LECTURE SERIES - CYTOKINE TESTING

 

Caveat: This section contains slides that are part of Malcolm Hooper's educational lecture series.

This section and other sections on Cytokines is not the practise of medicine.

 CYTOKINE PANELS - 'PREDICTIVE AND PREVENTATIVE ANALYSIS'

CYTOKINE GENE BIOMARKERS ARE A MEASURE TO 'QUALITY OF LIFE'

 

Cytokine Gene Expression Testing is at the forefront of medical advances and immunotherapy interventions however not entirely new. 

The first member of the cytokine storm family to be recognized by physicians was sepsis. The appreciation that the consequences of sepsis are a result not of the pathogen, but rather the immune response to the pathogen, dates back to observations made by William Osler in 1904 in his book, The Evolution of Modern Medicine. Accordingly, the idea that sepsis might be most effectively treated by immunomodulation is not new. With the identification of tumor necrosis factor (TNF) and interleukin-1b (IL-1b) as major inflammatory cytokines in models of sepsis in the last part of the twentieth century, trials were undertaken to block these cytokines to treat septic cytokine storm.

  • Type into Google your "health condition" and "cytokines" and you will be overwhelmed with the "evidence".

Cytokine Storm Syndrome 

  • A diverse set of (autoimmune) conditions unified by a clinical phenotype of systemic inflammation, multi‐organ failure, hyperferritinemia and if untreated, often death. This "clinical constellation" (cytokine storm) is caused by the elaboration of extreme amounts of circulating pro-inflammatory cytokine mediators (IL1, IL6, TNFa, IL8) resulting from unchecked feedforward immune activation and amplification. The initiating factors leading to the end state of cytokine storm are heterogeneous and derive from rheumatologic, oncologic, and infectious origins. 

Pro-inflammatory Cytokines

Adv Drug Deliv Rev. 2017 Feb 15. pii: S0169-409X(17)30028-5. doi: 10.1016/j.addr.2017.02.001. [Epub ahead of print]

Mimicking Oxygen delivery and waste removal functions of blood.

Zhang H1, Barralet JE2.

Author information

Abstract

In addition to immunological and wound healing cell and platelet delivery, ion stasis and nutrient supply, blood delivers oxygen to cells and tissues and removes metabolic wastes. For decades researchers have been trying to develop approaches that mimic these two immediately vital functions of blood. Oxygen is crucial for the long-term survival of tissues and cells in vertebrates. Hypoxia (oxygen deficiency) and even at times anoxia (absence of oxygen) can occur during organ preservation, organ and cell transplantation, wound healing, in tumors and engineering of tissues. Different approaches have been developed to deliver oxygen to tissues and cells, including hyperbaric oxygen therapy (HBOT), normobaric hyperoxia therapy (NBOT), using biochemical reactions and electrolysis, employing liquids with high oxygen solubility, administering hemoglobin, myoglobin and red blood cells (RBCs), introducing oxygen-generating agents, using oxygen-carrying microparticles, persufflation, and peritoneal oxygenation. Metabolic waste accumulation is another issue in biological systems when blood flow is insufficient.

Metabolic wastes change the microenvironment of cells and tissues, influence the metabolic activities of cells, and ultimately cause cell death. This review examines advances in blood mimicking systems in the field of biomedical engineering in terms of oxygen delivery and metabolic waste removal.

OXYMED Cytokines & Athletes

The Global Challenge

GlycA – The Biomarker GlycA (Glycoprotein Acetylation) is associated with chronic inflammation and predicts long term risk of severe infection.

Ritchie et al 2015 Cell Systems P293-301 Elsevier 2015.

  • Elevated GlycA in apparently healthy patients – for greater than 10-years is associated with chronic low grade inflammation.

  • Elevated GlycA was stable for up to a decade.

  • GlycA marked the levels of myriad inflammatory cytokines in circulation – Cytokine Storm.

  • A gene network enriched for neutrophil functions was associated with GlycA.

  • GlycA strongly predicted future risk of infection; increased hospitalisation and death from diverse infections within 14-years – particularly septicaemia and pneumonia – that persists over a decade.

  • Healthy individuals with elevation GlycA have persistent but clinically silent - low grade chronic inflammation that mimics an antimicrobial response and myriad cascade of inflammatory cytokines.

J Interferon Cytokine Res. 2018 Jun 11. doi: 10.1089/jir.2018.0019. [Epub ahead of print]

Cytokines in the Treatment of Cancer.

Conlon KC1, Miljkovic MD1, Waldmann TA1.

Author information

Abstract

Cytokines are major regulators of innate and adaptive immunity that enable cells of the immune system to communicate over short distances. Cytokine therapy to activate the immune system of cancer patients has been an important treatment modality and continues to be a key contributor to current clinical cancer research.

 - Interferon alpha (IFNα) is approved for adjuvant treatment of completely resected high-risk melanoma patients and several refractory malignancies.

 - High-dose interleukin-2 (HDIL-2) is approved for treatment of metastatic renal cell cancer and melanoma, but both agents are currently less commonly used with the development of newer agents.

 - Granulocyte-macrophage colony-stimulating factor (GM-CSF), IFN gamma (IFNγ), IL-7, IL-12, and IL-21 were evaluated in clinical trials and remain part of certain investigational trials.

 - The initial single-agent clinical trials with the long-awaited IL-15 have been completed and combination trials with antitumor antibodies or checkpoint inhibitors (CPIs) have been initiated. However, cytokines in monotherapy have not fulfilled the promise of efficacy seen in preclinical experiments. They are often associated with severe dose-limiting toxicities that are manageable with appropriate dosing and are now better understood to induce immune-suppressive humoral factors, suppressive cells, and cellular checkpoints, without consistently inducing a tumor-specific response.

To circumvent these impediments, cytokines are being investigated clinically with new engineered cytokine mutants (superkines), chimeric antibody-cytokine fusion proteins (immunokines), anticancer vaccines, CPIs, and cancer-directed monoclonal antibodies to increase their antibody-dependent cellular cytotoxicity or sustain cellular responses and anticancer efficacy. In this review, we summarize current knowledge and clinical application of cytokines either as monotherapy or in combination with other biological agents. We emphasize a discussion of future directions for research on these cytokines, to bring them to fruition as major contributors for the treatment of metastatic malignancy.

Zhonghua Nan Ke Xue. 2018 Jun;24(6):491-498.

[Fosfomycin tromethamine inhibits the expressions of TNF-α, IL-8 and IL-6 in the prostate tissue of rats with chronic bacterial prostatitis].

OBJECTIVE:

To investigate the effects of fosfomycin tromethamine (FT) on the expressions of tumor necrosis factor-α (TNF-α), interleukin-8(IL-8), and interleukin-6 (IL-6) in the prostate tissue of the rats with chronic bacterial prostatitis (CBP).

METHODS:

We randomly divided 70 male SD rats into 7 groups of equal number: blank control, CBP model control, positive control, 14 d low-dose FT, 7 d low-dose FT, 14 d high-dose FT, and 7 d high-dose FT. The CBP model rats in the latter five groups were treated intragastrically with levofloxacin at 100 mg/kg/d for 30 days and FT at 200 mg/kg/d for 14 and 7 days and at 300 mg/kg/d for 14 and 7 days, respectively. Then we collected the prostate tissue from the animals for determination of the levels of TNF-α, IL-8 and IL-6 by ELISA.

RESULTS:

Compared with the blank controls, the CBP model rats showed significantly increased levels of TNF-α ([19.83 ± 6.1] vs [32.93 ± 6.21] ng/g prot, P <0.01), IL-8 ([8.26 ± 0.52] vs [16.2 ± 2.84] ng/g prot, P <0.01) and IL-6 ([1.55 ± 0.11] vs [2.51 ± 1.06] ng/g prot, P <0.05) in the prostate tissue. In comparison with the CBP model controls, the levels of TNF-α and IL-8 were remarkably decreased in the groups of positive control ([20.54 ± 5.78] ng/g prot, P <0.01; [12.43 ± 4.02] ng/g prot, P <0.05), 14 d low-dose FT ([21.95 ± 6.48] ng/g prot, P <0.01; [11.11 ± 2.86] ng/g prot, P <0.01), 7 d low-dose FT ([23.8 ± 6.93] ng/g prot, P <0.05; [12.43 ± 4.02] ng/g prot, P <0.05), 14 d high-dose FT ([19.97 ± 2.58] ng/g prot, P <0.01; [8.83 ± 1.32] ng/g prot, P <0.01), and 7 d high-dose FT ([21.97 ± 3.38] ng/g prot, P <0.01; [12.68±1.97] ng/g prot, P <0.05). No statistically significant differences were observed between the positive control and FT groups in the contents of TNF-α, IL-8 or IL-6 (P >0.05). The expression of IL-6 was markedly reduced in the 14 d high-dose FT group as compared with the model controls ([1.76 ± 0.46] vs [2.51 ± 1.06] ng/g prot, P<0.05) but exhibited no significant difference between the CBP model control and the other groups (P >0.05).

CONCLUSIONS:

Fosfomycin tromethamine inhibits the expressions of TNF-α, IL-8 and IL-6 in the prostate tissue, suppresses its inflammatory reaction, promotes the repair of damaged prostatic structure, and thus contributes to the treatment of chronic bacterial prostatitis in rats.

Minocycline modulates cytokine and gene expression profiles in the brain after whole-body exposure to radiation.

Mehrotra S1, Pecaut MJ, Gridley DS.

Author information - Chan Shun Pavilion, Room A-1010, 11175 Campus Street, Loma Linda University, Loma Linda, CA 92354, U.S.A. dgridley@llu.edu.

Abstract

An effective countermeasure against radiation damage to normal tissues is urgently needed. The major goal of the present study was to determine if minocycline could modify the immunomodulatory effects of radiation on the brain. C57BL/6 mice were treated with minocycline intraperitoneally for 5 days beginning immediately before total-body exposure to 0, 1, 2 and 3 Gray (Gy) (60)Co γ-rays. Brains were collected on days 4 and 32 post-irradiation for cytokine and gene analyses.

  • Minocycline treatment significantly increased the levels of interleukin (IL)-10, IL-15 and vascular endothelial growth factor (VEGF) in the brain on day 4 in one or more irradiated groups compared to radiation-alone (p<0.05). 

  • IL-10 is anti-inflammatory,

  • IL-15 can prevent apoptosis and VEGF is neuroprotective.

 

On day 32, the drug decreased IL-1β in the 2- Gy group (p<0.05 vs. 2-Gy alone); this cytokine is implicated in immune-related central nervous system pathologies. Microarray analysis of brains on day 32 showed that while radiation increased expression of inflammatory genes such as Il1f10, Il17, Tnfrsf11b, Tnfsf12, Il12b and Il1f8, these were no longer up-regulated in the minocycline-treated groups.

Similarly, the pro-apoptotic gene Bik and nitric oxide synthase producer (Nostrin) were no longer up-regulated in the drug-treated groups. Pathway analysis based on gene data suggested that catenin-β1 and tumor suppressor-related transcription regulation were significantly activated by radiation and/or minocycline (activation z-score >2.0).

Overall, the data warrant further testing of minocycline as a potential neuroprotectant against radiation-induced damage.

Cytokine Growth Factor Rev​ . 2006 Oct;17(5):325-37. doi: 10.1016/j.cytogfr.2006.07.002. Epub 2006 Aug 22.

Cytokines in Breast Cancer

A Nicolini 1A CarpiG Rossi

Abstract

In recent decades many advances have occurred in the understanding of the role of cytokines in breast cancer. New signalling pathways of interleukin (IL)-1 family, IL-6, IL-11, IL-18, interferons (IFNs) and interferon regulatory factors 1 (IRF-1) and 2 (IRF-2) have been found within tumour microenvironments and in metastatic sites. Some cytokines (IL-1, IL-6, IL-11, TGFbeta) stimulate while others (IL-12, IL-18, IFNs) inhibit breast cancer proliferation and/or invasion. Similarly, high circulating levels of some cytokines seem to be favourable (soluble IL-2R) while others are unfavourable (IL-1beta, IL-6, IL-8, IL-10, IL-18, gp130) prognostic indicators. So far IL-2, IFNalpha, IFNbeta and occasionally IFNgamma, IL-6, IL-12 have been the cytokines used for anti tumour treatment of advanced breast cancer either to induce or increase hormone sensitivity and/or to stimulate cellular immunity. Disappointing results occurred in most trials; however, two long-term pilot studies suggest that IL-2 and IFNbeta, when used appropriately can have a positive effect on clinical benefit and overall survival of patients with minimal residual disease after chemotherapy or with disseminated disease controlled by conventional endocrine therapy.

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