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Thiomersal in influenza vaccine: more than just an adjuvant


Thiomersal (THI), an ethylmercury releasing compound, is added to some vaccines as an adjuvant, mainly to hinder bacterial growth in multidose vials. Ethylmercury (EtHg+) is a potent neurotoxin, with a toxicity resembling that of methylmercury in some aspects but with different toxicokinetics. Introduced in 1931, THI never went through a safety evaluation, but can be used in vaccines because of its introduction before the establishment of such regulation. Banned in the EU since 2001, THI-containing vaccines (TCV) are still in use in the USA and in developing countries.

Mercury species show a high affinity to sulphur and tend to form adducts with biomolecules having thiol groups, such as glutathione (GSH) or cysteine-containing proteins.  So far, adduct formation of organic mercury species was observed with GSH, human serum albumin (HSA), hemoglobin and carbonic anhydrase. Previous studies have also shown that adducts of mercury and xenobiotic proteins can lead to a formation of mercury-specific antibodies. This observation leads to the conclusion that adducts of EtHg+ and antigens are not only important to investigate for understanding toxicokincetics, but may also induce a mercury-specific immune response. Anyhow, such unwanted reaction by the use of TCV has not been reported before.

The new study:
The researchers from the University of Münster studied the interaction of THI with the seasonal tetravalent influenza vaccine "Vaxigrip tetra" (Sanofi, Paris) containing inactivated split virions of two influenza A and two influenza B strains (each 30 μg mL−1 hemagglutinin (HA), propagated in chicken eggs). After adding different amounts of the adjuvant THI to the vaccine, the vaccine was stored for 20 days in a refrigerator to allow incubation under storage conditions of vaccines.

The incubated samples were analyzed by LC-ICP-MS after desalting via ultrafiltration. Separations were performed by size-exclusion chromatography. The corresponding chromatographs are shown in figure 1.

Figure 1: SEC-ICP-MS chromatograms (m/z 202) for influenza vaccine, which was incubated with different concentrations of THI in a refrigerator for 20 d.

It can be seen that adduct formation increases with higher concentrations of THI and with the highest concentration used, adduct formation is still on the rise. Although binding positions on the protein seem not to be saturated, most of the EtHg+ occurs freely in solution. Mercury binds on a protein, that is larger than 133 kDa, indicating adduct formation with HA. External calibration using THI was possible using meso-2,3-Dimercaptosuccinic acid (DMSA) as complexing agent and a mercury concentration of 141 μg Hg L−1 (SD: 6 μg Hg L−1) was found to bind to the high-molecular fraction of the vaccine. These results cleary show that thiomersal is not only a preservative but reacts with the proteins of the vaccine forming adducts.

The original study:

Philipp Strohmidel, Michael Sperling, Uwe Karst, Investigations on the binding of ethylmercury from thiomersal to proteins in influenza vaccines, J. Trace Elem. Med. Biol., 50 (2018) 100-104. DOI: /10.1016/j.jtemb.2018.06.011

Used techniques and instrumentation:

Related studies (newest first)

J. Dórea, Low-dose Thimerosal (ethyl-mercury) is still used in infants' vaccines: Should we be concerned with this form of exposure ?, J. Trace Elem. Med. Biol., 49 (2018) 134-139. DOI: 10.1016/j.jtemb.2018.05.010.

M. Davidson, Vaccination as a cause of autism-myths and controversies, Dialogues Clin. Neurosci. 19 (2017) 403–407. DOI: 10.1080/02699930302286

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J. Hogeback, M. Schwarzer, C.A. Wehe, M. Sperling, U. Karst, Investigating the adduct formation of organic mercury species with carbonic anhydrase and hemoglobin from human red blood cell hemolysate by means of LC/ESI-TOF-MS and LC/ICP-MS, Metallomics 8 (2016) 101–107. DOI: 10.1039/C5MT00186B.

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May 5, 2014: Global policy on the use of mercury as a preservative in vaccine called discriminatory
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June 19, 2012: Vaccine ingredient causes brain damage; some nutrients prevent it
October 28, 2011: WHO worries mercury treaty could affect costs and availability of vaccines 
August 8, 2011: UNEP Global Mercury Treaty May Include Ban on Mercury in Medicine
March 17, 2011: Researchers Urge the Removal of Mercury From Flu Shots
August 16, 2010: Methylmercury: What have we learned from Minamata Bay?
March 31, 2010: Researcher Behind CDC Vaccine Safety Claims Disappears with $2M
September 24, 2009: Huge field experiment for assessing human ethylmercury risk starting in october
July 15, 2009: New Study Finds: Thimerosal Induces Autism-like Neurotoxicity
March 24, 2006: American lawmakers initiate mercury probe for vaccines
April 4, 2005: New results about toxicity of thimerosal
February 11, 2005: New findings about Thimerosal Neurotoxicity

last time modified: July 2, 2017

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