estimated to affect up to one-third of the general population, and new therapies are urgently required. Our laboratory previously developed a controlled-release mitochondrial protonophore (CRMP) that is functionally liver-targeted and promotes oxidation of hepatic triglycerides. Although we previously demonstrated that CRMP safely reverses hypertriglyceridemia, fatty liver, hepatic inflammation, and fibrosis in diet-induced rodent models of obesity, there remains a critical need to assess its safety and efficacy in a model highly relevant to humans. Here, we evaluated the impact of longer-term CRMP treatment on hepatic mitochondrial oxidation and on the reversal of hypertriglyceridemia, NAFLD, and insulin resistance in high-fat, fructose-fed cynomolgus macaques (n = 6) and spontaneously obese dysmetabolic rhesus macaques (n = 12). Using positional isotopomer nuclear magnetic resonance tracer analysis (PINTA), we demonstrated that acute CRMP treatment (single dose, 5 mg/kg) increased rates of hepatic mitochondrial fat oxidation by 40%. Six weeks of CRMP treatment reduced hepatic triglycerides in both nonhuman primate models independently of changes in body weight, food intake, body temperature, or adverse reactions. CRMP treatment was also associated with a 20 to 30% reduction in fasting plasma triglycerides and low-density lipoprotein (LDL)–cholesterol in dysmetabolic nonhuman primates. Oral administration of CRMP reduced endogenous glucose production by 18%, attributable to a 20% reduction in hepatic acetyl–coenzyme A (CoA) content [as assessed by whole-body β-hydroxybutyrate (β-OHB) turnover] and pyruvate carboxylase flux. Collectively, these studies provide proof-of-concept data to support the development of liver-targeted mitochondrial uncouplers for the treatment of metabolic syndrome in humans.

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An observational study of innate immune responses in patients with acute appendicitis

Scientific Reports ◽

10.1038/s41598-020-73798-3 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Toon Peeters ◽

Sandrina Martens ◽

Valentino D’Onofrio ◽

Mark H. T. Stappers ◽

Jeroen C. H. van der Hilst ◽

...

Keyword(s):

Acute Appendicitis ◽

Immune Responses ◽

Complicated Appendicitis ◽

Healthy Controls ◽

Innate Immune Responses ◽

Serum Cytokine ◽

Post Surgery ◽

Tnf Α ◽

Cytokine Levels ◽

Recognition Receptor

Abstract Acute appendicitis is a common surgical emergency worldwide. Exaggerated immune responses could be associated with appendicitis. This study aimed at characterizing immune responses towards a large variety of gut commensals and pathogens, and pattern recognition receptor (PRR) ligands, and investigating the course of systemic inflammation in a prospective cohort of acute appendicitis patients. PBMC responses of 23 patients of the cohort and 23 healthy controls were characterized more than 8 months post-surgery. Serum cytokine levels were measured in 23 patients at the time of appendicitis and after one month. CRP, WBC and percentage of neutrophils were analyzed in the total cohort of 325 patients. No differences in PBMC responses were found between patients and controls. Stronger IL-10 responses were found following complicated appendicitis. A trend towards lower IL-8 responses was shown following gangrenous appendicitis. Serum IL-10 and IL-6 were significantly elevated at presentation, and IL-6, IL-8 and TNF-α levels were higher in complicated appendicitis. Routine biomarkers could predict severity of appendicitis with high specificities, but low sensitivities. Cytokine responses in patients following acute appendicitis did not differ from healthy controls. Higher serum cytokine levels were found in acute complicated and gangrenous cases. Further research into discriminative biomarkers is warranted.

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Effect of peanut oil and nandrolone on serum cytokines in pubertal term rats

Indian Journal of Animal Research ◽

10.18805/ijar.v0iof.7819 ◽

2017 ◽

Author(s):

Erdal Tasgin ◽

Seyfullah Haliloglu

Keyword(s):

Serum Levels ◽

Peanut Oil ◽

Male Rats ◽

Control Group ◽

Nandrolone Decanoate ◽

Serum Cytokine ◽

Blood Samples ◽

Serum Cytokines ◽

Cytokine Levels ◽

Cytokine Synthesis

The purpose of this study was to determine the effect of nandrolone on cytokine levels in pubertal term female and male rats. Totally 60 pubertal term Spraque Dawley rat (30/30 female/male) were divided into 3 equal groups. Control group received no treatment. PO group received intraperitoneally peanut oil every 5 days with an intervening gap of 2 days where no treatment was given. ND group received intraperitoneally nandrolone decanoate (10 mg/kg) within peanut oil like PO group. At the end of the four weeks, blood samples were obtained. Serum cytokine levels were measured by ELISA. Peanut oil significantly (P less than 0.05) increased serum levels of interleukin-1â and interleukin-6 in pubertal term male rats, while nandrolone decanoate had no any effect on the cytokine levels compared to experimental groups. It may be stated that nandrolone decanoate has no excessive inducer effect on the cytokine synthesis in pubertal male and female term.

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SARS-CoV-2 infection protects against rechallenge in rhesus macaques

Science ◽

10.1126/science.abc4776 ◽

2020 ◽

Vol 369 (6505) ◽

pp. 812-817 ◽

Cited By ~ 199

Author(s):

Abishek Chandrashekar ◽

Jinyan Liu ◽

Amanda J. Martinot ◽

Katherine McMahan ◽

Noe B. Mercado ◽

...

Keyword(s):

Immune Responses ◽

Protective Immunity ◽

Nonhuman Primates ◽

Rhesus Macaques ◽

Viral Loads ◽

Macaque Model ◽

Cellular Immune Responses ◽

Immunologic Control ◽

Cellular Immune ◽

Public Health Strategies

An understanding of protective immunity to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is critical for vaccine and public health strategies aimed at ending the global coronavirus disease 2019 (COVID-19) pandemic. A key unanswered question is whether infection with SARS-CoV-2 results in protective immunity against reexposure. We developed a rhesus macaque model of SARS-CoV-2 infection and observed that macaques had high viral loads in the upper and lower respiratory tract, humoral and cellular immune responses, and pathologic evidence of viral pneumonia. After the initial viral clearance, animals were rechallenged with SARS-CoV-2 and showed 5 log10 reductions in median viral loads in bronchoalveolar lavage and nasal mucosa compared with after the primary infection. Anamnestic immune responses after rechallenge suggested that protection was mediated by immunologic control. These data show that SARS-CoV-2 infection induced protective immunity against reexposure in nonhuman primates.

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Cytokine release syndrome-like serum responses after COVID-19 vaccination are frequent but clinically inapparent in cancer patients under immune checkpoint therapy

10.1101/2021.12.08.21267430 ◽

2021 ◽

Author(s):

Thomas Walle ◽

Sunanjay Bajaj ◽

Joscha A. Kraske ◽

Thomas Rösner ◽

Christiane S. Cussigh ◽

...

Keyword(s):

Immune Responses ◽

Cancer Patients ◽

Immune Checkpoint ◽

Elevated Serum ◽

Cytokine Release ◽

Cytokine Release Syndrome ◽

Serum Cytokine ◽

Short Term ◽

Cytokine Levels ◽

Tumor Types

AbstractCancer patients frequently receive immune checkpoint therapies (ICT) which may modulate immune responses to COVID-19 vaccines. Recently, cytokine release syndrome (CRS) was observed in a cancer patient who received the BTN162b2 vaccine under ICT. Here, we analyzed adverse events (AEs) in patients of various solid tumor types undergoing (n=64) or not undergoing (n=26) COVID-19 vaccination under ICT as an exploratory endpoint of a prospectively planned cohort study. We did not observe clinically relevant CRS after vaccination (95% CI [0,0.056]). Short term (<4 weeks) serious AEs were rare (12.5%) and overall AEs under ICT were comparable to unvaccinated patients. Despite the absence of CRS symptoms, we observed a pairwise-correlated set of CRS-associated cytokines upregulated in 42% of patients after vaccination and ICT (>1.5fold). Hence, clinically meaningful CRS appears to be rare in cancer patients under ICT and elevated serum cytokine levels are common but not sufficient to establish CRS diagnosis.

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Cytokine and Chemokine Profile in Patients with Multiple Myeloma Treated with Bortezomib

Mediators of Inflammation ◽

10.1155/2020/1835836 ◽

2020 ◽

Vol 2020 ◽

pp. 1-13 ◽

Cited By ~ 1

Author(s):

Paweł Robak ◽

Edyta Węgłowska ◽

Izabela Dróżdż ◽

Damian Mikulski ◽

Dariusz Jarych ◽

...

Keyword(s):

Multiple Myeloma ◽

Refractory Disease ◽

Serum Cytokine ◽

Single Measurement ◽

Early Relapse ◽

Serum Cytokines ◽

Cytokines And Chemokines ◽

Cytokine Levels ◽

Normal Calcium ◽

Human Cytokine

The aim of the study was to determine the levels of selected cytokines and chemokines in the serum of multiple myeloma (MM) patients treated with bortezomib-based regimens. A total of 71 MM patients were examined: 41 with primary refractory disease (17) or early relapse (28), and 30 who were bortezomib sensitive with no progression for at least six months. Patients who demonstrated CR or PR after bortezomib-based therapies longer than six months after treatment discontinuation were designated bortezomib sensitive. Serum cytokine levels were assayed with Bio-Rad Bio-Plex Pro Human Cytokine 27-Plex Assay on the MAGPIX Multiplex Reader and the Bio-Plex® 200 System (Bio-Rad). Higher levels of MIP-1α and lower levels of MIP-1β and IL-9 were associated with better responses to bortezomib-based treatment, and higher levels of IL-1ra and IL-8 were associated with bone involvement. MCP-1 was elevated in patients with hemoglobin<10 g/dl compared to those without anemia. The levels of IL-8, MIP-1α, and TNF-α were significantly higher in patients with renal insufficiency. Only MIP-1α was elevated in patients with hypercalcemia compared to patients with normal calcium levels. In conclusion, distinct cytokines are involved in the pathogenesis of MM and may play a prominent role in the prediction of treatment response. However, a single measurement of serum cytokines should be interpreted with caution and further studies are needed.

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Safety and Pharmacokinetics of Intravesical Chitosan/Interleukin-12 Immunotherapy in Murine Bladders

Bladder Cancer ◽

10.3233/blc-211542 ◽

2021 ◽

pp. 1-11

Author(s):

Khue G. Nguyen ◽

Ethan S. Wagner ◽

Maura R. Vrabel ◽

Siena M. Mantooth ◽

Danielle M. Meritet ◽

...

Keyword(s):

Bladder Cancer ◽

Immune Responses ◽

Acute Inflammation ◽

Inflammatory Responses ◽

Interleukin 12 ◽

Laboratory Mice ◽

Serum Cytokine ◽

Preclinical Models ◽

Intravesical Administration ◽

Cytokine Levels

BACKGROUND: Intravesical administration of interleukin 12 (IL-12) co-formulated with the biopolymer, chitosan (CS/IL-12), has demonstrated remarkable antitumor activity against preclinical models of bladder cancer. However, given historical concerns regarding severe toxicities associated with systemic IL-12 administration in clinical trials, it is important to evaluate the safety of intravesical CS/IL-12 prior to clinical translation. OBJECTIVE: To evaluate the pharmacokinetics as well as the local and systemic toxicities of intravesical CS/IL-12 immunotherapy in laboratory mice. METHODS: Local inflammatory responses in mouse bladders treated with intravesical IL-12 or CS/IL-12 were assessed via histopathology. Serum cytokine levels following intravesical and subcutaneous (s.c.) administrations of IL-12 or CS/IL-12 in laboratory mice were compared. Systemic toxicities were evaluated via body weight and liver enzyme levels. RESULTS: Intravesical IL-12 and CS/IL-12 treatments did not induce significant local or systemic toxicity. IL-12 dissemination and exposure from intravesical administration was significantly lower compared to s.c. injections. Weekly intravesical CS/IL-12 treatments were well-tolerated and did not result in blunted immune responses. CONCLUSIONS: Intravesical CS/IL-12 is safe and well-tolerated in mice. In particular, the lack of cystitis and acute inflammation justifies continued investigation of intravesical CS/IL-12 immunotherapy in larger animals and patients with bladder cancer.

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Protection of rhesus macaques against vaginal SHIV challenges by VRC01 and an anti-α4β7 antibody

10.1101/365551 ◽

2018 ◽

Author(s):

Giulia Calenda ◽

Ines Frank ◽

Géraldine Arrode-Brusés ◽

Amarendra Pegu ◽

Keyun Wang ◽

...

Keyword(s):

T Cell ◽

Immune Responses ◽

Rhesus Macaques ◽

Cell Loss ◽

High Dose ◽

Cell Responses ◽

Cell Counts ◽

Siv Infection ◽

Shiv Infection ◽

The Impact

ABSTRACTVRC01 protects macaques from vaginal SHIV infection after a single high-dose challenge. Infusion of a simianized anti-α4β7 mAb (Rh-α4β7) just prior to, and during repeated vaginal exposures to SIVmac251 partially protected macaques from vaginal SIV infection and rescued CD4+ T cells. To investigate the impact of combining VRC01 and Rh-α4β7 on SHIV infection, 3 groups of macaques were treated with a suboptimal dosing of VRC01 alone or in combination with Rh-α4β7 or with control antibodies prior to the initiation of weekly vaginal exposures to a high dose (1000TCID50) of SHIVAD8-EO. The combination Rh-α4β7-VRC01 significantly delayed SHIVAD8-EO vaginal infection. Following infection, VRC01-Rh-α4β7-treated macaques maintained higher CD4+ T cell counts and exhibited lower rectal SIV-DNA loads compared to the controls. Interestingly, VRC01-Rh-α4β7-treated macaques had less IL-17 producing cells in the blood and the gut during the acute phase of infection. Moreover, higher T cell responses to the V2-loop of the SHIVAD8-EO envelope in the VRC01-Rh-α4β7 group inversely correlated with set point viremia. The combination of suboptimal amounts of VRC01 and Rh-α4β7 delayed infection, altered anti-viral immune responses and minimized CD4+ T cell loss. Further exploration of the effect of combining bNAbs with Rh-α4β7 on SIV/HIV infection and anti-viral immune responses is warranted and may lead to novel preventive and therapeutic strategies.Short summaryA combination of VRC01 and Rh-α4β7 significantly delayed SHIV acquisition, protected CD4 counts, decreased gut viral load and modified the immune response to the virus.

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