Endotoxin Inhalation Hazard

Smoking Cannabis or Tobacco, Vaping, Farm work and other industrial exposure to Endotoxin can create Chronic Disease with Fatal outcomes

In a recent article I discussed the exposure of Mice to nebulized Endotoxin to deliberately inflame their Lungs.¹

Thought I should throw a few references together on Inhalation risks of these bacterial cell wall fragments.

A review from 2008 pointed to the inadequacy of the Horseshoe LAL test to evaluate effects of inhaled Endotoxin.² It covered studies showing genetic and gender variations in allergic response, the receptors known at the time, exposures in cotton, livestock, paper, timber and other industries. Cannabis is a known Endotoxin hazard.

Photo credit.³

An interesting 2013 Human study⁴ (with many references to earlier work) exposed healthy volunteers to repeated nebulized Endotoxin and also treated those exposed with Roflumilast, a very nasting looking organoChloroFluorine compound with this structure.⁵

That is of great interest because Pfizer suggested that Roflumilast could be responsible for some Anaphylaxis cases observed in its Process 2 Periodic Safety Update Report.

But, let’s return to the Endotoxin nebulization study that delivered 2 μg GMP-grade LPS (manufactured under Good Manufacturing Practice standards) to the Human volunteers airways. The 12 volunteers (3 female and 9 male) were challenged with Methacholine to pre-qualify sputum production. They were subjected to FEV₁ = lung function measurement, EBT = exhaled breath temperature, BT = body temperature, and gave blood samples. One subject dropped out.

Interleukin-8 and MyeloPerOxidase (MPO) were analyzed in sputum. Different types of cells in sputum investigated were CD4 (FITC)/8 (PE), CD86 (PE-Cy7), HLA-DR (PE)) and Beckman Coulter (CD14 (APC)).

Endotoxin inhalation

“induced a massive influx of neutrophils into the airways”. Treatment with the PDE4-inhibitor Roflumilast for 5 days changed the expression of HLA-DR and CD86 on sputum macrophages, but did not result in a significant attenuation of the inflammatory cell influx.

Comparison of sputum of 11 subjects collected on average more than 3 months (median 111, minimum 56, maximum, 157 days) after the last LPS inhalation revealed a higher mean neutrophil count as compared to the baseline visit of this study

Janssen and coworkers saw signs of Endotoxin Tolerance developing in their subjects.

A 2021 review covered inhalation as a component of Endotoxin hazard from Nanoparticles.⁶

Pfizer excluded Vaping or Smoking subjects

The Pfizer trials using low Endotoxin mRNA generated by RT-PCR excluded subjects with a history of Vaping or Smoking.⁷

In 2006 it was shown that a Nicotine patch attenuates the Endotoxin poisoning of Human volunteers given dose of 2 ng/kg over a 5-min period through the intravenous catheter.⁸ As expected TNF-α, IL-6, IL-8 and sE-selectin⁹ increased on Endotoxin injection. Nicotine increased anti-inflammatory IL-10.

A Harvard study¹⁰¹¹ found 23% of 75 popular e-cigarette products - cartridges (single use) and e-liquids (refillable material) contained Endotoxin.

Byssinosis linked to Endotoxin

Byssinosis¹² is an occupational lung disease caused by inhalation of cotton, hemp or jute dust. It has been linked to Endotoxin.¹³

Cannabis user allergies have been linked to Endotoxin.¹⁴

Particulate Air Pollution Endotoxin source

A 2013 study found course particulate air pollution is a significant source of Endotoxin chronic exposure and illness.¹⁵ Thirty-five healthy adults were deliberately exposed and within 3 hours increased blood leukocytes was measured.

An earlier study¹⁶ found airborne particulate Endotoxin using Mouse monocytic-macrophagic RAW264.7 cells, finding increased Interleukin-6 and Tumor Necrosis Factor-α (TNFα).

Lipid A is known Inhalation Hazard

Avanti Polar Lipids, supplier of lipids to Pfizer for its LNPs, also sells the Supertoxin Lipid A Kdo2 for US$259 per milligram.

In the Material Safety Data sheet they state:

"If breathed in, move person into fresh air. If not breathing, give artificial respiration".

Reassuring, isn’t it!

Pfizer Protocol Amendment 31 March 2021

Thanks to Brook Jackson, I learned that when Pfizer launched its Booster trial, only clinical sites with laminar flow hood or biosafety cabinets were continued.

I find that most intriguing.

Question

Are Cannabis users and investors blind to Endotoxin hazard in jabs as well as the plants?

February 2024 update

My very good friend invested many $$, time and energy measuring Endotoxin in Cannabis using a Fluorescence technique. He sent me this picture before he went on holidays late 2023 showing the distinctive colour developed by a modified LAL test. A spectrometer can quantify the Endotoxin by comparing with a standard curve.

And in recent days Kevin has been reporting on detection of Single Nucleotide Polymorphisms (SNPs) that appear on Chromosomes 9 and 12 when he incubates Human Ovarian Cancer Cells with Pfizer Jabs.¹⁷

Of particular interest to me was this figure¹⁸ of known Diseases associated with gene variations on Chromosome 9.

I spotted Endotoxin Hyporesponsivenes.

A quick google brought up a lovely paper by Nancy Arbour and coworkers¹⁹ published in 2000, showing that Endotoxin Hyporesponsivenes is caused by a SNP on Chromosome 9 in the gene that codes for TLR4 Endotoxin receptor.

In this Figure they show the dramatic difference in effects Primary Human Epithelial cells stimulated release of IL-1α. Note the scale is in Picogram/ml.

1.

2. Liebers V, et al. Health effects due to endotoxin inhalation (review). https://link.springer.com/article/10.1007/s00204-008-0290-1

3. https://commons.wikimedia.org/wiki/File:Hampa_Cannabis_sativa_L._(n%C3%A4rbild).jpg

4. Janssen O, et al. 2013. Low-dose endotoxin inhalation in healthy volunteers - a challenge model for early clinical drug development. https://bmcpulmmed.biomedcentral.com/articles/10.1186/1471-2466-13-19

5. https://en.wikipedia.org/wiki/Roflumilast

6. Hannon G and Prina-Mello A. 2021. Endotoxin contamination of engineered nanomaterials: Overcoming the hurdles associated with endotoxin testing. https://wires.onlinelibrary.wiley.com/doi/10.1002/wnan.1738

7. Pfizer Interim Clinical Study Report Protocol C4591001. December 2020. Final Analysis Interim Report: A Phase 1/2/3, Placebo- Controlled, Randomized, Observer-Blind, Dose-Finding Study to Evaluate the Safety, Tolerability, Immunogenicity, and Efficacy of SARS-COV-2 RNA Vaccine Candidates Against COVID-19 in Healthy Individuals

8. Wittebole X, et al. Nicotine exposure alters in vivo human responses to endotoxin. https://academic.oup.com/cei/article/147/1/28/6458870

9. https://en.wikipedia.org/wiki/E-selectin

10. https://www.hsph.harvard.edu/news/press-releases/microbial-contaminants-found-in-popular-e-cigarettes/

11. Lee M-S et al. 2019. Endotoxin and (1→3)-β-D-Glucan Contamination in Electronic Cigarette Products Sold in the United States. https://ehp.niehs.nih.gov/doi/10.1289/EHP3469

12. https://en.wikipedia.org/wiki/Byssinosis

13. Er M, et al. 2016. Byssinosis and COPD rates among factory workers manufacturing hemp and jute. https://pubmed.ncbi.nlm.nih.gov/26489943/

14. Skypala IJ, et al. 2022. Cannabis-related allergies: An international overview and consensus recommendations. https://onlinelibrary.wiley.com/doi/full/10.1111/all.15237

15. Behbod B, et al. 2013. Endotoxin in concentrated coarse and fine ambient particles induces acute systemic inflammation in controlled human exposures. https://pubmed.ncbi.nlm.nih.gov/24143017/

16. Guastadisegni C, et al. 2010. Determinants of the Proinflammatory Action of Ambient Particulate Matter in Immortalized Murine Macrophages. https://ehp.niehs.nih.gov/doi/10.1289/ehp.1002105

17. Vaccine targeted qPCR of Cancer Cell Lines treated with BNT162b2

18. https://doe-humangenomeproject.ornl.gov/wp-content/uploads/2022/08/Chrom09.jpg

19. Nancy C. Arbour, Eva Lorenz, Brian C. Schutte, Joseph Zabner, Joel N. Kline, Michael Jones, Kathy Frees, Janet L. Watt and David A. Schwartz. 2000. TLR4 mutations are associated with endotoxin hyporesponsiveness in humans. https://www.nature.com/articles/ng0600_187